Cha, In Jun;Lee, Davin;Park, Sung Soon;Chung, Chang Geon;Kim, Seung Yeon;Jo, Min Gu;Kim, Seung Yeol;Lee, Byung-Hoon;Lee, Young-Sam;Lee, Sung Bae
Molecules and Cells
/
v.43
no.10
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pp.870-879
/
2020
Dendrites require precise and timely delivery of protein substrates to distal areas to ensure the correct morphology and function of neurons. Many of these protein substrates are supplied in the form of ribonucleoprotein (RNP) complex consisting of RNA-binding proteins (RBPs) and mRNAs, which are subsequently translated in distal dendritic areas. It remains elusive, however, whether key RBPs supply mRNA according to local demands individually or in a coordinated manner. In this study, we investigated how Drosophila sensory neurons respond to the dysregulation of a disease-associated RBP, Ataxin-2 (ATX2), which leads to dendritic defects. We found that ATX2 plays a crucial role in spacing dendritic branches for the optimal dendritic receptive fields in Drosophila class IV dendritic arborization (C4da) neurons, where both expression level and subcellular location of ATX2 contribute significantly to this effect. We showed that translational upregulation through the expression of eukaryotic translation initiation factor 4E (eIF4E) further enhanced the ATX2-induced dendritic phenotypes. Additionally, we found that the expression level of another disease-associated RBP, fragile X mental retardation protein (FMRP), decreased in both cell bodies and dendrites when neurons were faced with aberrant upregulation of ATX2. Finally, we revealed that the PAM2 motif of ATX2, which mediates its interaction with poly(A)-binding protein (PABP), is potentially necessary for the decrease of FMRP in certain neuronal stress conditions. Collectively, our data suggest that dysregulation of RBPs triggers a compensatory regulation of other functionally-overlapping RBPs to minimize RBP dysregulation-associated aberrations that hinder neuronal homeostasis in dendrites.
Background: This study was performed to investigate the impact of exogenous ghrelin on the pancreatic ${\alpha}$-amylase outputs and responses of pancreatic proteins to ghrelin that may relate to pancreatic exocrine. Methods: Sprague-Dawley male rats (9 weeks old, $300{\pm}10g$) were injected with ghrelin via intraperitoneal (i.p.) infusion at dosage of 0, 0.1, 1.0 and $10.0{\mu}g/kg$ body weight (BW), respectively. The plasma ghrelin and cholecystokinin (CCK) level were determined using enzyme immunoassay kit; the mRNA expression of ghrelin receptor ($GHSR-1{\alpha}$) and growth hormone (GH) receptor were assessed by reverse transcription PCR; the expressions of pancreatic ${\alpha}$-amylase activity, extracellular-signal-regulated kinases (ERK), phosphorylated extracellular-signal-regulated kinases (pERK) and c-Jun N-terminal kinase (JNK) were evaluated by western blotting; moreover the responses of pancreatic proteins to ghrelin were analyzed using the two-dimensional gel electrophoresis system. Results: The exogenous ghrelin (1.0 and $10.0{\mu}g/kg\;BW$) elevated the level of plasma ghrelin (p < 0.05), and suppressed the expression of pancreatic ${\alpha}$-amylase at a dose of $10.0{\mu}g/kg\;BW$ (p < 0.05). No difference in the level of plasma CCK was observed, even though rats were exposed to any dose of exogenous ghrelin. In addition, a combination of western blot and proteomic analysis revealed exogenous ghrelin ($10.0{\mu}g/kg\;BW$) induced increasing the JNK and ERK expressions (p < 0.05) and four proteins such as Destrin, Anionic trypsin-1, Trypsinogen, and especially eukaryotic translation initiation factor 3 in rat pancreas. Conclusions: Taken together, exogenous ghrelin by i.p. infusion plays a role in the pancreatic exocrine secretion via mitogen-activated protein kinase signaling pathway.
Paik Soon-Young;Ra Kyung Soo;Cho Hoon Sik;Koo Kwang Bon;Baik Hyung Suk;Lee Myung Chul;Yun Jong Won;Choi Jang Won
Journal of Microbiology
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v.44
no.1
/
pp.64-71
/
2006
To investigate the effects of the nucleotide sequences in Shine-Dalgarno (SD) and the spacer region (SD-ATG) on bovine growth hormone (bGH) gene expression, the expression vectors under the control of the T7 promoter (pT7-7 vector) were constructed using bGH derivatives (bGH1 & bGH14) which have different 5'-coding regions and were induced in E. coli BL21 (DE3). Oligonucleotides containing random SD sequences and a spacer region were chemically synthesized and the distance between the SD region and the initiation codon were fixed to nine bases in length. The oligonucleotides were annealed and fused to the bGH1 and bGH14 cDNA, respectively. When the bGH gene was induced with IPTG in E. coli BL21(DE3), some clones containing only bGH14 cDNA produced considerable levels of bGH in the range of $6.9\%\;to\;8.5\%$ of total cell proteins by SDS-PAGE and Western blot. Otherwise, the bGH was not detected in any clones with bGH1 cDNA. Accordingly, the nucleotide sequences of SD and the spacer region affect on bGH expression indicates that the sequences sufficiently destabilize the mRNA secondary structure of the bGH14 gene. When the free energy was calculated from the transcription initiation site to the +51 nucleotide of bGH cDNA using a program of nucleic acid folding and hybridization prediction, the constructs with values below -26.3 kcal/mole (toward minus direction) were not expressed. The constructs with the original sequence of bGH cDNA also did not show any expression, regardless of the free energy values. Thus, the disruption of the mRNA secondary structure may be a major factor regulating bGH expression in the translation initiation process. Accordingly, the first stem-loop among two secondary structures present in the 5'-end region of the bGH gene should be disrupted for the effective expression of bGH.
