Superoxide dismutase is a family of important antioxidant metalloenzymes and catalyzes the dismutation of toxic superoxide anions into dioxygen and hydrogen peroxide. A recent study identified the partial superoxide dismutase (SOD) gene in olive flounder (Paralichthys olivaceus). The same study reported that it strongly induced benzo[a]pyrene and that it was an indicator of aquatic oxidative stress responses. However, its transcriptional response against viral infection has not been investigated. In the present study, the spatial and temporal expression profiles were analyzed to investigate the function of Of-SOD in the antiviral response. The Of-SOD transcripts were ubiquitously detected at various levels in diverse tissues in a real-time PCR. The expression of Of-SOD was significantly higher in the muscles, liver, and brain but extremely low in the stomach and spleen. Following a VHSV challenge, the expression of Of-SOD increased within 3 h in the kidneys and decreased to the original level 2 days postchallenge. In muscle, liver, and brain, Of-SOD mRNA was similarly up-regulated at 3-6 h postchallenge and then decreased to the basal level. Although the expression pattern and induction time differed slightly depending on the tissue, the transcript of Of-SOD consistently increased in the acute infection response, but the expression was low in the chronic response. The expression of Of-SOD was induced after the VHSV infection, and Of-SOD was probably involved in the immune response against the viral challenge. These results suggest that SOD may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in olive flounder.
We evaluated the growth performance, biochemical characteristics, and immune responses in weaning pigs given a diet containing MR-1 (0.2%/feed) or antibiotics (0.1%/feed) for 45 days. In vitro study showed that MR-1 has antibacterial activity against a variety of strains of pathogenic bacteria, especially a strain of cattle-derived Escherichia coli K99 (E. coli K99) by agar diffusion assay. In the in vivo model, 0.2% MR-1-given group clearly ameliorated the weight gain and feed efficiency in the growth performance of weaning pigs compared to the basal diet group (p<0.05). Additionally, 0.2% MR-1 induced an elevation in the levels of mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) and showed a similar pattern ($TNF{\alpha}$ and $IFN{\gamma}$ production) to the antibiotic treated pigs. Taken together, we suggest that 0.2% MR-1 makes probiotics an alternative to antibiotics in weaning pigs.
An, Ji-Hong;Park, Hwan-Jun;Nam, Gi-Heum;Lee, Byoung-Yoon;Park, Chan-Ho;Kim, Jung-Hyun
Korean Journal of Ecology and Environment
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v.50
no.4
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pp.381-402
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2017
In order to investigate distribution of vascular plants along elevational gradient in the Nodong valley of Gyebangsan, vascular plants of eight sections with 100-meter-high were surveyed from the Auto-camping site (800 m) to the top of a mountain (1,577 m). There were a total of 382 taxa: 89 families, 234 genera, 339 species, 7 subspecies, 34 varieties, and 2 forms. As a result of analyzing the pattern of species richness, it showed a reversed hump-shaped with minimum richness at mid-high elevation. As a result of analyzing habitat affinity types, the proportion of forest species increased with increasing elevation. But, the ruderal species decreased with increasing elevation, and then increased at the top of a mountain. As for the proportion of life forms, the annual herbs gradually decreased with increasing elevation, but it did not appear between 1,300 m and 1,500 m and then increased at the top of a mountain. The trees gradually increased with elevation and decreased from 1,300~1,400 m. The vascular plants divided into four groups by using DCA. The arrangement of each stands was arranged in order from right to left on the I axis according to the elevation. The distribution of vascular plants is determined by their own optimal ranges of vegetation. Also, rise in temperature due to climate change affects the distribution of vascular plants, composition, and diversity. Therefore, continuous monitoring is necessary to confirm ecological and environmental characteristics of vegetation, distribution ranges, changes of habitat. Furthermore, plans for conservation and management based on these data should be prepared according to climate change.
The root of Stnchys Sieboldif MIQ was extracted three times with methanol and extract was found to contain 3.02% of polyphenols and 1.97% of flavonoids. DPPH radical scavenging method, ferric thiocyanate method, and nitrite scavenging ability method were employed to investigate the constituents of the extract and to measure their activity on antioxidation. The fraction extracted by ethylacetate showed higher anti oxidation value than that of $\alpha-tocopherol$, butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) at the same concentration. UV-VIS spectral data of the extract by ethylacetate that was isolated on a silica gel column proved adsorption maxima in the range of 280∼330 nm. The fraction ES-RS that has $\lambda_{max}(nm)$ of band 1, 325nm and band II, 289nm exhibitd the strongest activity on antioxidation. ES-R5 fraction showed similar pattern to flavones by the analysis of UV-VIS spectral data.
