The residues of abamectin 1.8% EC, resisted for control of pine wood nematode, Bursaphelenchus xylophilus in pine tree were surveyed in tissue of Pinus thunbergii and P. koraiensis after injection of a liquid formulation. Limits of detection of abamectin in tissue of P. thunbergii were $0.05\;mg\;kg^{-1}$ and mean recoveries at $0.5\;mg\;kg^{-1}$ trunk injection were 90.9% and 93.1% respectively in stem and trunk of P. thunbergii. Abamectin 1.8% EC, trunk injected in 15 m height P. thunbergii were detected in all stem (edible part of carrier insect of pine wood nematode, Monochamus alternatus) from 0.29 to $0.73\;mg\;kg^{-1}$ after 150 days injection. Amount of residue of abamectin 1.8% EC in 12.6 cm mean breast height diameter (DBH) P. thunbergii were variable depending on individual trees in natural forest. Amount of residues in lower and middle part of trunk were reduced with the passage of the injection time. In upper part of trunk were detected $1.84\;mg\;kg^{-1}$ on 30 days after injection however $0.65\;mg\;kg^{-1}$ on 15 days after injection and under detection limit on 100 and 180 days after injection in P. thunbergii. Bottom and middle parts of crown were detected $0.183\;mg\;kg^{-1}$ and $0.173\;mg\;kg^{-1}$ respectively on 180 days after injection in P. thunbergii. Mean residues of abamectin in crown and trunk were $0.80\;mg\;kg^{-1}$ and $0.30\;mg\;kg^{-1}$ on 170 days after trunk injection in 20 cm DBH and 9 m height P. koraiensis. Mean residues of abamectin in crown and trunk were $0.67\;mg\;kg^{-1}$ and $0.36\;mg\;kg^{-1}$ on 170 days after trunk injection in 15 cm DBH and 6 m height P. koraiensis.
A total of 15 different residues of polycyclic aromatic hydrocarbons (PAHs) in each 20 samples of Pacific oysters, dried laver and rockfish obtained from seafood markets were analyzed. The prevalence of samples in which more than one PAH residues were found was 75% in oyster, 35% in rock fish hepatopancreas, 0% in rockfish muscle and laver, respectively. To estimate factors contributing to this residue level difference among organisms, tissue concentrations were analyzed after exposing three organisms to phenanthrene, a representative PAH, with concentration of 0.01 or $0.1{\mu}g/mL$ for 2 weeks. Phenanthrene levels after exposure were higher in the oyster digestive gland, laver and rockfish hepatopancreas, but were lower in the oyster whole meat or rockfish muscle. This finding disproved that any close relationship between the residue difference of market samples and concentrating properties of PAHs. The second possible factor analyzed was total lipid contents in the three organisms. Although higher lipid level in hepatopancreas of rockfish may contribute accumulation of PAH residues in the rockfish, lipid factor did not affect to PAH levels in other organism samples. Activity of 7-ethyoxyresorufin-O-deethylase (EROD), a kind of cytochrome $P_{450}$ enzyme, was measured to evaluate the eliminated amount of PAHs through metabolism. The higher EROD activity in rockfish, compared to that in oyster, was likely to contribute to the lower PAH residues in the rockfish. More factors, such as different exposure history, organisms' ability to escape, ingestion through prey organisms, and post-harvest loss, should be studied in the future.
