• Parasites, Hosts and Diseases
• /
• 제53권3호
• /
• pp.265-270
• /
• 2015

Hemoglobinopathy and malaria are commonly found worldwide particularly in malaria endemic areas. Thalassemia, the alteration of globin chain synthesis, has been reported to confer resistance against malaria. The prevalence of thalassemia was investigated in 101 malaria patients with Plasmodium falciparum and Plasmodium vivax along the Thai-Myanmar border to examine protective effect of thalassemia against severe malaria. Hemoglobin typing was performed using low pressure liquid chromatography (LPLC) and ${\alpha}$-thalassemia was confirmed by multiplex PCR. Five types of thalassemia were observed in malaria patients. The 2 major types of thalassemia were Hb E (18.8%) and ${\alpha}$-thalassemia-2 (11.9%). There was no association between thalassemia hemoglobinopathy and malaria parasitemia, an indicator of malaria disease severity. Thalassemia had no significant association with P. vivax infection, but the parasitemia in patients with coexistence of P. vivax and thalassemia was about 2-3 times lower than those with coexistence of P. falciparum and thalassemia and malaria without thalassemia. Furthermore, the parasitemia of P. vivax in patients with coexistence of Hb E showed lower value than coexistence with other types of thalassemia and malaria without coexistence. Parasitemia, hemoglobin, and hematocrit values in patients with coexistence of thalassemia other than Hb E were significantly lower than those without coexistence of thalassemia. Furthermore, parasitemia with coexistence of Hb E were 2 times lower than those with coexistence of thalassemia other than Hb E. In conclusion, the results may, at least in part, support the protective effect of thalassemia on the development of hyperparasitemia and severe anemia in malaria patients.

• 대한두경부종양학회지
• /
• 제14권2호
• /
• pp.199-205
• /
• 1998

Objective: Telomerase, a specialized ribonucleoprotein polymerase associated with cellular immortality, is expressed by most malignant cells and is inactive in most normal somatic cells. The assays of telomerase activity in various tumors have provided both diagnostic and prognostic information. This study was carried out to determine whether telomerase activity could be useful in distinguishing benign and malignant thyroid diseasees. Materials & Methods: Telomerase activity was determined using Oncor $TRAP_{EZE}^{TM}ELISA$ Telomerase Detection Kit for performing PCR-based telomeric repeat amplification protocol (TRAP) assay followed by ELISA detection in both normal and tumor tissues of 23 adenomatous hyperplasias, 12 follicular adenomas, 4 follicular carcinomas, 16 papillary carcinomas, 4 Hashimoto's thyroiditises and 3 malignant lymphomas. We also examined all cases microscopically to review the status of lymphoid infiltrate. Results: Of the 62 cases, extensive lymphoid infiltrates were contained in 20 tumor tissues(4 Hashimoto's thyroiditises, 3 malignant lymphomas, 6 adenomatous hyperplasias and 7 papillary carcinomas), all of which showed positive telomerase activity. All the normal tissues without lymphoid infiltrates(n=43) did not express telomerase activity. Of 42 tumor tissues without lymphoid infiltrates, 37(88.0%) showed positive telomerase activity: 13 of 17 adenomatous hyperplasias(76.5%), 11 of 12 follicular adenomas(91.7%), 4 of 4 follicular carcinomas(100.0%) and 9 of 9 papillary carcinomas(100.0%). Conclusions: Our methods showed high sensitivity in the detection of telomerase activity and the exclusion of lymphoid infiltrates may be important in telomerase assay. In our work, the measurement of telomerase activity was not useful in distinguishing benign and malignant thyroid diseases.

