Phylogenetic analysis was conducted to monitor transmission of HIV and to investigate the genetic structure of primary isolates from 12 HIV-1 subtype A infected Koreans. The individuals infected with subtype A viruses had been diagnosed as HIV-1 seropositives during the period 1987 to 1995 and blood samples have been collected from 1991 to 1997. DNA of each individual was isolated from uncultured or cultured peripheral blood mononuclear cells. V3-V5 (0.7 kb) fragment of HIV-1 env gene was amplified by nested polymerase chain reaction and the PCR products were sequenced. The mean value of the divergence of nucleotide of HIV-1 env V3-V5 fragment was $17.0{\pm}4.06%$ ($8.6{\sim}25.8%$) within HIV-1 subtype A isolates from Koreans. This diversity was higher than those of African isolates ($13.7{\pm}2.66%$). In the phylogenetic tree, Korean subtype A isolates were not grouped together, but intermingled into African isolates. The results of this study suggested that HIV-1 subtype A variants be introduced from multiple sites of Africa into Korea and the big genetic diversity of Korea HIV-1 subtype A isolates may be further influenced by the range of geographic locations in which the infection occurred rather than the elapsed time between infection and collection of samples and the disease progression.
A VP6 fragments was subcloned with BamHI in the binary pMBP-1 vector under Califlower Mosaic Virus (CaMV) 355 promoter and neomycin phosphotransferase II (npt II) gene. The recombinant binary vector was mobilized into Agrobacterium-tumefaciens LBA4404 by the freeze-thaw method and potato (Solanum tubensum L. cv Desiree) was transformed by modified leaf-disc cocultivation. Shoots were induced on MS medium with 0.01 mg/L NAA, 0.1 mg/L GA$_3$, 2.0 mg/L Zeatin, 100.0 mg/L kanamycin, 500.0 mg/L carbenicillin. In order to identify the copy number of VP6 into potato plant, total genomic DNA was isolated from transgenic potato and analysed by Southern blotting. Genomic DNA and total mRNA analysis demonstrated the incorporation of the foreign gene into the potato genome, as well as their transcription.
Lee, Kyunghee;Han, Ki Soo;Kwon, Young Sang;Lee, Jung Han;Kim, Sun Ho;Chung, Woo Sik;Kim, Yujung;Chun, Sung-Sik;Kim, Hee Kyu;Bae, Dong-Won
Molecules and Cells
/
v.28
no.4
/
pp.383-388
/
2009
The dehydration responsive element binding protein 2C (DREB2C) is a dehydration responsive element/C-repeat (DRE/CRT)-motif binding transcription factor that induced by mild heat stress. Previous experiments established that overexpression of DREB2C cDNA driven by the cauliflower mosaic virus 35S promoter (35S:DREB2C) resulted in increased heat tolerance in Arabidopsis. We first analyzed the proteomic profiles in wild-type and 35S:DREB2C plants at a normal temperature ($22^{\circ}C$), but could not detect any differences between the proteomes of wild-type and 35S: DREB2C plants. The transcript level of DREB2C in 35S: DREB2C plants after treatment with mild heat stress was increased more than two times compared with expression in 35S:DREB2C plants under unstressed condition. A proteomic approach was used to decipher the molecular mechanisms underlying thermotolerance in 35S:DREB2C Arabidopsis plants. Eleven protein spots were identified as being differentially regulated in 35S:DREB2C plants. Moreover, in silico motif analysis showed that peptidyl-prolyl isomerase ROC4, glutathione transferase 8, pyridoxal biosynthesis protein PDX1, and elongation factor Tu contained one or more DRE/CRT motifs. To our knowledge, this study is the first to identify possible targets of DREB2C transcription factors at the protein level. The proteomic results were in agreement with transcriptional data.
