• Korean Journal of Veterinary Research
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• v.44 no.3
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• pp.433-439
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• 2004

This study was carried out to investigate the efficacy of rifampin with or without streptomycin in male Sprague-Dawley (SD) rats experimentally inoculated with Brucella abortus. Thirty rats were intraperitoneally inoculated with $1.0{\times}10^9$colony-forming units of B. abortus. They were divided into 3 groups by treatment with antibiotic. 10 rats in Group A were orally administrated with rifampin, 10 rats in Group B with rifampin orally and with streptomycin intramuscularly over 12 weeks starting at 1 week post infection (PI). A placebo recipient in Group C was inoculated with sterile saline without antibiotics. All animals were monitored by tube agglutination test (TAT) and AMOS-PCR to evaluate the efficiency of the antibiotics to B. abortus infection. The antibody titers in Groups A, B and C were 1:400, 1:400 and 1:800 as measured by TAT at the first week PI, respectively. The antibody titer in Group A decreased to 1:100 by the 13th week PI. That in the control group was observed as high antibody titer until 13th weeks PI, but the antibody response in Group B was low(1:50) from the 5th week to the 13th week PI. AMOS-PCR there was evidence of relapse of B. abortus in group A in liver and spleen specimens at the 13th week PI. B. abortus DNA was detected in Group C in liver and spleen specimens from the 1st week to 13th week PI by AMOS-PCR. However AMOS-PCR could not detect any organism in Group B from the 3rd week PI until the end of the study. This study demonstrated that administration of a combination of rifampin and streptomycin was more efficacious than administration of rifampin alone. A significant reduction in antibody titer was observed when a combination of 15 mg/kg/day of rifampin per os and 15 mg/kg/day streptomycin intramuscularly was used in comparison with the antibody of control group.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.4
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• pp.476-482
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• 2013

The present study makes an investigation into expression of genes related to cardiac development in chicken, quail and chicken-quail hybrids during the early stage of embryogenesis. Real-time PCR was used to detect mRNA expressions of Nkx2-5, GATA4 and TBX5 in the heart of chicken, quail and chicken-quail hybrids embryos during the 3rd to 7th days of incubation. Results showed that NKX2-5 mRNA displayed a similar expression trend in chicken, quail and chicken-quail hybrids. The initial and highest expression of Nkx2-5 was focused on the 3rd day of incubation, then it declined till 5th day of incubation, thereafter, it fluctuated. Expression of Nkx2-5 gene in quail was significantly higher than in chicken and chicken-quail hybrids, and no significant difference was observed between the two latter species. GATA4 mRNA showed a similar expression trend between chicken and quail, which displayed a steady increase from 3rd to 6th d, then, the expression level decreased. However, GATA4 mRNA expression in chicken-quail hybrids was significantly higher than that in chicken and quail from 3rd to 5th d (p<0.01), but significantly lower than that in chicken and quail during the later stage of the experiment (p<0.05), due to the dramatic drop from 5th d onwards (p<0.01). TBX5 mRNA expression in chicken and quail showed the same trend as GATA4 expressed in the two species. Furthermore, TBX5 expression in chicken-quail hybrids was significantly higher than that in chicken and quail during the whole course of experiment, although relatively lower TBX5 expression was detected in the early stage. In conclusion, Nkx2-5, GATA4 and TBX5 genes showed dynamic changes during the process of cardiac development in chicken, quail and their hybrids embryos. In addition, the expression trend in chicken was similar to that in quail, and there was no significant difference for gene expression level, except NKX2-5. However, expression of these genes in chicken-quail hybrids was significantly different from their parents, the difference mechanism needs to be further explored.

