• Parasites, Hosts and Diseases
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• v.37 no.3
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• pp.163-169
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• 1999

House dust mite allergens have been well established as sensitizing agents that are important in the induction of allergic diseases. In order to analyze epitopes of the allergen and to develop a quantitative method of the allergen exposure, monoclonal antibodies against a recombinant Der p 2 (rDer p 2), one of the major allergens of Dermatophogoides pteronyssinus, were produce. Four monoclonal antibodies produced wee species-specific and did not cross-react to the D. farinae crude extract. Two of the monoclonal antibodies were found to be IgG1 and the others were IgM. For the analysis of epitopes, a Der p 2 cDNA encoding 126 amino acids (aa) was dissected into three fragments with several overlapping peptides, A (aa residues 1-49), B (44-93), and C fragment (84-126). Three monoclonal antibodies showed reactivities to the recombinant B fragment and to the full-length rDer p 2, but one monoclonal antibody reacted only with the full-length rDer p 2. Two-site capture ELISA was developed using two different monoclonal antibodies for quantitating Der p2 in house dust. The sensitivity limit was 4ng/ml with rDer p2 and $8{\;}\mu\textrm{g}/ml$ with the d. pteronyssinus crude extract. The result suggested that the assay using monoclonal antibodies against rDer p2 could be useful for the environmental studies and for the standardization of mite allergen extracts.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.4
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• pp.2439-2442
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• 2013

We investigated the possible association of interleukin-10 (IL-10) single nucleotide polymorphisms (SNPs) and susceptibility to acute myeloid leukemia (AML) in 115 patients and 137 healthy controls. Genetic analysis of IL-10 SNPs at -819 and -592 was carried out with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) approach. The IL-10 mRNA expression of AML patients and controls with different genotype was detected by real-time quantitative polymerase chain reaction (RT-PCR). Genetic analysis of IL-10 revealed that the -819AA genotype frequencies and the -819A allele frequencies in the AML group were higher than in the controls (59.1% vs 40.9%; 75.6% vs 63.9%, respectively); there were remarkable differences in -819T/C and -592A/C gene distribution (P<0.05) and the TA haploid frequencies were higher in the AML group (75.6% vs 63.9%, P<0.05). IL-10 mRNA expression in incipient AML patients was obvious higher than the non-tumor group and the remission group ($7.78{\times}10^{-3}$ vs $2.43{\times}10^{-3}$, $3.64{\times}10^{-3}$, P<0.05).The study suggested that the haploid TA and genotype TA/TA may be associated with AML in Han people in Hunan province.The IL-10 SNPs at -819 and -592 sites were associated with AML and may affect IL-10 mRNA expression in AML patients.

• International Journal of Industrial Entomology and Biomaterials
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• v.36 no.2
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• pp.25-30
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• 2018

The larvae of Hermetia. illucens have a high probability of coming into contact with microorganisms such as bacteria and fungi. Therefore, the survival of H. illucens is primarily the protection of their own against microbial infection. This effect depends on the development of the innate immune system. Antimicrobial Peptides (AMPs) exhibit antimicrobial activity against other bacterial strains and can provide important data to understand the basis of the innate immunity of H. illucens. In this study, we injected larvae with Enterococcus. faecalis (gram-positive bacteria) and Serratia. marcescens as (gram-negative bacteria) to test the hypothesis that H. illucens is protected from infection by its immune-related gene expression repertoire. To identify the inducible immune-related genes, we performed and cataloged the transcriptomes by RNA-Seq analysis. We compared the transcriptomes of whole larvae and obtained a DNA fragment of 465 bp including the poly (A) tail by RACE as a novel H. illucens immune-related gene against bacteria. A novel target mRNA expression was higher in immunized larvae with E. faecalis and S. marcescens groups than non-immunized group. We expect our study to provide evidence that the global RNA-Seq approach allowed for the identification of a gene of interest which was further analyzed by quantitative RT-PCR, together with genes chosen from the available literature.