Kim, In-Wha;Lee, Sang-Hoon;Kim, Young-Min;Jung, Sung-Youb;Kwon, Se-Chang;Lee, Gwan-Sun;Chung, Suk-Jae;Shim, Chang-Koo
Journal of Pharmaceutical Investigation
/
v.31
no.2
/
pp.89-94
/
2001
The pharmacokinetics of recombinant human granulocyte colony stimulating factor (rhG-CSF) following intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) administration of HM1041l-lyo and HM10411-liq (lyophilized and liquid formulations of rhG-CSF, recently under development by Hanmi Pharmaceutical Company) were studied in rats, and compared with that of Filgrastim (conventional formulation of rhG-CSF on market). The plasma concentration of rhG-CSF was quantified using a specific ELISA. The pharmacokinetic parameters of rhG-CSF, after i.v., i.m. and s.c. administration of Filgrastim, HM1041l-lyo and HM1041l-liq to rats at a rhG-CSF dose of $10\;{\mu}g/kg$, were almost identical among the three formulations. No dose-dependency was observed in the pharmacokinetic parameters of rhG-CSF following i.v. administration in the dose range of $5{\sim}100\;{\mu}g/kg$. rhG-CSF, after i.v. administration of the three preparations at a dose of $10\;{\mu}g/kg$ to rats, was detected at low levels in all of the body tissues with highest tissue/plasma ratio of $0.46{\sim}0.51$ for the kidney at 30 min after the administration. The pharmacokinetics of rhG-CSF, after i.v. administration to mice at a dose of $10\;{\mu}g/kg$, were comparable among the three formulations. In conclusion, HM10411-lyo and HM10411-liq exhibited similar pharmacokinetics for rhG-CSF with Filgrastim regandless of animal species. Considering the fact that HM10411 series, contrary to Filgrastim, are proteins lacking a methionine residue, the methionine moiety in rhG-CSF molecule does not appear to influence the pharmacokinetics of the protein significantly.
Proceedings of the Plant Resources Society of Korea Conference
/
2010.05a
/
pp.17-17
/
2010
Ethylene, known as a stress hormone regulate wide developmental processes including germination, root hair initiation, root and shoot primordial formation and elongation, leaf and flower senescence and abscission, fruit ripening. The acceleration of ethylene biosynthesis in plant associated with environmental and biological stresses. 1-Aminocycloprophane-1-carboxlyate deaminase(ACCD) is an enzyme that cleaves ACC into and ammonia, a precursor of the plant hormone ethylene. Plant growth-promoting rhizobacteria (PGPR) having ACCD can decrease endogenous ACC level of tissue, resulting in reduced production of ethylene in plants. ACC deaminse was a key enzyme for protect stressed plants from injurious effects of ethylene. ACCD gene was encoded from Pseudomonas flourescens, PGPR and was cloned in Escherichia coli. We expressed the recombinant ACCD(rACCD) containing 357 amino acids with molecular weight 39 kDa that revealed by SDS-PAGE and western blot. The rACCD was purified by Ni-NTA purification system. The active form of rACCD having enzyme activity converted ACC to a-ketobutyrate. The optimal pH for ACC deaminase activity was pH 8.5, but no activity below pH 7.0 and a less severe tapering activity at base condition resulting in loss of activity at over pH 11. The optimal temperature of the enzyme was $30^{\circ}$ and a slightly less severe tapering activity at 15 - 30$^{\circ}$, but no activity over $35^{\circ}$. P. flourescens ACC deaminase has a highly conserved residue that plays in allowing substrate accessibility to the active sites. The enzymatic properties of this rACCD will provide an important reference for analysis of newly isolated ACCD and identification of newly isolated PGPR containing ACCD.
The residue depletion of ampicillin was investigated in the olive flounder (Paralichthys olivaceus), rockfish (Sebastes schlegeli), and red sea bream (Pagrus major) after 5 days treatment with medicated feed at a dose of 100 mg/kg bw/day. Fishes were sampled for muscle on 1st, 2nd, 3rd, and 4th day after treatment. Ampicillin concentrations were determined by high performance liquid chromatography after SPE column extraction. The recovery rates of ampicillin in muscle samples ranged 94-98% and 83-88% for the concentration of 0.05 mg/kg and 0.1 mg/kg, respectively. Ampicillin concentrations detected on 1st day after treatment were 0.143, 0.138, and 0.187 mg/kg in the muscle of olive flounder, rockfish, and red sea bream, respectively. After a withdrawal of 3 days, muscle concentrations were 0.016, 0.012, and 0.021 mg/kg in the olive flounder, rockfish, and red sea bream, respectively. Ampicillin was not detectable in muscle samples on 4 days following withdrawal of the medicated feed. From results of the present study, a withdrawal period of ampicillin is proposed on 5 days after 5 days treatment with medicated feed at a dose of 100 mg/kg bw/day to avoid the presence of excessive residues of the edible muscles of olive flounder, rockfish, and red sea bream.