• The Journal of Internal Korean Medicine
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• v.27 no.2
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• pp.379-393
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• 2006

Objectives: We are aimed to identify anti-tumor effects of Curcuma aromatics on some kinds of cancer cells through molecular biologic methods. Materials & Methods: We used 4 kinds of cancer cell lines such as lung cancer cells(AS49), cervical cancer cells(HeLa), glioma cancer cells(A172) and prostate cancer cells(PC3). We treated the boiled extract of Curcuma aromatica $5{\mu}g,\;10{\mu}g$ to cultural media(ml) for 24 hours. We measured the cytotoxicitv on 4 kinds of cancer cells through tryphan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which are genes related to apoptosis. We examined the effect on the revelation of Bcl-2 Protein and Bar protein by western blot analysis. Results : In the experiment of tryphan blue exclusion test, the extract of Curcuma aromatica showed more significant killing effect on AS49, HeLa than the control group with density dependent manner, which was statistically significant. In the experiment of MTT assay the extract of Curcuma aromatica showed more suppressive effect on viability of A549, HeLa than the control group with density dependent manner, which was statistically significant. Curcuma aromatica induced apoptosis by decreasing the membrane potential of mitochondria in A549, HeLa. In the experiment of the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A549, HeLa treated with Curcuma aromatica with dose dependent manner. In the experiment of the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in AS49, HeLa treated with Curcuma aromatica with dose dependent manner. Conclusions: From this study, we can infer that Curcuma aromatica has anti-tumor effect on lung cancer cells and uterine carcinoma cells but not on glioma cells and prostate cancer cells.

• The Journal of Internal Korean Medicine
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• v.27 no.2
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• pp.429-443
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• 2006

Objectives : The Purpose of this study was to identify anti-tumor effects of Curcuma longa L. on some kinds of cancer cells through molecular biologic methods. Materials & Methods : We used 4 kinds of cancer cell lines such as glioma cells(A172), cervical cancer cells(HeLa), Prostate cancer cells(PC3), lung cancer cells(A549). We injected the boiled extract of Curcuma longa L. $5{\mu}g,\;10{\mu}g$ to culture media(ml) for 24 hours. We measured the cytotoxic effect on 4 kinds of cancer cells through trypan blue exclusion test and the suppressive effect on viability of 4 kinds of cancer cells via MTT assay. We measured the change of mitochondria membrane potential via flow cytometry. The quantitative RT-PCR was used to examine the effect on the revelation of Bcl-2 and Bax which genes are related to apoptosis. We examined the effect on the revelation of Bcl-2 protein and Bax protein by western blot analysis. Results: 1. Extract of Curcuma longa L. showed significant cytotoxic effect on A172, HeLa, PC3 compared to the control group with density dependent manner. 2. Extract of Curcuma longs L. showed significant suppressive effect on viability of A172, HeLa, PC3 compared to the control group with density dependent manner. 3. Curcuma longs L. induced apoptosis by decreasing the membrane potential of mitochondria in A172, HeLa, PC3. 4. In the test about the revelation of genes related to apoptosis, the revelation of Bcl-2 decreased and the revelation of Bax increased in A172. HeLa, PC3 treated with Curcuma longa L. with density dependent manner. 5. In the test about the revelation of protein related to apoptosis, the protein levels of Bcl-2 decreased and the protein levels of Bax increased in A172, HeLa, PC3 treated with Curcuma longa L. Conclusions: This experiment shewed that Curcuma longs L. has anti-tumor effect on glioma, cervical, Prostate cancer cells except on lung cancer. We hope that anti-tumor effects of Curcuma longa L. will be more Practically identified.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.37-37
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• 2015

