Resveratrol (RES) and genistein (GEN) are the dietary natural products known to possess chemopreventive property and also the ability to repair DNA damage induced by mutagens/carcinogens. It is believed that the therapeutic activity of these compounds could be primarily due to their interaction with nucleic acids but detailed reports are not available. We here explore the interaction of these drugs with nucleic acids considering DNA and RNA as a potential therapeutic target. The interaction of RES and GEN has been analysed in buffered solution with DNA [saline sodium citrate (SSC)] and RNA [tris ethylene diammine tetra acetic acid (TE)] using UV-absorption and Fourier transform infrared (FTIR) spectroscopy. The UV analysis revealed lesser binding affinity with nucleic acids at lower concentration of RES (P/D = 5.00 and 10.00), while at higher drug concentration (P/D = 0.75, 1.00 and 2.50) hyperchromic effect with shift in the ${\lambda}_{max}$ is noted for DNA and RNA. A major RES-nucleic acids complexes was observed through base pairs and phosphate backbone groups with K = $35.782\;M^{-1}$ and K = $34.25\;M^{-1}$ for DNA-RES and RNA-RES complexes respectively. At various concentrations of GEN (P/D = 0.25, 0.50, 0.75, 1.00 and 2.50) hyperchromicity with shift in the ${\lambda}_{max}$ from 260 $\rightarrow$ 263 om and 260 $\rightarrow$ 270 nm is observed for DNA-GEN and RNA-GEN complexes respectively. The binding constant (from UV analysis) for GEN-nucleic acids complexes could not be obtained due to GEN absorbance overlap with that of nucleic acids at 260 nm. Nevertheless a detailed analysis with regard to the interaction of these drugs (RES/GEN) with DNA and RNA could feasibly be understood by FTIR spectroscopy. The NH band of free DNA and RNA which appeared at $3550-3100\;cm^{-1}$ and $3650-2700\;cm^{-1}$ shifted to $3450-2950\;cm^{-1}$ and $3550-3000\;cm^{-1}$ in DNA-RES and RNA-RES complexes respectively. Similarly shifts corresponding to $3650-3100\;cm^{-1}$ and $3420-3000\;cm^{-1}$ have been observed in DNA-GEN and RNA-GEN complexes respectively. The observed reduction in NH band of free nucleic acids upon complexation of these drugs is an indication of the involvement of the hydroxyl (OH) and imino (NH) group during the interaction of the drugs and nucleic acids (DNA/RNA) through H-bonded formation. The interaction of RES and GEN with bases appears in the order of G $\geq$ T > C > A and A > C $\geq$ T > G. Further interaction of these natural compounds with DNA and RNA is also supported by changes in the vibrational frequency (shift/intensity) in symmetrical and asymmetrical stretching of aromatic rings of drugs in the complex spectra. No appreciable shift is observed in the DNA and RNA marker bands, indicating that the B-DNA form and A-family conformation of RNA are not altered during their interaction with RES and GEN.
Objective : The purpose of this study was to evaluate the effects of alcohol on neurocognitive function, psychomotor performance and subjective response in healthy Korean adults with different ALDH2 genotypes. Method : A total of 24 males, half with active $ALDH2^*1/2^*1$ and the other with inactive $ALDH2^*1/2^*2$, was selected through genotyping using restriction fragment length polymorphism. In a double-blind, placebo-controlled cross-over design, each subject consumed 0.5g/kg dose of alcohol, given as a mixture of 40% vodka and orange juice, and placebo(orange juice) on two separate occasions on an average of weekly intervals. The blood alcohol concentrations(BACs) were measured using a breath analyzer at baseline and at 30, 60 minutes after drinking. P300s were measured at baseline and at 30 minutes after alcohol and placebo intake. Vital signs and psychomotor performance[Critical Flicker Fusion Threshold(CFFT), Choice Reaction Time (CRT), Digit Symbol Substitution(DSS)] were measured at baseline and at 60 minutes after alcohol and placebo intake. Subjective responses were measured at the end of the study. The statistical analysis focused on whether there were any differences between groups with different ALDH2 genotypes. Results : The major results are as follows. 1) BACs in the inactive group were overall equivalent to those in the active group. Only in terms of time, BACs were significantly higher overall at 30 minutes than at 60 minutes after alcohol intake. 2) Pulse rates were significantly increased after alcohol intake compared with placebo, and the increase was greater in the inactive than in the active group. 3) P300 latencies in leads Fz(frontal), Cz(cental) and Pz(parietal) were significantly increased after alcohol intake compared to placebo, and the increase was greater in the inactive than in the active group. P300 amplitudes in leads Cz and Pz were significantly decreased overall after alcohol intake compared to placebo. 4) Compared with placebo, alcohol produced significant effect on the psychomotor performance : impairment in the inactive group, improvement in the active group. 5) Compared with placebo, alcohol significantly induced a negative or an intense effect on the subjective responses in the inactive group, but little negative and even a somewhat positive effect in the active group. Conclusions : These results suggest that ALDH isozyme variance might be an important factor to determine the effects of acute dose of alcohol on the various psychobehavioural functions and also to determine the alcohol use pattern and to predict the future development of alcohol overuse and/or abuse.
