• 대한수의학회지
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• 제50권3호
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• pp.213-220
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• 2010

Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.

• 한국수산과학회지
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• 제49권4호
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• pp.454-459
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• 2016

Pre- or post-harvest processing is required to mitigate the risk of norovirus infection mediated by shellfish or seafood. We investigated the environmental resistance of human norovirus (HuNoV) under various conditions of temperature, salinity, and pH in seawater. Male-specific coliphage (MSC) was as the reference virus for all tests. At 4℃, HuNoV GII4 spiked into seawater was continually detected by RT-PCR for 35 days, regardless of salinity or pH level. It maintained nearly stable concentrations, meaning HuNoV can sustain a viral population in seawater long enough to be accumulated by shellfish and other filter feeders during winter. MSC was also stable at 4℃ although viral infectivity dropped sharply after 28 days. The effects of salinity and pH on MSC were indistinct. At 25℃ the detectable period of HuNoV GII4 by RT-PCR in seawater decreased to about one-third or half of the period at 4℃. High salinity (32 psu) and alkaline pH (8.5) were also unfavorable for sustaining HuNoV abundance at 25℃ in seawater. The resistance patterns of MSC to high temperature, high salinity, and alkaline pH were more dramatic and viral infectivity decreased over time, almost in direct proportion to experimental days. MSC was undetectable after 12 days under all salinities and pH levels at 25℃.

• Clinical and Experimental Pediatrics
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• 제52권11호
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• pp.1241-1248
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• 2009

목 적:가와사끼병은 소아의 후천성 심질환의 가장 흔한 원인이나, 그 원인은 아직 밝혀지지 않았다. 최근 급성 호흡기 감염증의 원인 중 하나인 코로나바이러스(human coronavirus NL63)와 가와사끼병 발병과의 연관성이 제기된 바 있다. 이에 저자들은 급성 호흡기 바이러스 감염증과 가와사끼병의 발병 및 임상적 특징과의 연관성을 연구하고자 하였다. 방 법:2003년 10월부터 2006년 9월까지 서울대학교병원과 분당서울대학교병원에서 가와사끼병으로 진단받은 환자 54명으로부터 비인두 흡인물을 채취하여 총 11가지 호흡기 바이러스(호흡기 세포융합 바이러스, 아데노바이러스, 라이노바이러스, 파라인플루엔자 바이러스 1형 및 3형, 인플루엔자 바이러스 A형 및 B형, 메타뉴모바이러스, 보카바이러스, 코로나바이러스 OC43/229E 및 NL63)를 바이러스 배양검사 혹은 다중 역전사 중합 효소 반응(multiplex RT-PCR)로 검출하였다. 대상 환자들의 임상 양상은 후향적으로 검토하였다. 결 과:대상 환자들의 중앙 연령은 32개월(범위 6개월-10.4세)이었으며, 37례(69%)에서 발병 전에 호흡기 증상이 동반되었다. 호흡기 바이러스는 12례(22%)에서 검출되었으며, 라이노바이러스 4례, 파라인플루엔자 바이러스 3형 2례, 보카바이러스 2례, 그리고 아데노바이러스, 호흡기 세포융합 바이러스, 파라인플루엔자 바이러스 1형, 인플루엔자바이러스 A형, 코로나바이러스 NL63 각각 1례씩 검출되었다. 이 중 파라인플루엔자 바이러스 3형과 라이노바이러스가 중복 검출된 경우가 1례 있었다. 발병 전 호흡기 증상이 동반된 경우는 바이러스 양성군이 7례(58.3%), 음성군이 30례(71.4%)이었고, 정맥내 면역글로불린 1회 치료 후 해열된 경우는 바이러스 양성군이 8례(67%), 음성군이 36례(86%)로 나타났으며, 관상동맥 병변이 2개월 이상 지속된 경우는 바이러스 양성군이 2례(17%), 음성군이 4례(9.5%)이었으나, 모두 통계적으로 유의한 차이는 없었다. 결 론:본 연구에서는 가와사끼병 환자에서 호흡기 증상이 동반되는 경우가 흔하였으나, 특정 호흡기 바이러스와의 인과적 관계는 규명되지 않았다. 본 연구는 소규모 단일기관 연구로서, 향후 대규모 다기관 연구를 통하여 호흡기 바이러스 감염증과 가와사끼병의 연관성 및 예후에 미치는 영향이 검토되어야 할 것으로 보인다.

