Mutations in Fused in Sarcoma (FUS) have been identified in patients with amyotrophic lateral sclerosis (ALS) or Frontotemporal Dementia (FTD). Pathological FUS is mis-localized to cytosol and forms aggregates associated with stress granules (SG), while FUS is normally localized to nucleus. However, it is largely unknown how pathological FUS forms SG-aggregates and which domains are responsible for this process. In this study, we examined cellular localization and aggregation of ALS-linked FUS missense mutants (P525L, R521C, R521H, R521G), analyzed the domains responsible for cytosolic FUS aggregation in HEK293T cells, and confirmed this in cultured mouse neurons. To do this, we firstly generated missense mutants of FUS and then examined their cellular localization. We found that P525L was mostly mis-localized to cytosol and formed FUS-positive SG aggregates while R521C, R521H, or R521G was localized to both nucleus and cytosol. To further characterize the domains required for aggregate formation of cytosolic FUS, we generated different domain-deletion mutants using FUS-∆17 which has a deletion of nuclear localization signal. Interestingly, cytosolic FUS without SYGQ and RGG1 domain or cytosolic FUS without RGG2-ZnF-RGG3 domain did not form FUS-positive SG aggregates, while cytosolic FUS without RRM domain generated more aggregates compared to FUS-∆17. Taken together, these data suggest that SYGQ-RGG1 or RGG2-ZnF-RGG3 domain contributes to formation of cytosolic aggregate, while RRM domain might interfere with FUS aggregation. Therefore, our studies will provide important insight for understanding cellular pathogenesis of neurodegeneration associated with FUS aggregate as well as finding therapeutic targets for ALS or FTD.
Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORTTM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.
Kim, Sung-Hee;Lee, Woo-Chun;Kim, Soon-Oh;Na, So-Young;Kim, Hyun-A;Lee, Byung-Tae;Lee, Byoung-Cheun;Eom, Ig-Chun
Journal of Korean Society of Environmental Engineers
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v.35
no.11
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pp.835-860
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2013
Recently, the consumption of nanomaterials has been significantly increased in both industrial and commercial sectors, as a result of steady advancement in the nano-technologies. This ubiquitous use of nanomaterials has brought up the concern that their exposure to environments may cause detrimental effects on human health as well as natural ecosystems, and it is required to characterize their behavior in various environmental media and to evaluate their ecotoxicity. For the sake of accomplishing those assessments, the development of methods to effectively separate them from diverse media and to quantify their properties should be requisitely accompanied. Among a number of separation techniques developed so far, this study focuses on Field-Flow Fractionation (FFF) because of its strengths, such as relatively less disturbance of samples and simple pretreatment, and we review overseas and domestic literatures on the separation of nanomaterials using the FFF technique. In particular, researches with Flow Field-Flow Fractionation (FlFFF) are highlighted due to its most frequent application among FFF techniques. The basic principle of the FlFFF is briefly introduced and the studies conducted so far are classified and scrutinized based on the sort of target nanomaterials for the purpose of furnishing practical data and information for the researchers struggling in this field. The literature review suggests that the operational conditions, such as pretreatment, selection of membrane and carrier solution, and rate (velocity) of each flow, should be optimized in order to effectively separate them from various matrices using the FFF technique. Moreover, it seems to be a prerequisite to couple or hyphenate with several detectors and analyzers for quantification of their properties after their separation using the FFF. However, its application has been restricted regarding the types of target nanomaterials and environmental media. Furthermore, domestic literature data on both separation and characterization of nanomaterials are extremely limited. Taking into account the overwhelmingly increasing consumption of nanomaterials, the efforts for the area seem to be greatly urgent.
