• Toxicological Research
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• v.23 no.4
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• pp.353-362
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• 2007

Deoxynivalenol (DON) is a common food borne mycotoxin and occurs predominantly in grains such as wheat, barley, oats, etc. DON induces systemic health problems such as loss of appetite, emesis and diarrhea in both human and farm animals. Reliable diagnostic parameters for DON intoxication are needed to prevent deep health impact. In order to establish useful diagnostic parameters, we investigated clinical signs, hematological values, serum biochemical values, gross-, histo- and toxico-pathological findings in B6C3F1 male mice after oral administration of DON (0.83, 2.5 and 7.5 mg/kg) for 8 days. Body weight gain was significantly decreased at the highest dose of DON. Anorexia, ataxia, for crudness and lack of vigor were observed at the highest dose DON group. In hematological values, the numbers of WBC and platelets and hemoglobin content were reduced with decreased neutrophil and monocytes by 7.5 mg/kg DON. Prothrombin time (PT) and activated partial thromboplastin time (aPTT) were prolonged in a dose-dependent manner and the content of fibrinogen was elevated at high dose of DON. Of serum biochemical values, total protein, globulin, BUN, cholesterol and test-osterone were reduced but total bilirubin and albumin/globulin ratio increased. The enzyme activity of alkaline phosphatase was decreased while that of alanine aminotransferase was elevated. Relative organ weights of thymus, seminal vesicle/prostate and testes were dose-dependently reduced but those of liver and left adrenal gland increased with dose dependency. As for pathological findings, atrophy of thymus, seminal vesicle/prostate and testes and submucosal edema and ulceration in stomach and depletion of lymphocytes in thymus cortex were observed. In conclusion, these clinical, hematological, blood biochemical and patholgical parameters obtained in the present studies can be used for diagnosis of DON-mycotoxicosis, especially, low WBC, platelets, protein, BUN and testosterone and delayed prothrombin time can be available as for reliable diagnostic parameters.

• Biomedical Science Letters
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• v.6 no.1
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• pp.11-17
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• 2000

SNAP-25 was first investigated as a neuron-specific protein preferentially expressed in CA3 pyramidal neurons of mouse hippocampus. It is a presynaptic plasma membrane protein in the nerve cell and plays an important role in the synaptic vesicle membrane docking and fusion pathway. We have recently isolated SNAP-25 cDNA from a rat brain cDNA library using a probe of Z2 cDNA. It consisted of 2,101 bp and an open reading frame (ORF) was identified between nucleotides (nt) 209 and 827. The AUG codon (nt 209∼211) was surrounded by CTACCATGG, which corresponded to the consensus sequence of ribosomal binding site. The ORF was terminated by TAA (nt 827∼829) to encode a polypeptide of 206 amino acid residues. The 3'-untranslated region contained two extensive stretches of repeated (CA)28 and (CA)19 at positions 925∼980 and 1645∼1682. It is noteworthy that cysteine residues were clustered in the span of amino acid residues 84∼991 : Cys-Gly-Leu-Cys-Val-Cys-Pro-Cys. Rat SNAP-25 showed 88% and 97% identity in nucleotide sequences to that of human and mouse, respectively. Amino acid sequence of rat SNAP-25 showed 100% identity to that of mouse and human SNAP-21.

• The Korean Journal of Pharmacology
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• v.32 no.1
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• pp.57-66
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• 1996

To investigate properties of Ca-release channel in the reptile skeletal muscle, electrophoretical analysis, purification of RyR, $[^3H]ryanodine$binding study, and $^{45}Ca-release$ were carried out in the SR vesicles prepared from the snake skeletal muscle. The snake SR vesicle has the single high molecular weight protein band on SDS-PAGE, and its mobility was similar with that of rat skeletal SR vesicles. The high molecular weight band on SDS-PACE was found in the $[^3H]ryanodine$ peak fractions $(Fr_{5-7})$ obtained from the purification step of the RyR. Maximal binding site and Kd of the snake SR RyR were 6.36 pmole/mg protein and 17.62 nM, respectively. Specific binding of $[^3H]ryanodine$ was significantly increased by calcium and AMP (P<0.05), but not or slightly inhibited by tetracaine, ruthenium red (5.4%), or $MgCl_2$ (21%). $^{45}Ca-release$ from the SR vesicles loaded passively was significantly increased by the low concentration of calcium $(1{\sim}10{\mu}M)$ and AMP (5 mM)(P<0.05), but significantly decreased by the high concentration $(300{\mu}M)$ of calcium, tetracaine (1 mM), ruthenium red $(10{\mu}M)$, and $MgCl_2$ (2 mM)(P <0.05). From the above results, it is suggested that snake SR vesicles also have the RyR showing the similar properties to those of mammalian skeletal RyR with the exceptions of no or slight inhibition of $[^3H]ryanodine-binding$ by tetracaine, ruthenium red, or $MgCl_2$.

