RESUMEN
BACKGROUND: As a result of the reintroduction of malaria in the Republic of Korea (ROK) in 1993 and the threat to military and civilian populations, the Korea Military National Defense (MND) increased emphasis on vector control in 2012 at ROK Army (ROKA) installations located near the DMZ, while decreasing chemoprophylaxis, fearing potential drug resistance. Mosquito surveillance demonstrated a need for continuous monitoring of disease patterns among ROKA soldiers and vector malaria infection rates to ensure positive outcomes. METHODS: Anopheles spp. were collected from May-October at three ROKA installations in three locations near the DMZ. Each of the areas included one installation <2 km and two installations 11-12 km from the DMZ in Paju and Yeoncheon counties, Gyeonggi Province. Anopheles spp. were identified by polymerase chain reaction (PCR) techniques and then assayed for the presence of vivax malaria sporozoites. The ROK MND reported vivax malaria patients monthly to Korea Centers for Disease Control and Prevention. Correlations for the incidence of Plasmodium vivax patients and infected Anopheles species were analysed using the Wilcoxon rank sum test, Pearson correlation test and liner regression analysis. RESULTS: A total of 4282 Anopheles spp. were collected. Anopheles kleini (69.5 %) was the most commonly collected, followed by Anopheles pullus (17.3 %), Anopheles belenrae (4.5 %), Anopheles sineroides (4.2 %), Anopheles sinensis sensu stricto (2.7 %), and Anopheles lesteri (1.9 %). Overall, 21 malaria patients were reported by the ROK MND. The monthly incidence of the malaria patients correlated with the monthly number of Plasmodium vivax sporozoite positive Anopheles spp. The monthly numbers of An. kleini demonstrated the highest correlations to the numbers of ROKA malaria patients throughout the mosquito season (P < 0.01). Anopheles spp. positive for P. vivax and malaria patients at ROKA installations located <2 km from the DMZ were higher than for ROKA installations located 11-12 km from DMZ. CONCLUSION: The number of Anopheles spp. positive for P. vivax sporozoites correlated with the monthly number of malaria cases and exposure of ROKA soldiers from May-October to P. vivax malaria infections. Malaria vector surveillance and vector control is warranted as part of an effective malaria management program at ROKA installations located near DMZ.
Asunto(s)
Anopheles/crecimiento & desarrollo , Malaria Vivax/epidemiología , Personal Militar , Mosquitos Vectores/crecimiento & desarrollo , Plasmodium vivax/aislamiento & purificación , Animales , Anopheles/parasitología , Femenino , Humanos , Incidencia , Mosquitos Vectores/parasitología , República de Corea/epidemiología , EsporozoítosRESUMEN
Creutzfeldt-Jakob disease (CJD) is a representative human transmissible spongiform encephalopathy associated with central nervous system degeneration. Prions, the causative agents of CJD, are composed of misfolded prion proteins and are able to self-replicate. While CJD is a rare disease affecting only 1-1.5 people per million worldwide annually, it has attracted both scientific and public attention as a threatening disease since an epidemic of variant CJD (vCJD) cases appeared in the mid-1990s. Due to its unconventional transmission and invariable fatality, CJD poses a serious risk to public health. The hundreds of sporadic, genetic, and iatrogenic CJD cases as well as potential zoonotic transmission suggest that CJD is an ongoing concern for the field of medicine. Nevertheless, treatment aimed at clinical prevention and treatment that reverses the course of disease does not exist currently. Active surveillance and effective laboratory diagnosis of CJD are, therefore, critical. In this report, the surveillance systems and laboratory tests used currently to diagnose CJD in different countries are reviewed. The current efforts to improve surveillance and diagnosis for CJD using molecular and biochemical findings are also described.
Asunto(s)
Técnicas de Laboratorio Clínico/métodos , Síndrome de Creutzfeldt-Jakob/diagnóstico , Síndrome de Creutzfeldt-Jakob/epidemiología , Monitoreo Epidemiológico , HumanosRESUMEN
Apoptosis has been shown to be induced and downregulated by the Hantaan virus (HTNV) nucleocapsid (N) protein. To address these conflicting data, expression of the p53 protein, one of the key molecules involved in apoptosis, was assessed in the presence of the N protein in A549 and HeLa cells. The amount of p53, increased by drug treatment, was reduced when cells were infected with HTNV or transfected with an expression vector of the HTNV N protein. When cells were treated with a proteasome inhibitor (MG132) or an MDM2 antagonist (Nutlin-3), p53 expression was not reduced in N protein-overexpressed cells. We concluded that the HTNV N protein ubiquitinates and degrades p53 MDM2-dependently. Here we report downregulation of p53 expression through a post-translational mechanism: MDM2-dependent ubiquitination and degradation by the HTNV N protein. These results indicate that N protein-dependent p53 degradation through the ubiquitin proteasome system is one of the anti-apoptotic mechanisms employed by HTNV.