• Clinical and Experimental Pediatrics
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• 제53권3호
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• pp.373-379
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• 2010

목 적 : 급성 하기도 감염으로 입원하는 소아에서 최근 알려진 hRV C 및 hBoV를 포함하여 13종 호흡기 바이러스의 임상 양상 및 역학을 알아보고자 하였다. 방 법 : 2008년 5월부터 2009년 4월까지 충북대학교병원 소아과에 급성 하기도 감염으로 입원한 소아 중 325명을 대상으로 비인두 흡인물에서 multiplex RT-PCR법을 이용하여 총 13종의 바이러스를 검출하였으며, 대상 소아의 의무기록을 검토하였다. 결 과 : 대상 소아 중 270례(83.1%)에서 호흡기 바이러스가 검출되었으며, 혼합 감염은 71례(26.3%)에서 관찰되었다. 바이러스 검출 빈도는 RSV 108례(33.2%), hRV 62례(19.1%), Flu A 55례(16.9%), hMPV 50례(15.4%), PIV 27례(8.3%), hBoV 26례(8.0%), ADV 19례(5.8%), hCoV 7례(2.2%)였다. 임상진단은 세기관지염 37.5%, 폐렴 34.5%, 급성 천식 악화 20.9%, 크룹 7.1%이었으며, 세기관지염과 폐렴으로 진단된 소아에서 가장 높은 빈도로 검출된 호흡기 바이러스는 RSV, hRV, hMPV, Flu A였다. Flu A와 hRV는 3세 이상의 천식 악화로 진단된 소아에서 가장 높은 빈도로 검출되었다. hRV A와 hRV C는 각각 48명(14.8%)와 14명(4.3%)에서 검출되었으며, hRV C가 검출된 소아의 평균 연령은 $4.1{\pm}3.5$세로 hRV A가 검출된 소아에서의 $1.7{\pm}2.3$세에 비해 유의하게 높았다(P =0.009). hBoV는 세기관지염 또는 폐렴으로 진단된 소아에서 주로 검출되었고, 이들의 평균 연령은 $2.3{\pm}3.4$세였다. 결 론 : 본 연구에서는 한국 소아에서 13종의 바이러스에 의한 하기도 감염의 양상을 관찰하였다. 소아 하기도 감염에서 새로 알려진 바이러스들의 역할을 명확히 알기 위해서는 향후 지속적인 연구가 필요하다.

• Clinical and Experimental Pediatrics
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• 제48권11호
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• pp.1206-1211
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• 2005

목 적 : 현재 연구 개발되고 있는 PspA에 근거한 폐구균 단백 백신은 기존의 백신과 달리 여러 종류의 혈청형에 대해 동시에 광범위한 방어 면역을 유도할 가능성이 있다. 동물 실험에서 PspA는 폐구균 보균과 호흡기 감염 뿐 아니라 패혈증에 대해서도 방어 면역을 형성함이 보고되었다. 본 연구는 한국 소아와 어른에게서 분리된 폐구균에서 PspA 백신에 포함된 PspA family인 family 1과 family 2의 분포를 알기 위해 시행되었다. 방 법 : 총 89주의 폐구균을 대상으로 항혈청을 이용하여 슬라이드 응집 방법으로 폐구균 피막의 혈청형을 분류하였다. 또한 PCR을 이용하여 family 1과 family 2의 시발체를 사용하여 분리된 폐구균의 PspA family를 정하였다. 결 과 : 총 89주의 폐구균 분리주에서 17 종류의 폐구균 혈청형이 분류되었다. PspA 종류는 79주(88.8%)에서 결정되었는데 20주(22.5%)가 family 1, 59주(66.3%)가 family 2이었다. 아홉주(10.1%)에서 family 1과 2에 모두 양성 반응을 보였으므로 family 1과 2 단백을 포함하는 PspA 백신의 가능한 방어 범위는 98.9%이었다. 각 PspA family들은 균이 분리된 사람의 나이, 분리된 곳, 혹은 페니실린에 대한 감수성 등과 의미 있는 관련성이 없었으나 피막 혈청형에 따라서는 의미 있는 차이를 보였다. 결 론 : 본 연구에서 분리된 폐구균 분리주의 98.9%가 PspA family 1과 2에 속하였으므로 이를 포함한 백신은 폐구균에 대해 혈청형과 관련 없이 좀 더 광범위한 효과를 보일 것으로 추정된다. 향후 좀더 다양하고 많은 폐구균 분리주를 대상으로 PspA family의 연구가 이루어진다면 PspA 백신의 개발과 적용에 많은 도움이 될 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제24권8호
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• pp.1133-1142
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• 2014

