Using bioinformatic tools for searching the massive genome databases, it is possible to Identify new genes in few minutes for initial discoveries based on evolutionary conservation, domain homology, and tissue expression patterns, followed by further verification and characterization using the bench-top works. The development of high-density two-dimensional arrays has allowed the analysis of the expression of thousands of genes simultaneously in the humans, mice, rats, yeast, and bacteria to elucidate the genes and pathways involved in physiological processes. In addition, rapid and automated protein identification is being achieved by searching protein and nucleotide sequence databases directly with data generated from mass spectrometry. Recently, analysis at the bio-chemical level such as biochemical screening and metabolic profiling (Biochemical genomics) has been introduced as an additional approach for categorical assignment of gene function. To make advantage of recent achievements in computational approaches for facilitated gene discoveries in the avian model, chicken expression sequence tags (ESTs) have been reported and deposited in the international databases. By searching EST databases, a chicken heparanase gene was identified and functionally confirmed by subsequent experiments. Using combination of sub-tractive hybridization assay and Genbank database searches, a chicken heme -binding protein family (cSOUL/HBP) was isolated in the retina and pineal gland of domestic chicken and verified by Northern blot analysis. Microarrays have identified several host genes whose expression levels are elevated following infection of chicken embryo fibroblasts (CEF) with Marek's disease virus (MDV). The ongoing process of chicken genome projects and new discoveries and breakthroughs in genomics and proteomics will no doubt reveal new and exciting information and advances in the avian research.
Intestinal parasitic infections are one of the major causes of diarrhea in human immunodeficiency virus (HIV) seropositive individuals. Antiretroviral therapy has markedly reduced the incidence of many opportunistic infections, but parasite-related diarrhea still remains frequent and often underestimated especially in developing countries. The present hospital-based study was conducted to determine the spectrum of intestinal parasitosis in adult HIV/AIDS (acquired immunodeficiency syndrome) patients with or without diarrhea with the levels of $CD4^+$ T-cell counts. A total of 400 individuals were enrolled and were screened for intestinal parasitosis. Of these study population, 200 were HIV seropositives, and the remaining 200 were HIV uninfected individuals with or without diarrhea. Intestinal parasites were identified by using microscopy as well as PCR assay. A total of 130 (32.5%) out of 400 patients were positive for any kinds of intestinal parasites. The cumulative number of parasite positive patients was 152 due to multiple infections. A significant association of Cryptosporidium (P<0.001) was detected among individuals with $CD4^+$ T-cell counts less than $200cells/{\mu}l$.
Kim, Mun-Hyeon;Seo, Min-Goo;Lee, Hyang-Sim;Kim, Ji-Yeon;Kim, Ji-Ye;Kim, Yong-Sang;Kim, Yeon-Hee
Korean Journal of Veterinary Service
/
v.42
no.4
/
pp.285-288
/
2019
Foot-and-mouth disease (FMD) causes economic problems in livestock industry because of fast spread and inducing low productivity. FMD outbreaks occurred in South Korea over the period from 2000 to 2019. Vaccination is the most practical and effective means of controlling or preventing these outbreaks, and a national vaccination policy has been in place for all FMD-susceptible animals since 2010. To prevent and control of FMD, South Korea has been using vaccines imported from the United Kingdom, Argentina, and Russia. The Animal and Plant Quarantine Agency of South Korea oversees continuous quality control of imported FMD vaccines. FMD vaccines were evaluated characteristics, sterility, pH, inactivation, safety, potency test by Korean FMD vaccine standard assay (Test for National Lot Release). The 6 company vaccines (A~F) were used Test for National Lot Release by each method. We evaluated quality of each FMD vaccine from 2015 to 2019. All batch of vaccine showed good quality control and were passed the Test for National Lot Release. The serotypes of vaccine are increasingly changing to multiple vaccine because the FMD was outbreak by various serotype virus in South Korea. Furthermore, this data may be useful as a basis for ensuring the quality of FMD vaccines and for base data to manage them. Additional study is required to simple approach for rapid evaluation of quality and antigen content identification in vaccines.
Seo, Ji-Hyun;Yeom, Jung Sook;Youn, Hee-Shang;Han, Tae-Hee;Chung, Ju-Young
Clinical and Experimental Pediatrics
/
v.58
no.3
/
pp.102-107
/
2015
Purpose: Human parechovirus (HPeV) and enterovirus (EV) are causative agents of a sepsis-like illness in neonates and of infections of the central nervous system in young children. The objectives of this study were to assess the prevalence of HPeV3 and EV infection in young children with a sepsis-like illness or with meningitis in Jinju, Korea. Methods: Cerebrospinal fluid (CSF) samples were collected from 267 patients (age range, 1 day to 5 years) and assessed for HPeV and EV by performing reverse transcription polymerase chain reaction assay. Amplification products of the VP3/VP1 region of HPeV and of the VP1 region of EV were sequenced to identify the virus type. Results: HPeV and EV were detected in 3.4% and 7.5% of the total CSF samples assessed, respectively. The age distribution of EV-positive patients (median age, 1.4 months) had a significantly broader range than that of HPeV-positive patients (median age, 7.8 months). The peak seasons for HPeV and EV infection were spring and summer, respectively. The clinical symptoms for HPeV and EV infection were similar, and fever was the most common symptom. Pleocytosis was detected in 22.2% of HPeV-positive patients and 35.5% of EV-positive patients. The VP3/VP1 gene sequence of the nine Korean strains clustered most closely with the Japanese strain (AB759202). Conclusion: The data indicate that HPeV infection is predominant in young infants (<6 months) and that meningitis without pleocytosis was caused by both HPeV and EV infection in children.