Asunto(s)
Proteínas de la Cápside/metabolismo , Regulación de la Expresión Génica , Virus Hantaan/patogenicidad , Proteínas Proto-Oncogénicas c-mdm2/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Proteínas del Núcleo Viral/metabolismo , Apoptosis , Línea Celular , Regulación hacia Abajo , Genes p53 , Virus Hantaan/metabolismo , Células HeLa , Humanos , Complejo de la Endopetidasa Proteasomal , Proteínas Proto-Oncogénicas c-mdm2/genética , Proteína p53 Supresora de Tumor/genética , Ubiquitina/metabolismo , UbiquitinaciónRESUMEN
The complete genome sequence of the KGH strain of the first human rabies virus, which was isolated from a skin biopsy of a patient with rabies, whose symptoms developed due to bites from a raccoon dog in 2001. The size of the KGH strain genome was determined to be 11,928 nucleotides (nt) with a leader sequence of 58 nt, nucleoprotein gene of 1,353 nt, phosphoprotein gene of 894 nt, matrix protein gene of 609 nt, glycoprotein gene of 1,575 nt, RNA-dependent RNA polymerase gene of 6,384 nt, and trailer region of 69 nt. Sequence similarity was compared with 39 fully sequenced rabies virus genomes currently available, and the result showed 70.6-91.6 % at the nucleotide level, and 82.8-97.9 % at the amino acid level. The deduced amino acids in the viral protein were compared with those of other rabies viruses, and various functional regions were investigated. As a result, we found that the KGH strain only had a unique amino acid substitution that was identified to be associated either with host immune response and pathogenicity in the N protein, or with a related region regulating STAT1 in the P protein, and related to pathogenicity in G protein. Based on phylogenetic analyses using the complete genome of 39 rabies viruses, the KGH strain was determined to be closely related with the NNV-RAB-H strain and transplant rabies virus serotype 1, which are Indian isolates, and was confirmed to belong to the Arctic-like 2 clade. The KGH strain was most closely related to the SKRRD0204HC and SKRRD0205HC strain when compared with Korean animal isolates, which was separated around the same time and place, and belonged to the Gangwon III subgroup.
Asunto(s)
Virus de la Rabia/aislamiento & purificación , Rabia/veterinaria , Rabia/virología , Anciano , Animales , Gatos , Perros , Hurones , Zorros , Humanos , Masculino , Datos de Secuencia Molecular , Filogenia , Virus de la Rabia/clasificación , Virus de la Rabia/genética , Mapaches , República de Corea , Proteínas Virales/genéticaRESUMEN
Misfolding of prion protein (PrP to PrPSc) can cause neurodegenerative prion diseases. As a glycosylphosphatidylinositol (GPI)-anchored membrane protein, the normal form of PrP (PrPC) can function as a receptor for ligands in the extracellular space. PrPC was suggested to be involved in memory, synaptic neuronal communication, and anti-oxidation as a neuroprotective agent. The recently identified interaction between PrPC and Aß(1-42) oligomers suggested another role for PrP as a receptor for Aß(1-42) oligomers, thereby influencing cytotoxicity and apoptosis. Here, the association between PrPC and Aß(1-42) oligomers was investigated by visualizing protein localization in neuronal cells by immunocytochemistry. Aß(1-42) oligomer-induced cytotoxicity was tested in respective expressions of PrPC by using mouse neuroblastoma-2a (N2a) cells, the prion protein overexpressed cells (L2-2B1), and a Prnp-null mouse hippocampal cell line (HpL 3-4). Moreover, apoptotic proteins such as caspase-8 were used to assess the effect of PrPC on Aß(1-42) oligomer-mediated apoptosis. In L2-2B1 and HpL 3-4 cells, the difference in the cytotoxicity of Aß(1-42) oligomers could be clearly distinguished. In addition, Aß(1-42) oligomers induced mitochondria dysfunction, reactive oxygen species (ROS) generation, and calcium influx PrPC-dependently. Apoptosis, related to mitochondria dysfunction, was further investigated to determine the cytotoxic pathway; the results suggest that PrPC could be involved in both the intrinsic and extrinsic apoptotic pathways. Finally, cells with abundant PrPC expression seemed to be more susceptible to Aß(1-42) oligomer toxicity, suggesting the importance of the level of PrPC expression in the induction of apoptosis.