Interleukin-32 (IL-32) is a cytokine and inducer of various proinflammatory cytokines such as $TNF{\alpha}$, IL-$1{\beta}$, and IL-6 as well as chemokines. There are five splicing variants (${\alpha}$, ${\beta}$, ${\gamma}$, ${\delta}$, and ${\varepsilon}$) and IL-$32{\gamma}$ is the most active isoform. We generated human IL-$32{\gamma}$ transgenic (IL-$32{\gamma}$ TG) mice to express high level of IL-$32{\gamma}$ in various tissues, including immune cells. The pathology of sepsis is based on the systemic inflammatory response that is characterized by upregulating inflammatory cytokines in whole body, particularly in response to gram-negative bacteria. We investigated the role of IL-$32{\gamma}$ in a mouse model of experimental sepsis by using lipopolysaccharides (LPS). We found that IL-$32{\gamma}TG$ mice resisted LPS-induced lethal endotoxemia. IL-$32{\gamma}$ reduced systemic cytokines release after LPS administration but not the local immune response. IL-$32{\gamma}TG$ increased neutrophil influx into the initial foci of the primary injected site, and prolonged local cytokines and chemokines production. These results suggest that neutrophil recruitment in IL-$32{\gamma}TG$ occurred as a result of the local induction of chemokines but not the systemic inflammatory cytokine circulation. Together, our results suggest that IL-$32{\gamma}$ enhances an innate immune response against local infection but inhibits the spread of immune responses, leading to systemic immune disorder.

• Asian Pacific Journal of Cancer Prevention
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• 제14권12호
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• pp.7045-7056
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• 2013

Since the first report of RNA interference (RNAi) less than a decade ago, this type of molecular intervention has been introduced to repress gene expression in vitro and also for in vivo studies in mammals. Understanding the mechanisms of action of synthetic small interfering RNAs (siRNAs) underlies use as therapeutic agents in the areas of cancer and viral infection. Recent studies have also promoted different theories about cell-specific targeting of siRNAs. Design and delivery strategies for successful treatment of human diseases are becomingmore established and relationships between miRNA and RNAi pathways have been revealed as virus-host cell interactions. Although both are well conserved in plants, invertebrates and mammals, there is also variabilityand a more complete understanding of differences will be needed for optimal application. RNA interference (RNAi) is rapid, cheap and selective in complex biological systems and has created new insight sin fields of cancer research, genetic disorders, virology and drug design. Our knowledge about the role of miRNAs and siRNAs pathways in virus-host cell interactions in virus infected cells is incomplete. There are different viral diseases but few antiviral drugs are available. For example, acyclovir for herpes viruses, alpha-interferon for hepatitis C and B viruses and anti-retroviral for HIV are accessible. Also cancer is obviously an important target for siRNA-based therapies, but the main problem in cancer therapy is targeting metastatic cells which spread from the original tumor. There are also other possible reservations and problems that might delay or even hinder siRNA-based therapies for the treatment of certain conditions; however, this remains the most promising approach for a wide range of diseases. Clearly, more studies must be done to allow efficient delivery and better understanding of unwanted side effects of siRNA-based therapies. In this review miRNA and RNAi biology, experimental design, anti-viral and anti-cancer effects are discussed.

• 대한수의학회지
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• 제8권2호
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• pp.125-146
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• 1968

Throughout the studies the following experimental results were summarized. 1. It was impossible to infect and kill the mice, weighing 10 to 12 gm, by inoculating 0.2ml of virulent Cl. chauvoei, diluted 1 to 10 with physiological saline, via subcutaneous, intramuscular, intraperitoneal or intraveonus, route. 2. The mice which were inoculated in brain with 0.03ml of Cl. chauvoei diluted 1 : 5120 with physiological saline were resulted in all death after infection, but not in case of attenuated strain even in dilution of one to five. 3. Virulent Cl. chauvoei were diluted with each of those of whole blood, erythrocytes and serum of horse, calf, swine, sheep, rabbit, guinea pig, chicken and duck, human plasma and 2% CaCl solution, and inoculated subcutaneously 0.25 to 0.5ml in mice, weighing 12 to 15gm. It was resulted in significant increase in virulence as comparing with the case of physiological saline solution except when horse and pig sera were used. Such a phenomena were not seen in attenuated strain. 4. Virulence of virulent Cl. Chauvoei could be increased significantly in rat, as the procedures used in mice, by suspending in whole blood, erythrocytes, serum, or plasma of various animals, or 2% $CaCl_2$ solution and by inoculating subcutaneously 0.5 to 10ml in rat, weighing 30 to 60 gm, as compared with those of control group which used physiological saline solutionos diluent. 5. Mice resisted 100 and 80 percent against challenge of $10^3$ and $10^4$ M.L.D.. respectively, 24 hours after inoculation of 0.5ml black leg antiserum. 6. Immune response to the black leg living vaccine in mice could be obtained more favorably in the group of respected vaccination rather than those of single inoculation and the most profitable inoculm size of the vacine was 0.5 to 1.0ml. 7. Challenge for the immunized mice could be carried out effectively 3 weeks after first vaccination. 8. Satisfactory results could be obtained by inoculating subcutaneously for the immunization and intracerebrally or subcutaneously for the challenge. 9. Mice which were inoculated with 0.5ml of black leg living vaccine via subtaneucously two times at seven days interval and 21 days after first inoculation and challenged with 5 and 10 M.L.D. of virulent strain, resited 100 and 70 to 80 percent respectively. Same results were obtainable in black leg killed vaccine as the procedures used in living vaccine. 10. There were significantly different resistances against the definite challenge does between the mice groups which were immnuized with the living vaccine diluted five or 10 times and the undiluted. 11. For the biological assay of black leg living vaccine and antiserum, satisfactory results could be obtained using mice.

