As a series of studies on the suitability as a second intermediate host of Clonorchis sinensis, artificial infection experiments were applied to Tilapia mossambica. And then, in order to elucidate the defence mechanism of the fish to Clonorchis, clonorchicidal substance in the epidermal mucus of the fish was isolated by silica gel column and thin layer chromatography and analyzed for its chemical structure by UV, IR and NMR-spectroscopy. The results obtained are summarized as follows: 1. The cercariae which attempted to contact with the fish in the water were observed under stereomicroscope. After contact, the cercariae began to separate their tail from the body after several minutes and then the number increased to $80\%$ more than 10 minutes after the encounter. But very few cercariae could actually invade the epidermis of the fish. 2. The fish were reared with Parafossarulus manchouricus which were shedding numerous cercariae of Clonorchis in the aquarium for 24 hours. Only a few cercariae could invade the epidermis but most of the invaded cercariae died out before forming their cysts. Very few number of the remaining encysted cercariae were also found to be in a state of suspended animation within 42 hours. 3. In the cases of the control fish, Pseudorasbora parva, numerous cercariae of Clonorchis were found to invade the fish through the epidermis under stereomicroscope. Then many metacercariae of Clonorchis were also found in the fish while they were kept in the aquarium. 4. A sample of the epidermal mucus of Tilapia mossambica was extracted with ethly ether 6 times repeatedly. In silica gel column chromatography, using petroleum ether: chloroform/30:70(v/v) as a first solvent and MeOH as a second solvent, the extract was fractionated into the yellow and brownish red solutions in the first solvent and the clonorchicidal brownish yellow solution in the second sovent. 5. The clonorchicidal brownish yellow solution was added to petroleum ether, and the mixture was stored for 5 days at $5^{\circ}C$ and was, then, separated into supernatant fraction and precipitate. Ten mg/ml of the supernatant fraction killed, in vitro, the excysted metacercariae in 45 minutes but the precipitate in 600 minutes. 6. In silica gel column chromatography, using acetone: benzene/10:90(v/v) as a solvent, the more clonorchicidal supernatant fraction was fractionated into the first fraction with Rf. 0.2966 and the second fraction with Rf. 0.072. In vitro, 10mg/ml of the first fraction killed the excysted metacercariae in 28 minutes, the second fraction in 80 minutes and the first fraction was, therefore, determined to be a final clonorchicidal substance. 7. By this purification procedures, the most clonorchicidal substance from the epidermal mucus of Tilapia mossambica was purified 71-folds with $0.2075\%$ yield. Infra red, nuclear magnetic resonance and ultraviolet spectrometric analysis of the purified substance revealed that the substance is linoleic acid. According to the results of the present studies it seemed that this species can not serve as a proper intermediate host of Clonorchis sinensis, and that defence mechanism to the fluke seems to be correlated with linoleic acid in the epidermal mucus of this species.
The appearance of the primordial germ cells (PGC's), early gonadogenesis, sex differentiation and sex ratio of the embryo in the viviparous teleost, Ditrema temmincki were investigated by using photomicroscopy. The PGC's were first observed in the fibrous mesenchymal tissue located between the early alimentary tract and the dorsal body wall in the late embryonic development stage. During the period from the hatching to the individual total length (TL) of 4.0 mm the PGC's were evenly distributed in the fibrous mesenchymal tissue between alimentary tract and body wall. But the period of TL 5.0 mm mesenchymal tissue separated from the dorsal body wall, the PGC's moved to the posterior mesenchymal tissue and formed the primitive gonad. During the early gonadogenesis, differentiation of the testis and ovary were distinguished from the arrangement of the germ cells and somatic cells. Gonad of embryo in TL 10.0 mm can be separated into the ovary and testis by external morphology. The testis had a separated form which was consisted with two lobes, and the ovary had a fused form in half-posterior part. In the testicular differentiation of the embryo, histological pattern of the seminiferous tubule appeared when TL of the embryo was to be 25.0 mm, for the seminal vesicle was formed In the individual TL of 30.0mm. The testis of the embryo with TL of 45.0 mm was similar to that of the adult fish in the external and internal structures. In the ovarian differentiation, formation of the ovigerous folds and the ovarian cavity were clearly observed when the TL reached to 30.0 mm. The ovary from the individual with TL of 60.0 mm was differentiated into a similar ovary as seen in the adult fish in the external and internal structure. Right before parturition the total length of the individual was approximately 63.0 mm of which the individual embryo has an ovary containing the oocytes in the chromatin nucleolus stage, or a testis containing the spermatogonia, respectively. And the embryonic sex ratio of female to male was 1.65 : 1. Ditrema temmincki is dioecism and the pattern of sex differentiation is belonged to a differentiation type.
