This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.
Park, Jin-Kyu;Kang, Jeong-Hee;Chong, Yong-Gil;Lee, Nam-Hoon
Journal of the Korea Organic Resources Recycling Association
/
v.20
no.2
/
pp.66-75
/
2012
The objective of this study was to assess the effects of methanogenic bacteria-activated leachate recirculation method for enhancing waste stabilization and landfill gas production from a solid waste landfill. To simulate a conventional landfill (Lys-A), a landfill recirculated only fresh leachate (Lys-B), and two landfills recirculated leachate after pretreating with ASBR (Lys-C and Lys-D), four lysimeters were operated over a period of 4 years. Lys-D was recirculated two times of pretreated leachate volume than that of Lys-C. In the case of the landfill recirculated only fresh leachate and the landfill recirculated leachate after pretreating with ASBR, methane productions were increased until about 600 days, but there were not effect of leachate recirculation for enhancing methane production after about 600 days. It was assumed that leachate recirculation into fewer biodegradable organic wastes had not effect to enhance landfill gas production. Lys-C and Lys-D showed the highest performance for enhancing cumulative methane yield as well as acceleration waste stabilization. In cumulative methane yield, Lys-C (35.51 mL $CH_4/g$ VS) and Lys-D (36.12 mL $CH_4/g$ VS) were much higher than Lys-A (28.37 mL $CH_4/g$ VS) and Lys-B (30.07 mL $CH_4/g$ VS). In case of between Lys-B and Lys-C with the same recirculation rate, COD concentration in Lys-C was more rapidly decreased compared with that in Lys-B. This was attributed to the presence of methanogenic bacteria as well as dilution of inhibitory substances by the methanogenic bacteria-activated leachate recirculation. Therefore, the landfill recirculated leachate after pretreating with ASBR was found to be the most appropriate operating techniques for enhancing waste stabilization and landfill gas production.
Kim, Yong-Doo;Choi, Ok-Ja;Shim, Ki-Hoon;Cho, In-Kyung
Journal of the Korean Society of Food Science and Nutrition
/
v.35
no.3
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pp.366-372
/
2006
This study is to investigate the physicochemical properties of differently pretreated chestnut starches during starch isolation and to examine their gelatinization properties by both heat and alkali treatments. One kind is starch A made by alkali method from peeled chestnut. The other is starch B made from chestnut with the outer layer. The results are as follows. Starch A has higher water binding capacity of 86.9% than starch B with 80.66%. Swelling powers of both starch A and B increased rapidly from $60^{\circ}C\;to\;80^{\circ}C$ in both, and since then it has changed a bit. Both began to show their solubility at $60^{\circ}C$ and increased continuously as the temperature went up. Starch A has higher swelling power and solubility than starch B. In iodine reaction, starch A has higher ${\lambda}max$ and absorbance at ${\lambda}max$ than starch B. X-ray diffraction patterns showed that starch A is type $C_b$ and that starch B is type B. Starch B has higher relative crystallinity of 37.0% than starch A with 36.2%. The results by differential scanning calorimetry revealed that starch A gelatinized from $66.95^{\circ}C$ to $77.5^{\circ}C$ and its enthalpy is 2.04 cal/g. And starch B gelatinized from $67.09^{\circ}C\;to\;77.5^{\circ}C$, and its enthalpy is 2.29 cal/g. Amylograms of chestnut starch at 6.5% concentration indicated that starch B needs higher onset temperature when beginning to gelatinize than starch A does. But starch A shows much higher peak viscosity, breakdown and setback than starch B does. Starch A shows higher viscosity, gel volume, and optical transmittance in gelatinization properties by alkali than starch B does.
Yang, In;Park, Dae-Hak;Choi, Won-Sil;Oh, Sei Chang;Ahn, Dong-uk;Han, Gyu-Seong;Oh, Seung Won
Korean Chemical Engineering Research
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v.56
no.4
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pp.577-584
/
2018
This study was conducted to investigate the potential of chicken feather (CF), which is a by-product in poultry industry, as a partial substitute of wood fiber in the production of wood-based fiberboard. Keratin-type protein constituted the majority of CF, and its appearance did not differ from that of wood fiber. When the formaldehyde (HCHO) adsorptivities of CF compared by its pretreatment type, feather meal (FM), which was pretreated CF with high temperature and pressure and then grounded, showed the highest HCHO adsorptivity. In addition, there was no difference between the adsorbed HCHO amounts, which was measured by dinitrophenylhydrazine method, of scissors-chopped CF and CF beated with an electrical blender. Mechanical properties and HCHO emission of medium-density fiberboards (MDF), which were fabricated with wood fiber and 5 wt% CF, beated CF or FM based on the oven-dried weight of wood fiber, were not influenced by the pretreatment type of CF. However, when the values compared with those of MDF made with just wood fiber, thickness swelling and HCHO emission of the MDF were improved greatly with the addition of CF, beated CF or FM. Based on the results, it might be possible to produce MDF with improved dimensional stability and low HCHO emission if CF, beated CF or FM is added partially as a substitute of wood fiber in the manufacturing process of MDF produced with the conventional urea-formaldehyde resin of $E_1$ grade. However, the use of CF or FM in the production of MDF has a low economic feasibility at the current situation due to the securing difficulty and high cost of CF. In order to enhance the economic feasibility, it requires to use CF produced at small to medium-sized chicken meat plants. More importantly, it is considered that the technology developed from this research has a great potential to make provision for the prohibition of animal-based feed and to dispose environmentally avian influenza-infected poultry.