Asthma is a chronic inflammatory disorder of the airways, which characterized by bronchial hyperresponsiveness, reversible airflow limitation and respiratory symptoms. Internationally, the prevalence of asthma has been increased over last 3 decades. Recently, several studies of asthma have been reported with gradually increasing importance. To tesify the hypothesis that interleukin (IL)-4 and IL-10 may be an important determinant of the severity of airway inflammation, their expression was studied in mouse model of asthma. BALB/c mouse, IL-4 Knockout (KO) mouse and IL-10 KO mouse were sensitized with intraperitoneal injection of ovalbumin adsorbed to aluminum potassium sulfate, followed by challenges with intranasal ovalbumin on 3 consecutive days. The severity of pulmonary inflammation was assessed by eosinophilia in BAL fluid, number of total BAL cells, histopathological changes in lung tissues, and immunohistochemical staining against IL-4 and IL-10. In BAL fluid, the number of total cells was significantly increased in asthma induced mouse compare to the control. In asthma induced mouse, eosinophil was increased to 56% and neutrophil was 0.2%. In H &E stains, eosinophilic infiltration and epithelium hyperplasia were clearly noticed in asthma induced mouse. In immunohistochemical staining for IL-4 and IL-10, there was no positive reaction in control group. However, very strong reactions were appeared in asthma induced group. In this research, IL-4 and IL-10, which seem to play a central role in allergic asthma, KO mouse was utilized to test the causative relationship between airway inflammation and role of specific cytokine. Asthma induced IL-4 and IL-10 KO mice showed much decreased inflammatory reactions in the number of total BAL cells, in eosinophilic infiltration, and in immunohistochemical stains against diverse inflammatory proteins. These results suggest that IL-4 and IL-10 increase the asthmatic reactions in vivo mice model.
This study attempted to investigate the developmental alterations of the cerebral cortex. The effects of EGEE on the developmental cerebral cortex in the prenatal, postnatal and adults were examined by lectin histochemical methods. To investigate the change of sugar residues of glycoconjugates, biotinylated lectins(DBA, SBA, PNA, BSL-1, RCA-1, sWGA, UEA-1, Con A and LCA) were detected with by IHC using ABC kit. In the case of injection of EGEE, the reactions to Con A and LCA were shown in binding phase in the cerebral cortex commonly, and the reactions to PNA, RCA-1 and LCA were shown partially, the number of lectins to be shown reaction were decreased, and there were no reactions to DBA, SBA, BSL-1, RCA-1 and UEA-1. The reaction to Con A was similar to control group during developmental phases. The reaction to LCA was increased in the fetal, suckling, and weanning phases compared with control group. But there were no reactions to SBA and sWGA, the reaction to PNA was decreased in the frontal and occipital cortex and no reaction to sWGA in the fetal phase. There were no reactions to sWGA and PNA in the suckling phase and, no reaction to PNA and sWGA. The reaction to Con A was decreased in the frontal, parietal and occipital cortexes and, the reaction to LCA was decreased in the frontal and occipital cortexes in adult phase. As the results, the effects of EGEE, environmental hormone on the each part of cerebral cortex have shown differences. But, It had deep effect on the differentiation and growth in the cerebral cortex pathologically. In particular, the effect was severe in suckling phase. $Galactosyl-({\beta}-1,3)-N-acetyl-D-galactosamine$,${\beta}-N-acetyl-D-galactosamine$ and ${\beta}-N-acetyl-D-glucosamine$ were decreased while ${\alpha}-D-mannose$ and ${\alpha}-D-glucose$ were increased. It affected the sugar metabloism, and it was severe in fetal and suckling phases.
Kang, Taekyeong;Kim, Sang Ho;Jung, Mi Ja;Cho, Yong Kweon
Journal of Life Science
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v.25
no.5
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pp.487-495
/
2015
We carried out pH stability, chemical inhibition, chemical modification, and pH-dependent kinetic parameter assessments to further characterize protocatechuate 3,4-dioxygenase from Pseudomonas pseudoalcaligenes KF707. Protocatechuate 3,4-dioxygenase was stable in the pH range of 4.5~10.5. L-ascorbate and glutathione were competitive inhibitors with $K_{is}$ values of 0.17 mM and 0.86 mM, respectively. DL-dithiothreitol was a noncompetitive inhibitor with a $K_{is}$ value of 1.57 mM and a $K_{ii}$ value of 8.08 mM. Potassium cyanide, p-hydroxybenzoate, and sodium azide showed a noncompetitive inhibition pattern with $K_{is}$ values of 55.7 mM, 0.22 mM, and 15.64 mM, and $K_{ii}$ values of 94.1 mM, 8.08 mM, and 662.64 mM, respectively. $FeCl_{2}$ was the best competitive inhibitor with a $K_{is}$ value of $29{\mu}M$. $FeCl_{3}$, $MnCl_{2}$, $CoCl_{2}$, and $AlCl_{3}$ were also competitive inhibitors with $K_{is}$ values of 1.21 mM, 0.85 mM, 3.98 mM, and 0.21 mM, respectively. Other metal ions showed noncompetitive inhibition patterns. The pH-dependent kinetic parameter data showed that there may be at least two catalytic groups with pK values of 6.2 and 9.4 and two binding groups with pK values of 5.5 and 9.0. Lysine, cysteine, tyrosine, carboxyl, and histidine were modified by their own specific chemical modifiers, indicating that they are involved in substrate binding and catalysis.