Asunto(s)
Péptidos beta-Amiloides/metabolismo , Apoptosis , Fragmentos de Péptidos/metabolismo , Proteínas PrPC/metabolismo , Péptidos beta-Amiloides/toxicidad , Animales , Calcio/metabolismo , Caspasa 8/metabolismo , Línea Celular Tumoral , Supervivencia Celular , Ratones , Fragmentos de Péptidos/toxicidad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
We determined for the first time the complete genome sequences of two Korean strains of the tick-borne encephalitis virus (TBEV), designated KrM 93 and KrM 213, isolated from the lung tissues of wild rodents in 2006. The genomes are 11,097 nucleotides (nt) in length and consist of a 132 nt 5'-noncoding region (NCR), a 10,245 nt open reading frame (ORF) containing 10 viral protein-coding regions (3,415 amino acids), and a 720 nt 3'-NCR. Compared with the 31 fully sequenced TBEV strains currently available, KrM 93 and KrM 213 show genomic nucleotide (and deduced amino acid) sequence divergences ranging from 1.8 (0.7) to 19.2 (26.6)% and 1.9 (0.8) to 19.3 (26.7)%, respectively. Phylogenetic and recombination analyses based on the complete genome sequence were performed to identify genetic variations and relationships between the TBEV strains. These showed that the Korean TBEV strains clustered with the Western subtype rather than with Far-Eastern or Siberian subtypes, and phylogenetic trees derived from capsid (C), envelope (E), nonstructural (NS) 4B and NS5 regions represented the same branching pattern shown by the complete genome-based tree. Although no recombination events were identified in these two Korean strains, 11 putative recombination events were identified within the NS5 regions or in the 3'-NCRs of TBEV strains in general. The results provide insight into the genetics of TBEV strains to understand the molecular epidemiology, genetic diversity, and evolution of TBEV.
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Virus de la Encefalitis Transmitidos por Garrapatas/genética , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Encefalitis Transmitida por Garrapatas/veterinaria , Genoma Viral , Enfermedades de los Roedores/virología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Línea Celular , Virus de la Encefalitis Transmitidos por Garrapatas/clasificación , Encefalitis Transmitida por Garrapatas/virología , Genómica , Ratones , Datos de Secuencia Molecular , Filogenia , República de Corea , Roedores , Proteínas Virales/genéticaRESUMEN
We sequenced the envelope (E) gene of 17 strains of the Japanese encephalitis virus (JEV) isolated in South Korea in 1983-2005 and compared the sequences with those from previously reported strains. Our results show the remarkable genetic stability of the E gene sequence in Korean JEV strains. Five pairs of E gene sequences from 10 Korean strains were identical, despite geographical differences and a maximum five-year time span. Sequence comparisons with other Asian strains revealed that the Korean strains are closely related to those from China, Japan, and Vietnam. Genotype 3 strains were predominant in Korea before 1993, when genotype 1 strain K93A07 was first isolated. The two genotypes were detected simultaneously in 1994 but since then, only genotype 1 has been isolated in South Korea. Thus, the genotype change occurred according to the year of isolation rather than the geographical origin.