As facilities and equipments for learning activities in college campuses are handled by mass public, their contact surfaces may function as major routes of cross-infection of microbial pathogens. However, unlike items in cafeteria which is the typical target for campus hygiene, those surfaces are not under regular surveillance or sanitary maintenance. In this study, I made a quantitative comparison of the risk of being exposed to microbial pathogens from use of learning facilities such as classrooms and library to the risk from use of cafeteria, for about 1,500 students in a college. Regarding total coliforms as surrogate model of bacterial pathogens, exposure rates were estimated for each item in learning facilities and cafeterias by devising deterministic exposure algorithms based on bacterial abundance, contract rates and transfer rates. The exposure rate in cafeterias was 1.0 CFU/day while learning facilities imposed the rate of 0.5 CFU/day, which reaches a half of the exposure rate in cafeterias. However, 70% of students were exposed more in learning facilities than cafeteria because individuals had different frequencies in using cafeteria. Based on the results, some human-contact surfaces of learning facilities, including elevator buttons, may require regular sanitary maintenance. An efficient sanitary maintenance considering seasonality in diversity of pathogens involved with cross-infections is suggested besides improvement of personal hygiene among students.
The correlations between environmental influences on microorganisms in soil and its effects on disease development in ginseng field were studied to obtain some useful data for increasing ginseng production and effective preventive measures against the root rot caused by soil-borne pathogens. The diseased replanted ginseng fields were selected as the diseased field and the healthy plot in first planted field selected as control in three major Korean ginseng producing areas such as Kumsan, Goesan and Poonggi. The physicochemical characteristics of the soil were analyzed and microorganisms susceptible for root rot of ginseng, such as Fusarium spp. and general fungi were investigated for their population density in various soil conditions. Correlations between soil microbial populations and environmental factors were investigated. The numbers of Fusarium spp. propagules were abundant in fall in both soil conditions. The numbers of Fusarium spp. were 1.9 to 2.6 times higher in replanted field than first planted field except Goesan area. Relative ratio of Fusarium spp. to total fungi propagules in replanted field was 1.6 times higher in replanted field than first planted field indicating higher numbers of Fusarium spp. distributed in replanted field of soil. The numbers of propagules of total fungi were increased in June and July and there was no sensitive variation according to the temperature. There was no significant difference in vertical distributions of total fungi according to soil depth, while the total fungi were abundant in the surface layer and $10{\sim}15\;cm$ layer. The contents of organic matter and phosphate in healthy field were somewhat high, and phosphate/organic matter ratio and Mg contents were high in diseased field. All of the soils showed a weak acidic pH of 4.5 to 5.7. Soil moisture contents were increased during winter season, but did not show any significant changes during the growing periods, showing 24.6% in healthy field and 19.5% in diseased field respectively. Soil temperature was the highest in July and August and the lowest in January and February.
