• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.474-481
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• 1996

Since the commercially available rabbit anti-cyclin D3, generated from c-terminal 16 amino acid residues which are common to human and murine cyclin D3, is highly cross-reactive with many other cellular proteins of mouse, a new rabbit polyclonal anti-cyclin D3 has been raised by using murine cyclin D3 protein expressed at a high level in Escherichia coli as the immunogen. To express murine cyclin D3 protein in E. coli, the cyclin D3 cDNA fragment encoding c-terminal 236 amino acid residues obtained by polymerase chain reaction (PCR) was inserted into the NcoI/BamHI site of protein expression vector, pET 3d. Molecular mass of the cyclin D3 overexpressed in the presence of IPTG (Isopropyl $\beta$-D-thiogalactopyranoside) was approximately 26 kDa as calculated from the reading frame on the DNA sequence, and the protein was insoluble and mainly localized in the inclusion bodies that could be easily purified from the other cellular soluble proteins. When renaturation was performed following denaturation of the insoluble cyclin D3 protein in the inclusion bodies using guanidine hydrochloride, 4.4 mg of soluble form of cyclin D3 protein was produced from the transformant cultured in 100ml of LB media under the optimum conditions. Four-hundred micrograms of the soluble form of cyclin D3 protein was used for each immunization of a rabbit. When the antiserum obtained 2 weeks after tertiary immunization was applied to Western blot analysis, it was able to detect 33 kDa cyclin D3 protein in both murine lymphoma cell line BW5147.G.1.4 and human Jurkat T cells at 3,000-fold dilution with higher specificity to murine cyclin D3, demonstrating that the new rabbit polyclonal anti-murine cyclin D3 generated against c-terminal 236 amino acid residues more specifically recognizes murine cyclin D3 protein than does the commercially available rabbit polyclonal antibody raised against c-terminal 16 amino acids residues.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.326-336
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• 2016

Saccharomyces cerevisiae is one of the most employed cell factories for the production of bioproducts. Although monomeric hexose sugars constitute the preferential carbon source, this yeast can grow on a wide variety of nitrogen sources that are catabolized through central nitrogen metabolism (CNM). To evaluate the effects of internal perturbations on nitrogen utilization, we characterized strains deleted or overexpressed in GLT1, encoding for one of the key enzymes of the CNM node, the glutamate synthase. These strains, together with the parental strain as control, have been cultivated in minimal medium formulated with ammonium sulfate, glutamate, or glutamine as nitrogen source. Growth kinetics, together with the determination of protein content, viability, and reactive oxygen species (ROS) accumulation at the single cell level, revealed that GLT1 modulations do not significantly influence the cellular physiology, whereas the nitrogen source does. As important exceptions, GLT1 deletion negatively affected the scavenging activity of glutamate against ROS accumulation, when cells were treated with H₂O₂, whereas Glt1p overproduction led to lower viability in glutamine medium. Overall, this confirms the robustness of the CNM node against internal perturbations, but, at the same time, highlights its plasticity in respect to the environment. Considering that side-stream protein-rich waste materials are emerging as substrates to be used in an integrated biorefinery, these results underline the importance of preliminarily evaluating the best nitrogen source not only for media formulation, but also for the overall economics of the process.

• Applied Microscopy
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• v.28 no.2
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• pp.225-236
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• 1998

