Kim, In-Woo;Markkandan, Kesavan;Lee, Joon Ha;Subramaniyam, Sathiyamoorthy;Yoo, Seungil;Park, Junhyung;Hwang, Jae Sam
Journal of Microbiology and Biotechnology
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제26권11호
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pp.1863-1870
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2016
Antimicrobial peptides/proteins (AMPs) are present in all types of organisms, from microbes and plants to vertebrates and invertebrates such as insects. The grasshopper Oxya chinensis sinuosa is an insect species that is widely consumed around the world for its broad medicinal value. However, the lack of available genetic information for this species is an obstacle to understanding the full potential of its AMPs. Analysis of the O. chinensis sinuosa transcriptome and expression profile is essential for extending the available genetic information resources. In this study, we determined the whole-body transcriptome of O. chinensis sinuosa and analyzed the potential AMPs induced by bacterial immunization. A high-throughput RNA-Seq approach generated 94,348 contigs and 66,555 unigenes. Of these unigenes, 36,032 (54.14%) matched known proteins in the NCBI database in a BLAST search. Functional analysis demonstrated that 38,219 unigenes were clustered into 5,499 gene ontology terms. In addition, 26 cDNAs encoding novel AMPs were identified by an in silico approach using public databases. Our transcriptome dataset and AMP profile greatly improve our understanding of O. chinensis sinuosa genetics and provide a huge number of gene sequences for further study, including genes of known importance and genes of unknown function.
Md. Samiul Haque;Mohammad Saiful Islam;Myung-Jo You
Parasites, Hosts and Diseases
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제61권4호
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pp.439-448
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2023
Tick infestation causes a significant threat to human and animal health, requiring effective immunological control methods. This study aimed to investigate the potential of recombinant Haemaphysalis longicornis enolase protein for tick vaccine development. The exact mechanism of the recently identified enolase protein from the H. longicornis Jeju strain remains poorly understood. Enolase plays a crucial role in glycolysis, the metabolic process that converts glucose into energy, and is essential for the motility, adhesion, invasion, growth, and differentiation of ticks. In this study, mice were immunized with recombinant enolase, and polyclonal antibodies were generated. Western blot analysis confirmed the specific recognition of enolase by the antiserum. The effects of immunization on tick feeding and attachment were assessed. Adult ticks attached to the recombinant enolase-immunized mice demonstrated longer attachment time, increased bloodsucking abilities, and lower engorgement weight than the controls. The nymphs and larvae had a reduced attachment rate and low engorgement rate compared to the controls. Mice immunized with recombinant enolase expressed in Escherichia coli displayed 90% efficacy in preventing tick infestation. The glycolytic nature of enolase and its involvement in crucial physiological processes makes it an attractive target for disrupting tick survival and disease transmission. Polyclonal antibodies recognize enolase and significantly reduce attachment rates, tick feeding, and engorgement. Our findings indicate that recombinant enolase may be a valuable vaccine candidate for H. longicornis infection in experimental murine model.
In this study, we analyzed the use of general radiography imaging and effective dose in inpatients. Our aim is to help reduce national medical radiation exposure doses and develop rational health-care financial policies. The effective dose for each general radiography was calculated using the ALARA-GR program for 53 types (total: 260 codes) general radiography codes selected from 'National Health Insurance Care Benefit Cost'. The usage of general radiography was analyzed in the 2018 inpatient patient data of the Health Insurance Review and Assessment Service, and the effective dose for each general radiography was analyzed. 89.00% of inpatients undergo general radiography imaging at least once, with an average of 12.63 scans per person and an effective dose of 1.00 mSv. Those who received support from Medical Aid showed a higher value compared to those who were insured by National Health Insurance, with 17.39 cases and 1.43 mSv (p<.001). Chest had the highest usage rate at 23.12% for general radiography imaging, while L-spine had the highest effective dose at 24.53%. It is estimated that 420 inpatients patients undergo 121 to 820 general radiography imaging procedures per year, and 233 inpatients are estimated to have an annual effective dose of >20.00~58.25 mSv. Rational use of health-care finances and the practice of medical radiation safety management are essential for the well-being of individuals, the enhancement of quality of life, and the improvement of health-care quality.