$TiO_2$-coated bamboo activated carbon has been prepared and utilized under UV irradiation as a pretreatment method for an effective biodegradation of the recalcitrant polyaromatic hydrocarbons (PAHs). The anatase $TiO_2$ was successfully coated on the bamboo activated carbon (AC) and it showed the highest photoactivity against methylene blue. In the absence of the PAHs-degrading bacteria PAHs having low molecular weight (i.e., naphthalene, acenaphthylene, acenaphthene, and fluorene) were degraded by 9.8, 76.2, 74.1, and 40.5%, respectively. Higher molecular weight PAHs, however, maintained high residual concentrations of PAHs (400-1,000 ${\mu}g$/L) after the same treatment. On the other hand, the overall concentrations of PAHs became lower than 340 ${\mu}g$/L when the pretreated PAHs were subjected to biodegradation by a PAH-degrading consortium for a week. Herein, phenanthrene, anthracene, fluoranthene, and pyrene were removed by 29.3, 61.4, 27.0, and 44.3%, respectively, indicating the facilitated potential biodegradation of PAHs. Activated carbon coated with $TiO_2$ appeared to inhibit growth of PAH degraders on the surface of AC, indicating planktonic degraders were dominantly involved in the PAH biodegradation in presence of the $TiO_2$-coated bamboo AC. It was proposed that an effective remediation technology for the recalcitrant PAHs could be developed when an optimum pretreatment process is further established.
Quality changes in carrot during storage were studied to investigate the efficiency, cooling properties and the washing and storage effects of hydrocooling method. As a result of plotting the nondimensionalized carrot temperature versus cooling time, its cooling rate coefficient was shown $-0.0171min^{-1}{\sim}-0.0121min^{-1}\;(R^{2}=0.99{\sim}0.95)$, and $-0.141min^{-1}{\sim}-0.0038min^{-1}\;(R^{2}=0.98{\sim}0.92)$ in package condition7. Rate of weight loss and change in moisture content of carrot were not significantly different by treatment conditions during storage at $5^{\circ}C$. During storage at $15^{\circ}C$, however, weight loss of hydrocooled carrot was lower than that of non-treated carrot from the 30th to 40th day. Especially, PE was more effective than tray for packaging hydrocooled carrots. Carrot pretreated wish sterilizing agent, packed with PE vinyl film and with residual water removed after hydrocooling showed a lower decaying-rate than any other carrots. Changes in Hunter L and b values of hydrocooled carrot were slower than those of non-treated one. The carotenoids contents of stored carrot $(0.736{\sim}0.780mg%)$ decreased to $9{\sim}43%$ after 40 days of storage at $5^{\circ}C$ and before 20 days of storage at $15^{\circ}C$. It could be presumed that the addition of sterilizing agent reduced the initial level of overall microbial and coliform counts and their growth rate during storage.
Background: The up-regulation of the nitric oxide (NO)-cGMP pathway might be involved in the change of vascular reactivity in rats 3 days after they suffer acute myocardial infarction. However, the underlying mechanism for this has not been clarified. Material and Method: Acute myocardial infarction (AMI) was induced by occluding the left anterior descending coronary artery (LAD) for 30 min (Group AMI), whereas the sham-operated control rats were treated similarly without LAD occlusion (Group SHAM), The concentration-response relationships for phenylephrine (PE), KCl, acetylcholine (Ach) and sodium nitroprusside (SNP) were determined in the endothelium intact E(+) and endothelium denuded E(-) thoracic aortic rings from the rats 3 days after AMI or a SHAM operation. The concentration-response relationships of PE in the E(+) rings from the AMI rats were compared with those relationships in the rings pretreated with nitric oxide synthase (NOS) inhibitor $N{\omega}$-nitro-L-arginine methyl ester (L-NAME) or the cyclooxygenase inhibitor indomethacin. The plasma nitrite/nitrate concentrations were checked via a Griess reaction. The cyclic GMP content in the thoracic aortic rings was measured by radioimmunoassay and the endothelial nitric oxide synthase (eNOS) mRNA expression was assessed by real time PCR. Result: The mean infarct size (%) in the rats with AMI was $21.3{\pm}0.62%$. The heart rate and the systolic and diastolic blood pressure were not significantly changed in the AMI rats. The sensitivity of the contractile response to PE and KCl was significantly decreased in both the E(+) and E(-) aortic rings of the AMI group (p<0.05). L-NAME completely reversed these contractile responses whereas indomethacin did not (p<0.05). Moreover, the sensitivity of the relaxation response to Ach was also significantly decreased in the AMI group (p<0.05). The plasma nitrite and nitrate content (p<0.05), the basal cGMP content (p<0.05) and the eNOS mRNA expression (p=0.056) in the AMI rats were increased as compared with the SHAM group. Conclusion: Our findings indicate that the increased eNOS activity and the up-regulation of the NO-cGMP pathway can be attributed to the decreased contractile or relaxation response in the rat thoracic aorta 3 days after AMI.