Adriamycin is a one of anthracyclin antibiotics isolated from the culture media of Streptomyces peucetius var casius. The formation of reactive oxygen metabolite by redox cycling during the metabolism and the inhibition of DNA synthesis results in antineoplastic effects of adriamycin. The authors have demonstrated the effects of SOD(superoxide dismutase) or DMTU (dimethyl thiourea), which are used as an antioxidant, on the ultrastructural changes of the gastric chief cells after the administration of adriamycin in the rat. Adriamycin (30 mg/kg) was administered intraperitoneally to the Sprague-Dawley rats weighing about 220 gm and SOD (15000 unit/kg) or DMTU (500 mg/kg) were administered intraperitoneally to the rats 30 minutes after the administration of adriamycin. The gastric chief cells 24, 48 and 72 hours after the administration of adriamycin were observed with Hitachi-600 electron microscope. The results were as follows. 1. SOD or DMTU alone did not affect the ultra structures of the gastric chief cells in the rat. 2. Dilation, sacculation and segmentation of the cisternae of rough endoplasmic reticulum, dilation of the saccules of Golgi complex and dilated mitochondria with electron lucent matrix were seen in the adriamycin treated rats. In the course of time, the ultrastructures of the chief cell changed markedly. 72 hours after drug administration, severely dilated cisternae of rough endoplasmic reticulum, with clumping of chromatin around the nuclear envelope and mitochondria with electron lucent matrix and dilated cristae were seen in the chief cell. 3. The treatment of SOD is more effective than DMTU to attenuated the ultrastructural changes of the chief cells in the adriamycin administered rat. Consequently it is suggested that adriamycin would induce the degenerative changes of the organelles of the chief cell. The treatment of SOD is more effective than DMTU to attenuate the adriamycin induced damage.

• Journal of Embryo Transfer
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• v.31 no.1
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• pp.39-46
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• 2016

Cryopreservation has been applied successfully in many mammalian species. Nevertheless, pig embryos, because of their greater susceptibility to cryoinjuries, have shown a reduced developmental competence. The aim of this study was to evaluate the survival status of vitrified-warmed porcine embryos. Forced blastocoele collapse (FBC) and non-FBC blastocysts are vitrified and concomitantly cultured in culture media which were supplemented with/without fetal bovine serum (FBS). Porcine vitrified-warmed embryos were examined in four different methods: group A, non-FBC without FBS; group B, non-FBC with FBS; group C, FBC without FBS; group D, FBC with FBS. After culture, differences in survival rates of blastocysts derived from vitrified-warmed porcine embryos were found in group A~D (39.5 (A) vs 52.5 (B) and 54.8 (C) vs 66.7% (D), respectively, p<0.05). Reactive oxygen species (ROS) level of survived blastocysts was lower in group D than that of another groups (p<0.05). Moreover, total cell number of survived blastocysts was higher in group D than that of other groups (p<0.05). Otherwise, group D showed significantly lower number of apoptotic cells than other groups ($2.0{\pm}1.5$ vs $3.2{\pm}2.1$, $2.8{\pm}1.9$, and $2.7{\pm}1.6$, respectively, p<0.05). Taken together, these results showed that FBS/FBC improves the developmental competence of vitrified porcine embryos by modulating intracellular levels of ROS and the apoptotic index during the vitrification/warming procedure. Therefore, we suggest that FBS and FBC are effective treatment techniques during the vitrification/warming procedures of porcine blastocysts.

• Journal of Life Science
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• v.12 no.3
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• pp.325-331
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• 2002

There is growing interest ill understanding the role and mechanisms of flavonoid as antioxidant on LDL. The antioxidative activity of flavonoid typically present in Oenothers odorate Jacquin was investigated in vitro using a human LDL oxidation assay. In present work, LDL was incubated with increasing concentrations of extracts of extracts of Oenothers odorate Jacquin and LDL oxidation was started by adding CuSO$_4$to the media. Substances in leaves extracts of Oenothers odorate Jacquin are capable of inhibiting the initiation and the propagation of LDL oxidation. They inhibit LDL oxidation, monitored by thiobarbituric acid-reactive substances(TBARS), as well as modification as shown through direct measurement of electrophoretic mobility, diene conjugates. Inhibition is a dose dependent effect that becomes already apparent at concentration of extracts as low as 40$\mu\textrm{g}$/mL. Inhibition is almost complete at 80$\mu\textrm{g}$/mL. Extracts of Oenothers odorate Jacquin were more potent antioxidative activity than either ascorbic acid and dl-a -tocopherol.