Asunto(s)
Virus de la Encefalitis Japonesa (Especie)/genética , Encefalitis Japonesa/epidemiología , Encefalitis Japonesa/virología , Secuencia de Aminoácidos , Secuencia de Bases , Virus de la Encefalitis Japonesa (Especie)/química , Virus de la Encefalitis Japonesa (Especie)/clasificación , Virus de la Encefalitis Japonesa (Especie)/aislamiento & purificación , Genotipo , Humanos , Epidemiología Molecular , Datos de Secuencia Molecular , Filogenia , República de Corea/epidemiología , Alineación de SecuenciaRESUMEN
As noted in other flaviviruses, the envelope (E) protein of Japanese encephalitis virus (JEV) interacts with a cellular receptor and mediates membrane fusion to allow viral entry into target cells, thus eliciting neutralizing antibody response. The formation of the flavivirus prM/E complex is followed by the cleavage of precursor membrane (prM) and membrane (M) protein by a cellular signalase. To test the effect of prM in JEV biology, we constructed JEV-MuLV pseudotyped viruses that express the prM/E protein or E only. The infectivity and titers of JEV pseudotyped viruses were examined in several cell lines. We also analyzed the neutralizing capacities with anti-JEV sera from JEV-immunized mice. Even though prM is crucial for multiple stages of JEV biology, the JEV-pseudotyped viruses produced with prM/E or with E only showed similar infectivity and titers in several cell lines and similar neutralizing sensitivity. These results showed that JEV-MuLV pseudotyped viruses did not require prM for production of infectious pseudotyped viruses.
Asunto(s)
Virus de la Encefalitis Japonesa (Especie)/fisiología , Proteínas del Envoltorio Viral/fisiología , Ensamble de Virus , Animales , Anticuerpos Neutralizantes/inmunología , Anticuerpos Antivirales/inmunología , Línea Celular , Chlorocebus aethiops , Perros , Virus de la Encefalitis Japonesa (Especie)/inmunología , Humanos , Ratones , Proteínas del Envoltorio Viral/inmunologíaRESUMEN
BACKGROUND: Polymorphisms of the human prion protein gene (PRNP) contribute to the genetic determinants of Creutzfeldt-Jakob disease (CJD). Numerous polymorphisms in the promoter regions as well as the open reading frame of PRNP were investigated. Greater than 90% of Korean, Chinese, and Japanese carry the homozygote 129 MM codon. In Korea, polymorphisms have not been comprehensively studied, except codons 129 and 219 in PRNP among Korean CJD cases. Although polymorphisms at codons 129 and 219 play an important role in susceptibility to sporadic CJD, patients with other polymorphisms in PRNP exhibited critical distinctions of clinical symptoms. METHODS: The genetic analyses of PRNP were carried out among probable CJD patients in comparison with the results from magnetic resonance imaging (MRI) and electroencephalogram (EEG). RESULTS: The molecular analyses revealed that three mutations at codons D178N, E200K, and M232R in heterozygosity. Patients with the D178N and M232R mutations had a 129MM codon, whereas the patient with the E200K mutation showed 129MV heterozygosity. They all revealed strong 14-3-3 positive signals. The 67-year-old patient with the D178N-129M mutation showed progressive gait disturbance and dysarthria was in progress. The 58-year-old patient with the E200K mutation coupled to the 129MV codon had gait disturbance, dysarthria, agitation, and ataxic gait, and progressed rapidly to death 3 months from the first onset of symptoms. The 65-year-old patient with the M232R mutation showed rapidly progressive memory decline and gait disturbance, and died within 16 months after onset of symptoms. CONCLUSION: Despite differences in ethnicity, the clinical and pathological outcomes were similar to the respective mutations around the world, except absence of insomnia in D178N-129M subject.
Asunto(s)
Codón/genética , Síndrome de Creutzfeldt-Jakob/genética , Predisposición Genética a la Enfermedad , Priones/genética , Proteínas 14-3-3/líquido cefalorraquídeo , Anciano , Pueblo Asiatico/genética , Línea Celular , Análisis Mutacional de ADN , Electroencefalografía , Genotipo , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Mutación , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , Proteínas PriónicasRESUMEN
To determine whether the tick-borne encephalitis virus (TBEV) is present in vector ticks and mammalian hosts in Korea, we examined two tick species, Haemaphysalis longicornis (n = 548) and Ixodes nipponensis (n = 87), and the lungs or spleens of rodents Apodemus agrarius (n = 24) and wild boars (n = 16). Tick-borne encephalitis virus was detected in samples by reverse transcriptase (RT)-nested polymerase chain reaction (PCR), after which TBEV-positive samples were inoculated into BHK-21 cells and suckling mice. Tick-borne encephalitis virus genes were detected in 4 of 38 tick pools and 5 of 24 wild rodents. Suckling mice inoculated intracerebrally with TBEV-positive rodent samples showed signs of encephalitis at six days post-inoculation. The isolation of TBEV was confirmed by inoculating samples obtained from the brains of sick mice in cell culture. Phylogenetic analysis showed that the E genes of the TBEV isolates were clustered with the Western subtype (98% identity). This study suggests the possible occurrence of tick-borne encephalitis in Korea.