dsRNA was found in malformed cultures of Lentinula edodes strain FMRI0339, one of the three most popular sawdust cultivated commercial strains of shiitake, and was also found in healthy-looking fruiting bodies and actively growing mycelia. Cloning of the partial genome of the dsRNA revealed the presence of the RdRp sequence of a novel L. edodes mycovirus (LeV), and sequence comparison of the cloned amplicon showed an identical sequence to known RdRp genes of LeV found in strain HKA. The meiotic stability of dsRNA was examined by measuring the ratio of the presence of dsRNA among sexual monokaryotic progeny. More than 40% of the monokaryotic progeny still contained the dsRNA, indicating the persistence of dsRNA during sexual reproduction. Comparing the mycelia growth of monokaryotic progeny suggested that, although variations in the growth rate existed among progeny and virus infection was observed in highly actively growing progeny, there appeared to be a tendency toward a lower frequency of virus incidence in actively growing progeny. This study attempted to cure the edible mushroom L. edodes strain FMRI0339 of the L. edodes mycovirus (LeV) in order to obtain an isogenic virus-free fungal strain as well as a virus-infected strain for comparison. Mycelial fragmentation, followed by being spread on a plate with serial dilutions resulted in a virus-free colony. Viral absence was confirmed with gel electrophoresis after dsRNA-specific virus purification, Northern blot analysis, and PCR using reverse transcriptase (RT-PCR). Once cured, all of fungal cultures remained virus-free over the next two years. Interestingly, the viral titer of LeV varied depending on the culture condition. The titer from the plate culture showed at least a 20-fold higher concentration than that grown in the liquid culture. However, the reduced virus titer in the liquid culture was recovered by transferring the mycelia to a plate containing the same medium. In addition, oxygen-depleted culture conditions resulted in a significant decrease of viral concentration, but not to the extent seen in the submerged liquid culture. Although no $discernable phenotypic changes in colony morphology were observed, virus-cured strains showed significantly higher growth rates and mycelial mass than virus-infected strains. We were also explored effects of LeV on fruiting body formation and mushroom yield. The fruiting body formation yield of virus-free L. edodes was larger than virus-infected L. edodes. These results indicate that LeV infection has a deleterious effect on mycelial growth and fruiting body formation. In addition, we have been investigated host-parasite interaction between L. edodes and its mycovirus interaction to study viral mechanism by establishment of proteomics.

• 보존과학연구
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• s.20
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• pp.5-20
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• 1999

We have analyzed the allele and genotype frequencies from 10 fractions of ancient human skeleton in 3 pieces of Jar coffin excavated from Naju Bokamni3rd tumulus by PCR amplification, high resolution polyacrylamide gelelectorphoresis and silver staining. We could isolate human genomic DNA from 3 bone fractions but the rest of them could not be used as materials due to being decayed. We could detect sex determination as male and 3 genotypes of STR system, HUMTHO1, HUMTPOX and HUMC5F1PO from the bone fraction of left side in Jar coffin 3 and see the slightly reaction suggesting the sex as male from the bone fraction of the left side in Jar coffin 2 and female from the right side in Jar coffin 3.We have also analyzed the genotype frequencies of mitochondria from the bone fractions of the left side and the right side in jar coffin 3, respectively. From the result of indetifiying at nucletide position between 16018 and 16378of the base of hyper variable region(HV1) in the control region, We can presume that the both bones have the same maternal inheritance.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.4
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• pp.261-270
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• 2004

The purpose of this experiment was to examine the histological changes and the pattern of expression of type I, II collagen in the elongated area by distraction osteogenesis in the rabbit mandible. Sixteen rabbits weighing 2.5kg-3kg were used for this experiment. Experimental group was distracted at the rate of 0.7mm, twice/day for 7days, and control group was only osteotomized. After 5 days latency, osteotomic site is distracted for 7days. Consolidation period is 28days. The animal was sacrificed at the 3rd, 7th, 14th, 28th day after the operation. The distracted bone was examined by histological analysis and RT-PCR analysis. The results were summarized as follows: 1. Experimental group was observed that the gaps between the distracted bone edges were occupied by new bone. 2. Expression of Type I collagen were detected throughout the experiment in both groups and Expression of Type I collagen were markedly increased during distraction and consolidation period in experimental group than control group. 3. Expression of Type II collagen were detected throughout the experiment in both groups and expression of Type II collagen were maintained at high level during distraction and consolidation period in experimental group than control group. From these results, in contrast to type II collagen, type I collagen seemed to be more expressed by mechanical stimuli during distraction and consolidation period. The predominent mechanism of new bone formation in the distraction gap was intramembranous bone formation, but some of the regenerated bone was formed by endochondral ossification.