Eun, Byung Wook;Chung, Yoo Mi;Kang, Hee Gyung;Ha, Il Soo;Cheong, Hae Il;Lee, Hoan Jong;Choi, Yong
Clinical and Experimental Pediatrics
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v.46
no.3
/
pp.265-270
/
2003
Purpose : To characterize the infants under 3 months of age with urinary tract infections(UTIs), and especially patients with bacteremia or meningitis Methods : Hospital records of all the infants under 3 months of age discharged from our hospital for 69 consecutive months with the diagnosis of initial episode of UTI were reviewed. UTI was defined when patients had fever with pyuria, and had urine culture results of ${\geq}10^5$ colony forming units/mL from a bag specimen. Patients with previously known urologic abnormality or immunodeficiency were excluded. Nosocomial infections were also excluded from the study. Results : The male:female ratio was 35 : 6. Of the urine cultures, 40(97.6%) yielded single pathogen, one yielded two pathogens. Escherichia coli was the predominant isolate from the urine. Five patients(12%) also had bacteremia. Pathogens isolated from the blood cultures were E. coli(4) and Enterococcus faecalis(1). No patient had culture-positive meningitis or cerebrospinal fluid pleocytosis. Clinical or laboratory findings between patients with and without bacteremia were not different significantly. The rate of vesicoureteral reflux(VUR) was 44%. The sensitivity of ultrasound for detection of VUR was 38%; specificity was 50%. Conclusion : Clinical and laboratory data were not helpful for identifying patients with bacteremia at the time of presentation. Consequently, blood cultures need to be obtained from all febrile infants under 3 months of age with UTIs. A large-scale study including the indication of lumbar puncture for infants with a febrile UTI and study of evaluation and treatment of infants under 3 months of age with UTIs are required.
Ko, Jeong Hee;Lee, Jee Hyun;Sim, Eun Jung;Cho, Do Jun;Min, Ki Sik;Yoo, Ki Yang;Lee, Dae Hyoung;Kang, Hee Jung
Clinical and Experimental Pediatrics
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v.52
no.10
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pp.1147-1152
/
2009
Purpose:To characterize the pathogens and their antibiotic susceptibilities in more than 24-month-old children with urinary tract infection (UTI) and to study the Escherichia coli antimicrobial susceptibility trend. Methods:We retrospectively reviewed the record of more than 24-month-old children with UTI between January 2003 and December 2008. Positive results for 1 bacterial species with a colony count of ${\geq}10^5CFU/mL$ was considered statistically significant. We analyzed uropathogens and their antibiotic susceptibilities. To investigate E. coli antibiotic susceptibility trend, we compared 2 study periods (group A: 2003-2005 versus group B: 2006-2008) using the chi-square test for trend. Results:In all, 63 bacterial isolates were identified in children with febrile UTI. The most common pathogen was E. coli (77.8%). There was no difference in the resistance patterns of uropathogens during the 2 study periods (P>0.05). Antibiotic susceptibility of the E. coli isolates to aztreonam, cefotetan, cefotaxime, ceftriaxone, cefepime, amikacin, and imipenem was >90% to trimethoprim/sulfamethoxazol, 49% and to ampicillin and ampicillin/sulbactam, 20-25%. Over the 2 study period, the E. coli susceptibilities to most antibiotics did not change: the susceptibility to cefuroxime increased from 74.1% to 95.5% (P=0.046) and that to ciprofloxacin increased from 59.3% to 86.4% (P=0.039). Conclusion:Empirical treatment with trimethoprim/sulfamethoxazole, ampicillin, and ampicillin/sulbactam alone appeared to be insufficient in childhood UTI because of the high resistance of E. coli and other gram-negative uropathogens. Antibiotics for empirical therapy should be selected based on the sensitivity and resistance pattern of uropathogens found in a particular region.
