• Title/Summary/Keyword: double infection.

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Development of a diagnostic method for human enteric Adenovirus-41 with rapid, specific and high sensitivity using the loop-mediated isothermal amplification assay

  • Lee, Jin-Young;Rho, Jae Young
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.673-681
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    • 2020
  • Human enteric Adenovirus 41 (HueAdV-41) is a major waterborne virus that causes human gastroenteritis and is classified as a viral group I double-strand DNA virus, Adenoviridae. HueAdV-41 has been detected with the polymerase chain reaction (PCR) in various samples such as ground water. However, the PCR-based diagnostic method has problems such as reaction time, sensitivity, and specificity. Thus, the loop-mediated isothermal amplification (LAMP) assay has emerged as an excellent method for field applications. In this study, we developed a LAMP system that can rapidly detect HueAdV-41 with high specificity and sensitivity. HueAdV-41 specific LAMP primer sets were tested through a specific, non-specific selection and sensitivity test for three prepared LAMP primer sets, of which only one primer set and optimum reaction temperature were selected. The developed LAMP primer set condition was confirmed as 63℃, and the sensitivity was 1 copy. In addition, to confirm the system, a LAMP positive reaction was developed with the restriction enzyme Taq I (T/GCC). The developed method in this study was more specific, rapid (typically within 2 - 3 hours), and highly sensitive than that of the conventional PCR method. To evaluate and verify the developed LAMP assay, an artificial infection test was done with five cDNAs from groundwater samples, and the results were compared to those of the conventional PCR method. We expect the developed LAMP primer set will be used to diagnose HueAdV-41 from various samples.

Inhibitory effect of Korean Red Ginseng extract on DNA damage response and apoptosis in Helicobacter pylori-infected gastric epithelial cells

  • Kang, Hyunju;Lim, Joo Weon;Kim, Hyeyoung
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.79-85
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    • 2020
  • Background: Helicobacter pylori increases reactive oxygen species (ROS) and induces oxidative DNA damage and apoptosis in gastric epithelial cells. DNA damage activates DNA damage response (DDR) which includes ataxia-telangiectasia-mutated (ATM) activation. ATM increases alternative reading frame (ARF) but decreases mouse double minute 2 (Mdm2). Because p53 interacts with Mdm2, H. pylori-induced loss of Mdm2 stabilizes p53 and induces apoptosis. Previous study showed that Korean Red Ginseng extract (KRG) reduces ROS and prevents cell death in H. pylori-infected gastric epithelial cells. Methods: We determined whether KRG inhibits apoptosis by suppressing DDRs and apoptotic indices in H. pylori-infected gastric epithelial AGS cells. The infected cells were treated with or without KRG or an ATM kinase inhibitor KU-55933. ROS levels, apoptotic indices (cell death, DNA fragmentation, Bax/Bcl-2 ratio, caspase-3 activity) and DDRs (activation and levels of ATM, checkpoint kinase 2, Mdm2, ARF, and p53) were determined. Results: H. pylori induced apoptosis by increasing apoptotic indices and ROS levels. H. pylori activated DDRs (increased p-ATM, p-checkpoint kinase 2, ARF, p-p53, and p53, but decreased Mdm2) in gastric epithelial cells. KRG reduced ROS and inhibited increase in apoptotic indices and DDRs in H. pylori-infected gastric epithelial cells. KU-55933 suppressed DDRs and apoptosis in H. pylori-infected gastric epithelial cells, similar to KRG. Conclusion: KRG suppressed ATM-mediated DDRs and apoptosis by reducing ROS in H. pylori-infected gastric epithelial cells. Supplementation with KRG may prevent the oxidative stress-mediated gastric impairment associated with H. pylori infection.

