• Journal of the Korea Institute of Information and Communication Engineering
• /
• v.21 no.6
• /
• pp.1212-1217
• /
• 2017

We constructed a population of myelinated cells with co-culture of neuronal cells and Schwann cells from DRG. Schwann cells and neuronal cells were isolated from dorsal root ganglion (DRG) in embryos of rat in vitro respectively. The cultured Schwann cells and cultured neuronal cells, respectively were co-cultured in a same plate. This procedure contains following four steps: first step of suspension of the embryonic dorsal root ganglion cells, second step of addition of anti-mitoticcocktail, third step of purification of dorsal root cells, and fourth step of addition of Schwann cells to dorsal root ganglion cells. These cells were performed accomplishment of myelination. This myelinated co-culture system was infected by Semliki forest virus and then induced demyelination processing in this myelinated co-culture. We identified myelination and demyelination processing using antibody of peripheral myelin protein 22 (PMP 22) meaning presence of myelinated neuron.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.2
• /
• pp.217-224
• /
• 2014

Chicken are considered as the most important source of human infection by Campylobacter jejuni, which primarily arises from contaminated poultry meats. However, the genes expressed in vivo of the interaction between chicken and C. jejuni have not been screened. In this regard, in vivo-induced antigen technology (IVIAT) was applied to identify expressed genes in vivo during interaction between chicken and C. jejuni, a prevalent foodborne pathogen worldwide. Chicken sera were obtained by inoculating C. jejuni NCTC 11168 into Leghorn chickens through oral and intramuscular administration. Pooled chicken sera, adsorbed against in vitro-grown cultures of C. jejuni, were used to screen the inducible expression library of genomic proteins from sequenced C. jejuni NCTC 11168. Finally, 28 unique genes expressed in vivo were successfully identified after secondary and tertiary screenings with IVIAT. The genes were implicated in metabolism, molecular biosynthesis, genetic information processing, transport, regulation and other processes, in addition to Cj0092, with unknown function. Several potential virulence-associated genes were found to be expressed in vivo, including chuA, flgS, cheA, rplA, and Cj0190c. We selected four genes with different functions to compare their expression levels in vivo and in vitro using real-time RT-PCR. The results indicated that these selected genes were significantly upregulated in vivo but not in vitro. In short, the expressed genes in vivo may act as potential virulence-associated genes, the protein encoded by which may be meaningful vaccine candidate antigens for campylobacteriosis. IVIAT provides an important and efficient strategy for understanding the interaction mechanisms between Campylobacter and hosts.

• Food Science of Animal Resources
• /
• v.43 no.2
• /
• pp.305-318
• /
• 2023

This study investigated the protein digestibility of chicken breast and thigh in an in vitro digestion model to determine the better protein sources for the elderly in terms of bioavailability. For this purpose, the biochemical traits of raw muscles and the structural properties of myofibrillar proteins were monitored. The thigh had higher pH, 10% trichloroacetic acid-soluble α-amino groups, and protein carbonyl content than the breast (p<0.05). In the proximate composition, the thigh had higher crude fat and lower crude protein content than the breast (p<0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of myofibrillar proteins showed noticeable differences in the band intensities of tropomyosin α-chain and myosin light chain-3 between the thigh and breast. The intrinsic tryptophan fluorescence intensity of myosin was lower in the thigh than in the breast (p<0.05). Moreover, circular dichroism spectroscopy of myosin revealed that the thigh had higher α-helical and lower β-sheet structures than the breast (p<0.05). The cooked muscles were then chopped and digested in the elderly digestion model. The thigh had more α-amino groups than the breast after both gastric and gastrointestinal digestion (p<0.05). SDS-PAGE analysis of the gastric digesta showed that more bands remained in the digesta of the breast than that of the thigh. The content of proteins less than 3 kDa in the gastrointestinal digesta was also higher in the thigh than in the breast (p<0.05). These results reveal that chicken thigh with higher in vitro protein digestibility is a more appropriate protein source for the elderly than chicken breast.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.2
• /
• pp.207-212
• /
• 2004