• Journal of Microbiology
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• v.41 no.2
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• pp.148-151
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• 2003

Waterborne parasite infections are considered a reemerging threat. Most studies on the epidemiology of human cryptosporidiosis, giardiasis, and amebiasis have been carried out in developed countries, and there is little data on the occurrence of these infections in other areas. The objective of this study was to investigate the presence of waterborne parasites such as Cryptosporidium parvum, Giardia lamblia and Entamoeba histolytica in various water samples in Ankara, turkey. A total of 85 samples were examined, 43 from the municipal water supply, 34 from wells, 6 from the Ankara River, and 2 from two untreated dams; by conventional microscopy, immunologically and by polymerase chain reaction (PCR). Oocysts of C. parvum and cysts of G. lamblia were detected by using an indirect fluorescence (antigen) assay, whereas an enzyme linked immunosorbent assay was used to detect the cysts of E. histolytica and E. dispar. In addition, PCR was used for E. histolytica, E. dispar, C. parvum and G. lamblia detection. G. lamblia was found in 2 of the 34 well water samples, and parasites were found in 3 of the 6 Ankara River samples. The 1$\^$st/ contained E. histolytica cysts and Strongyloides stercoralis larvae. the 2$\^$nd/ E. histolytica cysts, and Trichuris trichiura eggs, and the 3$\^$rd/ C. parvum oocysts only. No parasite was observed in the municipal water samples and untreated dam water samples. These results extend our knowledge on waterborne parasites, such occurrence information on waterborne pathogens assists the management and treatment of municipal water.

• Macromolecular Research
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• v.13 no.4
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• pp.285-292
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• 2005

We developed alginate beads loaded with transforming growth $factor-{\beta}_{1}(TGF-{\beta}_{1})$ to examine the possible application of the scaffold and cytokine carrier in tissue engineering. In this study, bone marrow stromal cells (BMSCs) and $TGF{\beta}_{1}$ were uniformly encapsulated in the alginate beads and then cultured in vitro. The cell morphology and shape of the alginate beads were observed using inverted microscope, scanning electron microscope (SEM), histological staining and RT-PCR to confirm chondrogenic differentiation. The amount of the $TGF{\beta}_{1}$ released from the $TGF-{\beta}_{1}$ loaded alginate beads was analyzed for 28 days in vitro in a phosphate buffered saline (pH 7.4) at $37^{\circ}C$. We observed the release profile of $TGF-{\beta}_{1}$ from $TGF-{\beta}_{1}$ loaded alginate beads with a sustained release pattern for 35 days. Microscopic observation showed the open cell pore structure and abundant cells with a round morphology in the alginate beads. In addition, histology and RT-PCR results revealed the evidence of chondrogenic differentiation in the beads. In conclusion, these results confirmed that $TGF-{\beta}_{1}$ loaded alginate beads provide excellent conditions for chondrogenic differentiation.

• Journal of Veterinary Science
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• v.22 no.1
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• pp.1.1-1.12
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• 2021

Background: Goslings in several Taiwanese farms experienced gosling feather loss disease (GFL) at 21-35 days and goose broke feather disease (GBF) at 42-60 days. The prevalence ranges from a few birds to 500 cases per field. It is estimated that about 12,000 geese have been infected, the morbidity is 70-80% and the mortality is 20-30%. Objectives: This study aims to investigate the pathogens that cause GFL and GBF. Focus on the study of the correlation between goose circovirus (GoCV) and goose parvovirus (GPV) with the goose feather loss in southern Taiwan. Furthermore, a phylogenetic tree was established to align the differences between southern and northern Taiwan and compare with virus strains from China and Europe. Methods: Samples were collected from animal hospitals. Molecular and microscopy diagnostics were used to examine 92 geese. Specific quantitative polymerase chain reaction (Q-PCR) assays are performed to evaluate GPV and GoCV viral loads and simultaneously evaluated the feather loss conditions in geese with the scoring method. Results: High prevalence of GoCV and GPV infection in geese showing signs of GFL and GBF. Inclusion body was detected in the feather follicles and Lieberkuhn crypt epithelial cells. The Q-PCR showed the high correlation between feather loss and viruses during 3rd-5th week. However, the infection was not detected using the same test in 60 healthy geese. Conclusions: Thus, GFL and GBF appear to be significantly closely related to GoCV and GPV. The geese feathers showed increasing recovery after being quarantined and disinfected.