The Bric-a-brac, Tramtrack, Broad-complex (BTB) domain is a protein-protein interaction domain that is found in many zinc finger transcription factors. BTB containing proteins play important roles in a variety of cellular functions including regulation of transcription, regulation of the cytoskeleton, protein ubiquitination, angiogenesis, and apoptosis. Here, we report the cloning and characterization of a novel human gene, KLHL31, from a human embryonic heart cDNA library. The cDNA of KLHL31 is 5743 bp long, encoding a protein product of 634 amino acids containing a BTB domain. The protein is highly conserved across different species. Western blot analysis indicates that the KLHL31 protein is abundantly expressed in both embryonic skeletal and heart tissue. In COS-7 cells, KLHL31 proteins are localized to both the nucleus and the cytoplasm. In primary cultures of nascent mouse cardiomyocytes, the majority of endogenous KLHL31 proteins are localized to the cytoplasm. KLHL31 acts as a transcription repressor when fused to GAL4 DNA-binding domain and deletion analysis indicates that the BTB domain is the main region responsible for this repression. Overexpression of KLHL31 in COS-7 cells inhibits the transcriptional activities of both the TPA-response element (TRE) and serum response element (SRE). KLHL31 also significantly reduces JNK activation leading to decreased phosphorylation and protein levels of the JNK target c-Jun in both COS-7 and Hela cells. These results suggest that KLHL31 protein may act as a new transcriptional repressor in MAPK/JNK signaling pathway to regulate cellular functions.
This study has researched the national expansion, dispersion of nutria and investigated its inhabitation status for the past 3 years. The report has shown that the number of nutria habitat, reported to be distributed in 9 cities or districts in 2010, has been increased to 13 in 2012. In the research of 629 nutria habitats, 95.9% of habitat traces were found in Busan-Kyungnam area. From the research of relative density by location type for the 6 survey areas in these 5 areas above, it was shown 3.98(${\pm}2.56$)ind./100m in 2010, 2.90(${\pm}2.69$)ind./100m in 2011 and 1.39(${\pm}0.66$)ind./100m in 2012. From the research of relative density by habitat types, it was shown 3.48(${\pm}2.15$)ind./100m in palustrine wetland area, $1.01{\pm}(0.25)ind.$/100m in river area and $3.69{\pm}(2.83)ind.$/100m in riverine wetland area. It was shown that the annual average density in the areas has slightly been decreased for the past 3 years between 2010 and 2012. It also reported that the habitat density in the river area is a bit lower than that in the wetland area. Currently it can be determined that the nutria has fully adapted themselves to the various local environments of wetland, river, stream, and so on in Korea and the Relative density could be remarkably increased especially in the favorable condition like a wetland. As the ones living in the area with over-density could be spread out to other areas, the efficient management plan to control should be prepared considering ecological conditions.
The study on the atomic structure of iron-bearing silicate glasses has significant geological implications for both diverse igneous processes on Earth surface and ultra-low velocity zones at the core-mantle boundary. Here, we report experimental results on the effect of iron content on the atomic structure in iron-bearing alkali silicate glasses ($Na_2O-Fe_2O_3-SiO_2$ glasses, up to 16.07 wt% $Fe_2O_3$) using $^{29}Si$ and $^{17}O$ solid-state NMR spectroscopy. $^{29}Si$ spin-lattice ($T_1$) relaxation time for the glasses decreases with increasing iron content due to an enhanced interaction between nuclear spin and unpaired electron in iron. $^{29}Si$ MAS NMR spectra for the glasses show a decrease in signal intensity and an increase in peak width with increasing iron content. However, the heterogeneous peak broa-dening in $^{29}Si$ MAS NMR spectra suggests the heterogeneous distribution of $Q^n$ species around iron in iron-bearing silicate glasses. While nonbridging oxygen ($Na-O-Si$) and bridging oxygen (Si-O-Si) peaks are partially resolved in $^{17}O$ MAS NMR spectrum for iron-free silicate glass, it is difficult to distinguish the oxygen clusters in iron-bearing silicate glass. The Lorentzian peak shape for $^{29}Si$ and $^{17}O$ MAS NMR spectra may reflect life-time broadening due to spin-electron interaction. These results demonstrate that solid-state NMR can be an effective probe of the detailed structure in iron-bearing silicate glasses.