Kim, Deok-Jin;Kim, Mi-Sun;Lee, Ye-Seul;Sohn, Ho-Yong
Microbiology and Biotechnology Letters
/
v.41
no.3
/
pp.311-319
/
2013
In an effort to characterize the physicochemical properties and microbial risks associated with the soy sauce jangachi (Korean traditional pickle), 15 different home-made products, which were prepared from medicinal plants and wild edible vegetables, from head-families of Andong, Kyungsangbuk-do Province in Korea, and 6 different commercial products sold at supermarket, were investigated. The average pH of the mature soaking solutions and plants soaked in the 21 jangachi were $3.99{\pm}0.38$ and $3.51{\pm}0.41$, and the average acidity of the mature soaking solutions and soaked plants were $1.59{\pm}0.54$ and $1.65{\pm}0.76$, respectively. The average brix of the mature soaking solutions and plants soaked were $27.67{\pm}8.38$ and $25.61{\pm}6.60$, respectively. In salinity, which is a major factor in jangachi industry production, the average salinity of the mature soaking solutions and soaked plants were $7.55{\pm}3.26$ and $5.75{\pm}2.23$, respectively. In particular, the hot-peppers, eusuri, du-rup, kaet-ip, kuji-ppong, myeng-i and sancho jangachi were amongst the home-made products, and the salinity was above 8.8%, which was 2 folds-higher than that of the commercial sterilized products, and 1/3-lower than commercial non-sterilized products. The color difference and turbidity of jangachi were dependent on the plant parts used. In microbial risk assessment, the microorganisms related with food-borne disease, such as Escherichia coli, Salmonella sp, and Shigella sp., were not detected. After some time, total cell count analysis revealed that the commercial products sold at supermarkets were more vulnerable than the home-made products.
Kim, Eunhye;Hwang, Yon-Jin;Kim, Suhee;Lee, Hyeri;Hong, Soonsung;Park, Kyung-Hun;Kim, Jeong-Han
The Korean Journal of Pesticide Science
/
v.16
no.4
/
pp.343-349
/
2012
Exposure and risk assessments were conducted to evaluate the relative safety of mixing/loading work of indoxacarb between wettable powder (WP) and water dispersible granule (WG). Hand exposure was monitored using cotton gloves while inhalation exposure was measured using personal air monitor. Method validation for the exposure monitoring was established successfully through several experiments. Limit of determination and limit of quantitation were 0.25 and 1 ng, respectively. $R^2$ of calibration curve linearity was more than 0.9999 and reproducibility was 0.7-6. Recovery of indoxacarb from gloves, solid sorbent and glass fiber filter at three different levels was 81.5-108.8%. Trapping efficiency and breakthrough tests gave 981.5-108.8% of recovery. During mixing/loading procedure, hand exposure amount (75 percentile of 30 repetitions) for indoxacarb WP was 6 folds (459.8 mg/kg a.i) than that of WG (81.4 mg/kg a.i). This result indicates that WG has less drift than WP thanks to its granular type of formulation. Inhalation amount was $10^{-8}-10^{-7}%$ of spray mixture prepared and $10^{-4}-10^{-3}%$ of hand exposure. In inhalation case, no significant differences were observed between two formulations. Margin of safety was calculated for risk assessment using male Korean average body weight and acceptable operator exposure level as the important exposure factors. Mixing/loading procedures for both of the formulations were considered to be of least risk because calculated MOS values were more than 1.
Pollen germination viability is an essential factor to produce seeds from pollination and fertilization, which are required to maintain plant generation. In this study, we tried to identify the effect of boric acid on pollen germination and tube grouch in non-transgenic and transgenic plants expressing monoclonal antibodies (anti-colorectal cancer mAb CO17-1A, anti-breast cancer mAb BR55, and anti-rabies virus mAb57). The pollen of non-transgenic plant was treated with different concentration of boric acid (0, 5, 10, 15, 20, $40{\mu}g/mL$) in germination buffer to investigate its effect on in vitro pollen germination. At $20{\mu}g/mL$ of boric acid, tile pollen germination rate was the highest (49.5%) compared to other concentrations. In general, the germination rate significantly increased 3-10 folds in boric acid ($20{\mu}g/mL$) treated group in non-transgenic and transgenic plants. Also, the pollen tube length increased in boric acid ($20{\mu}g/mL$) treated groups. In the treated group, the pollen tube length increased until 3 h boric acid treatment and decreased after the 3 h, indicating that the 3 h is the most appropriate incubation time period. Western blot analysis showed that the mAb transgene expression was more stable in leaf than pollen in transgenic plants. This study suggested that $20{\mu}g/mL$ of boric acid is ideal concentration to induce in vitro pollen germination of transgenic plants expressing therapeutic monoclonal antibodies, indicating stable pollination and fertilization in transgenic plants.