Ruminal digestion of dry matter (DM) and neutral detergent fiber (NDF) of processed (ensiled, deepstacked or composted) broiler litter (BL) was determined in situ and in vitro, and compared with rice straw (RS). DM disappearances at 24 and 48 h and digestion of differently processed BL were higher than those of RS. Compared with RS, processed BL was low in NDF disappearance at 72 h incubation, digestion rate ($K_dB$) and digestibility at 0.025 of passage rate; however, deepstacked BL was similar in these NDF characteristics. Processing of BL affected ruminal digestion of nutrients such as DM and NDF adversely. NDF of composted BL, especially, was the most indigestible. This in situ nutritional evaluation indicated that deepstacked BL, the most widely used form of BL, was superior in DM characteristics (fractions, ruminal disappearance and digestibility) and similar in NDF characteristics (ruminal disappearance and digestibility) to RS.

• Journal of Microbiology and Biotechnology
• /
• v.11 no.5
• /
• pp.887-889
• /
• 2001

The gene encoding yeast pro-carboxypeptidase Y (pro-CPY) has been cloned and expressed in Escherichia coli. Most of the expressed pro-CPY was accumulated as cytoplasmic insoluble aggregates. In our previous study, active CPY was obtained by renaturation of entirely denatured pro-CPY followed by in vitro proteolytic processing with proteinase K along with the activation process. The same refolding process was performed to produce an active CPY from pro-CPY inclusion bodies with renaturation buffers containing proteinase K at different concentrations. The refolding efficiency decreased from $25\%\;to\;2\%$ in the renaturation buffers containing proteinase K at concentrations of $60{\mu}g/ml\;and\;0.6{\mu}g/mi$, respectively. In an attempt to increase the refolding efficiency with a lesser amount of proteinase K, a novel fed-batch refolding process was developed. In a fed-batch refolding, 99 ml of the renaturation buffer containing pro-CPY was gradually added into 1 ml of the renaturation buffer containing $60{\mu}g/ml$ of proteinase K to give a final proteinase K concentration of $0.6{\mu}g/ml$. The fed-batch refolding process resulted in a refolding efficiency of $18\%$, which corresponded to a 9-fold increase over that ($2\%$) in the batch process.

• Plant Resources
• /
• v.8 no.1
• /
• pp.52-59
• /
• 2005

A cDNA, PSOD1, encoding cytosolic copper/zinc superoxide dismutase (CuZn-SOD) was cloned and characterized from a full length cDNA library prepared from Populus alba${\times}$Populus glandulosa cultured in vitro. A PSOD1, is 725 nucleotides long and has an open reading frame of 459 bp with 152 amino acid residues (pI 5.43). The deduced amino acid sequence of PSOD1 perfect matched to the previously reported CuZn-SOD (CAC33845.1). Consensus amino acid residues (His-45, -47, -62, -70, -79, -119) were involved in Cu-, Cu/Zn-, and Zn- binding ligands. The deduced amino acid sequence of PSOD1 exhibited the high level of similarity from 100 to $85\%$ among previously registered SOD genes. The expression of PSOD1 in poplar increased at the 1 mM $H_{2}O_2$ and drought stress during 30 min and 60 min, but the ozone treated poplar increased at 30 min in the early time and then decreased at 60 min.

• Asian-Australasian Journal of Animal Sciences
• /
• v.8 no.6
• /
• pp.641-645
• /
• 1995