Scrub typhus, a mite-borne infectious disease, is caused by Orientia tsutsugamushi. Despite many attempts to develop a protective strategy, an effective preventive vaccine has not been developed. The identification of appropriate Ags that cover diverse antigenic strains and provide long-lasting immunity is a fundamental challenge in the development of a scrub typhus vaccine. We investigated whether this limitation could be overcome by harnessing the nanoparticle-forming polysorbitol transporter (PST) for an O. tsutsugamushi vaccine strategy. Two target proteins, 56-kDa type-specific Ag (TSA56) and surface cell Ag A (ScaA) were used as vaccine candidates. PST formed stable nano-size complexes with TSA56 (TSA56-PST) and ScaA (ScaA-PST); neither exhibited cytotoxicity. The formation of Ag-specific IgG2a, IgG2b, and IgA in mice was enhanced by intranasal vaccination with TSA56-PST or ScaA-PST. The vaccines containing PST induced Ag-specific proliferation of CD8+ and CD4+ T cells. Furthermore, the vaccines containing PST improved the mouse survival against O. tsutsugamushi infection. Collectively, the present study indicated that PST could enhance both Ag-specific humoral immunity and T cell response, which are essential to effectively confer protective immunity against O. tsutsugamushi infection. These findings suggest that PST has potential for use in an intranasal vaccination strategy.
The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.
Vaccination with tumor peptide epitopes associated with MHC class I molecules is an attractive approach directed at inducing tumor-specific CTLs. However, challenges remain in improving the therapeutic efficacy of peptide epitope vaccines, including the low immunogenicity of peptide epitopes and insufficient stimulation of innate immune components in vivo. To overcome this, we aimed to develop and test an innovative strategy that elicits potent CTL responses against tumor epitopes. The essential feature of this strategy is vaccination using tumor epitope-loaded nanoparticles (NPs) in combination with polyinosinic-polycytidylic acid (poly-IC) and anti-PD1 mAb. Carboxylated NPs were prepared using poly(lactic-co-glycolic acid) and poly(ethylene/maleic anhydride), covalently conjugated with anti-H-2Kb mAbs, and then attached to H-2Kb molecules isolated from the tumor mass (H-2b). Native peptides associated with the H-2Kb molecules of H-2Kb-attached NPs were exchanged with tumor peptide epitopes. Tumor peptide epitope-loaded NPs efficiently induced tumor-specific CTLs when used to immunize tumor-bearing mice as well as normal mice. This activity of the NPs significantly was increased when co-administered with poly-IC. Accordingly, the NPs exerted significant anti-tumor effects in mice implanted with EG7-OVA thymoma or B16-F10 melanoma, and the anti-tumor activity of the NPs was significantly increased when applied in combination with poly-IC. The most potent anti-tumor activity was observed when the NPs were co-administered with both poly-IC and anti-PD1 mAb. Immunization with tumor epitope-loaded NPs in combination with poly-IC and anti-PD1 mAb in tumor-bearing mice can be a powerful means to induce tumor-specific CTLs with therapeutic anti-tumor activity.
The role of macrophages in the resistance of ICR mice to Candida albicans and Salmonella typhimurium was assessed using silica, agent which selectively inactivates macrophages or poly-2-vinylpyridine-N-oxide(PVNO), a lysosomal stabilizing agent. In addition, effect of silica on humoral and cellular immune responses to sheep red blood cells(SRBC) or polyvinylpyroridone(PVP) was examind. Colonyforming units(CFU) of C. albicans or S. typhimurium in the spleen, livers and kidneys of silica-treated or diluent-treated mice were enumerated at various times after infection. Silica was given i. v. to mice at 4 days or 1 day before infection. Although there was no apparent differences in the number of CFU of C. albicans cultured from the spleens or livers of silica-treated and control mice at every assay period, significant differences in the number of CFU of C. albicans in the kidneys of silica-treated and control mice. Namely, silica given to mice 1 day before infection significantly increased the number of CFU of C. albicans in the kidneys at 2, 4 and 6 days after infection, but did not change the number of CFU at 8 days after infection. Silica given to mice at 4 days before infection significantly increased the number CFU in the kidneys at 2 and 4 days after infection, but rather decreased the number of CFU at 8 days after infection. The number of CFU of C. albians cultured from the kidneys of splenectomized which were experimentally infected mice was similar to the number recovered from sham-operated mice at 4 and 8 days postinfection irrespective of time of infection relative to operation. The pretreatment of mice with PVNO appeared to abrogate the silica-induced susceptibility of mice to C. albicans. PVNO alone showed somewhat protective effect against challenge with C. albicans. In contrast, silica treatment did not alter the number of CFU of S. typhimurium recovered from the spleens and kidneys of mice. The administration of silica to mice at 4 days or 1 day before SRBC immunization significantly suppressed delayed-type hypersensitivity(DTH) reactions to SRBC and antibody production to SRBC, a thymus-dependent antigen and PVP, a thymus-independent antigen. These results provide evidence that macrophages play an important role in susceptibility to Candida infection and strongly demonstrated that macrophages play an essential role in the induction of immune responses in mice.