Kim, Won-Joon;Lee, Kwang-Youn;Ha, Jeoung-Hee;Kwon, Oh-Cheol
The Korean Journal of Pharmacology
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v.28
no.2
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pp.147-162
/
1992
The objectives of the present experiments were to characterize the effects of the peptides belonging to the pancreatic polypeptide family on the contractility of cerebral arteries and to observe the interactions of these peptides with the cyclic nucleotide activators and the potassium channel openers. Dogs of either sex, $20{\sim}30\;Kg$ in weight, were sacrificed. Basilar and middle cerebral arteries from brain were isolated and prepared for myography in the PSS equilibrated with 95% $O_2$ and 5% $CO_2$ at $37^{\circ}C$. The endothelial cells of the spiral strips were removed by CHAPS solution (0.3% w/v, 15 seconds). 1. PP, PYY and NPY contracted the arterial strips concentration-dependently with a rank order of potency of PYY>NPY>PP. These peptides were 20 to 200 times more potent than norepinephrine, and only PYY showed a greater potency than 5-HT. 2. Cyclic nucleotide activators, forskolin (for cAMP) and sodium nitroprusside (for cGMP) reduced the basal tone and inhibited the PP-, PYY- and NPY- induced contractions by concentration-dependent manners. Forskolin was more potent in reducing basal tone than sodium nitroprusside. 3. Potassium channel openers, RP 49356, P 1060 and BRL 38227 reduced the basal tone concentration-dependently and tended to inhibit the PP-, PYY- and NPY- induced contractions. Notably, BRL 38227 with low concentration $(0.1\;{\mu}M)$ enhanced the contractions induced by those peptides while P 1060 inhibited the contractions concentration-dependently. 4. The combinations of the cyclic nucleotide activators and the potassium channel openers were slightly additive in reducing the basal tone. P 1060 and BRL 38227 enhanced the relaxant effect of sodium nitroprusside significantly. On the PYY-induced contration $(0.1\;{\mu}M)$, $K^+$ channel openers tended to inhibit the inhibitory actions of forskolin and sodium nitroprusside. P 1060 and BRL 38227 antagonized the inhibitory action of sodium nitroprusside significantly. The results of the present study may be summarized that in canine cerebral arteries, not only NPY but also PYY may play a role in a cerebrovascular spasm, and intracellular concentration of either cAMP or cGMP may be involved in the mechanism of vasoconstrictive actions of these peptides, which may be affected either positively or negatively by a $K^+$ channel opener.
During the past few years it has been proposed that lithium clearance can be used as a reliable measure for the outflow of tubular fluid from the proximal tubule. This study was aimed to characterize the inflow dependent reabsorption of Na in renal tubule beyond the proximal tubule. For this purpose, lithium clearance was used as a measure for the inflow from the proximal tubule and the changes in reabsorption fraction of Na and water were determined in rabbits. Rabbits were pretreated with hypotonic saline solutions for an hour (50 mM/L NaCl, 20 ml/hr/kg). And then a hypertonic solution of 500 mM/L NaCl (20 ml/kg) was administered intraperitoneally in conjunction with a bolus of LiCl solution (2 mM/kg, i.v.) for conditioning the $C_{Li}$ and urine flow rate. To rule out the effect of $Li^+$ on tubular functions, a bolus of NaCl solution (2 mM/kg, i.v.) was administered. Fifteen, thirty, and sixty minutes after injection of hypertonic saline arterial blood and urine samples were taken. Urinary and plasma concentrations as well as urinary output of $Li^+,\;Na^+\;and\;K^+$ were measured. From these $C_{Li},\;C_{Na}$ and the reabsorption fraction of Na and water $(Fr_{Na}\;&\;FrH_2O)$ were calculated. These results were compared with those from control groups in which the same amount of isotonic saline (145 mM/L NaCl) and of 15% dextran solution were administered in the same way as that in experimental group. Followings are the results obtained. 