Asunto(s)
Vectores Arácnidos/virología , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Encefalitis Transmitida por Garrapatas/transmisión , Murinae/virología , Sus scrofa/virología , Garrapatas/virología , Animales , Animales Salvajes/virología , Secuencia de Bases , Encefalitis Transmitida por Garrapatas/epidemiología , Encefalitis Transmitida por Garrapatas/virología , Ixodes/virología , Corea (Geográfico) , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedades de los Roedores/epidemiología , Enfermedades de los Roedores/transmisión , Enfermedades de los Roedores/virología , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/transmisión , Enfermedades de los Porcinos/virologíaRESUMEN
Zika virus (ZIKV) is an arthropod-borne virus mainly transmitted by Aedes species. A total of nine of the 16 imported ZIKV reported cases during the mosquito season in the Republic of Korea (ROK), following the return of local nationals from foreign ZIKV endemic countries, were surveyed for Aedes albopictus. Surveillance and vector control of Ae. albopictus, a potential vector of ZIKV, and related species are critical for reducing the potential for autochthonous transmission in the ROK. Surveillance and vector control were coordinated by Korean Centers for Disease Control & Prevention (KCDC) and conducted by local health authorities within 200 m of imported ZIKV patients' residences. After diagnosis, thermal fogging (3 × week × 3 weeks), residual spray for homes and nearby structures (1 × week × 3 weeks), and larval control (3 × week × 3 weeks) were conducted in accordance with national guidelines developed by KCDC in early 2016. Of the nine residences surveyed using BG Sentinel traps, Ae. albopictus trap indices (TIs) for the three (3) patients' residences located near/in forested areas were significantly higher than the six patients' residences located inside villages/urban areas or low-lying farmland without trees. Overall, Ae. albopictus TIs in forested areas decreased by 90.4% after adult and larval control, whereas TIs decreased by 75.8% for residences in nonforested areas. A total of 3,216 Aedes and Ochlerotatus spp. were assayed by real-time polymerase chain reaction for ZIKV, dengue, and chikungunya virus. Both species collected before and after vector control were negative for all viruses. Vector control within 200 m of residences of imported ZIKV patients, conducted in accordance with established guidelines, may have effectively reduced human-mosquito-human transmission cycle by competent vectors in South Korea.
Asunto(s)
Aedes/virología , Insectos Vectores/virología , Control de Mosquitos , Infección por el Virus Zika/prevención & control , Virus Zika , Animales , Humanos , Larva/virología , Control de Mosquitos/métodos , República de Corea/epidemiología , Infección por el Virus Zika/epidemiología , Infección por el Virus Zika/transmisiónRESUMEN
Understanding the mechanisms of insecticide resistance to vector mosquitoes is critical for the implementation of effective control measures. A nulliparous susceptible Culex pipiens pallens (KSCP) laboratory colony and two field strains from Paju (PAJ) and Jeonju (JEO) Korea were evaluated for susceptibility to five pesticides by microapplication techniques. Unfed PAJ and JEO females demonstrated increased resistance compared to unfed KSCP females, respectively. While blood-fed KSCP females demonstrated <10-fold decreased susceptibility to pesticides compared to unfed KSCP females, blood-fed PAJ and JEO females demonstrated 25.0-50.0- and 16.0-38.6-fold increased resistance compared to unfed PAJ and JEO females, respectively. Unfed and blood-fed groups were assayed for α- and ß-esterase, glutathione S-transferases, and cytochrome P-450 (P450) enzyme activity assays. P450 activity was 58.8- and 72.8-fold higher for unfed PAJ and JEO females, respectively, than unfed KSCP females. P450 enzyme activity of KSCP females assayed 1 and 7 days after a blood meal increased by 14.5- and 11.8-fold, respectively, compared to unfed KSCP females, while PAJ and JEO females demonstrated 164.9- and 148.5- and 170.7- and 160.4-fold increased activity, respectively, compared to unfed females of each population. However, other three resistance-related metabolic enzymes showed low activation at <10-fold after a blood meal. The data demonstrate that P450 acts on elevated insecticide resistance after blood meals in resistant field populations. Our findings might reveal that suppressing of the P450 protein by artificial gene mutation increases insecticidal susceptibility of Cx. pipiens and will promise effective vector mosquito control.