Accurate vital statistics are essential for a national health planning and evaluation. Among various vital statistics, birth and death rates, and infant and matemal mortality rates together with the causes of death are the very basic ones for above purposes as well as for the maternal and child health management. These statistics are based on the birth and death reports. It is required by law to report every birth and death within one month after its occurrence. However, in case of a neonatal death occurring prior to the birth report, most of the birth and death are not reported. Thus accurate infant and maternal mortality rates are not available yet in Korea. The main objective of this study is to develop a birth and infant death reporting system via computer network. We designed a new birth report form based on the current form and data from the analysis of medical record forms of 14 hospitals. A new form is basically addition of essential medical information to the current birth report form. Since a revision of the rules and regulations related wtih the birth report is necessary to use a new form, we kept the current from intact to make it acceptable to the government office for a field trial. We also developed computer programs for data input for birth and death reports at a medical faciltiy, data processing for production of maternal and child health indices at a health center, and management of maternal and child health services including immunization and postantal care at health center. The birth certificate and birth report can be printed out at a medical facility. The computer packages were programmed by Borland Delphi 3.0 and can be run under Windows 95 system. We proposed a new birth and death reporting system via computer network after a field trial for data input, transmission, and processing. The medical and demographic data o birth and death at medical facilities will be sent to health centers directly via computer network. The health center will retain the medical data for analysis and forward only the data for birth and death reports required by current regulations to the Dong, Up, or Myun Office. Once the birth or death is reported via computer network to the Dong Office, then the Dong Office will notify the baby's mother of the birth report and request to submit the baby's name by mail. When the baby's name its submitted. the Dong Office will forward the birth reports to the Common Court and Statistics Agency in the same way as the current system, Upon the completion of birth registration of the Common Court, the court will issue the birth certificate to mother which will be used in lieu of the family record. The advantages of proposed birth and death reporting system via computer network ar as follows ; I) The accuracy, timing, and completeness of reporting will be improved and more accurate maternal and child health indices can be obtained, ii) The maternal and child health services of health center will be obtained, iii) Epidemiologic data for pregnancy and birth can be obtained, iv) Manpower for birth and death reporting will be saved.
In this county, the gap between the urban 'haves' and the rural 'have-nots' continues to be an increasing problem. WHO and UNICEF see primary health care(PHC) as the key to achieving an acceptable level of health throughout the world as a community development. PHC is essential health care made accessible to individuals and families in the community by means acceptable to them. It is the first level of contact of individual, the family, and community with the national health system. It includes at least education on health system. It includes at least education on health problems, promotion of food supply, MCH including family planning, immunization against infectious diseases, control of endemic diseases, treatment of common diseases and injuries, promotion of mental health, and provision of essential drugs. However, of the aboves, education concerning of mental health problems and the methods to identify, prevent, and control them is the principal step of establishment. In Korea, the category of PHC worker includes the physician as public doctor and nurse as primary health care practitioner and community health leader as village health worker. PHC workers of the aboves will thus function best if they are appropriately trained to respond to the health needs of the community. However in this country, since the national PHC service project launched in 1980, the government has not developed and performed appropriate and enough education and training activities. In light of above reasons, several categories of health education activities had been planned and performed being aimed at above specific target groups and the main focus was on the village health workers for about one year from July 1991 to July 1992 in Yeoju Kun of Kyonki Province. At the end of the period, evaluation of education input was carried out to measure the improvement of healthful life of people in terms of awareness, attitude, and practice. At the end of the period, evaluation of education input was carried out to measure the improvement of healthful life of people in terms of awareness, attitude, and practice. The totals of 80 village health workers, 13 public health practitioners and 9 public docters took in the course of health education for a few hours at every month and the evaluation works of educational effect were taken. The results the study were as follows. 1) Number of persons who realized the maxim "health care of the people is a duty of the government" increased after the education course, On the other hand, the rate of satisfaction on the effort of government for health promotion of the people decreased. 2) Public doctors and primary health care practitioners(nurses) liked and enjoyed the education schedule as a meeting of peer group. It provided chances of communication with staffs of Korea University Hospital. It was said that lectures covered great deal of knowledge and technic they urgently needed in the field. 3) After finishing the education course, more of village health workers(VHW) thought they adapted themselves to their roles and functions showing increased number of home visit and contact with primary health care practitioners by month. 4) In case of patient refer, VHW preferred primary health care practitioners to public doctors. 5) Capability of VHWs in most of their functions increased dramatically after when the education course finished except tuberculosis control.
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