1) The plasma concentration of $Na^+$ in rabbits injected with hypertonic saline reached the peak value after 15 min and thereafter no significant change was observed. Hematocrit values did not show any change, while urinary excretion of $Na^+$ increased markedly during the first 15 min and decreased thereafter. These results were not affected by an injection of a small amount of LiCl. 2) The clearances of $Li^+,\;Na^+\;and\;K^+$ in rabbits injected with hypertonic saline and LiCl solution decreased. 3) In spite of the variation in $C_{Li},\;Fr_{Na}$ did not show any significant change while $FrH_2O$ increased gradually. 4) $C_{Li}$ decreased also in rabbits received isotonic saline. $Fr_{Na}$ tended to be higher than that in hypertonic saline group, while $FrH_2O\;and\;Fr_{Na}$ did not associated with the decrease in $C_{Li}$. 5) $C_{Li}$ of the rabbits received dextran solution fluctuated persistently and $Fr_{Na}\;and\;FrH_2O$ did not change in along with $C_{Li}$ although $Fr_{Na}$ had a tendency to be higher than that in hypertonic saline group. 6) From the above results it was concluded that: (a) In rabbits with normal body store of $Na^+$, the $Fr_{Na}$ of renal tubule beyond proximal tubule. calculated from $C_{Li}$ as a measure of inflow from proximal tubule is constant in spite of variations in $C_{Li}$. (b) The $FrH_2O$ calculated from $C_{Li}$ is dependent largely upon ADH rather than inflow from proximal tubule. (c) When there is a decrease in plasma $Na^+$ concentration or ineffective body fluid. $Li^+$ reabsorption may occur in the thick segnent of Henle's loop and hence the determination of $Fr_{Na}$ and $FrH_2O$ will not be easy one, but $Fr_{Na}$ is constant under the same experimental conditions.
Kang Chang Keun;Baik Myung Sun;Kim Jeong Bae;Lee Pil Yong
Korean Journal of Fisheries and Aquatic Sciences
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v.35
no.1
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pp.35-45
/
2002
Seasonal quantitative van Veen grab sampling was conducted to characterize the composition and structure of the benthic polychaete community inhabiting the shellfish farming ground of a coastal bay system of Jiniu Bay (Korea). A total of 132 polychaete species were identified and the polychaetes accounted for about $80\%$ of overall abundance of benthic animals. There was little significant seasonal difference in densities (abundances) of polychaetes, Maximum biomass was obseued in summer (August) and minimum value was recorded in winter (February) and spring (May). Conversely, diversity and richness were lowest in summer, indicating a seasonal variability in the polychaetous community structure, The cluster analysis indicated that such a seasonal variability resulted mainly from the appearance of a few small, r-selected opportunists in spring and the tubiculous species of the family Maldanidae in summer. On the other hand, several indicator species for the organically enriched environments such as Capitelia capitata, Notoniashs Jatericeus and hmbrineris sp. showed high densities during all the study period. Density and biomass of univariate measures of community structure were significantly lower in the arkshell-farming ground of the southern area than in the non-farming sites of the bay, A similar general tendency was also found in the spatial distributions of species diversity and richness. Principal component analysis revealed the existence of different groups of benthic assemblages between the arkshell-farming ground and non-farming sites, The lack of colonization of r-selected opportunists and/or tubiculous species in the former ground seemed to contribute to the spatial differences in the composition and structure of the polychaetous communities. Although finer granulometric composition and high sulfide concentration in sediments of the arkshell-farming ground and low salinity in the northern area were likely to account for parts of the differences, other environmental variables observed were unlikely. The spatial distribution of polychaetes in Jiniu Bay may be rather closely related to the sedimentary disturbance by selection of shells for harvesting in spring.