Biochemical Quantitation of PM2 Phage DNA as a Substrate for Endonuclease Assay

  • Joo, Yoo-Jin;Kim, Hee-Ju;Lee, Jae-Yung;Kim, Joon
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.99-102
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    • 2004
  • Bacteriophage PM2 has a closed circular form of double stranded DNA as a genome. This DNA from the phage is a useful source for nick-circle endonuclease assay in the fmol range. Due to difficulties in the maintenance of viral infectivity, storage conditions of the phage should be considered for the puri-fication of PM2 DNA. The proper condition for a short-term storage of less than 2 months is to keep the PM2 phage at 4$^{\circ}C$; whereas the proper condition for a long-term storage of the PM2 phage for over 2 months is to keep it under liquid nitrogen in 7.5 % glycerol. The optimal conditions for a high yield of phage progeny were also considered with the goal to achieve a successful PM2 DNA preparation. A MOI(Multiplicity Of Infection) of 0.03, in which the OD$\sub$600/ of the host bacteria was between 0.3 and 0.5, turned out to be optimal for the mass production of PM2 phage with a burst size of about 214. Considerations of PM2 genome size, and the concentrations and radiospecific activities of purified PM2 DNA, are required to measure the endonuclease activity in the fmol range. This study reports the proper quantitation of radioactivity and the yield of purified DNA based on these conditions.

Reduction of the Isolated Anterior Wall of the Maxillary Sinus Fracture with Double Urinary Balloon Catheters and Fibrin Glue

  • Kim, Jaehee;Yang, Ho Jik;Kim, Jong Hwan;Kim, Su Jin
    • Archives of Craniofacial Surgery
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    • v.18 no.4
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    • pp.238-242
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    • 2017
  • Background: Conservative treatment is performed for isolated anterior wall of the maxillary sinus fractures, in many cases when the fracture is clinically not severe and asymptomatic. Despite the absence of symptoms, complications such as sinusitis, rhinitis, and chronic purulent secretion may develop; therefore, successful reduction is required. We attempted to reduce the risk of complications using an alternative technique: reduction of the fracture with two urinary balloon catheters inserted through the maxillary ostium and fixation using fibrin glue, which minimizes the damage to the bony fragments and sinus mucosa. Methods: In this study, 38 patients who were diagnosed with an isolated anterior wall of the maxillary sinus fracture at our hospital between January 2014 and January 2017 were enrolled. The fracture site was exposed via the Caldwell-Luc approach followed by reduction through the insertion of two urinary balloon catheters using a nasal endoscope and fixation with fibrin glue. The sex, cause of fracture, physical examination, and presence of complications were examined and patient's medical records and facial bone computed tomography scans were analyzed. Results: Radiological evaluation showed that there was no evidence of collapsed reduction fragments. Although some patients had remaining symptoms of hypoesthesia (15%; 3 patients), there were no complications such as infection, rhinitis, sinusitis, and chronic purulent secretion at the surgical site. Conclusion: In this study, we present an alternative surgical technique using two urinary balloon catheters and fibrin glue for the successful reconstruction of an isolated anterior wall of the maxillary sinus fracture. This technique enables precise restoration with a reduced risk of complications.

Convenient Suture Technique for Pediatric Facial Lacerations (소아 안면열상 환자의 치료에 있어서 유용한 봉합술)

  • Kim, Jun-Hyung;Kwon, Soon-Beom;Eo, Su-Rak;Cho, Sang-Hun;Markowitz, Bernard L.
    • Archives of Plastic Surgery
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    • v.37 no.4
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    • pp.496-498
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    • 2010
  • Purpose: Lacerations requiring formal wound closure compose a significant number of all childhood injuries presenting to the emergency department. The problem with conventional suture technique are that suture removal is quite cumbersome, especially in children. Unwanted soft tissue damage can result in the process of suture removal, which calls for sedation, stressful for both medical personnel and child. The purpose of this study is to introduce the convenient suture technique for pediatric facial lacerations. Methods: Children under the age of four, presenting to the emergency department with facial lacerations were enrolled in the study. From March 2008 to June 2009, 63 patients (41 males and 22 females) with an average age of 1.4 years were treated with our convenient suture technique using utilized a loop suspended above a double, flat tie. Clean, tension free wounds were treated with our technique, wounds with significant skin defect and concomitant fractures were excluded. Results: The Patients were followed-up in 1, 3 and 5 days postoperatively. On the third hospital visit, suture removal was done by simply cutting the loop suspended above the wound margin and gently pulling the thread with forceps. There were no significant differences in the rates of infection and dehiscence compared with conventional suture technique. Conclusion: The use of our technique was to be simple with similar operative time compared with conventional suture technique. Removal of suture materials were easy without unwanted injuries to the surrounding tissue which resulted in less discomfort for the patient and greater parental satisfaction, minimized the complications. It can be considered as a viable alternative in the repair of pediatric facial lacerations.