In April 2014, Fusarium wilt was found on kohlrabi seedlings in Iksan, Korea. Symptoms included wilting of foliage, drying and withering of older leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelium. Macroconidia were falcate to almost straight, thin walled and usually 3-septate. Microconidia were usually formed abundantly in false-heads on short monophialides on the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $4-11{\times}2.5-5{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers (internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$), the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of Fusarium oxysporum on kohlrabi in Korea.
Objective: This study evaluated the effects of different levels of protein concentrate supplementation on the growth performance of yak calves, and correlated the growth rate to changes occurring in the plasma- amino acids, -insulin profile, and signaling activity of mammalian target of rapamycin (mTOR) cascade to characterize the mechanism through which the protein synthesis can be improved in early weaned yaks. Methods: For this study, 48 early (3 months old) weaned yak calves were selected, and assigned into four dietary treatments according to randomized complete block design. The four blocks were balanced for body weight and sex. The yaks were either grazed on natural pasture (control diet) in a single herd or the grazing yaks was supplemented with one of the three protein rich supplements containing low (17%; LP), medium (19%; MP), or high (21%; HP) levels of crude proteins for a period of 30 days. Results: Results showed that the average daily gain of calves increased (0.14 vs 0.23-0.26 kg; p<0.05) with protein concentrates supplementation. The concentration of plasma methionine increased (p<0.05; 8.6 vs $10.1-12.4{\mu}mol/L$), while those of serine and tyrosine did not change (p>0.05) when the grazing calves were supplemented with protein concentrates. Compared to control diet, the insulin level of calves increased (p<0.05; 1.86 vs $2.16-2.54{\mu}IU/mL$) with supplementation of protein concentrates. Addition of protein concentrates up-regulated (p<0.05) expression of mTOR-raptor, mammalian vacuolar protein sorting 34 homolog, the translational regulators eukaryotic translation initiation factor 4E binding protein 1, and S6 kinase 1 genes in both Longissimus dorsi and semitendinosus. In contrast, the expression of sequestosome 1 was down-regulated in the concentrate supplemented calves. Conclusion: Our results show that protein supplementation improves the growth performance of early weaned yak calves, and that plasma methionine and insulin concentrations were the key mediator for gene expression and protein deposition in the muscles.
CodY is a highly conserved protein in low G+C gram-positive bacteria that regulates genes involved in sporulation and stationary-phase adaptation. Bacillus thuringiensis is a grampositive bacterium that forms spores and parasporal crystals during the stationary phase. To our knowledge, the regulatory mechanism of CodY in B. thuringiensis is unknown. To study the function of CodY protein in B. thuringiensis, BMB171codY- was constructed in a BMB171 strain. A shuttle vector containing the ORF of cry1Ac10 was transformed into BMB171 and BMB171codY-, named BMB171cry1Ac and BMB171codY-cry1Ac, respectively. Some morphological and physiological changes of codY mutant BMB171codY-cry1Ac were observed. A comparative proteomic analysis was conducted for both BMB171codY-cry1Ac and BMB171cry1Ac through two-dimensional gel electrophoresis and MALDI-TOF-MS/MS analysis. The results showed that the proteins regulated by CodY are involved in microbial metabolism, including branched-chain amino acid metabolism, carbohydrate metabolism, fatty acid metabolism, and energy metabolism. Furthermore, we found CodY to be involved in sporulation, biosynthesis of poly-β-hydroxybutyrate, growth, genetic competence, and translation. According to the analysis of differentially expressed proteins, and physiological characterization of the codY mutant, we performed bacterial one-hybrid and electrophoretic mobility shift assay experiments and confirmed the direct regulation of genes by CodY, specifically those involved in metabolism of branched-chain amino acids, ribosomal recycling factor FRR, and the late competence protein ComER. Our data establish the foundation for in-depth study of the regulation of CodY in B. thuringiensis, and also offer a potential biocatalyst for functions of CodY in other bacteria.