• 한국동물위생학회지
• /
• 제35권3호
• /
• pp.175-182
• /
• 2012

Rotaviruses have been known to be a major etiological agent of gastroenteritis in both infants and young animals. Subsequently new rotaviruses, which were morphologically indistinguishable but antigenically and electrophoretically distinct with each other, were reported from several animals throughout world including Korea. These new rotaviruses were named as non-group A or group B or group C rotaviruses and so on. It has been very difficult to isolate and grow the non-group A rotaviruses in vitro, and this has greatly limited the characterizations of non-group A rotaviruses and serological studies. In this study, monoclonal antibodies (MAbs) against porcine non-group A rotavirus were produced and characterized. The VP6 gene of porcine group C rotavirus Korean isolate(#06-52-1) was cloned and expressed. For expression of VP6 gene, baculovirus expression system was applied. The VP6 gene and expressed protein in the recombinant virus were confirmed by polymerase chain reaction (PCR), indirect fluorescence antibody (IFA) test and Western blot, respectively. The expressed VP6 was used for MAbs production. The MAbs produced in this study would be promising as diagnostic reagents for detection of group C rotavirus infection.

• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2016년도 춘계학술대회 및 임시총회
• /
• pp.33-33
• /
• 2016

Mycotoxins are toxic secondary metabolites produced by fungi. They can be present in where agricultural-based commodities are contaminated with toxigenic fungi. These mycotoxins cause various toxicoses in human and livestock when consumed. Small grains including corn, barley, rice or wheat are frequently contaminated with mycotoxins due to infection mainly by toxigenic Fusarium species and/or under environment favorable to fungal growth. One of the most well-known Fusarium toxin groups in cereals is trichothecenes consisting of many toxic compounds. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin, and various derivatives belong to this group. Zearalenone and fumonisin (FB) are also frequently produced by many species of the same genus. In order to monitor Korean cereals for contamination with Fusarium and other mycotoxigenic fungal species as well, barley, corn, maize, rice grains, and soybean were collected from fields at harvest or during storage for several years. The fungal colonies outgrown from the grain samples were identified based on morphological and molecular characteristics. Trichothecene chemotypes of Fusarium species or presence of FB biosynthetic gene were determined using respective diagnostic PCR to predict possible toxin production. Heavy grain contamination with fungi was detected in barley, rice and wheat. Predominant fungal genus of barley and wheat was Alternaria (up to 90%) while that of rice was Fusarium (~40%). Epicoccum also appeared frequently in barley, rice and wheat. While frequency of Fusarium species in barley and wheat was less than 20%, the genus mainly consisted of Fusarium graminearum species complex (FGSC) which known to be head blight pathogen and mycotoxin producer. Fusarium composition of rice was more diverse as FGSC, Fusarium incarnatum-equiseti species complex (FIESC), and Fusarium fujikuroi species complex (FFSC) appeared all at considerable frequencies. Prevalent fungal species of corn was FFSC (~50%), followed by FGSC (<30%). Most of FFSC isolates of corn tested appeared to be FB producer. In corn, Fusarium graminearum and DON chemotype dominate within FGSC, which was different from other cereals. Soybeans were contaminated with fungi less than other crops and Cercospora, Cladosporium, Alternaria, Fusarium etc. were detected at low frequencies (up to 14%). Other toxigenic species such as Aspergillus and Penicillium were irregularly detected at very low frequencies. Multi-year survey of small grains revealed dominant fungal species of Korea (barley, rice and wheat) is Fusarium asiaticum having NIV chemotype.

• Journal of Microbiology and Biotechnology
• /
• 제21권7호
• /
• pp.719-727
• /
• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

• Journal of Microbiology and Biotechnology
• /
• 제31권4호
• /
• pp.529-539
• /
• 2021