RESUMEN
We investigated the 47-kDa outer membrane protein (OMP), which is a periplasmic serine protease and an antigenic major surface protein of Orientia tsutsugamushi, as a vaccine candidate. We developed a conventional subunit vaccine expressing recombinant 47-kDa OMP (rec47) and a DNA vaccine (p47). In mouse immunization experiments, intranasal immunization with rec47 alone or with rec47 plus heat-labile enterotoxin B subunit from Escherichia coli or plus cholera toxin (CT) as adjuvants induced a higher amount of rec47-specific antibodies than intramuscular immunization with p47 alone or with p47 plus pBOOST2-samIRF7/3 (pB) as adjuvant. Moreover, the combination of rec47 and CT induced a strong cellular immune response to 47-kDa OMP, as demonstrated by a spleen cell proliferation assay, and also induced Th1- and Th2-type cytokine production, as demonstrated by a cytokine enzyme-linked immunosorbent assay. Intranasal immunization with rec47 plus CT was the most effective method for the induction of humoral and cell-mediated immune responses. Furthermore, relatively strong protection against homologous O. tsutsugamushi strain Boryong challenge was observed in mice immunized with rec47 plus CT. Therefore, 47-kDa OMP is an attractive candidate for developing a prophylactic vaccine against scrub typhus by O. tsutsugamushi infection.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas de la Membrana Bacteriana Externa/aislamiento & purificación , Vacunas Bacterianas/inmunología , Orientia tsutsugamushi/inmunología , Proteínas Recombinantes/inmunología , Tifus por Ácaros/prevención & control , Adyuvantes Inmunológicos , Animales , Antígenos Bacterianos/inmunología , Vacunas Bacterianas/genética , Ensayo de Inmunoadsorción Enzimática , Femenino , Inmunidad Celular , Ratones , Ratones Endogámicos BALB C , Orientia tsutsugamushi/genética , Proteínas Recombinantes/genética , República de Corea , Tifus por Ácaros/inmunologíaRESUMEN
The compounds 1,8-cineole and zerumbone (ZER) from the Cyperus rotundus rhizome along with another 11 previously identified rhizome essential oil constituents and α-humulene, which lacks the only carbonyl group present in ZER, as well as binary mixtures of ZER and seven active compounds were tested for repellency to male B. germanica. The results were compared to N,N-diethyl-3-methylbenzamide (deet). In filter-paper choice tests, ZER was the most repellent compound, and α-humulene was ineffective, which indicates that the α,ß-unsaturated carbonyl group of ZER is a prerequisite component for repellency. At 81.5 µg cm-2, enhanced repellency was produced by binary mixtures of ZER and 1,8-cineole, (+)-dihydrocarvone or (R)-(+)-limonene (70:30, 50:50 and 30:70 ratios by weight). These mixtures were very effective against male B. germanica within 24 h and were more repellent than a single compound or deet alone. The optimum ZER content was determined to be more than 50%. In Ebeling choice box tests at 652.4 µg cm-2, these compounds and deet resulted in complete repellency to intact male B. germanica, while they exhibited 35-47% repellency to antennectomized male one. Mixtures formulated from the active constituents of the C. rotundus rhizome could be useful as potential repellents for controlling B. germanica.
Asunto(s)
Blattellidae/efectos de los fármacos , Cyperus/química , Repelentes de Insectos/farmacología , Rizoma/química , Sesquiterpenos/farmacología , Animales , Conducta Animal/efectos de los fármacos , Fraccionamiento Químico , Repelentes de Insectos/química , Repelentes de Insectos/aislamiento & purificación , Masculino , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Sesquiterpenos/química , Sesquiterpenos/aislamiento & purificaciónRESUMEN
The fluorescent antibody test (FAT) is the most commonly used method for detection of the rabies virus (RV). The plaque assay can only be applied to fixed RVs, and cannot be used for street RVs. In this study, plaque formation allowing the determination of both fixed and street RVs was achieved using the immune plaque assay. The immune plaque assay carried out using both fixed and street RVs showed the formation of clear and countable plaques after immunostaining with anti-RV P monoclonal antibody and HRP-conjugated anti-mouse IgG. Plaque size increased with incubation time, and the plaque morphology differed according to viral strain. Fixed RVs had the dot-shaped regular plaque morphology and street RVs had the small irregular-shape plaque morphology. In addition, no significant differences were observed between the growth kinetics of the KGH strain when the virus was titrated using the FAT and the immune plaque assay. It allowed the successful detection and quantification of both street and fixed RVs through the production of clear, countable plaques, making it easy to obtain objective results. The assay is an applicable tool for the detection of RVs in various investigations, including virus neutralizing antibody testing, cell-to-cell spread, and viral drug sensitivity testing.