This study was conducted to investigate the present situation and problems related to the use of nursing diagnosis in practice. The data were obtained from 332 subjects (27 director of nursing service, 302 staff nurses) who worked in university hospitals in Korea from July through August 1988 using a mailed questionnaire. Data were analyzed by frequency, X$^2$ test and t-test. The findings were as follows ; 1, Clinical use of nursing diagnosis by directors of nursing service and staff nurses. 1) The majority of the nursing departments (88.9%) conducted group education on nursing diagnosis during the last 5 years and 81.5% of them kept a record format for nursing diagnosis : 88.9% of them had had prior experience with the nursing diagnosis. 2) Most of nurses (97.0%) had received education on nursing diagnosis. 2. Factors related to the clinical use of nursing diagnosis in nursing service departments and by staff nurses. 1) The one factor related to the use of nursing diagnosis in the nursing service department was the existence of a record. 2) Factors related to the use of nursing diagnosis by the staff nurses were the organization style of the nursing service department, group education during the last 5 years, existence of a record, the attitude of the director of nursing service, and prior experience of the use of the nursing diagnosis as characteristics of nursing service department and educational experience of nursing diagnosis as a character of nurse. 3. Problems with the use of nursing diagnosis. 1) The primary problem was the lack of time and personnel (mean : 3.757) ; the second problem was the lack of knowledge and will to use nursing diagnosis in practice by the staff nurses(mean : 3.546). 2) There was no significant difference in problems expressed by the director of nursing services and the nurses. The majority of nurses who worked in the university hospitals expressed interest in and concern about the use of nursing diagnosis. Most of the nurses had had education about on nursing diagnosis but use in practice was limited. The primary problem was lack of time and manpower. Strategies for improving use of the nursing diagnosis in practice : 1) Strengthening the education about nursing diagnosis and a holistic approach to understanding human beings. 2) Develop protocols for the use of nursing diagnosis. 3) Eliminate the language barrier regarding nursing diagnosis by translation into in Korean. 4) Decentralization of the nursing service to promote accountability by individual nurses for use of nursing diagnosis.
The purpose of this study was to develop and validate Korean version of Deaf Acculturation Scales(DAS). Pilot items were made a faithful translation of the Acculturation Scales of Maxwell-McCaw and Zea (2011) and were modified for Korean Deaf people. The Scale involves two dimensions, in order to measure the acculturation of Deaf people; Deaf acculturation and hearing acculturation. Using factor analysis, we developed a Korean version of DAS consisted of twenty-five items for Deaf acculturation dimension and twenty-five items for hearing acculturation dimension. These analysis supported the four factors of Deaf acculturation dimension and the five factors of hearing acculturation dimension. Reliability, assessed by Cronbach's ${\alpha}$, was .93 for Deaf acculturation and .93 for hearing acculturation, respectively, which confirm the Koran version of DAS. Construct validity was demonstrated through correlation with Deaf acculturation-related variables: age, age of Deafness, Degree of hearing loss, American Sign Language ability, and lip-reading ability. Criterion validity was supported by correlation with Collective Self-Esteem Scale. Limitation and implication of this study and direction for future research were discussed.
From 2014 to 2016, Fusarium wilt disease was found on fassionfruit in Iksan and Jeju, Korea. Symptoms included wilting of foliage, drying and withering of leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelia with 35 mm after one week. Macroconidia were falcate to almost straight, thin-walled and usually 2-3 septate. Microconidia were usually formed on monophialides of the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $3-12{\times}2.5-6{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of two molecular markers, internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$, the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of F. oxysporum on fassionfruit in Korea.
Fusarium wilt caused by Fusarium oxysporum f. sp. fragariae (Fof) is the most important diseases of a strawberry field in Korea. We surveyed phylogenetic analysis, pathogenicity test, and fungicide response about Fof isolates isolated from Korea. Twenty-seven isolates of F. oxysporum isolated from strawberry plants were conducted in this study. Specific amplification by Fof specific primer was confirmed in all 26 isolates except Fo080701 isolate. The nuclear ribosomal intergenic spacer region and the translation elongation factor EF-lα gene sequences of isolates revealed three main lineages. Most of all isolates were contained DNA lineage group 1, but 2 and 3 group was shown only one and three isolates, respectively. All isolates were shown in pathogenicity with cv. Seolhyang. The EC50 mean values of prochloraz ranged 0.02-0.1 ㎍/ml except for Fo080701 and effectively inhibited mycelial growth at low concentrations. The EC50 value of metconazole was also 0.04-0.22 ㎍/ml, showing a similar inhibitory effect to that of prochloraz. The EC50 value of pyraclostrobin was 0.23-168.01 ㎍/ml, which was different according to the strain. In the field trial, boscalid+fludioxonil, fluxapyroxad+pyraclostrobin, and prochloraz manganese were selected as the effective fungicides for controlling Fusarium wilt.
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