Park, Kyung-Ho;Cho, Hwee-Dong;Yang, Nam-Gil;Ahn, E-Tay;Ko, Jeong-Sik;Kim, Jin-Gook
Applied Microscopy
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v.24
no.2
/
pp.78-92
/
1994
Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.
Background: We tested the effect of indomethacine and total spinal anesthesia on the improvement of placental flow during cardiopulmonary bypass on fetal lamb. Material and Method: Twenty fetuses at 120 to 150 days of gestation were subjected to bypass via trans-sternal approach with a 12 G pulmonary arterial cannula and 14 to 18 F venous cannula for 30 minutes. All ewes received general anesthesia with ketamine. In all the fetuses, no anesthetic agents were used except muscle relaxant. Ten served as a control group in which placenta was worked as an oxygenator during bypass (Control group). The remainder worked as an experimental group in which pretreatment with indomethacine and total spinal anesthesia was performed before bypass with the same extracorporeal circulation technique as control group (Experimental group). Observations were made every 10 minutes during a 30-minute bypass and 30-minute post bypass period. Result: Weights of the fetuses ranged from 2.2 to 5.2 kg. In Control group, means of arterial pressure decreased from 44.7 to 14.4 mmHg and means of Pa$CO_2$ increased from 61.9 to 129.6 mmHg at each time points during bypass. Flow rate was suboptimal (74.3 to 97.0 $m\ell$/kg/min) during bypass. All hearts fibrillated immediately after the discontinuation of bypass. On the contrary, in Experimental group, means of arterial pressure reamined higher (45.8 to 30 mmHg) during bypass (p<0.05). Means of Pa$CO_2$ were less ranging from 59.8 to 79.4 mmHg during bypass (P<0.05). Flow rates were higher (78.8 to 120.2 $m\ell$/kg/min) during bypass (p<0.05). There were slower deterioration of cardiac function after cessation of bypass. Conclusion: In this study, we demonstrated that the placental flow was increased during fetal cardiopulmonary bypass in the group pretreated with indomethacine and total spinal anesthesia. However, further studies with modifications of the bypass including a creation of more concise bypass circuit, and a use of axial pump are mandatory for the clinical application.
Objectives : The purpose of the present study was to examine the effect of subacute treatment with the selective serotonin reuptake inhibitors(paroxetine and sertraline) on immobility in the forced swim test(FST) and on FST-induced changes in immune parameters of the mice. Methods : Authors applied a modified method of FST by Porsolt et al. Over 5 BALB/c mice were used for each group of experiments. To explore the changes in immune parameters by FST, authors investigated the production of anti-rat RBC antibody, concanavalin A(ConA)- or lipopolysaccharide(LPS)-stimulated splenocytes proliferation assay and cytokine gene expression. Results : Both paroxetine and sertraline decreased the duration of immobility in a dose-related manner. FST-performed mice showed a significant decrease in mitogenic responses of splenocytes and a slight increasing tendency in anti-rat RBC antibody response. All these responses were attenuated significantly by paroxetine and attenuated nearly nominal significance level by sertraline. The cytokine profiles of ConA-stimulated splenocytes from FST-performed mice showed stronger expression of IL-4 and weaker expression of IL-2 than control mice, and no changes in the expressions of IFN-$\gamma$ and lymphotoxin. IL-6 and IL-10 were not expressed in both group of mice. The pretreatment of paroxetine and sertraline attenuated the altered cytokine expressions in FST-performed mice to some extent. Some alterations of the expressions of IL-6 and IL-10 were observed in the mice which the selective serotonin reuptake inhibitors had been pretreated. Conclusion : The subacute treatment of paroxetine and sertraline attenuated the FST-induced behavioral and immune changes, and these serotonin reuptake inhibitors may exert some modulating effects on the immune system by the induction of cytokine gene expression, especially IL-6 and IL-10.
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