• Parasites, Hosts and Diseases
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• v.33 no.1
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• pp.1-8
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• 1995

Gymnophalkides seoi has drawn medical attentions since the discovery of the first human case and a highly endemic area on a southwestern coastal island of Shinan- gun, Korea. Marine bivalves especially oysters were strongly suspected as the source of infection. In this study the oysters, Crassostrea girns, naturally produced (rom the endemic area were examined whether they contain gymnophallid metacercariae. All of 50 oysters examined were infected with the metacercaviae of a gymnophallid, with the metacercarial density per oyster of 610 on average (2-4,792 in range). Later they were identified as G. seoi by obtaining adult worms from experimental mice. The metacercariae were unencysted, and firmly attached on the mantle surface of the oysters with their oral sucker. In sectioned specimens they were equipped with the ventral pit, a peculiar organ of the genus Gymophalloides, and non-muscular genital pore which was connected dorsally to the seminal vesicle. The seminal vesicle was in a great majority mono-sac. By this study, it has been confirmed that the oyster is a 2nd intermediate host of G. seoi as well as the major source of human infection with this fluke.

• Journal of Pharmaceutical Investigation
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• v.25 no.3
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• pp.249-264
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• 1995

Although liposome has many advantages as a pharmaceutical dosage form, its application in the industrial field has been limited because of some problems such as preparation method, reproducibility, scale-up, stability and sterilization etc. Liposomes prepared by microemulsification method had defined size, narrow size distribution, reproducibility and high entrapment efficiency. For enhancing the stability, the dry form of liposome was recommended. These types of liposome are proliposome and freeze-dried liposome. The liposome must have some properties for preparing of freeze-dried liposome; small size $(50{\sim}200\;nm)$, narrow size distribution and cryoprotectant. In this experiment, the liposomes containing 5-Fluorouracil(5-FU) and its prodrug(pentyl-5-FU-1-acetate; PFA, hexyl-5-FU-1-acetate; HFA) were made with soybean phosphatidylcholine, cholesterol, stearylamine(SA) and dicetyl phosphate(DCP) employing hydration method or microemulsification method using $Microfluidizer^{TM}$. Both or liposome types were MLV and MEL. After preparation, freeze drying and rehydration were performed. In the process of freezing, trehalose(Tr) was added as a cryoprotectant. Their evaluation methods were as follows; entrapment efficiency, mean particle size and size distribution, dissolution test, retain of entrapment efficiency and turbidity after freeze-drying. The results are summarized as belows. The entrapment efficiency of 5-FU was dependent on total lipid concentration and cholesterol content but that of PFA and HFA was decreased when cholesterol was added. When DCP and SA were added, entrapment efficiency was decreased. As the partition coefficient of drug was increased, entrapment efficiency was increased. Under the same condition, entrapment efficiency of MEL is similar to that of MLV. The mean particle size and size distribution of MEL were smaller than those of MLV. Dissolution rates of drug from both liposome types were comparatively similar. Dissolution rates of drugs with serum and liver homogenate were faster than without these material. After preparation of liposome, free drug was removed efficiency by Dowex 50W-X4. When liposome was freeze-dried and then rehydrated in the presence of Tr, characteristics of liposome were maintained well in MEL than MLV. Tr Was used successfully as a cryoprotectant in the process of freeze drying and the optimal ratio of Tr:Lipid was 4:1(g/g).

• Korean Journal of Environmental Biology
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• v.32 no.4
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• pp.353-362
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• 2014

In this study, we investigated the effect of geographical variations on Sargassum thunbergii morphology to make a taxonomic reconsideration about infraspecific taxa in this species. In order to examine the morphological characteristics of S. thunbergii, total 27 matured and morphologically intact thalli were collected from the east, west and south coast in spring 2011. Interestingly, it was observed that the species populations on the west coast were characterized by short, thin and coarse thallus, and soft texture. However, the populations on the east coast showed thicker thallus, larger leaf and vesicle, and tougher texture. Thallus height of S. thunbergii was found to be similar at both east and west coast. Further, the height of the thallus and lateral branch of the species populations residing south coast were highest whereas the size of leaf and vesicle are shortest, in comparison with east and west coast species. Although morphological characteristics of the north east coast populations corresponded to the original description of S. thunbergii f. latifolium, we could not find exact morphological features and diagnostic characters to distinguish form in S. thunbergii. These results indicated that it is not the optimal characteristics to identify infraspecific form in this species. In contrast, morphological variations may signify the adaptation of this species to local environmental factors. Thus, we recommend that intraspecific morphological variation of S. thunbergii should be carefully used to identify infraspecific taxa.