Sardine, Sardinops melanosticta, has been caught more than fifty thousand metric tons every year in adjacent sea of Korea, but most of them used for uneatable fish meal because of their rapid spoilage. Usually it is known that the main reason of putrefaction of foods is caused by the maicro-organisms included in them. Therefore, this experiment was carried out to identify the micro-organisms isolated from the intestine of fresh sardine and characterize their proteolytic enzymes produced from them. Aerobic cell count ranged from $1.7{\times}10^4\;to\;3.6{\times}10^5/g$, while anaerobic cell count, from $2.9{\times}10^4\;to\;5.5{\times}10^5/g$. Most of the isolated strains were psychrotrophic mesophiles. Among the two hundred and eighty strains isolated from the fresh samples, fifty-six strains ($20.0\%$) were proteolytics, one hundred and seventy-five strains ($62.5\%$) were lipolytics and tenty-nine strains ($10.5\%$) had the ability to produce hydrogn sulfide. The most predominantly isolated microbial groups from the fresh sardine were Moraxella ($31.4\%$) and Pseudomonas sup. ($28.6\%$). Flavobacterium-Cytophaga, Vibrio, Acinetobacter, Micrococcus spp. and Enterobacteriaceae appeared from $7.9\%\;to\;5\%$ out of total tested strains. The average bacterial count in the spoiled samples (stored at about $18^{\circ}C$ for 48 hours) was increased to the level of $2.9{\times}10^8/g$ for aerobes, $1.5{\times}10^8/g$ for anaerobes, then one hundred and ten strains, corresponding to $52\%$, out of two hundred and thirteen strains submitted to the test were proteolytics. The strongest proteolytic bacterium among the two hundred and eighty strains was identified as Pseudomonas 101 which grew best at $25^{\circ}C$. The optimum condition for the activity of the proteolytic enzyme produced by Pseudomonas 101 appeared $35^{\circ}C$ and pH 9.0, but the activity was relatively unchanged between 5.0 and 11.0 of pH and between $30^{\circ}C\;and\;50^{\circ}C$ of temperature.
Makoto Shinkai, an animated film maker in Japan, has been featured as a one-person production system and as a 'writer of light', but his 2016 release of "Your Name" was a departure from the elements that characterize his existing works. At the same time, by the combination of the traditional musubi(むすび) story, ending these, it was a big hit due to its rich narratives and attraction of open interpretation possibility. As it can be guessed from the title of this work, this work shows the encounter between the Japanese ancient language and the modern language in relation to the 'name', and presents the image that the role of the name(language) is repeatedly emphasized with various variations in events for the perfect 'encounter'. In this work, the interpretations of $Signifi\acute{e}$ for characters and objects are extended and reserved as a metaphorical role of the similarity, depending on the meaning of the subject which they touch. The relationship between words and objects analyzed through the structure of Signifiant and $Signifi\acute{e}$ is an epoch-making ideological discovery of modern times revealed through F. Saussure. Focusing on "the difference" between being this and that from the notion of Saussure, Derrida dismissed logocentrism, rationalism that fully obeyed the order of Logos. Likewise, dismissing the center, or dismissing the owner had emerged after the exclusive and closed principle of metaphysics in the west was dismissed. Derrida's definition of 'deconstruction' is a philosophical strategy that starts with the insight on the nature of language. 'Dissemination,' a metaphor that he used as a methodological concept to read texts acts as interpretation and practice (or play), but does not pursue an ultimate interpretation. His 'undecidability' does not start with infinity, but ends with infinity. The researcher testifies himself and identifies that we can't be an interpreter of the world because we, as a human are not the subject of language but a user. Derrida also interpreted the world of things composed of Signifiant and $Signifi\acute{e}$ as open texts. In this respect, this study aimed to read Makoto's works telling about the meeting of a thing and a thing with name as a guide, based on Derrida's frame of 'deconstruction' and 'dissemination.' This study intends to re-consider which relationship the Signifiant and $Signifi\acute{e}$ have with human beings who live in modern times, examine the relationship between words and objects presented in this work through Jacques Derrida's destruction and dissemination concepts, and recognize that we are merely a part of Signifiant and $Signifi\acute{e}$. Just as Taki and Mitsuha confirm the existence by asking each other, we are in the world of things, expecting musubi that a world of names calls me.
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