Alagozlu, Hakan;Ozdemir, Ozturk;Koksal, Binnur;Yilmaz, Abdulkerim;Coskun, Mahmut
Asian Pacific Journal of Cancer Prevention
/
v.14
no.9
/
pp.5489-5494
/
2013
In the current study we aimed to show the common YMDD motif mutations in viral polymerase gene in chronic hepatitis B patients during lamivudine and adefovir therapy. Forty-one serum samples obtained from chronic hepatitis B patients (24 male, 17 female; age range: 34-68 years) were included in the study. HBV-DNA was extracted from the peripheral blood of the patients using an extraction kit (Invisorb, Instant Spin DNA/RNA Virus Mini Kit, Germany). A line probe assay and direct sequencing analyses (INNO-LIPA HBV DR v2; INNOGENETICS N.V, Ghent, Belgium) were applied to determine target mutations of the viral polymerase gene in positive HBV-DNA samples. A total of 41 mutations located in 21 different codons were detected in the current results. In 17 (41.5%) patients various point mutations were detected leading to lamivudin, adefovir and/or combined drug resistance. Wild polymerase gene profiles were detected in 24 (58.5%) HBV positive patients of the current cohort. Eight of the 17 samples (19.5%) having rtM204V/I/A missense transition and/or transversion point mutations and resistance to lamivudin. Six of the the mutated samples (14.6%) having rtL180M missense transversion mutation and resistance to combined adefovir and lamivudin. Three of the mutated samples (7.5%) having rtG215H by the double base substituation and resistance to adefovir. Three of the mutated samples (7.5%) having codon rtL181W due to the missense transversion point mutations and showed resistance to combined adefovir and lamivudin. Unreported novel point mutations were detected in the different codons of polymerase gene region in the current HBV positive cohort fromTurkish population. The current results provide evidence that rtL180M and rtM204V/I/A mutations of HBV-DNA may be associated with a poor antiviral response and HBV chronicity during conventional therapy in Turkish patients.
Juglans mandshurica belongs to the family Juglandaceae is known to contain a wide range of pharmacological activities including anti-cancer, anti-inflammation, astringent, and anti-human immunodeficiency virus-type 1 (HIV-1). Melanogenesis refers to the biosynthesis of melanin pigment in melanocytes. In this study, to investigate the whitening activity of the extracts from Juglans mandshurica, we measured effects on a tyrosinase activity, a melanogenesis, and a tyrosinase synthesis in the B16/BL6 melanoma cells and an antioxidant activity. The extracts significantly scavenged a 1,1-diphenyl-2-picrylhydrazyl (DPPH) and a superoxide anion radicals in a dose-dependent manner with a $SC_{50}$ value of $20\;{\mu}g/mL$ and $25\;{\mu}g/mL$, respectively. Also, the tyrosinase activity and melanogenesis were significantly inhibited by the extracts. Furthermore, the synthesis of tyrosinase protein was significantly decreased by the extracts in enzyme-linked immunosorbent assay. Double blind study on the clinical efficacy of a cream containing 2% of the extracts showed that the extracts have a significant skin whitening effect. Therefore, this study demonstrates that the extracts from Juglans mandshurica may be useful as a potential agent for skin whitening.
Purpose : Hepatitis B virus DNA transfected cell line(HepG2.2.15) was cultured to evaluate the effect of herbs on the expression of HBeAg and the replication of HBV. HepG2.2.15 produces HBV particles as well as viral proteins into cell culture media. Methods : Extracts of herbs were adminitered to the cells on the proper concentration. Culture media was collected 48 hours after the herbal administration and HBeAg level in the media was examined by ELISA method. To confirm that the anti-viral effect was not due to direct cytotocixity of the extracts, normal cell proliferation was shown by cell counting. And as of the interference in protein synthesis of HepG2.2.15 by herb-extracts, we used the result of study that we performed before by ${\alpha}FP$ assay using EIA method. Results& Conclusion : Herb medicines like 地楡(Sanguisorbae Radix) and 覆盆子(Rubi Frusctus) showed significant inhibitory effect on HBeAg expression at p<0.01 and 五味子(Acanthopanacis Cortex) at p<0.05. Whereas, though some herbs such as ?草根(Rubiae Radix), 山査(Crataegii Fructus), 白芍藥(Paeoniae Radix Alba), and 大黃(Rhei Radix et Rhizoma) showed the tendecy to suppress HBeAg. most of them were not significant statistically. From the above, we could conclude that those herb medicines can be applied to patients effectively and further studies on effective fraction of some herbs are thought to be needed.