In order to anticipate disease pattern and health problems of Koreans in the 1st part of 21st century (by the year 2020), transition of population characteristics, mortality and morbidity data during the last 30 years Koreans have experienced were reviewed. On the actual basis of epidemiologic transition process that has undergone during last 30 years since 1960 along with socioeconomic development and successful implementation of selective national health policies (family planning, medical insurance and etc.), following changes can be expected in the 21st century in Korea, under the assumption that the current rate of progress is maintained. The population of South Korea alone will be doubled the population of 1960 by the year 2013 : aged Population older than 65 years will be increased from 3.3% in 1960 to 11.4% in 2020 with increased average age of the population from 23.6 year in 1970 to 39.2 year in 2020; urban population from 28% in 1960 to 83% in 2005. GNP/capita has increased tremendously from U.S. $120 in 1970 to $6,749 in 1992, and the government estimated it would be 519,350 in 2010 and $29,460 in 2020. Growth and developmental indices of children, educational achievement and social status of women also showed a remarkable improvement and anticipated to make futher progress. Leading causes of mortality and morbidity have shown a striking change during the last 30 years, from infectious diseases to chronic degenerative diseases and man-made injuries. Occurrence of communicable diseases may become minimal although viral hepatitis, venereal diseases Including AIDS, and well adapted herpes virus infections will maintain their endemic level. Newly evolving infectious agents, however, should be carefully monitored because of rapidly changing environments and human behaviours. Tuberculosis may increase up to the epidemic level when AIDS prevails. Ischemic heart diseases may increase steadily with increasing occurrence of hypertension and diabetes mellitus whereas cerebrovascular diseases may be decreased slowly. Musculaskeletal diseases which contribute a lot to the disability of aged people may be a major health problems due to increased aged population. Mental diseases, particularly that caused by alcohol and drug abuse, and senile dementia may become a prominent health problem. On the other hand injuries caused by traffic and industrial accidents that have shown most striking increase till now may be decreased considerably by intensive intervention. The health policies in the 21st century will be oriented to the health promotion for good quality life rather than life-savings.
World-wide crop loss caused by insect pest and nematode reaches critical level. In Korea, similar crop loss has been gradually augmented in the field and greenhouse due to continuous crop rotation. The current methods on controlling herbivorous insects and plant parasitic nematodes are mostly depended on agro-chemicals that have resulted additional side-effect including occurrence of resistant insects and nematodes, environmental contamination, and accumulation in human body. To overcome the pitfalls, microbe-based control method have been introduced and applied for several decades. Here, we revised biological control using by the bacteria, fungi, and virus in order to kill insect and nematode and to attenuate its virulence mechanism. The introduced microbes mainly secreted out the hydrolysing enzymes and toxic compounds to target host membrane or cell wall directly. Indirectly, the microbe-triggered plant innate immunity against insects and nematodes was also reported. In conclusion, we provide a new frontier of microbe-based environmentally friendly procedure and effective methods to manage insects and nematodes.
Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained Tissue distribution of RHDV in rabbits were examined by immunofluorescence and ABC methods. Tissues including liver, spleen, kidneys, lungs and brain were frozen, cut in a crycut, and fixed in 10% buffered formalin, embedded in paraplast, and cut $5{\sim}7{\mu}m$ thickness. Sections were immunostained with primary antiserum and conjugated second antibodies as recommended by manufacturer. None of the cultures tested showed virus-induced phenomena. Immunoreactive products were commonly found in the liver, in some cases there were also positive staining in the spleen and kidneys. Other organs showed weak or insignificant immunoreactions. By ABC method on the formalin-fixed, paraffin-embedded liver tissues, strong immunoreactivity was found in the periportal triad lesions and peripheral lesions of the hepatic lobules. Immunoreactive products showed diffuse fine granular in the cytoplasm of hepatocytes and sinusoidal cells. In some cells, immunoproducts marginate at the periphery of the cells. The intensive staining of the cytoplasm of infected cells allowed their exact differentiation from surrounding uninfected cells. The positive area involved coincided with histopathological lesion on serial liver sections. In conclusion, liver was proved to be a consistent target organ in RHD, and the immunoperoxidase method in the section of formalin-fixed, paraffin-embedded hepatic tissue could be broadly used for the routine diagnosis of the disease.
Choi, Sook-Kyung;Kim, Tai Jeon;Hong, Seung Bok;Lee, Hun Taeck
Korean Journal of Clinical Laboratory Science
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v.36
no.2
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pp.178-184
/
2004
This research focuses on the overall evaluation of tumor protein (p53) and cell growth marker (Ki-67) in their functions as carcinogenic factors in both a HPV-infected group and in a HPV-noninfected group with the precancerous dysplasia of uterine cervix. Histological grades were determined by the H&E staining and the expression level of p53 and Ki-67 were tested by the immunohistochemistry method. The results were as follows. Among the total of 66 cases, p53 (+) was observed in 19 cases (29.0%) from the mild grade group, 22 cases (33.0%) from the moderate grade group, and 19 cases (29.0%) from the severe grade group. The values correlate with the increase of dysplasia intensity in HPV-noninfected group and showed significant correlation (p<0.05), but there were no significant difference from the HPV-infected group. Among a total of 66 cases, the mitotic index of Ki-67 (+) were observed in 19 cases (29.0%) from the mild grade group, 22 cases (33.0%) from the moderate grade group, and 19 cases (29.0%) from the severe grade group. The values were significantly different against dysplasia intensity (p<0.05), but showed no significant difference from HPV infection. After cross comparing the statistical parameters of p53 and ki-67 in their significance, p53 was shown to be statistically significant with Ki-67 while there was no statistically significant difference with Ki-67 (p<0.05). Taken together, tumor protein (p53) and an index of Ki-67 observed in cervical dysplasia and in HPV related dysplasia of cervix uterine did not have any notable significance with HPV infection. The incidence rate of p53, however, had some significant correlation with dysplasia while Ki-67 had no particular statistical significance. As a result, p53 and Ki-67 can be considered as effective diagnostic markers in predicting the disease progression of dysplasia to cervical cancer.