• Journal of the Korean Magnetics Society
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• v.5 no.5
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• pp.874-879
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• 1995

Magnetic properties of FeMoN, FeMoTaN, FeTaN and FeTaC films deposited by DC magnetron reactive sputter were investigated, and correlated with their microstructure. FeMoN films were not showen the soft magnetic prop¬erties, because of generated $Fe_{2}Mo$, $Fe_{3-2}N$ and $Fe_{4}N$ phases. Ta added films, however, effectivly retarded the $\alpha$-Fe grain growth and suppressed the generation of Fe nitrides or carbides during heat treatement. The soft magnetic properties of $B_{s}:15\;kG,\;H_{e}:0.25\;Oe,\;\mu':4000(at\;5\;MHz),\;and\;B_s:14.5\;kG,\;He:0.25\;Oe,\;\mu':2700(5MHz)$ were observed in $Fe_{78.8} Ta_{8.5}N_{12.7}\;and\;Fe{75.6}Ta_{8.1}C_{16.3}$ films, respectively.

• Journal of Animal Reproduction and Biotechnology
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• v.39 no.1
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• pp.48-57
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• 2024

Background: Cadmium (Cd) is toxic heavy metal that accumulates in organisms after passing through their respiratory and digestive tracts. Although several studies have reported the toxic effects of Cd exposure on human health, its role in embryonic development during preimplantation stage remains unclear. We investigated the effects of Cd on porcine embryonic development and elucidated the mechanism. Methods: We cultured parthenogenetic embryos in media treated with 0, 20, 40, or 60 µM Cd for 6 days and evaluated the rates of cleavage and blastocyst formation. To investigate the mechanism of Cd toxicity, we examined intracellular reactive oxygen species (ROS) and glutathione (GSH) levels. Moreover, we examined mitochondrial content, membrane potential, and ROS. Results: Cleavage and blastocyst formation rates began to decrease significantly in the 40 µM Cd group compared with the control. During post-blastulation, development was significantly delayed in the Cd group. Cd exposure significantly decreased cell number and increased apoptosis rate compared with the control. Embryos exposed to Cd had significantly higher ROS and lower GSH levels, as well as lower expression of antioxidant enzymes, compared with the control. Moreover, embryos exposed to Cd exhibited a significant decrease in mitochondrial content, mitochondrial membrane potential, and expression of mitochondrial genes and an increase in mitochondrial ROS compared to the control. Conclusions: We demonstrated that Cd exposure impairs porcine embryonic development by inducing oxidative stress and mitochondrial dysfunction. Our findings provide insights into the toxicity of Cd exposure on mammalian embryonic development and highlight the importance of preventing Cd pollution.

• Development and Reproduction
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• v.10 no.2
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• pp.105-113
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• 2006

Reactive oxygen species(ROS) generated in cellular metabolism have an effect on cell maturation and development. In human reproductive tract, oxidative injury by ROS may induce female infertility. Also, oxidative injury may be responsible for developmental retardation and arrest of mammalian preimplantation embryos. Activating transcription factor 4(ATF4) is a member of the cyclic-AMP response element-binding(CREB) familiy of basic region- leucine zipper(bZip). ATF4 is known to regulate stress response to protect cell from various stress factors and inducer of apoptisis. The purpose of this study was to investigate whether ATF4 is involved in the defensive mechanism in oxidative stress condition during the development of mouse preimplantation embryos. To verify the expression of ATF4 in oxidative stress condition, 2-cell stage embryos were cultured in HTF media containing 0.1mM, 0.5mM or 1mM hydrogen peroxide($H_2O_2$) for 1hr(2-cell), 8hr(4-cell), 17hr(8-cell), 24hr(morula), 48hr(early blastocyst) or 64hr(late blastocyst). The developmental rate decreased in the 0.1mM $H_2O_2$ treated group compared with control group. In embryos treated with 0.5mM and 1mM $H_2O_2$ showed 2-cell block. As a results of the semi-quantitative RT-PCR analysis of SOD1, ATF4 and Bax gene expression, SOD1, ATF4 and Bax genes were increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. In 2-cell embryos, expression of SOD1, ATF4 and Bax genes were notably increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. Immunofluorescence analysis showed that ATF4 protein was localized at the cytoplasm of preimplantation embryos. The increase in ATF4 immunoreactivety was observed in the 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. It suggests that oxidative stress by $H_2O_2$ induces expression of ATF4 and may be involved in protection mechanism in preimplantation embryos from oxidative injury.