• The Plant Pathology Journal
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• v.40 no.1
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• pp.59-72
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• 2024

A comprehensive survey of mungbean-growing areas was conducted to observe leaf spot disease caused by Alternaria alternata. Alternaria leaf spot symptoms were observed on the leaves. Diversity of 50 genotypes of mungbean was assessed against A. alternata and data on pathological traits was subjected to cluster analysis. The results showed that genotypes of mungbean were grouped into four clusters based on resistance parameters under the influence of disease. The principal component biplot demonstrated that all the disease-related parameters (% disease incidence, % disease intensity, lesion area, and % of infection) were strongly correlated with each other. Alt a 1 gene that is precisely found in Alternaria species and is responsible for virulence and pathogenicity. Alt a 1 gene was amplified using gene specific primers. The isolated pathogen produced similar symptoms when inoculated on mungbean and tobacco. The sequence analysis of the internal transcribed spacer (ITS) region, a 600 bp fragment amplified using specific primers, ITS1 and ITS2 showed 100% identity with A. alternata. Potato virus X (PVX) -based silencing vector expressing Alt a 1 gene was constructed to control this pathogen through RNA interference in tobacco. Out of 50 inoculated plants, 9 showed delayed onset of disease. Furthermore, to confirm our findings at molecular level semi-quantitative reverse transcriptase polymerase chain reaction was used. Both phenotypic and molecular investigation indicated that RNAi induced through the VIGS vector was efficacious in resisting the pathogen in the model host, Tobacco (Nicotiana tabacum). To the best of our knowledge, this study has been reported for the first time.

• Clinical and Experimental Reproductive Medicine
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• v.50 no.4
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• pp.253-261
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• 2023

Objective: Azoospermia (the total absence of sperm in the ejaculate) affects approximately 10% of infertile males. Despite diagnostic advances, azoospermia remains the most challenging issue associated with infertility treatment. Our study evaluated transition nuclear protein 2 (TNP2) and synaptonemal complex protein 3 (SYCP3) polymorphisms, azoospermia factor a (AZFa) microdeletion, and gene expression levels in 100 patients with azoospermia. Methods: We investigated a TNP2 single-nucleotide polymorphism through polymerase chain reaction (PCR) restriction fragment length polymorphism analysis using a particular endonuclease. An allele-specific PCR assay for SYCP3 was performed utilizing two forward primers and a common reverse primer in two PCR reactions. Based on the European Academy of Andrology guidelines, AZFa microdeletions were evaluated by multiplex PCR. TNP2, SYCP3, and the AZFa region main gene (DEAD-box helicase 3 and Y-linked [DDX3Y]) expression levels were assessed via quantitative PCR, and receiver operating characteristic curve analysis was used to determine the diagnostic capability of these genes. Results: The TNP2 genotyping and allelic frequency in infertile males did not differ significantly from fertile volunteers. In participants with azoospermia, the allelic frequency of the SYCP3 mutant allele (C allele) was significantly altered. Deletion of sY84 and sY86 was discovered in patients with azoospermia and oligozoospermia. Moreover, SYCP3 and DDX3Y showed decreased expression levels in the azoospermia group, and they exhibited potential as biomarkers for diagnosing azoospermia (area under the curve, 0.722 and 0.720, respectively). Conclusion: These results suggest that reduced SYCP3 and DDX3Y mRNA expression profiles in testicular tissue are associated with a higher likelihood of retrieving spermatozoa in individuals with azoospermia. The homozygous genotype TT of the SYCP3 polymorphism was significantly associated with azoospermia.

• Food Science of Animal Resources
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• v.35 no.6
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• pp.738-747
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• 2015