This study was carried out to classify the Hanjis into three groups that were indigenous Hanji, traditional Hanji, and improved Hanji handmade by paper making method according to the physical properties of each paper sheet such as tensile, bursting and tearing strength, folding endurance and fiber orientation in each layer. The results obtained were summarized as follows: 1. The multi-layered Hanjis made by "Oebal" Hanji making method in different direction of fiber orientation have good properties in tearing resistance. 2. The multi-layered Hanji in different direction of fiber orientation has good properties in the tearing resistance, but the burst index and the breaking length results were lower than the single layered Hanjis. 3. The different fiber orientation and multi-layered method didn't increase, the three indexes(burst index, tear index, breaking length). Only, the different direction of fiber orientation decreased the difference of width and length strength (tensile, tear) of the Hanji. 4. "Dochim"(Korean finishing touch process for indigenous Hanji by fulling round sticks) greatly increase folding endurance(double folds, not $log_{10}$) and good effect to tensile strength and burst strength. 5. The today's Oebal Hanji were the maximum of 2 layers and the indigenous Oebal Hanji were 16 layers the maximum. In addition, average of the indigenous Oebal Hanji was 4 layers(all 4-layer Hanji were the different fiber orientation of each layer). 6, The indigenous Hanji(multi-layered, and different fiber orientation) was good condition with "Dochim". Dochim increased tensile strength and burst strength of the indigenous Hanji. So the three-strength indexes were similar level("--"). 7. When the number of layer which were same fiber orientation increase, the increased Hanji became similar strength pattern("V", breaking length and burst index was higher than tear index) with "Ssangbal" Hanji. 8. The single layered papers that made by "Oebal" Hanji making method were similar strength pattern with Ssangbal Hanji. 9. There was no way to find the width and length direction of multi-layered Hanji by comparison between the difference of tensile strength and the difference of tearing resistance. 10. The compared pattern of tensile strength and tearing resistance of indigenous Oebal Hanji was different from today's Oebal Hanji. Especially, the tearing resistance of all indigenous Oebal Hanji(16 samples) was stronger on width of tearing resistance. And in the half of indigenous Oebal Hanji samples, the width of tensile strength and tearing resistance was stronger than length strength (Indigenous Oebal: '$\ulcorner\lrcorner$' 50%, '$\bigcup$' 50% $\leftrightarrow$ Today's Oebal: '$\ulcorner\lrcorner$' 12%, '$\bigcup$'6%, '$\llcorner\urcorner$'17%, '$\bigcap$'65%). In 65% today's Oebal, the length direction of tensile strength and tearing resistance was stronger than the width direction.
This study was to investigate of characteristics and antioxidant function of yuza from 4 different area. The hot-water extracts of yuza peel extracts from Geoje, Goseong, Goheung and Namhae(Changseon, Seolcheon) were tested for antioxidant activity in different reaction systems. Contents of total phenolic compounds and flavonoids were $122.18{\pm}1.44$ mg/100 g and $114.39{\pm}0.94$ mg/100 g in hot water extracts from Namhae-Seolcheon, respectively. The highest contents of hesperidin and naringin were $55.45{\pm}1.36$ mg/100 g and $28.41{\pm}0.64$ mg/100 g in hot water extracts from Geoje, respectively. Antioxidant activity of yuza peel hot-water extracts were significantely increased as the increament of sample adding concentration ($500{\sim}10,000{\mu}g$/ml). Reducing power was $6{\sim}9$ folds higher in 10,000 ${\mu}g$/ml concentration than 500 ${\mu}g$/ml and it's O.D. value showed $0.68{\pm}0.012{\sim}0.97{\pm}0.021$. ABTs scavenging activity was more than 80% in 10,000 ${\mu}g$/ml concentration samples, except from Goseong ($78.13{\pm}1.30%$). Hydroxyl radical scavenging activity was higher in Namhae-Seolcheon ($31.36{\pm}1.36%$) and Namhae-Changseon ($30.28{\pm}1.60%$) at 10,000 ${\mu}g$/ ml concentration, others activity were below 30%. At 10,000 ${\mu}g$/ml concentration, NO radical scavenging activity and antioxidant activity in ${\beta}$-carotene linoleic acid system were $26.49{\pm}1.77{\sim}40.85{\pm}0.95%$ and $24.40{\pm}1.91{\sim}38.17{\pm}0.56%$, respectively.