BACKGROUND: Plant factory system of an artificial light type using Light-Emitting Diodes (LEDs), fluorescent light, or metal halide lamp instead of sun light is an ultimated method for plant production without any pesticides regardless of seasonal changes. The plant factory is also completely isolated from outside environmental conditions such as a light, temperature, or humidity compared to conventional greenhouse. Light-environment control such as a quality or quantity in the plant factory system is essential for improving the growth and development of plant species. However, there was little report that the effects of various light qualities provided by LEDs on Ledebouriella seseloides growth under the plant factory system. METHODS AND RESULTS: Ledebouriella seseloides seedlings transplanted at urethane sponge were grown in the plant factory system of a horizontal type with LED artificial lights for 90 days. Yamazaki solution for hydroponic culture of the seedlings was regularly irrigated by the deep flow technique (DFT) system on the culture gutters. Electrical Conductivity (EC) and pH of the solution was recorded at 1.4 ds/m and 5.8 in average, respectively during the experimental period. Number of unfolded leaves, leaf length, shoot fresh and dry weight of the seedlings were three times measured in every 30 days after beginning of the experiment. Blue LEDs, red LEDs, and fluorescent lights inside the plant factory were used as light sources. Conventional fluorescent lamps were considered as a control. In all the treatment, light intensity was maintained at $100{\mu}mol/m^2/s$ on the culture bed. Fresh weight of the seedlings was 3.7 times greater in the treatment with the mixture radiation of fluorescent light and blue+red LEDs (1:3 in energy ratio; Treatment FLBR13) than in fluorescent light treatment (Treatment FL). In FLBR13 treatment, dry weight per seedling was two times greater than in FL or BR11 treatment of blue+red LEDs (1:3 in energy ratio; Treatment BR11) during the culture period. Increasing in number of unfolded leaves was also significantly affected by the FLBR13 treatment comparing with BR11 treatment. CONCLUSION(S): Hydroponic culture of Ledebouriella seseloides seedlings was successfully achieved in the plant factory system with mixture lights of blue, red LEDs and fluorescent lights. Shoot growth of the seedlings was significantly promoted by the FLBR13 with the mixture radiation of fluorescent light, blue, and red LEDs under 1:3 mixture ratio of blue and red LEDs during the experimental period compared to conventional light conditions.
Dopamine(DA), norepinephrine(NE), and epinephrine(E) belong to a class of neurotransmitters known as catecholamine (CA) which are synthesized and secreted by mammalian brain and adrenal medulla. CA regulate several behavior patterns connected with breeding, and regulate GnRH-gonadotropin hormone axis' vitality between hypothalamus-pituitary gland linking with reproduction freeze. The present study examined effects of sex steroid hormone on the transcriptional activities of CA biosynthesis enzymes, tyrosine hydroxylase(TH), dopamine $\beta$ -hydroxylase(DBH), and phenylethaolamine-N-methyl transferase(PNMT). Mature female rats were ovariectomized(OVX) and implanted with 17 $\beta$-estradiol(E$_2$: 500 $\mu\textrm{g}$/ml) or sesame oil. Forty-eight hours after implantation all the animals were sacrificed. Total RNAs were extracted immediately and were applied to semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR). The expression level of TH was appeared by hypothalamus > SNc> adrenal medulla orders in OVX+Oil group, and by SNc > hypothalamus) adrenal medulla orders in OVX+E$_2$ group. Treatment with E$_2$ significantly increased TH expression in SNc and adrenal medulla but in hypothalamus, the reduced TH expression was observed. The expression level of DBH was appeared by adrenal medulla > SNc > hypothalamus orders in OVX+Oil group and in OVX+E$_2$ group. Administration of E$_2$ significantly reduced DBH expression in SNc, and increased in adrenal medulla. Two cDNA products, large(PNMT1) and small(PMNTs) species of 110bp difference, were amplified in SNc and hypothalamus, but only PNMTs was observed in adenal medulla. The PNMTs expression level was in the order of adrenal medulla > hypothalamus > SNc in both OVX+Oil and OVX+E$_2$ group. The PNMTs expression in SNc and adrenal medulla was significantly increased byE$_2$. The present report demonstrated that estrogen effects on transcriptional activities for CA biosynthethic enzymes were tissue specific in adrenal medulla as well as different region of brain. These results suggest that it might be crucial relationship between the type of estrogen receptor and CA enzyme gene expression.