Giardia intestinalis infections arise primarily from contaminated food or water Zoonotic transmission is possible, and at least 7 major assemblages including 2 assemblages recovered from humans have been identified. The determination of the genotype of G. intestinalis is useful not only for assessing the correlation of clinical symptoms and genotypes, but also for finding the infection route and its causative agent in epidemiological studies. In this study, methods to identify the genotypes more specifically than the known 2 genotypes recovered from humans have been developed using the intergenic spacer (IGS) region of rDNA. The IGS region contains varying sequences and is thus suitable for comparing isolates once they are classified as the same strain. Genomic DNA was extracted from cysts isolated from the feces of 5 Chinese, 2 Laotians and 2 Koreans infected with G. intestinalis and the trophozoites of WB, K1, and GS strains cultured in the laboratory, respectively. The rDNA containing the IGS region was amplified by PCR and cloned. The nucleotide sequence of the 3' end of IGS region was determined and examined by multiple alignment and phylogenetic analysis. Based on the nucleotide sequence of the IGS region, 13 G. intestinalis isolates were classified to assemblages A and B, and assemblage A was subdivided into A1 and A2. Then, the primers specific to each assemblage were designed, and PCR was peformed using those primers. It detected as little as 10 pg of DNA, and the PCR amplified products with the specific length to each assemblage (A1, 176bp; A2, 261 bp; B, 319 bp) were found. The PCR specific to 3 assemblages of G. intestinalis did not react with other bacteria or protozoans, and it did not react with G. intestinalis isolates obtained from dogs and rats. It was thus confirmed that by applying this PCR method amplifying the IGS region, the detection of G. intestinalis and its genotyping can be determined simultaneously.
This study was carried out to investigate the distribution status and UI(urbanization index) of the naturalized plants and exotic plants. For this purpose 7 cities and counties in the northern South Jeolla Province were selected and investigations were conducted on a total of 14 sites, two sites from each city or county, during the period of March to November 2006. The exotic plants were identified as 111 taxa; 26 families, 79 genera, 105 species, and 6 varieties. As for urbanization index, Hampyeong County had the highest level of urbanization(21.95%) and Gokseong County the lowest level of urbanization(16.38%). Naturalized plants classified as family were as follows: Compositae was 36 kinds(32.43%) Graminae 17 kinds(15.31%) and Leguminosae 7 kinds(6.31%). Major communities of naturalized plants consisted of 21 communities including Robinia pseudoacacia-Amorpha fruticosa community, Erigeron annuus community, Erigeron canadensis community etc. Invasive naturalized plants consisted of 7 taxa: Paspalum disticum var. indutum, Paspalum distichum, Rumex acetocella, Solanum carolinense, Aster pilosus, Ambrosia artemisiifolia var. elatior and Ambrosia trifida. The study shows that the integrated management of exotic plants on a national scale is needed. It also recommend that a systematic and integrated management of exotic plants in the northern South Jeolla Province is crucial to preserve and protect natural species.
Acidic chitinases from the gizzards of a broiler were purified to homogeneity, using precipitation with $(NH_{4})_{2}SO_{4}$, ion exchanger chromatography, gel filtration, chromatofocusing and hydrophobic interaction chromatography. The enzymes, GAC1 and GAC2, were purified 180- and 194- folds with a recovery of 4.9% and 2.7%, respectively. The molecular mass of GAC1 and GAC2 were 48.2 kDa and 57.8 kDa, respectively. Chromatofocusing resulted in a pI of 3.1 for both enzymes. The purified enzymes were endochitinases that were devoid of ${\beta}-N-acetylglucosaminidase$ and lysozyme activity. Kinetic studies using $[^3H]chitin$ indicate that GAC1 has a $K_m$ and $V_{max}$ of 1.97 mg/ml and 185 mg/mg protein/h, respectively. The GAC2 has a $K_m$ and $V_{max}$ of 0.42 mg/ml and 92.3 mg/mg protein/h, respectively at optimal pH and temperature (pH 5.0 and $60^{\circ}C$). When the pentamer and hexamer of N-acetylglucosamine (GlcNAc) were used as a substrate, the major product by GAC1 was the dimer of GlcNAc with a differential accumulation of the monomer and trimer, depending upon the substrate. However, the GAC2 produced the dimer and trimer in an equal quantity, regardless of the substrate used. The first 9 $NH_2-terminal$ amino acid residues of the purified gizzard chitinase GAC1 and GAC2 shared a 100% homology. The first 25 $NH_2-terminal$ amino acid residues of GAC1 also shared 55-60% homology with animal chitinases and some animal proteins, such as whey protein and oviduct-specific proteins. However, little homology was found with either microbial and plant chitinases, or egg white lysozyme.