The bifunctional PheA protein, having chorismate mutase and prephenate dehydratase (CMPD) activities, is one of the key regulatory enzymes in the aromatic amino acid biosynthesis in Escherichia coli, and is negatively regulated by an end-product, phenyalanine. Therefore, PheA protein has been thought as useful for protein engineering to utilize mass production of essential amino acid phenylalanine. To obtain feedback resistant PheA protein against phenylalanine, we mutated by using random mutagenesis, extensively screened, and obtained $pheA^{FBR}$ gene encoding a feedback resistant PheA protein. The mutant PheA protein contains substitution of Leu to Phe at the position of 118, displaying that higher affinity (about $290{\mu}M$) for prephenate in comparison with that (about $850{\mu}M$) of wild type PheA protein. Kinetic analysis showed that the saturation curve of $PheA^{FBR}$ against phenyalanine is hyperbolic rather than that of $PheA^{WT}$, which is sigmoidal, indicating that the L118F mutant enzyme has no cooperative effects in prephenate binding in the presence of phenylalanine. In vitro enzymatic assay showed that the mutant protein exhibited increased activity by above 3.5 folds compared to the wild type enzyme. Moreover, L118F mutant protein appeared insensitive to feedback inhibition with keeping 40% of enzymatic activity even in the presence of 10 mM phenylalanine at which the activity of wild type $PheA^{WT}$ was not observed. The substitution of Leu to Phe in CMPD may induce significant conformational change for this enzyme to acquire feedback resistance to end-product of the pathway by modulating kinetic properties.
Human umbilical cord blood(HUCB) contains a rich source of hematopoietic stem cells, mesenchymal stem cells and endothelial cell precursors. Mesenchymal stem cells(MSCs) in HUCB are multipotent stem cells, differ from hematopoietic stem cells and can be differentiated into neural cells. We studied on transdifferentiation-promoting conditions in neural cells and cholinergic neuron induction of HUCB-derived MSCs. Neural differentiation was induced by addingdimethyl sulphoxide(DMSO) and butylated hydroxyanisole(BHA) in Dulbeco's Modified Essential Medium(DMEM) and fetal bovine serum(FBS). Differentiation of MSCs to cholinergic neurons was induced by combined treatment with basic fibroblast growth factor(bFGF), retinoic acid(RA) and sonic hedgehog(Shh). MSCs treated with DMSO and BHA rapidly assumed the morphology of multipolar neurons. Both immunocytochemistry and RT-PCR analysis indicated that the expression of a number of neural markers including $\beta$-tubulin III, GFAP and MBP, was markedly elevated during this acute differentiation. The differentiation rate was about $32.3{\pm}2.9%$ for $\beta$-tubulin III-positive cells, $11.0{\pm}0.9%$ for GFAP, and $9.4{\pm}1.0%$ for Gal-C. HUCB-MSCs treated combinatorially with bFGF, RA and Shh were differentiated into cholinergic neurons. After cholinergic neuronal differentiation, the $\beta$-tubulin III-positive cell population of total cells was $31.3{\pm}3.2%$ and of differentiated neuronal population, $70.0{\pm}7.8%$ was ChAT-positive showing 3 folds higher in cholinergic population than neural induction. Conclusively, HUCB-derived MSCs can be differentiated into neural and cholinergic neurons and these findings suggest that HUCB are alternative cell source of treatment for neurodegenerative diseases such as Alzheimer's disease.
To develope a process by which liquid foods can be stored in the liquid state at the frozen storage temperature, suitable cryoprotectants were selected. Orange juice, chosen as an example of liquid foods, was stored with combined cryoprotectants at $-15^{\circ}C$, and quality changes of orange juice during storage were evaluated. Among 7 cryoprotectants tested, NaCl solution had lower initial freezing point than others, and initial freezing points of glucose, fructose, glycerol, propylene glycol and citric acid were close to each other. Considering flavor quality of orange juice, cryoprotectants suitable for reducing freezing point of orange juice were glucose, fructose, glycerol, and citric acid. Combined cryoprotectants for reducing freezing point of 3 and 4 folds concentrated orange juice to $-15^{\circ}C$ consisted of 10% glucose, 8% frutose, 4.6% glycerol and 3% citric acid, and 5.5% glucose, 4.5% fructose, 4.6% glycerol, and 3% cirtric acid, respectively. When destruction of ascorbic acid, sedimentation volume and sensory flavor score of orange juice stored with combined cryoprotectants at $-15^{\circ}C$ and the control stored at $-18^{\circ}C$ were compared, there were no significant differences. These results indicated that liquid foods with suitable combined cryoprotectants could be stored at $-15^{\circ}C$ or below in the liquid state without adverse effect on quality of the stored products.
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