Angiotensin-converting enzyme (ACE) inhibitory activity was evaluated for the low-molecular-weight fraction (<3 kDa) obtained from milk fermentation by Bifidobacterium longum KACC91563. The ACE inhibitory activity in this fraction was 62.3%. The peptides generated from the <3 kDa fraction were identified by liquid chromatography-electrospray ionization quantitative time-of-flight mass spectrometry analysis. Of the 28 peptides identified, 11 and 16 were identified as β-casein (CN) and α_s1-CN, respectively. One peptide was identified as κ-CN. Three peptides, YQEPVLGPVRGPFPIIV, QEPVLGPVRGPFPIIV, and GPVRGPFPIIV, from β-CN corresponded to known antihypertensive peptides. We also found 15 peptides that were identified as potential antihypertensive peptides because they included a known antihypertensive peptide fragment. These peptides were as follows: RELEELNVPGEIVE (f1-14), YQEPVLGPVRGPFP (f193-206), EPVLGPVRGPFPIIV (f195-206), PVLGPVRGPFPIIV (f196-206), VLGPVRGPFPIIV (f197-206), and LGPVRGPFPIIV (f198-206) for β-CN; and APSFSDIPNPIGSENSEKTTMPLW (f176-199), SFSDIPNPIGSENSEKT- TMPLW (f178-199), FSDIPNPIGSENSEKTTMPLW (f179-199), SDIPNPIGSENSEKTTMPLW (f180-199), DIPNPIGSENSEKTTMPLW (f181-199), IPNPIGSENSEKTTMPLW (f182-199), PIGSENSEKTTMPLW (f185-199), IGSENSEKTTMPLW (f186-199), and SENSEKTTMPLW (f188-199) for α_s1-CN. From these results, B. longum could be used as a starter culture in combination with other lactic acid bacteria in the dairy industry, and/or these peptides could be used in functional food manufacturing as additives for the development of a product with beneficial effects for human health.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.23
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• pp.10299-10306
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• 2015

The esophageal squamous cell carcinoma (ESCC) is thought to develop through a multi-stage process. Epigenetic gene silencing constitutes an alternative or complementary mechanism to mutational events in tumorigenesis. Posttranscriptional regulation of human leukocyte antigen class I (HLA-I) and antigen processing machinery (APM) proteins expression may be associated with novel epigenetic modifications in cancer development. In the present study, we determined the expression levels of HLA-I antigen and APM components by immunohistochemistry. Then by a bisulfite-sequencing PCR (BSP) approach, we identified target CpG islands methylated at the gene promoter region of APM family genes in a ESCC cell line (ECa109), and further quantitative analysis of CpG site specific methylation of these genes in cases of Kazakh primary ESCCs with corresponding non-cancerous esophageal tissues using the Sequenom MassARRAY platform. Here we showed that the development of ESCCs was accompanied by partial or total loss of protein expression of HLA-B, TAP2, LMP7, tapasin and ERp57. The results demonstrated that although no statistical significance was found of global target CpG fragment methylation level sof HLA-B, TAP2, tapasin and ERp57 genes between ESCC and corresponding non-cancerous esophageal tissues, there was significant differences in the methylation level of several single sites between the two groups. Of thesse only the global methylation level of LMP7 gene target fragments was statistically higher ($0.0517{\pm}0.0357$) in Kazakh esophageal cancer than in neighboring normal tissues ($0.0380{\pm}0.0214$, p<0.05). Our results suggest that multiple CpG sites, but not methylation of every site leads to down regulation or deletion of gene expression. Only some of them result in genetic transcription, and silencing of HLA-B, ERp57, and LMP7 expression through hypermethylation of the promoters or other mechanisms may contribute to mechanisms of tumor escape from immune surveillance in Kazakh esophageal carcinogenesis.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1688-1696
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• 2020

Soil physical and chemical characteristics, soil potential denitrification rates (PDR), community composition and nirK-, nirS- and nosZ-encoding denitrifiers were studied by using MiSeq sequencing, quantitative polymerase chain reaction (qPCR), and terminal restriction fragment polymorphism (T-RFLP) technologies base on short-term (5-year) tillage field experiment. The experiment included four tillage treatments: conventional tillage with crop residue incorporation (CT), rotary tillage with crop residue incorporation (RT), no-tillage with crop residue retention (NT), and rotary tillage with crop residue removed as control (RTO). The results indicated that soil organic carbon, total nitrogen and NH₄⁺-N contents were increased with CT, RT and NT treatments. Compared with RTO treatment, the copies number of nirK, nirS and nosZ in paddy soil with CT, RT and NT treatments were significantly increased. The principal coordinate analysis indicated that tillage management and crop residue returning management were the most and the second important factors for the change of denitrifying bacteria community, respectively. Meanwhile, this study indicated that activity and community composition of denitrifiers with CT, RT and NT treatments were increased, compared with RTO treatment. This result showed that nirK, nirS and nosZ-type denitrifiers communities in crop residue applied soil had higher species diversity compared with crop residue removed soil, and denitrifying bacteria community composition were dominated by Gammaproteobacteria, Deltaproteobacteria, and Betaproteobacteria. Therefore, it is a beneficial practice to increase soil PDR level, abundance and community composition of nitrogen-functional soil microorganism by combined application of tillage with crop residue management.