Kim, Mi Young;Cho, Kang Hee;Chun, Jae An;Park, Seo Jun;Kim, Se Hee;Lee, Han Chan
Journal of Plant Biotechnology
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v.44
no.4
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pp.401-408
/
2017
Herein, we report the meristem-tip culture from dormant buds of grape 'Kyoho' single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2-5 leaf primordia, and 1-4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at $4^{\circ}C$). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.
This is preliminary report on anthrax epidemic occurred in an island with about 100 residents. Since 1982 there had been sudden deaths among all kinds of domestic animals including cattle, dogs, ducks, chicken and goat but only a few among cats in an isolated island about three hours distance away by ferry boat from Mokpo city. From 1986 through 1988 nine human deaths and four patients occurred, which made the government intervene for investigation on June 25 1988. The epidemiological investigation consisted of interview survey and medical examination, medical record analysis, laboratory work to isolate the pathogens under the direction of hypothesis derived from the study and further confirmation of the pathogens by international institute. The summarized results are as followings: 1. According to the interview survey there were many deaths among domestic animals usually in cold and dry season such as January through March and September through November; 36 heads of cattle leaving one head, more than 40 hogs(all), hundreds of chicken leaving few alive, goats that had taken home from mountain and two or three cats out of around 40 had sudden deaths from 1982 till 1985, when the residents stopped to purchase and take them into the island anymore. Also there were eleven persons who had experienced the similar syndrome complex to those of admitted and expired patients and four of them revealed typical chest X-ray findings; from one of these four patients(Rho) B. anthracis is isolated. 2. Medical record on patients who had been admitted, showed common characteristics of the disease course. On admission they had either gastrointestinal or upper respiratory infection symptoms which invariably progressed to septicemic nature with pulmonary interstitial infiltration and mediastinal widening/bulging, and then to deadly acute respiratory distress syndrome. At the end stage chest X-ray revealed multiple bullous emphysema. One of another characteristics was oral ulceration with bleeding occurred in about 50% of the patients. Laboratory test results in common were leukocytosis with left shift and abnormal liver and kidney functions, particularly at the later stage of the illness. 3. Epidemiological characteristics was striking in that both mortality and incidence rates were high: the mortality rate was 8.7% average, male being three times higher than females but there was no distictive clustering by age group. The incidence rate for both sexes was 28.2% and there was no sex difference although a tendency of higher incidence among older ages was noticed. The highest mortality and incidence were observed in Won village where the first death of animal occurred and with the highest frequency among three villages of the island. 4. Among twelve bacilli species isolated from various specimens, two strains, one from patient and the other from soil where the recently died cow is hurried, were confirmed as B. anthracis by Pasteur Institute and CDC of USA(strain from soil). CDC reported that the strain did not produce capsule in bicarbonate media but reacted with the bacteriophage and one of five sera taken from the patients. Mode of transmission as well as incubation period of the agent has not been established yet, which needs further investigation in relation to the antigenic structure of the variant when it is confirmed.
The release of hazardous waste materials into the environment poses serious risks in humans and ecosystems. The risk assessment of environmental pollutants including hazardous chemicals requires a comprehensive measurement of hazard and exposure of the chemicals that can be achieved by toxicity evaluation using a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed . In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 bp encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 Up of mRNA was consistently amplified in all the treated samples including control There were no significant differences in the TH expression level regardless of treating concentrations (1∼5,000 ppb) and time (0∼48 hr) The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue - specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard ofvarious environmental pollutants.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.29
no.5
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pp.272-281
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2003
Nontraditional or alternative medicine is becoming an increasingly attractive approach for the treatment of various inflammatory disorders and cancers. Curcumin is the major constitute of turmoric powder extracted from the rhizomes of the plant Curcuma longa. Resveratrol is a phytoalexin present in grapes and a variety of medicinal plants. In this report, We investigated the effect of curcumin and resveratrol on regulatory protein of cell cycle, induction of apoptosis and MMP activity. Treatment with 75 M curcumin for 24 hrs produced morphological changing in HN-4 cells. Curcumin and resveratrol inhibited the cellular growth in HN-4 cells. Inhibition of cell growth was associated with down-regulation of cell cycle regulatory proteins. Curcumin-induced caspase-3 activation and Bax degradation were dose-dependent with a maximal effect at a concentration of 100 M. The elevated caspase-3 activity in curcumin treated HN-4 cells are correlated with down-regulation of survivin and cIAP1, but not cIAP2. Curcumin induced a dose-dependent increase of cytochrome c in the cytosol. Curcumin induced-apoptosis was mediated through the release of cytochrome c. In addition, curcumin-induced apoptosis was caused by the generation of reactive oxygen species, which was prevented by antioxidant N-acetyl-cysteine (NAC). Cotreatment with NAC markedly prevented cytochrome c release, Bax cleavage and cell death. Also resveratrol-induced apoptosis was preceded by down-regulation of the anti-apoptotic Bcl-2, cIAP1, and caspase-3 activity. However, resveratrol-induced apoptosis was not prevented by antioxidant NAC. In addition, HN-4 cells release basal levels of MMP2 when cultured in serum-free medium. Treatment of the cells with various concentrations of PMA for 24 hr induced the expression and secretion of latent MMP9 as determined by gelatin zymography. HN-4 cells were treated with various concentrations of curcumin and resveratrol in the presence of 75 nM PMA, and MMP2 and 9 activities were inhibited by curcumin and resveratrol. These findings have implications for developing curcumin-based anticancer and anti-inflammation therapies.