Repair of Coarctation (including tubular hypoplasia) in Infancy and Children (영아 및 소아 연령에서의 대동맥 교약증의 교정 수술)

  • 한재진
    • Journal of Chest Surgery
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    • v.23 no.3
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    • pp.474-481
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    • 1990
  • We have experienced 44 cases of coarctation of aorta in the age of infancy and children from April 1986 to September 1989 at Seoul National University Children`s hospital. Patients were thirty males and fourteen females, and their age ranged from one month to ten years[mean 23.84 $\pm$33.06 months] with thirty-two infant cases. In the infantile age, congestive heart failure was the most common chief complaint[18/32], and above that age, frequent upper respiratory infection was most common[8/12]. We experienced thirteen cases of isolated COA, twenty-two cases of COA with VSD, eight cases of COA with VSD, eight cases of COA with intracardiac complex anomalies and one case of COA with atrial septal defect. The associated intracardiac complex anomalies were three Taussig-Bing type double outlet right ventricle, one single ventricle, one transposition of great arteries, one atrioventricular septal defect, one hypoplastic aortic arch with left heart hypoplasia, and one Tetralogy of Fallot. Operative techniques of COA were twenty-three subclavian flap arterioplasty, 12 resection and end to end anastomosis, eight onlay patch angioplasty, and I direct angioplasty after resection of web. Among the cases with other cardiac anomalies, staged operation was done in twenty-nine patients, and single stage total correction was performed only in three patients. There were seven operative mortality[15.9%], all being in infantile age group, and among fourteen cases associated with large VSD[Qp/Qs>2.0, mean pulmonary arterial pressure>50mmHg], four patients were died, but there was no mortality in patients with small VSD. With above results, we are intended to discuss about the interval between staged operation, the fate of VSD after coarctoplasty in case of COA with VSD, causes of death, complications etc.

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Morphological characterization of Vibrio alginolyticus specific bacteriophage isolated from fish farms on west coast of Korea (서해안 양식장에서 분리한 Vibrio alginolyticus의 특이 bacteriophage에 대한 구조적 특성)

  • Heo, Yong Ju;Lee, Chan Heun;Baek, Min Suk;Ahn, Hyun Mi;Hwang, Yo Sep;Park, Kwan Ha;Choi, Sang Hoon
    • Journal of fish pathology
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    • v.25 no.3
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    • pp.165-172
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    • 2012
  • Vibrio alginolyticus (V. alginolyticus), which is one of bacterial pathogens evoking severe infection in fish and shellfish as well as in human has been found at high frequency around all coast areas in Korea. Both V. alginolyticus and V. alginolyticus specific bacteriophage were isolated from sea water and various fishes from fish farms on west coast in Korea. In a morphological study based on electron microscope, the purified phage appeared to be composed of hexagon head of 60 nm and short tail of 20 nm. In the denatured SDS-PAGE analysis, the structural proteins of the phage were found to be 7 different protein fractions ranging from 37.8 to 198 kda. The kind of nucleic acid of the phage was ascertained to a double stranded DNA.