Kim, Ji-Woon;Lee, Soon-Young;Joo, So-Hyun;Song, Mi-Ryoung;Shin, Chan-Young
Biomolecules & Therapeutics
/
v.15
no.1
/
pp.16-26
/
2007
Tissue plasminogen activator (tPA) is a serine protease catalyzing the proteolytic conversion of plasminogen into plasmin, which is involved in thrombolysis. During last two decades, the role of tPA in brain physiology and pathology has been extensively investigated. tPA is expressed in brain regions such as cortex, hippocampus, amygdala and cerebellum, and major neural cell types such as neuron, astrocyte, microglia and endothelial cells express tPA in basal status. After strong neural stimulation such as seizure, tPA behaves as an immediate early gene increasing the expression level within an hour. Neural activity and/or postsynaptic stimulation increased the release of tPA from axonal terminal and presumably from dendritic compartment. Neuronal tPA regulates plastic changes in neuronal function and structure mediating key neurologic processes such as visual cortex plasticity, seizure spreading, cerebellar motor learning, long term potentiation and addictive or withdrawal behavior after morphine discontinuance. In addition to these physiological roles, tPA mediates excitotoxicity leading to the neurodegeneration in several pathological conditions including ischemic stroke. Increasing amount of evidence also suggest the role of tPA in neurodegenerative diseases such as Alzheimer's disease and multiple sclerosis even though beneficial effects was also reported in case of Alzheimer's disease based on the observation of tPA-induced degradation of $A{\beta}$ aggregates. Target proteins of tPA action include extracellular matrix protein laminin, proteoglycans and NMDA receptor. In addition, several receptors (or binding partners) for tPA has been reported such as low-density lipoprotein receptor-related protein (LRP) and annexin II, even though intracellular signaling mechanism underlying tPA action is not clear yet. Interestingly, the action of tPA comprises both proteolytic and non-proteolytic mechanism. In case of microglial activation, tPA showed non-proteolytic cytokine-like function. The search for exact target proteins and receptor molecules for tPA along with the identification of the mechanism regulating tPA expression and release in the nervous system will enable us to better understand several key neurological processes like teaming and memory as well as to obtain therapeutic tools against neurodegenerative diseases.
This study was carried out to establish a new breeding scheme which is connected with conventional breeding method and anther culture method. To develop a herbicide resistant and direct seeding rice, $F_1$ plants were subjected to anther culture and regenerated plants from 5 crosses were studied to confirm the introduction of bar gene. After PCR analysis, we selected 227 plants which were carrying herbicide resistance gene (bar) out of 1,508 regenerated plants from anther culture. Among 169 $A_2$ lines carrying herbicide resistant gene from 5 crosses including YR23235 (Dongjin Ds3(Ba $r^{R}$)/ Milyang165), 42 lines that had superior agronomic characters were selected for further research. Among them, YR23235Acp79 which showed herbicide resistance, direct seeding adaptability and superiority in major agronomic characters was named Milyang 204. This breeding scheme proved that the anther culture of $F_1$ plants crossed between transformant and cultivar or transform ant alone could be utilized in breeding programs for a rapid progeny fixation and development of a variety.y.
Lee Ji Won;Kang Han Seung;Gye Myung Chan;Hong Seok Ho;Shin Hyeonsang;Kang Soo Mnn;Lee Sung Eun;Kim Moon Kyoo
Development and Reproduction
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v.8
no.1
/
pp.49-55
/
2004
Aquaporins(AQPs) are a family of transmembrane water channel proteins that are widely distributed in various tissues throughout the body and play a major role in Oanscellular and Oansepithelial water movement. Uterine endometrium undergoes recurrent uterine stromal edema in response to hormonal stimuli, however, the mechanism regulating the fluid transport during the estrous cycle has not been fully understood. To investigate the possible role of AQPs in water movement in uterus during the estrous cycle, expression patterns of AQP -1, -3, -4, -5, -8, and -9 UMh in mouse uterus were analyzed by using semiquantitative reverse transcription- polymerase chain reaction(RT-nR). We employed a combination of laser capture microdissection(LCM) and RT-PCR to examine the expression patterns in specific uterine cell types luminal epithelial cells(LE) and stromal cells(S). Our results showed that the level of AQP-4 mRNA was significantly increased while the level of AQP-3 mRNA was significantly decreased during the proestous through the estrus stage. In addition LCM revealed that AQP-4 and -8 mRNAs were highly expressed in LE compared with S. Taken together, these results suggest that AQPs may have an important function in physiological changes of mouse uterus during the estrous cycle.
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