Differential display based on PCR was employed to identify genes expressed during tuber-developing stage of potato (Solanum tuberosum L. cv. Irish Cobbler). An eukaryotic initiation factor 5A (eIF-5A) clone isolated from a cDNA library constructed with developing micro-tuber using a probe of PCR fragment. We isolated three positive clones and ore of them contained open reading frame. This clone revealed high sequence similarity to tomato eIF 5A cDNA. At the DNA level, there is 94.8% identity with the tomato eIF-5A4, whereas at the protein level there is a high identity with 97.5%. The potato eIF 5A clone is 716 bp in length and contains a single open reading frame from 57 to 539 bp, a 56 bp 5'-untranslated region and a 177 bp 3'-untranslated region. The deduced protein composed of 160 amino acid residues, with a predicted molecular mass of 17.4 kD and an estimated pl of 5.5. The sequence of 12 (STSKTGKHGHAK) amino acids among eIF-5A proteins is perfectly conserved from yeast to human. That sequence in potato eIF-5A protein is also conserved at position 46 to 57 amino acid. This region embeds the post-translational modification site of the lysine residue (at the seventh K) to hypusine that is crucial to eIF-5A activity. The northern blot analysis of eIF5A has shown abundant expression, mainly in flower organs (stamen, ovary, petal, sepal), fruit and stolen.
The adoption of legume cover crops in no-tillage system can contribute to improve soil fertility by providing several benefits, including reduction in soil erosion, suppression of weed growth and N supply to subsequent crops. We conducted a field study to investigate the effect of cover crops and nitrogen fertilization rates on yield and nitrogen use efficiency of waxy corn (Zea mays L.) in no-tillage upland field. Two legume cover crops, hairy vetch (Vicia villosa Roth) and crimson clover (Trifolium incarnuturn L.) were mechanically terminated with roller in early June. For each cover crop treatment, nitrogen (N) fertilizer was applied at three different rates (145, 72.5 and $0kg\;N\;ha^{-1}$). The growth and yield characteristics of corn were significantly affected by the N fertilization rates in crimson clover plots, which suggest N mineralization from the cover crop residue was not sufficient. In contrast, N fertilization rates had no significant effect on growth and yield of corn in hairy vetch plots, indicating that the amount of N released from the cover crop is large enough to meet most of the N requirement of corn. However, the application of N fertilizer in hairy vetch cover plots resulted in slight increase of crop yield, though not statically significant, and high levels of N concentration in corn plant tissue possibly due to luxury consumption of N. Organic residues on the soil surface in hairy vetch cover plots had substantial amounts of N after harvest, ranging from 100 to $116kg\;N\;ha^{-1}$, which is presumably retained during winter season and released by microbial mineralization in subsequent year. The highest nitrogen yield efficiency was achieved in the plot with hairy vetch cover and no N fertilizer application, followed by the plot with hairy vetch cover and $72.5kg\;N\;ha^{-1}$ fertilization rate. In conclusion, hairy vetch showed better performance in corn productivity as compared with crimson clover. In addition, it was concluded that the application of N fertilizer between 0 and $72.5kg\;N\;ha^{-1}$ in combination with hairy vetch cover crop might be most efficient for corn yield under no-tillage system with climatic and soil characteristics similar to those of the experimental site.