HCV is a single-stranded RNA virus and more than 1 million new cases are reported annually worldwide. The six major HCV genotypes and numerous subtypes vary in their geographic distribution. It is thought that genetic heterogeneity of HCV may account for some of the differences in disease outcome and response to treatment observed in HCV infected persons. In this study, we determined HCV genotypes among chronic Korean HCV patients and evaluated direct sequence PCR protocols developed. For the study, 232 chronic HCV patient sera were used. HCV RNA was extracted and two pairs of consensus PCR primers were selected in 5'UTR region for amplification of HCV RNA. Amplification products obtained from the HCV positive cases were subjected to automatic sequencing. Sequences were compared with those in GenBank by using the BLAST program. From this study, five HCV genotypes, 1b, 2a, 2b, 2c and 3a were found. HCV genotypes 4, 5 and 6 were not determined. HCV genotype 1b (53.9%, 125/232) and 2a (35.8%, 83/232) were most frequently found. This group was followed by 2b (3.9%, 9/232), 3a (3.4%, 8/232) and 2c (3.0%, 7/232). The data presented here suggest a complex distribution of HCV types and they were well correlated with other reports on Koreans and will be helpful for type-specific follow-up of Korean HCV patients. This study showed that 5'UTR direct sequence analysis is a sensitive and rapid method to identify HCV genotypes.
Choi, Jae Chan;Kim, Young Chul;Choi, Hwan Jun;Park, Jeon Oh;Jeong, Hyun Do
Journal of fish pathology
/
v.28
no.1
/
pp.27-35
/
2015
VHSV is a major viral agent that affects freshwater and marine fish, causing serious economic losses in aquaculture in the world. Due to their filter-feeding activity, bivalve mollusks may act as viral transmitters after accumulation of the fish viruses released into seawater from infected fish. Amplification by RT-PCR was carried out to investigate the presence of VHSV in pacific oysters (Crassotrea gigas) and blue mussels (Mytilus edulis), inhabiting regions around aquatic farms in Korea. Primers designed from conserved regions of VHSVs allowed us to detect four different types of VHSV in a single PCR. Twenty two of the eighty four samples showed positive results of VHSV in a 2-step RT-PCR. Using six positive samples from three different regions in Korea, we cloned and sequenced the glycoprotein (G) gene (467-bp long) of VHSVs. Genetic analysis of the VHSVs detected in shellfish in various geographical areas of Korea showed highly restricted results to VHSV type Iva. This was in agreement with the reports showing only a single genotype of VHSV (Iva genotype) in outbreaks in cultured or wild fish in Korea. Consequently, we investigated VHSVs carried by bivalve mollusks inhabiting the vicinity of aquatic farms, and revealed correlationship between the type of viral accumulated in shellfish by filter-feeding, and those detected in disease outbreaks in fish.
This study was conducted to investigate the relationship between systemic diseases and oral malodor. The author measured the volatile sulfur compound(VSC) of the patients who visited Pusan National University Health Promote Center for a comprehensive medical testing. The patients were examined gingival bleeding on probing, CPI index, tongue coating. Their systemic diseases were diagnosed by the specialist. 182 patients consisted of 112 males and 70 females. In this study, Oral $Chroma^{(R)}$ was used to measure oral malodor. This equipment could measure the concentration of intraoral VSC (hydrogen sulfide, methyl mercaptan, dimethyl disulfide). All data were analylized using Statistical Package for the Social Science $12.0^{(R)}$. The result of this study was the followings. 1. There was significant difference of numbers of patient who visited health care center according to the VSC concentration level and the Community Periodontal Index, bleeding on probing, tongue coating. 2. The subjects with hyperlipidemia showed the high level of $CH_3SH$ concentration (p=0.036). The concentration of $H_2S$ tends to be high in the group with abnormal findings on pulmonary fuction test(p=0.086). The concentration of $CH_3SH$ in the groups with abnormal findings on lipid profile test(p=0.130) and bone mineral density test(p=0.099) and abdominal ultrasonograpy(p=0.088) tends to be higher than the other group. 3. The concentration of $(CH_3)_2S$ in the group with abnormal findings on blood pressure test(p=0.113), hepatitis B virus serology(p=0.069), Abdominal ultrasonograpy(p=0.091) tend to be higher than the other group.
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