Asunto(s)
Virus de la Rabia/crecimiento & desarrollo , Virus de la Rabia/aislamiento & purificación , Ensayo de Placa Viral/métodos , Animales , Anticuerpos Antivirales/sangre , Inmunoensayo/métodos , Ratones , Pruebas de Sensibilidad Microbiana , Pruebas de Neutralización/métodos , Rabia/diagnóstico , Virus de la Rabia/patogenicidadRESUMEN
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne infectious disease that is endemic to China, Japan, and the Republic of Korea (ROK). In this study, 8313 ticks collected from SFTS outbreak areas in the ROK in 2013 were used to detect the SFTS virus (SFTSV). A single SFTSV was isolated in cell culture from one pool of Haemaphysalis longicornis ticks collected from Samcheok-si, Gangwon Province, in the ROK. Phylogenetic analysis showed that the SFTSV isolate was clustered with the SFTSV strain from Japan, which was isolated from humans. To the best of our knowledge, this is the first isolation in the world of SFTSV in ticks collected from vegetation.
Asunto(s)
Ixodidae/virología , Phlebovirus/aislamiento & purificación , Animales , Brotes de Enfermedades , Humanos , Phlebovirus/genética , Filogenia , ARN Viral/genética , República de CoreaRESUMEN
After an extensive vaccination policy, Japanese encephalitis (JE) was nearly eliminated since the mid-1980s in South Korea. Vaccination in children shifted the affected age of JE patients from children to adults. However, an abrupt increase in JE cases occurred in 2010, and this trend has continued. The present study aimed to investigate the prevalence of neutralizing antibodies to the JE virus (JEV) among high-risk age groups (≥40 years) in South Korea. A plaque reduction neutralization test was conducted to evaluate the prevalence of neutralizing antibodies to JEV in 945 subjects within four age groups (30-39, 40-49, 50-59, and 60-69 years) in 10 provinces. Of the 945 enrolled subjects, 927 (98.1%) exhibited antibodies against JEV. No significant differences were found in the prevalence of neutralizing antibodies according to sex, age, or occupation. However, there were significant differences in the plaque reduction rate according to age and occupation; oldest age group had a higher reduction rate, and subjects who were employed in agriculture or forestry also had a higher value than the other occupations. We also found that three provinces (Gangwon, Jeonnam, and Gyeongnam) had a relatively lower plaque reduction rate than the other locations. In addition, enzyme-linked immunosorbent assays were conducted to determine recent viral infections and 12 (1.3%) subjects were found to have been recently infected by the virus [corrected]. In conclusion, the present study clearly indicated that the prevalence of neutralizing antibodies has been maintained at very high levels among adult age groups owing to vaccination or natural infections, or both. In the future, serosurveillance should be conducted periodically using more representative samples to better understand the population-level immunity to JE in South Korea.
Asunto(s)
Anticuerpos Neutralizantes/sangre , Virus de la Encefalitis Japonesa (Especie)/inmunología , Encefalitis Japonesa/epidemiología , Adulto , Anciano , Anticuerpos Neutralizantes/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pruebas de Neutralización , República de Corea , Factores de Riesgo , Estudios SeroepidemiológicosRESUMEN
Most reverse genetic (RG) systems for rabies viruses (RVs) have been constructed on the genome background of laboratory-adapted strains. In this study, we developed an RG system using a Korean wild type (KGH) strain to investigate the pathogenic potential of different strains. We developed a RG system with the KGH strain for the first time. Following the complete genome sequencing of the KGH strain, pKGH infectious clones were constructed using the CMV/T7 promoter, and HamRz and HdvRz were introduced to allow self-cleavage of the synthesized RNA. We successfully recovered the rescued virus by constructing chimeric RVs in which we replaced a part of the construct with the partial gene from the fixed RC-HL strain. The rescued viruses formed clearer and countable plaques in an immunostaining plaque assay, with a distinct plaque morphology. Furthermore, compared with the chimeric RVs, the pKGH/RCinsΔ4 strain containing the KGH strain G protein exhibited a decreased efficiency of cell-to-cell spreading in BHK-21 cells and significantly reduced (100-1000 fold) replication kinetics. However, pKGH/RCinsΔ4 strain-infected mice revealed 100% morbidity at 11days post-infection, whereas other chimeric RV strains showed no mortality. Our RG system is a useful tool for studying differences in the cell-to-cell spreading efficiency and replication with respect to the different internalization patterns of street and fixed laboratory-adapted viruses.