This study was conducted to develop a method for production of nuclear transplant bovine embryos using in vitro-matured (IVM) oocytes and to examine the effect of different conditions of electrofusion on fusion rate and developmental capacity of donor nucleus transplanted to enucleated oocytes. Eight- to sixteen-cell embryos derived from oocytes matured and fertilized in vitro used as donor blastomeres and IVM oocytes were used as recipient oocytes. Oocytes were enucleated immediately after 23-24 h IVM and then reconstituted with a donor blastomere in two different micromanipulation media. Fusion rate and subsequent development of the reconstituted oocytes was compared under the different electric stimuli and recipient oocyte ages. Success rate of enucleation was significantly higher in TCM-199 medium containing FCS than in DPBS. The high fusion rate(75-94%) and development (6.4-14.8%) to morulae and blastocyst (M + B) were obtained from 0.6-0.75 kV/cm DC voltage, although total cleavage was not different among the electric pulses. Most optimal condition of electric stimulation for fusion and development was 1 DC voltage of 0.75 kV/cm, in which 80.5% of oocytes were fused, 80.0% and 31.7% of which was cleaved and developed to M + B, respectively. No M + B was obtained from 1.2 kV/cm DC voltage regardless of pulse frequency. Recipint oocyte age at electrofusion greatly affected the cleavage and subsequent development to M + B, showing high rate at 40-41 h oocyte maturation. These results suggest that a suitable condition of electrofusion for donor nuclei derived from IVF may be 1-2 DC pulses of 0.7 kV/cm for $70{\mu}sec$ and that processing of a transplanted nucleus in IVM oocytes may be affected by maturation age of recipient oocytes.

• Journal of the Institute of Convergence Signal Processing
• /
• v.2 no.4
• /
• pp.46-52
• /
• 2001

In this paper it was examined both imaging system and processing algorithm for imaging's high speedization of fluorescent X-ray computed tomography using synchrotron radiation, The electronic system was used for dead time by about 6% but shortening of measure time was achieved by 2 seconds per 1 point. Also efficiency of reconstruction algorithm was proved, and memory and calculation amount was decreased by about 1/100 The fixed quantity was confirmed by physical phantom, and iodine distribution was presumed from image of thyroid gland in vitro These result shows realization possibility of fluorescent X-ray computed tomography measure in vivo.

• Clinical and Experimental Reproductive Medicine
• /
• v.21 no.2
• /
• pp.149-156
• /
• 1994

Failure of in vitro fertilization may occur even though oocyte and semen parameters seem satisfactory. Quantified ultrastructural study of spermatozoa was performed in three cases of failed in vitro fertilization. The results were compared to those of four fertile men. Quantification was achieved by cataloguing cell defects of the spermatozoon heads and mid/principal pieces of the flagella. Using the data from each specimen, the percentages of total cellular abnormalities in the head/mid/principal pieces were established. The percentages of anomalies of the midpiece and of the principal piece were not significantly different between failed cases and controls. The percentage of cell alterations of the head (96-100 vs 75${\pm}$3,4%), the percentage of combined anomalies of the head (80-86 vs 52.5${\pm}$1.9%), and the percentages of nuclear shape deformation (68-86 vs 47.5${\pm}$6.3%), acrosomal defects (86-96 vs 50${\pm}$4.3%), and postacrosomal sheath defects (78-88 vs 44.5${\pm}$7.2%) of the head were significantly different between failed cases and controls. Due to the cost and time involved in processing semen samples for electron microscopy, the widespread application of this technique to all couples presenting for IVF certainly is not warranted. However, in selected instances electron microscopy may play a crucial role in identifying an occult male factor.

• Nutrition Research and Practice
• /
• v.3 no.2
• /
• pp.149-155
• /
• 2009

In this study, we investigated the heat effect of digestion-resistant fraction (RF) from soybean on retarding bile acid transport in vitro. The RFs from soybean retarded bile acid transport. A raw, unheated RF of soybean (RRF-SOY) was significantly more effective than the heated RF of soybean (HRF-SOY). The RS1 which physically trapped in milled grains and inaccessible to digestive enzyme after 18 hrs incubation level of content in RRF-SOY was found to be as high as 24.1% and after heating the RS1 of HRF-SOY was significantly reduced to 16.8%. The X-ray diffraction pattern of RF from soybean was altered after heat treatment. The RFs from soybean were characterized by peak at diffraction angles of $12.0^{\circ}$ and $20.0^{\circ}$ corresponding to RS content. Cellulose contents of RRF-SOY was 5% higher than that of HRF-SOY and pentosan contents of RRF-SOY was 5% higher than that of HRF-SOY, too. Whereas the hemicellulose content of RRF-SOY was 13% lower than HRF-SOY.