• Applied Microscopy
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• v.32 no.3
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• pp.275-284
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• 2002

Granular gland regeneration in the toad after dorsal skin wound histologically was examined using scanning and transmission electron microscopy. After cutaneous wounds were induced by excision, animals were maintained in special cages for up to 20 days. In transmission electron microscopy (TEM), newly formed granular gland, though poorly developed, was seen on 4 day after injury. Epithelial cells moved toward apical region of newly formed gland. The cells had smooth surface and were not connected to other cells by desmosomes. Mitochondria rich cell (MRC) possessing long cytoplasmic processes formed a gland cavity and hemidesmosomes were found under the cell processes. Basal cavity of newly formed gland consisted of MRC, pro-granular producing cells (pGPC), and granular producing cell (GPC). Moreover it was observed that xanthophores moved to the base of the epithelial tissue on 10 day after the injury. These cells contained numerous pterinosomes and carotenoid vesicles. Immature pterinosomes were large and carotenoid vesicles were moderately electron dense. On 13 day after the injury, xanthophores contained abundant carotinoid vesicles and lammelated pterinosomes. Iridophores were also observed adjacent the developing xanthophores on 16 day post-injury. These observations indicated that regeneration of granular gland from glandular precursor cells during wound healing and subsequent expansion of the glandular cells might be dependent on maturation and proliferation of these newly formed cells.

• Applied Microscopy
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• v.31 no.1
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• pp.49-57
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• 2001

In this study, the distribution of lectin receptors in culutured fibroblast was explored using colloidal gold label complexed with lectin WGA purified from wheat germ (Triticum vulgare). The lectin WGA gold complex, shown to recognize GlcNAc (N-acetylgalactosamine) and NeuNAc (N-acetylneuraminic acid) regions, was applied to detect binding sites in Lowicryl HM 20 sections viewed under electron microscope Labeled sections of the culutured fibroblast revealed gold particles specifically distributed on the cytoplasm and cell surface of the fibroblast. Labeling of 24 hours culutured fibroblast was then quantified and compared to that of 72 hours culutured fibroblast. 24 hours culutured fibroblast sections resulted in specific gold particle distribution on the cytoplasmic vesicle of the culutured fibroblast. These results indicate that lectin WGA receptors are located in the cytoplasmic vesicle and cell surface of the 24 hours culutured fibroblast, and on the cell surface of the 72 hours culutured fibroblast. Therefore, the GlcNAc and NeuNAc regions on the cell surface appear to be functionally associated with cell-recognition and protection from other cell of the tissue, and linked with secretion and exocytosis of the fibroblast cytoplasm.

• Applied Microscopy
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• v.34 no.1
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• pp.71-81
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• 2004

When U. unicinctus mature oocytes were fertilized in vitro, germinal vesicle breakdown (GVBD) and meioses occurred and the zygotes entered cleavage stage. A modified pattern of spiral cleavages, suggestively based on behavior of mitotic spindles, have been observed in this work. The first and second cleavages were meridional and the third was equatorial, and then followed by repetitions of meridional-equatorial cleavage. The cleavage of the isolecithal egg were equal and holoblastic and its patterns were spiral. The anti-${\alpha}-,-{\beta}$- tubulin reactions and confocal microscopy revealed mitotic apparates tilted obliquely at each mitosis causing oblique displacements of the blastomeres. Despite isolecithal distribution of yolk, this observations implicated that tilting of mitotic apparates induced spiral cleavage and the displacements of blastomeres. However, these features would not be the typical spiral cleavage, but represented a modified pattern of known Spiralian s in the sense of the equal cleavage. During the first cleavage, heart-shaped eggs have been produced. Electron microscopies exhibited the first cleavage furrow extended with its membranous structure deeply into the cytoplasm. Contractile ring has not been observed.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.12-19
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• 1999

To improve biopolymer productivity and properties of Bacillus strains, protoplast fusion was performed between biopolymer producing Bacillus subtilis K-1 and lactose utilizing Bacillus coagulans. The results were as follows; Protoplasts mixed in fusion fluid containing 33% PEG 6000, 1% PVP and 10 mM $CaCl_2$ were reacted for 5 min at $37^{\circ}C$ and then centrifused protoplasts were directly overlaid on the selective media containing $100\;{\mu}g/ml$ antibiotics and incubated for 3 days. At this conditions, the frequency of protoplast fusion was generally in the range of $4.6{\times}10^{-5}\;to\;1.8{\times}10^{-7}$ in ratio. Segregation ratio was observed between 1 to 6% indicating genetic stability of all the fusants. Fusants growth were also observed on the media contained amino acid and antibiotics as required marked materials. DNA contents of the selected fusants were 1.6 to 2.7 times more than that of parent strains. With observation by TEM microscopy, spherical protoplasts were first released from the swollen parental cells and then contracted to fuse in the process of fusion. And fused cells were observed representative vesicle. Originally, the parental cells were observed as in the morphology of thick-walled and double membrane-surrounded rod shape with TEM microscopy.