• Journal of Korean Medicine Rehabilitation
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• v.27 no.2
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• pp.93-99
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• 2017

Objectives The purpose of this study is to understand the condition of the Ease-West Integrative Care system in one of Korean Medical hospital, to develop more effective system, and to collect advance information for future research. Methods We analyzed patient's status, patient's composition and the ranking of the major disease code. In addition, we investigated the operating system of how Ease-West Integrative Care in hospitals is operating in order to grasp the actual situation is being done. Results As a result of analyzing the status, there was a balanced cooperation between the Korean Medicine and Western Medicine with a ratio of 0.86:1. The disease status from Korean Medicine to Western Medicine were mostly occupied by stroke patients and from Western Medicine to Korean Medicine fragment were mostly by musculoskeletal pain patients. Conclusions The results of this study showed that the Ease-West Integrative Care system of surveyed Korean Medical hospital has more integrated medical characteristics than previous studies in terms of quality and quantitative. Future research based on detailed data collection and review for a longer period is expected in the further.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.52-67
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• 1998

The investigative research on the mammalian milk purely consisted of the physiological quality of lactoferrin was conducted to reveal the antimicrobial ativity of specifically functional foods with antibiotic characteristics as a basic data in food manufacturing. Bovine lactoferrin were isolated from raw milk samples, and was digested with pepsin, trypsin and chymotrypsin. It was necessary then to separate and purify lactoferrin from bovine raw milk, and in order to analyze the antimicrobial activity of the enzyme-treated bovine lactoferrin in their required quantitative fraction. Afterwards Escherichia coli and Staphylococcus aureus was incubated in it. It was that investigated to enzyme-treated fractions molecular weight and the peptide fragment with antimicrobial effect. 1. The purity of enzyme-treated bovine lactoferrin(BLF) was tested by SDS-PAGE. As a results of 12% SDS-PAGE assay, pepsin-treated LF did not exhibited band until if reaches 14 KDa, while trypsin and chymotrypsin treated LF, known to contain the non-digestive lactoferrin exhibited band at a molecular weight of 33 KDa. 2. Bovine lactoferrin was sucessfully purified through the use of Sephadex G-50 Column. In order to assay LF through the Sephadex G-50 column chromatography, the digestive bovine lactoferrin (BLFs) was eluted with a linear gradient of 0.05% Tris-HCl. When the gel-filtration analysis, pepsin, trypsin and chymotrypsin treatments of BLF fragments was showed 2, 3, and 2 peak, respectively. The results of the HPLC analysis confirmed that had a non-digestive lactoferrin receptor, and trypsin and chymotrypsin treated BLFs has an antimicrobial effect. 3. To measure the strength of the antimicrobial effect of enzyme treated lactoferrin it was compared to the antimicrobial activity taking place at the incubated Escherichia coli and Staphylococcus aureus. This might explain the resistance of the microorganisms for peptide fragment. The pepsin-treated of bovine lactoferrin was markedly reduced by incubation of the cells. Trypsin-treated of BLF was similar to chymotrypsin-treated of BLF. However, trypsin and chymotrypsin treatments of BLFs were showed the antimicrobial effect until eight hours incubation for native bovine lactoferrin. Therefore the enzyme-treated lactoferrin have an antimicrobial effect even non-digestive lactoferrin. 4. The digestive bovine lactoferrin fragments assay was carried out by the use of Sephadex G-50 column chromatography and SDS-PAGE. The pepsin and chymotrypsin-treated fragments has a low molecular weight and trypsin-treated lactoferrin was only showed a band. It was described that characteristics of digestive protein. It appeared that there may be a relation between virulence and resistance to enzyme-treated BLF.