PU.1, a tissue-specific transcription activator, binds to a purine-rich sequence(5'-GAGGAA-3') called PU box. The PU.1 cDNA consists of an open reading frame of 816 nucleotides coding for 272 amino acids. The amino terminal end is highly acidic, while the carboxyl terminal end is highly basic. Transcriptional activation domain is located at the amino terminal end, while DNA binding domain is located at the carboxyl terminal end. Activation of PU.1 transcription factor is supposed to be accomplished by the phosphorylation of serine residue(s). There exist 22 serines in the PU.1. Five(the 41, 45, 132$.$133, and 148th) of the serines(plausible phosphorylation site by casein kinase II), are the primary targets of interest in elucidating the molecular mechanism(s) of the action of the PU.1 gene. In this study, PU.1 cDNA coding for the five serine residues(41th AGC, 45th AGC, 132$.$133th AGC$.$TCA, and 148th TCT), was mutated to alanine codon(41th GCC, 45th GCC, 132$.$133th GCC$.$GCA, and 1481h GCT), respectively, by Splicing-Overlapping-Extension(SOE) using Polymerase Chain Reaction(PCR). And each mutated cDNA fragments was ligated into pBluescript KS+ digested with HindIII and Xba I, to generate mutant clones named pKKS41A, pRKS45A, pMKS132$.$133A, and pMKS148A. The clones will be informative to study the "Structure and Function" of the immu-nologically important gene, PU.1.
The residue depletion of amoxicillin was investigated in the olive flounder (Paralichthys olivaceus), rockfish (Sebastes schlegeli), and red sea bream (Pagrus major) after 7 days treatment with medicated feed at a dose of 400 mg/kg bw/day. Fishes were sampled for muscle on 1st, 2nd, 3rd, 4th, and 5th day after treatment. Amoxicillin concentrations were determined by high performance liquid chromatography with fluorescence detector. The recovery rates of amoxicillin in muscle samples ranged 84.3-101.3% and 75.0-91.5% for the concentration of 0.05 mg/kg and 0.1 mg/kg, respectively. Amoxicillin concentrations detected on 1st day after treatment were 0.137, 0.131, and 0.172 mg/kg in the muscle of olive flounder, rockfish, and red sea bream, respectively. After a withdrawal of 3 days, muscle concentrations were 0.012, 0.010, and 0.017 mg/kg in the olive flounder, rockfish, and red sea bream, respectively. Amoxicillin was not detectable in muscle samples on 4 days following withdrawal of the medicated feed. From results of the present study, a withdrawal period of amoxicillin is proposed on 4 days after 7 days treatment with medicated feed at a dose of 400 mg/kg bw/day to avoid the presence of excessive residues of the edible muscles of olive flounder, rockfish, and red sea bream.
Due to its biofilm formation and colonization of the osteocyte-lacuno canalicular network (OLCN), Staphylococcus aureus (S.aureus) implant-associated bone infection (SIABI) is difficult to cure thoroughly, and may occur recurrently subsequently after a long period dormant. It is essential to explore an alternative therapeutic strategy that can eradicate the pathogens in the infected foci. To address this, the polymethylmethacrylate (PMMA) bone cement and Fe3O4 nanoparticles compound cylinder were developed as implants based on their size and mechanical properties for the alternative magnetic field (AMF) induced thermal ablation, The PMMA mixed with optimized 2% Fe3O4 nanoparticles showed an excellent antibacterial efficacy in vitro. It was evaluated by the CFU, CT scan and histopathological staining on a rabbit 1-stage transtibial screw model. The results showed that on week 7, the CFU of infected soft tissue and implants, and the white blood cells (WBCs) of the PMMA+2% Fe3O4+AMF group decreased significantly from their controls (p<0.05). PMMA+2% Fe3O4+AMF group did not observe bone resorption, periosteal reaction, and infectious reactive bone formation by CT images. Further histopathological H&E and Gram Staining confirmed there was no obvious inflammatory cell infiltration, neither pathogens residue nor noticeably burn damage around the infected screw channel in the PMMA+2% Fe3O4+AMF group. Further investigation of nanoparticle distributions in bone marrow medullary and vital organs of heart, liver, spleen, lung, and kidney. There were no significantly extra Fe3O4 nanoparticles were observed in the medullary cavity and all vital organs either. In the current study, PMMA+2% Fe3O4+AMF shows promising therapeutic potential for SIABI by providing excellent mechanical support, and promising efficacy of eradicating the residual pathogenic bacteria in bone infected lesions.
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