Proceedings of the Korea Society of Poultry Science Conference
/
2000.11a
/
pp.81-84
/
2000
Effects of different photoperiod regimens on the cellular and humoral immunity in broiler chickens were studied(Exp 1). Total one hundred ninety two one-day-old commercial broiler chicks(Cobb$\times$Cobb) were raised between constant lighting(CL) and intermittent lighting (1h light: 3h darkness(IL; 1l; 3D) Body weight, feed intake and feed conversion were measured for seven week. Peripheral blood and splenic lymphocyte activities were tested at 3 and 5 wk of age by performing a mitogen cellproliferation assay with a polyclonal T-cell mitogen, concanavalin A (Con A), and B-cell mitogen, lipopolysaccharide (LPS). To investigate the effect of photoperiod on the humoral immunity, chicks were immunized with sheep red blood cell(SRBC) and iinactivated Newcastle disease virus(NDV) vaccine. Total immunoglobulin G(IgG) concentration was also determined. Diurnal change of melatonin was tested in sera. In experiment 2, 0.1ml melatonin were subcutaneously injected from three to five weeks old if immunomodulation effect of lighting regimen was due to the melatonin or not. Injections of melatonin were made at 0700h and the dosage was 10ng (M2), 100ng(M3), 1$\mu\textrm{g}$(M4) per bird daily, respectively. control were quivalent injections of vehicle(M1). Lymphocyte activities were tested and humoral immunities were examined at 5 weeks of age. Blood melatonin concentration was determined at 0h, 1, h, 2h, and 3h posterior to injection at five weeks old. It was higher in CL chicks than IL chickens during the subsequent period of 3 to 5 wk of age. However, weight gain of chicks raised IL were significantly higher at 6 wk of age than CL(P<0.05). Antibody response to NDV was not affected by both photoperiod regimens and melatonin injection, whereas anti-SRMB titer and IgG concentration were enhanced. Lymphocyte activity of chickens raised under IL was sighificantly higher than those of chickens raised under CL. Melatonin injection also increased lymphocyte activity. When peripheral blood lymphocytes were used, proliferation response to LPS and Con A were significantly increased in M2 and respectively. The results of this experiments suggest that IL improved host immune response and melatonin have immunomodulatory roles.
Hyun-Il Cho;Soon-Young Pail;Il-Hoan OH;Kyun-Jung Ahn;Dong-Wook Kim
Journal of Microbiology
/
v.39
no.4
/
pp.300-304
/
2001
Epstein-Barr Virus(EBV)-transformed lymphoblastoid B cell lines, BLCL which expresse antigens, are potential antigen-presenting cells(APCs) for the induction of CTL in vitro. However transfection of BLCLs with subsequent selection by antibiotics is notoriously difficult because plating efficiencies of BLCLsare reported to be 1% or less. To generated stable transfectants of BLCLs we produced high titers of retroviruess encoding pp 65 antigen of human cytomegalovirus of foreign antigens and trans-duced them of BLCLs. The pp 65 gene was cloned into the retroviral vector pLXSN. The recombinant retroviral vector was transfected to ecotropic packaging cell line, CP&E86, and this polyclonal recom-binant retrovirus was transduced to PA317 that is amphotropic pakaging cell line. The titers of colned PA317 amphotropic retroviruses ranged from 5 to $\times$10$^{6}$ colony forming units (CFU)per ml (CFU/ml) We performed three rounds of consecutive transductions to BLCLs in order to improve the clon-ing effieiencies. The expression of recombinant HCMV-pp65 antigen was more than 20% after the final transduction. THe third-transduced BLCLs were easily selected in optimal concentration of G418. BLCLs expressing foreign antigens could be used as target cells for CTL assay and/or as APCs for induction of in vitro CTL responses specific for viral and tumor antigens.
Mammary tissue-specificity of the caprine $\beta$-lactoglobulin promoter appears to be secured by repression in non-expressing cells. In order to identify the mechanism of the negative regulation, the upstream promoter sequence of the caprine $\beta$-lactoglobulin gene was analyzed in detail. The repression was mediated by the upstream flanking sequence from -47O to -205. The sequence could repress the promoter activity of $\beta$-lactoglobulin in either orientation. The effect of the putative negative regulation element of caprine $\beta$-lactoglobulin on heterlogous promoters, however, varied: the promoter activity of herpes simplex virus thimidine kinase was either repressed or activated by the sequence depending on its orientation, while the SV4O early promoter was activated rather than repressed. The regulatory sequence involving the putative negative regulatory element was strongly shifted with the nuclear extract from non-mammary HeLa and CV-1 cells, while only weak shift was observed with that of mammary HC11 cells. Such correlation between repression and factor binding suggests that the protected regions in foot-printing assay may be the negative regulatory elements of $\beta$-lactoalobulin that serve tissue-specific repression.
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