• Journal of the Korean Vacuum Society
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• v.22 no.2
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• pp.55-65
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• 2013

Sterilization of Neurospora crassa has been investigated in this research by using a surface air plasma with dielectric barrier discharged (DBD) structure under atmospheric pressure. The sinusoidal alternating current has been used in this experiment with discharge voltage of 1.4~2.3 kV. The phase difference between the voltage and current signals are found to be almost 80 degree due to the capacitive property of dielectric barrier. Temperature on the biomaterials has been minimized by radiating the heat with the air cooling system. It is noted that the substrate temperature remains under 37 degree for plasma exposure time of 10 minutes with operation of cooler system. It is found that the ozone, $O_3$, has been measured to be about 25~30 ppm within 1 cm region and to be about 5 ppm at the 150 cm downstream region away from the suface plasma. It is also noted that the nitric oxide, NO, and nitric dioxide, $NO_2$, are not nearly detected. Germination rate and mitochodrial activity of Neurospora crassa immersed in the deionized water have been found to be drastically decreased as the plasma treatment time and its electrical power are increased in this experiment. Here, the mitochondrial activity has been analyzed by MTT (3-(4,5-dimethy lthiazol-2yl)-2,5-diphenyl-2H-tetrazolium bromide) assay. However, sterilization of Neurospora crassa immersed in the Vogel's minimal media has been found to be low by plasma treatment, which is caused by surrounding background solution. This research shows the sterilization possibility of Neurospora crassa by using the noncontated surface DBD plasma, which is different from the plasma jet. This is mainly attibuted to the reactive species generated by the surface plasma, since they play a major role for inhibition of micobes such as Neurospora crassa.

• Pediatric Infection and Vaccine
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• v.31 no.1
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• pp.75-82
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• 2024

Purpose: As the coronavirus disease 2019 (COVID-19) pandemic ended, the number of patients with respiratory syncytial virus (RSV) pneumonia increased during the spring/summer of 2022. This study aimed to analyze the clinical features and antibiotic usage of children hospitalized for RSV pneumonia in a recently established general hospital in Sejong city. Methods: In this retrospective review, we included inpatients of the Pediatric Department of Chungnam National University Sejong Hospital diagnosed with RSV pneumonia between March 2022 and April 2023. Patients were divided into 2 groups: with and without antibiotic treatment. Demographic data, initial presentations, and clinical courses were reviewed. Results: A total of 116 patients with RSV pneumonia were hospitalized during this period, of which 102 were analyzed, excluding 14 with underlying diseases or who did not fall within the definition of pneumonia. The median age was 17 months. Diagnoses of bacterial infections (acute otitis media and sinusitis) were documented in 9.8% of cases. Intravenous (IV) antibiotics were administered in 46% of cases. The group receiving IV antibiotics showed higher inflammatory levels (C-reactive protein; CRP), more infiltration on initial chest X-rays, and longer fever duration. There was no difference in the length of hospitalization between the groups with and without IV antibiotics. Conclusions: This study showed a tendency for the attending physician to prescribe IV antibiotics to patients with longer fever duration, pulmonary infiltrations on the initial chest X-ray, and higher CRP levels. However, given the high rate of IV antibiotic usage compared to previous studies, care should be taken in antibiotic use.