Trichoplusia ni cells are used as a host permissive cell line in the baculovirus expression system, which is useful for large-scale production of human sugar transport proteins. However, the activity of endogenous sugar transport systems in insect cells is extremely high. Therefore, the transport activity resulting from the expression of exogenous transporters is difficult to detect. Furthermore, very little is known about the nature of endogenous insect transporters. To exploit the expression system further, the effect of D-fructose on 2-deoxy-D-glucose (2dGlc) transport by T. ni cells was investigated, and T. ni cell-expressed human transporters were photolabeled with [$^3H$] cytochalasin B to develop a convenient method for measuring the biological activity of insect cell-expressed transporters. The uptake of 1 mM 2dGlc by uninfected- and recombinant AcMPV-GTL infected cells was examined in the presence and absence of 300 mM of D-fructose, with and without $20{\mu}M$ of cytochalasin B. The sugar uptake in the uninfected cells was strongly inhibited by fructose but only poorly inhibited by cytochalasin B. Interestingly, the AcMPV-GTL-infected cells showed an essentially identical pattern of transport inhibition, and the rate of 2dGlc uptake was somewhat less than that seen in the non-infected cells. In addition, a sharply labeled peak was produced only in the AcMPV-GTL-infected membranes labeled with [$^3H$] cytochalasin B in the presence of L-glucose. No peak of labeling was seen in the membranes prepared from the uninfected cells. Furthermore, photolabeling of the expressed protein was completely inhibited by the presence of D-glucose, demonstrating the stereoselectivity of labeling.
Hox genes encode a highly conserved family of homeodomain-containing transcription factors controlling vertebrate pattern formation along the anteroposterior body axis during embryogenesis. Retinoic acid (RA) is a key morphogen in embryogenesis and a critical regulator of both adult and embryonic cellular activity. Specifically, RA regulates Hox gene expression in mouse- or human-derived embryonic carcinoma (EC) cells. Histone modification has been reported to play a pivotal role in the process of RA-induced gene expression and cell differentiation. As histone modification is thought to play an essential role in RA-induced Hox gene expression, we examined RA-induced initiation of collinear expression of Hox genes and the corresponding histone modifications in F9 murine embryonic teratocarcinoma (EC) cells. Hox expression patterns and histone modifications were analyzed by semiquantitative RT-PCR, RNA-sequencing, and chromatin immuno-precipitation (ChIP)-PCR analyses. The Hoxc4 gene (D0) was initiated earlier than the Hoxc5 to –c10 genes (D3) upon RA treatment (day 0 [D0], day 1 [D1], and day 3 [D3]). The Hox nonexpressing D0 sample had a strong repressive marker, H3K27me3, than the D1 and D3 samples. In the D1 and D3 samples, reduced enrichment of the H3K27me3 marker was observed in the whole cluster. The active H3K4me3 marker was closely associated with the collinear expression of Hoxc genes. Thus, the Hoxc4 gene (D1) and all Hoxc genes (D3) expressed H3K4me3 upon transcription activation. In conclusion, these data indicated that removing H3K27me3 and acquiring H3K4me3 regulated RA-induced Hoxc gene collinearity in F9 cells.
Familial hypokalemic periodic paralysis (HOKPP) is an autosomal dominant disorder characterized by reversible flaccid paralysis and intermittent hypokalemia. Although it has been reported that decreased activity in the $K_{ATP}$ channels of the skeletal muscle cell membrane plays a role in the pathogenesis of HOKPP, a clear mechanism has not yet been established. This study aimed to investigate the molecular biological mechanism underlying the decreased activity of $K_{ATP}$ channels in the skeletal muscles of familial HOKPP patients by studying the levels of the $K_{ATP}$ channel subunit Kir6.2. We found that when cells obtained from healthy individuals (normal cells) and HOKPP patients (patient cells) were treated with 4 mM potassium buffer, there was no quantitative change in the KCNJ11 mRNA levels and no difference in the Kir6.2 protein expression in the cytosol and cell membrane. On the other hand, when 1 mM potassium buffer was used, normal cells showed decreased expression of KCNJ11 mRNA as well as decreased expression of Kir6.2 protein in the cell membrane. However, patient cells treated with the same buffer showed no quantitative change in the levels of KCNJ11 mRNA or in the levels of Kir6.2 protein in the cytosol and cell membrane. Thus, in HOKPP patients, the Kir6.2 protein cannot be transported from the cell membrane to the cytosol, leading to closure of the $K_{ATP}$ channels, induction of depolarization, and subsequently, to the paralytic symptoms observed in the patient. Our findings thus provide new insights into the pathogenesis of HOKPP.
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