Survey on the Incidence of Homeless Pulmonary Tuberculosis Infection Rate through Chest X-ray Examination (흉부 X-선 검사를 통한 노숙인 폐결핵 감염률 현황조사)

  • Kim, Mi-Young;Shin, Sung-Rae;Ryu, Young-Hwan;Lim, Hwan-Yeal
    • Journal of Radiation Industry
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    • v.10 no.4
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    • pp.249-255
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    • 2016
  • This study, Seoul City shelter, you are trying to seek medical cooperation and cure rate increase proposal Yu findings's current situation and tuberculosis of homeless tuberculosis. Inspector, and has a total 591 people is targeted to implement an interview after acquiring utilization agreement in studies conducted chest X-ray photography. Of the interview questions, three or more protons, it is determined that the TB symptomatic conducted sputum examination, chest X-ray examination confirms the physician radiology, when sputum examination primarily chromatic findings the double implemented and conducted by requesting the ship inspection also said inspection sputum acid-fast bacteria if it is true one, respectively. confirmed case result of checking whether there is a difference due to risk factors(Jb) at the chi square black, it was found that there is no statistically significant difference at 95% confidence level. (${\chi}^2=0.276$, p>0.05), suspected case (Ac, Ae) results of examining whether there is a difference due to risk factors in chi square black, that there is a statistically significant difference at 99% confidence level is I found (${\chi}^2=9.414$, p<0.01). The nature of the homeless tuberculosis screening and directed to the distance homeless specific location are likely to evaluate the actual incidence low and aggressive or management needs, the rationale is allowed insufficient reality is. Through this research, future, for tuberculosis high risk tuberculosis patient, such as homeless to expand the tuberculosis screening of infectious tuberculosis patients in private medical institutions, and one-stop service that chest X-ray examination and sputum examination is carried out at the same time introduced immediately to prevent the inspection and examination, cure, and should establish a foundation that can be up to post administration.

Antiviral Treatment Reveals a Cooperative Pathogenicity of Baculovirus and Iflavirus in Spodoptera exigua, a Lepidopteran Insect

  • Roy, Miltan Chandra;Ahmed, Shabbir;Mollah, Md. Mahi Imam;Kim, Yonggyun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.529-539
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    • 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.

OAS1 and OAS3 negatively regulate the expression of chemokines and interferon-responsive genes in human macrophages

  • Lee, Wook-Bin;Choi, Won Young;Lee, Dong-Hyun;Shim, Hyeran;KimHa, Jeongsil;Kim, Young-Joon
    • BMB Reports
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    • v.52 no.2
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    • pp.133-138
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    • 2019
  • Upon viral infection, the 2', 5'-oligoadenylate synthetase (OAS)-ribonuclease L (RNaseL) system works to cleave viral RNA, thereby blocking viral replication. However, it is unclear whether OAS proteins have a role in regulating gene expression. Here, we show that OAS1 and OAS3 act as negative regulators of the expression of chemokines and interferon-responsive genes in human macrophages. Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein-9 nuclease (Cas9) technology was used to engineer human myeloid cell lines in which the OAS1 or OAS3 gene was deleted. Neither OAS1 nor OAS3 was exclusively responsible for the degradation of rRNA in macrophages stimulated with poly(I:C), a synthetic surrogate for viral double-stranded (ds)RNA. An mRNA sequencing analysis revealed that genes related to type I interferon signaling and chemokine activity were increased in $OAS1^{-/-}$ and $OAS3^{-/-}$ macrophages treated with intracellular poly(I:C). Indeed, retinoic-acid-inducible gene (RIG)-I- and interferon-induced helicase C domain-containing protein (IFIH1 or MDA5)-mediated induction of chemokines and interferon-stimulated genes was regulated by OAS3, but Toll-like receptor 3 (TLR3)- and TLR4-mediated induction of those genes was modulated by OAS1 in macrophages. However, stimulation of these cells with type I interferons had no effect on OAS1- or OAS3-mediated chemokine secretion. These data suggest that OAS1 and OAS3 negatively regulate the expression of chemokines and interferon-responsive genes in human macrophages.