Asunto(s)
Evolución Molecular , Virus de la Rabia/genética , Rabia/virología , Animales , Línea Celular , Clonación Molecular , ADN Complementario/genética , Modelos Animales de Enfermedad , Femenino , Orden Génico , Ingeniería Genética , Genoma Viral , Ratones , Plásmidos/genética , Virus de la Rabia/patogenicidad , Ensayo de Placa Viral , Virulencia , Replicación ViralRESUMEN
Inherited prion diseases (IPDs), including genetic Creutzfeldt-Jakob disease (gCJD), account for 10-15% of cases of prion diseases and are associated with several pathogenic mutations, including P102L, V180I, and E200K, in the prion protein gene (PRNP). The valine to isoleucine substitution at codon 180 (V180I) of PRNP is the most common pathogenic mutation causing gCJD in East Asian patients. In this study, we conducted follow-up analyses to identify candidate factors and their associations with disease onset. Whole-genome sequencing (WGS) data of five gCJD patients with V180I mutation and 145 healthy individuals were used to identify genomic differences. A total of 18,648,850 candidate variants were observed in only the patient group, 29 of them were validated as variants. Four of these validated variants were nonsense mutations, six were observed in genes directly or indirectly related to neurodegenerative disorders (NDs), such as LPA, LRRK2, and FGF20. More than half of validated variants were categorized in Gene Ontology (GO) terms of binding and/or catalytic activity. Moreover, we found differential genome variants in gCJD patients with V180I mutation, including one uniquely surviving 10 years after diagnosis of the disease. Elucidation of the relationships between gCJD and Alzheimer's disease or Parkinson's disease at the genomic level will facilitate further advances in our understanding of the specific mechanisms mediating the pathogenesis of NDs and gold standard therapies for NDs.
Asunto(s)
Síndrome de Creutzfeldt-Jakob/genética , Genómica , Mutación , Enfermedades Neurodegenerativas/genética , Proteínas Priónicas/genética , Anciano , Sustitución de Aminoácidos , Codón , Biología Computacional , Síndrome de Creutzfeldt-Jakob/diagnóstico , Síndrome de Creutzfeldt-Jakob/metabolismo , Progresión de la Enfermedad , Epistasis Genética , Femenino , Ontología de Genes , Redes Reguladoras de Genes , Genómica/métodos , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Masculino , Persona de Mediana Edad , Enfermedades Neurodegenerativas/metabolismo , Fenotipo , Proteínas tau/metabolismoRESUMEN
Tsutsugamushi disease is an infectious disease transmitted to humans through the bite of the Orientia tsutsugamushi-infected chigger mite; however, host-pathogen interactions and the precise mechanisms of damage in O. tsutsugamushi infections have not been fully elucidated. Here, we analyzed the global metabolic effects of O. tsutsugamushi infection on the host using 1H-NMR and UPLC-Q-TOF mass spectroscopy coupled with multivariate statistical analysis. In addition, the effect of O. tsutsugamushi infection on metabolite concentrations over time was analyzed by two-way ANOVAs. Orthogonal partial least squares-discriminant analysis (OPLS-DA) showed distinct metabolic patterns between control and O. tsutsugamushi-infected mice in liver, spleen, and serum samples. O. tsutsugamushi infection caused decreased energy production and deficiencies in both remethylation sources and glutathione. In addition, O. tsutsugamushi infection accelerated uncommon energy production pathways (i.e., excess fatty acid and protein oxidation) in host body. Infection resulted in an enlarged spleen with distinct phospholipid and amino acid characteristics. This study suggests that metabolite profiling of multiple organ tissues and serum could provide insight into global metabolic changes and mechanisms of pathology in O. tsutsugamushi-infected hosts.