Background: Sperm DNA damage is underlying aetiology of poor implantation and pregnancy rates but also affects health of offspring and may also result in denovo mutations in germ line and post fertilization. This may result in complex diseases, polygenic disorders and childhood cancers. Childhood cancer like retinoblastoma (RB) is more prevalent in developing countries and the incidence of RB has increased more than three fold in India in the last decade. Recent studies have documented increased incidence of cancers in children born to fathers who consume alcohol in excess and tobacco or who were conceived by assisted conception. The aetiology of childhood cancer and increased disease burden in these children is lin ked to oxidative stress (OS) and oxidative DNA damage( ODD) in sperm of their fathers. Though several antioxidants are in use to combat oxidative stress, the effect of majority of these formulations on DNA is not known. Yoga and meditation cause significant decline in OS and ODD and aid in regulating OS levels such that reactive oxygen speues meditated signal transduction, gene expression and several other physiological functions are not disrupted. Thus, this study aimed to analyze sperm ODD as a possible etiological factor in childhood cancer and role of simple life style interventions like yoga and meditation in significantly decreasing seminal oxidative stress and oxidative DNA damage and thereby decreasing incidence of childhood cancers. Materials and Methods: A total of 131 fathers of children with RB (non-familial sporadic heritable) and 50 controls (fathers of healthy children) were recruited at a tertiary center in India. Sperm parameters as per WHO 2010 guidelines and reactive oxygen species (ROS), DNA fragmentation index (DFI), 8-hydroxy-2'-deoxy guanosine (8-OHdG) and telomere length were estimated at day 0, and after 3 and 6 months of intervention. We also examined the compliance with yoga and meditation practice and smoking status at each follow-up. Results: The seminal mean ROS levels (p<0.05), sperm DFI (p<0.001), 8-OHdG (p<0.01) levels were significantly higher in fathers of children with RB, as compared to controls and the relative mean telomere length in the sperm was shorter. Levels of ROS were significantly reduced in tobacco users (p<0.05) as well as in alcoholics (p<0.05) after intervention. DFI reduced significantly (p<0.05) after 6 months of yoga and meditation practice in all groups. The levels of oxidative DNA damage marker 8-OHdG were reduced significantly after 3 months (p<0.05) and 6 months (p<0.05) of practice. Conclusions: Our results suggest that OS and ODD DNA may contribute to the development of childhood cancer. This may be due to accumulation of oxidized mutagenic base 8OHdG, and elevated MDA levels which results in MDA dimers which are also mutagenic, aberrant methylation pattern, altered gene expression which affect cell proliferation and survival through activation of transcription factors. Increased mt DNA mutations and aberrant repair of mt and nuclear DNA due to highly truncatred DNA repair mechanisms all contribute to sperm genome hypermutability and persistant oxidative DNA damage. Oxidative stress is also associated with genome wide hypomethylation, telomere shortening and mitochondrial dysfunction leading to genome hypermutability and instability. To the best of our knowledge, this is the first study to report decline in OS and ODD and improvement in sperm DNA integrity following adoption of meditation and yoga based life style modification.This may reduce disease burden in next generation and reduce incidence of childhood cancers.
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