The beet armyworm, Spodoptera exigua, is a serious insect pest infesting various vegetable crops. Two infectious insect viruses, baculovirus and iflavirus, are known to induce epizootics in S. exigua populations. Indeed, some laboratory colonies have appeared to be covertly infected by these viruses. Diagnostic PCR tests detected two different viruses: Spodoptera exigua multiple nucleopolyhedrosis virus (SeMNPV) and iflaviruses (SeIfV1 and SeIfV2). Viral extract from dead larvae of S. exigua could infect Sf9 cells and produce occlusion bodies (OBs). Feeding OBs to asymptomatic larvae of S. exigua caused significant viral disease. Interestingly, both SeIfV1 and SeIfV2 increased their titers at late larval stages. Sterilization of laid eggs with 1% sodium hypochloride significantly reduced SeMNPV titers and increased larval survival rate. Double-stranded RNA (dsRNA) specific to SeIfV1 or SeIfV2 significantly reduced viral titers and increased larval survival rate. To continuously feed dsRNA, a recombinant Escherichia coli HT115 expressing SeIfV1-dsRNA was constructed with an L4440 expression vector. Adding this recombinant E. coli to the artificial diet significantly reduced the SeIfV1 titer and increased larval survival. These results indicate that laboratory colony collapse of S. exigua is induced by multiple viral infections. In addition, either suppression of SeMNPV or SeIfV infection significantly increased larval survival, suggesting a cooperative pathogenicity between baculovirus and iflavirus against S. exigua.

• Journal of Ginseng Research
• /
• 제45권2호
• /
• pp.273-286
• /
• 2021

Background: Prostate carcinoma is the second most common cancer among men worldwide. Developing new therapeutic approaches and diagnostic biomarkers for prostate cancer (PC) is a significant need. The Chinese herbal medicine Panax quinquefolius saponins (PQS) have been reported to show anti-tumor effects. We hypothesized that PQS exhibits anti-cancer activity in human PC cells and we aimed to search for novel biomarkers allowing early diagnosis of PC. Methods: We used the human PC cell line DU145 and the prostate epithelial cell line PNT2 to perform cell viability assays, flow cytometric analysis of the cell cycle, and FACS-based apoptosis assays. Microarray-based gene expression analysis was used to display specific gene expression patterns and to search for novel biomarkers. Western blot and quantitative real-time PCR were performed to demonstrate the expression levels of multiple cancer-related genes. Results: Our data showed that PQS inhibited the viability of DU145 cells and induced cell cycle arrest at the G1 phase. A significant decrease in DU145 cell invasion and migration were observed after 24 h treatment by PQS. PQS up-regulated the expression levels of p21, p53, TMEM79, ACOXL, ETV5, and SPINT1 while it down-regulated the expression levels of bcl2, STAT3, FANCD2, DRD2, and TMPRSS2. Conclusion: PQS promoted cells apoptosis and inhibited the proliferation of DU145 cells, which suggests that PQS may be effective for treating PC. TMEM79 and ACOXL were expressed significantly higher in PNT2 than in DU145 cells and could be novel biomarker candidates for PC diagnosis.

• Parasites, Hosts and Diseases
• /
• 제60권2호
• /
• pp.127-131
• /
• 2022

Feline hemotropic mycoplasmosis (hemoplasmosis) is an infection of the red blood cells caused by the Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm), and Candidatus Mycoplasma turicensis (CMt). The existence of Mhf, CMhm, and CMt has been demonstrated in feral cats in Korea using molecular methods, but no clinical cases have yet been reported. This study reports 2 clinical cases of hemotropic mycoplasmosis caused by CMhm and CMt in 2 anemic cats. The first case was a client-owned intact female domestic shorthair cat that presented with fever, pale mucous membranes, and normocytic normochromic non-regenerative anemia. Prior to referral, an immunosuppressive prednisolone dose was administered at the local veterinary clinic for 1 month. The cat was diagnosed with high-grade alimentary lymphoma. Organisms were found on the surface of the red blood cells on blood smear examination. The second case was of a rescued cat that presented with dehydration and fever. The cat had normocytic normochromic non-regenerative anemia. Necropsy revealed concurrent feline infectious peritonitis. Polymerase chain reaction assay targeting 16S rRNA revealed CMhm infection in case 1 and dual infection of CMhm and CMt in case 2. Normocytic normochromic non-regenerative anemia was observed in both cats before and during the management of the systemic inflammation. This is the first clinical case report in Korea to demonstrate CMhm and CMt infections in symptomatic cats.