• Toxicological Research
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• 제34권3호
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• pp.221-229
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• 2018

Tenofovir nanoparticles are novel therapeutic intervention in human immunodeficiency virus (HIV) infection reaching the virus in their sanctuary sites. However, there has been no systemic toxicity testing of this formulation despite global concerns on the safety of nano drugs. Therefore, this study was designed to investigate the toxicity of Tenofovir nanoparticle (NTDF) on the liver and kidney using an animal model. Fifteen adult male Sprague-Dawley (SD) rats maintained at the animal house of the biomedical resources unit of the University of KwaZulu-Natal were weighed and divided into three groups. Control animals (A) were administered with normal saline (NS). The therapeutic doses of Tenofovir (TDF) and nanoparticles of Tenofovir (NTDF) were administered to group B and C and observed for signs of stress for four weeks after which animals were weighed and sacrificed. Liver and kidney were removed and fixed in formal saline, processed and stained using H/E, PAS and MT stains for light microscopy. Serum was obtained for renal function test (RFT) and liver function test (LFT). Cellular measurements and capturing were done using ImageJ and Leica software 2.0. Data were analysed using graph pad 6, p values < 0.05 were significant. We observed no signs of behavioural toxicity and no mortality during this study, however, in the kidneys, we reported mild morphological perturbations widening of Bowman's space, and vacuolations in glomerulus and tubules of TDF and NTDF animals. Also, there was a significant elevation of glycogen deposition in NTDF and TDF animals when compared with control. In the liver, there were mild histological changes with widening of sinusoidal spaces, vacuolations in hepatocytes and elevation of glycogen deposition in TDF and NTDF administered animals. In addition to this, there were no significant differences in stereological measurements and cell count, LFT, RFT, weight changes and organo-somatic index between treatment groups and control. In conclusion, NTDF and TDF in therapeutic doses can lead to mild hepatic and renal histological damage. Further studies are needed to understand the precise genetic mechanism.

• 대한의용생체공학회:의공학회지
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• 제35권5호
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• pp.160-168
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• 2014

Continuous body temperature monitoring is useful and essential in diverse medical procedures such as infection onset detection, therapeutic hypothermia, circadian rhythm monitoring, sleep disorder assessment, and gynecological research. However, the existing thermometers are too invasive or intrusive to be applied to long-term body temperature monitoring. In our previous study, we invented the bi-medium deep body thermometer which can noninvasively and continuously monitor deep tissue temperature. And the ratio of thermal resistances expressed as K-value should be obtained to estimate body temperature with the thermometer and it can be different under various measurement environments. Although the device was proven to be useful through preliminary simulation test and small group of human study, the experimental environment was restrictive in our previous approach. In this study, a finite element simulation was executed to obtain the K-value and evaluate the accuracy of bi-medium thermometer under various measurement environments. In addition, K-value estimation equation was developed by analyzing the influence of 5 measurement environmental factors (medium length, medium height, tissue depth, blood perfusion rate, and ambient temperature) on K-value. The results revealed that the estimation accuracy of bi-medium deep body thermometer based on computer simulation was very high (RMSE < $0.003^{\circ}C$) in various measurement environments. Also, bi-medium deep body thermometer based on K-value estimation equation showed relatively accurate results (RMSE < $0.3^{\circ}C$) except for one case. Although the K-value estimation technology should be improved for more accurate body temperature estimation, the results of finite element simulation showed that bi-medium deep body thermometer could accurately measure various tissue temperatures under diverse environments.