• Journal of Veterinary Science
• /
• 제25권4호
• /
• pp.45.1-45.12
• /
• 2024

Importance: Although the role of bovine coronavirus (BCoV) in calf diarrhea and respiratory disorders is well documented, its contribution to neurological diseases is unclear. Objective: This study conducted virological investigations of calves showing diarrhea and respiratory and neurological signs. Methods: An outbreak of diarrhea, respiratory, and neurological disorders occurred among the 12 calves in July 2022 in Istanbul, Türkiye. Two of these calves exhibited neurological signs and died a few days after the appearance of symptoms. One of these calves was necropsied and analyzed using molecular and histopathological tests. Results: BCoV RNA was detected in the brain, lung, spleen, liver, and intestine of the calf that had neurological signs by real-time reverse transcription polymerase chain reaction. Immunostaining was also observed in the intestine and brain. A 622 bp S1 gene product was noted on gel electrophoresis only in the brain. Phylogenetic analysis indicated that the BCoV detected in this study had a high proximity to the BCoV strain GIb with 99.19% nucleotide sequence homology to the strains detected in Poland, Israel, Türkiye, and France. No distinct genetic lineages were observed when the brain isolate was compared with the respiratory and enteric strains reported to GenBank. In addition, the highest identity (98,72%) was obtained with the HECV 4408 and L07748 strains of human coronaviruses. Conclusions and Relevance: The strain detected in a calf brain belongs to the GIb-European lineage and shares high sequence homology with BCoV strains detected in Europe and Israel. In addition, the similarity between the human coronaviruses (4408 and L07748) raises questions about the zoonotic potential of the strains detected in this study.

• Tuberculosis and Respiratory Diseases
• /
• 제55권1호
• /
• pp.41-58
• /
• 2003

연구배경 : 약제내성 결핵권의 조기 진단을 위해서, 최근 돌연변이 검출 및 질병의 진단 등에 새로운 기술로 대두되고 있는 올리고뉴콜레오티드 칩 기술을 이용하여 결핵균의 리팜핀, 아이소나아지드와 스트렙토마이신 내성과 관련된 rpoB, katG와 rpsL 유전자의 주요 돌연변이를 신속하고 정확하게 검출하고자 하였다. 방 법 : 리팜핀 내성 검출의 야생형 7개와 돌연변이형 13개, 아이소니아지드 내성 검출의 야생형 2개와 돌연변이형 3개, 그리고 스트렙토마이신 내성 검출을 위한 야생형과 돌연변이형 프로브 각 2종류를 고안한 후, 유리 슬라이드에 고정시켜 올리고뉴클레오티드 칩을 제작하였고, 약제내성을 가지고 있는 배양균주 55균주를 선택하여 PCR 증폭반응과 혼성화 반응을 실시한 후 비공초점 레이저 스케너를 이용하여 돌연변이를 검출하였다. 이를 염기서열방법으로 확인하여 돌연변이 다형성을 분석하였다. 결 과 : 리팜핀 내성은 코돈 531과 코돈 526에서 65%의 돌연변이를 검출하였고, 현재까지 보고되어 있지 않은 D516F의 새로운 돌연변이도 검출하였다. 아이소니아지드 내성은 S315T와 R463L 돌연변이가 45.2%로 검출되었고, 스트렙토마이신 내성은 K43R과 K88R 돌연변이가 78%로 검출되었다. 리팜핀 내성의 88%(35/40), 아이소니아지드 내성의 50%(20/42), 그리고 스트렙토마이신 내성의 78%(7/9)를 검출함으로써 현재까지 보고되어 있는 세가지 약제내성과 관련된 rpoB, katG와 rpsL 유전자의 주요 돌연변이는 대부분 검출할 수 있음을 확인할 수 있었고, 염기서열분석 결과와 비교하였을 때 모두 일치하는 결과를 얻음으로써 올리고뉴콜레오티드 칩의 유용성을 확인할 수 있었다. 결 론 : 따라서 본 연구에서 개발한 올라고뉴클레오티드 칩은 약재내성 결핵균의 조기 진단에 유용한 도구가 될 것으로 사료된다.