• Title/Summary/Keyword: cagA

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Approaches to Improving Production Efficiencies of Transgenic Animals

  • Tojo, Hideaki
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.7-8
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    • 2000
  • Transgenic animals are very useful for scientific, pharmaceutical, and agricultural purposes. In livestock, transgenic technology has been used forthe genetic alteration of farm animals, the production of human proteins inlarge quantities in the milk of transgenic farm animals, and the generation of animals with organs suitable for xenotransplantation. To date, the transfer of foreign genes into farm animals has been performed mainly by microinjection of DNA into the pronuclei of fertilized eggs. However, the overall success rate of transgenic animals in livestock so far has been disappointingly low, eg., the efficiency is 0∼5% in swine, and less than 1% in sheep and cattle, compared with the rate in mice where 5% microinjected develop into transgenic animals. Recently, McGreath et al. (2000) have succeeded in producing the gene targeted sheep by the use of nuclear transfer from cultured somatic cells transfected with a foreign gene in vitro. However, we may need plenty of time until currently employ this method for gene transfer to farm animals. We have been studying to exploit the method for improving production efficiencies of transgenic animals with emphasis of its application to farm animals. The present paper describes three approaches that we have made in our laboratory to improve production efficiencies of transgenic animals, based on the DNA microinjection method. 1. Co-injection of restriction enzyme with foreign DNA into the pronucleus for elevating production efficiencies of transgenic animals. 2. Efficient selection of transgenic mouse embryos using EGFP as a marker gene. 3. Phenotypes of tansgenic mice expressing WAP/hGH-CAG/EGFP fusion gene produced by selecting transgenic embryos. 4. Efficient site-specific integration of the transgene targeting an endogenous lox like site in early mouse embryos.

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Antigenic Proteins of Helicobacter pylori of Potential Diagnostic Value

  • Khalilpour, Akbar;Santhanam, Amutha;Lee, Chun Wei;Saadatnia, Geita;Velusamy, Nagarajan;Osman, Sabariah;Mohamad, Ahmad Munir;Noordin, Rahmah
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.3
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    • pp.1635-1642
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    • 2013
  • Helicobacter pylori antigen was prepared from an isolate from a patient with a duodenal ulcer. Serum samples were obtained from culture-positive H. pylori infected patients with duodenal ulcers, gastric ulcers and gastritis (n=30). As controls, three kinds of sera without detectable H. pylori IgG antibodies were used: 30 from healthy individuals without history of gastric disorders, 30 from patients who were seen in the endoscopy clinic but were H. pylori culture negative and 30 from people with other diseases. OFF-GEL electrophoresis, SDS-PAGE and Western blots of individual serum samples were used to identify protein bands with good sensitivity and specificity when probed with the above sera and HRP-conjugated anti-human IgG. Four H. pylori protein bands showed good (${\geq}$ 70%) sensitivity and high specificity (98-100%) towards anti-Helicobacter IgG antibody in culture-positive patients sera and control sera, respectively. The identities of the antigenic proteins were elucidated by mass spectrometry. The relative molecular weights and the identities of the proteins, based on MALDI TOF/TOF, were as follows: CagI (25 kDa), urease G accessory protein (25 kDa), UreB (63 kDa) and proline/pyrroline-5-carboxylate dehydrogenase (118 KDa). These identified proteins, singly and/or in combinations, may be useful for diagnosis of H. pylori infection in patients.

Progress in human ovarian rejuvenation: Current platelet-rich plasma and condensed cytokine research activity by scope and international origin

  • Sills, E. Scott;Wood, Samuel H.
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.4
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    • pp.311-315
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    • 2021
  • Objective: As clinicians and patients await consensus on intraovarian platelet-rich plasma (PRP) treatment, this project evaluated contemporary research trends in the literature. Methods: A PubMed/NLM search aggregated all ovarian PRP-related publications (n=54) to evaluate their scope, abstract utility, submission-to-publication interval, journal selected, article processing charge (APC), free reader access to full-text manuscripts, number and nationality of authors, and inclusion of international collaborators. The NIH Clinical Trials database was also audited. Results: Published output on intraovarian PRP has increased consistently since 2016, especially among investigators in Greece, Iran, USA, and Turkey. Between 2013 and 2021, 42 articles met the relevancy criteria, of which 40.5% reported clinical studies, small series, or case reports, 33% described experimental animal models, and 23.8% were opinion/review papers. Only two works included a placebo control group. The submission-to-publication interval (mean±standard deviation) was 130±96 days, there were 5.9±3.2 authors per project, and journals invoiced US $1,613±1,466 (range, $0-$3,860) for APCs. Conclusion: There was no correlation between APC and time to publish (Pearson's r=-0.01). Abstract content was inconsistent; sample size and patient age were often missing, yet free full-text "open access" was available for most publications (59.5%). The NIH Clinical Trials portal lists eight registered studies on "ovarian rejuvenation," of which two are actively recruiting patients, while four have been terminated or have an uncertain status. Two studies have concluded, with results from one posted to the NIH website. PRP and its derivatives for ovarian treatment show early promise, but require further investigation. Research is accelerating and should be encouraged, particularly placebo-controlled randomized clinical trials.

Evaluation of Radiation Dose to Patients according to the Examination Conditions in Coronary Angiography (심장동맥 조영 검사 시 검사 조건에 따른 환자 선량 평가)

  • Yong-In Cho
    • Journal of radiological science and technology
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    • v.46 no.6
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    • pp.509-517
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    • 2023
  • This study analyzed imaging conditions and exposure index through clinical information collection and dose calculation programs in coronary angiography examinations. Through this, we aim to analyze the effective dose according to examination conditions and provide basic data for dose optimization. In this study, ALARA(As Low As Reasonably Achievable)-F(Fluoroscopy), a program for evaluating the radiation dose of patients and the collected clinical data, was used. First, analysis of imaging conditions and exposure index was performed based on the data of the dose report generated after coronary angiography. Second, after evaluating organ dose according to 9 imaging directions during coronary angiography, with the LAO fixed at 30°, dose evaluation was performed according to tube voltage, tube current, number of frames, focus-skin distance, and field size. Third, the effective dose for each organ was calculated according to the tissue weighting factors presented in ICRP(International Commission on Radiological Protection) recommendations. As a result, the average sum of air kerma during coronary angiography was evaluated as 234.0±112.1 mGy, the dose-area product was 25.9±13.0 Gy·cm2, and the total fluoroscopy time was 2.5±2.0 min. Also, the organ dose tended to increase as the tube voltage, milliampere-second, number of frames, and irradiation range increased, whereas the organ dose decreased as the FSD increased. Therefore, medical radiation exposure to patients can be reduced by selecting the optimal tube voltage and field size during coronary angiography, maximizing the focal-skin distance, using the lowest tube current possible, and reducing the number of frames.

Effect of Attenuation Correction, Scatter Correction and Resolution Recovery on Diagnostic Performance of Quantitative Myocardial SPECT for Coronary Artery Disease (감쇠보정, 산란보정 및 해상도복원이 정량적 심근 SPECT의 관상동맥질환 진단성능에 미치는 효과)

  • Hwang, Kyung-Hoon;Lee, Dong-Soo;Paeng, Jin-Chul;Lee, Myoung-Mook;Chung, June-Key;Lee, Myung-Chul
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.5
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    • pp.288-297
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    • 2002
  • Purpose: Soft tissue attenuation and scattering are major methodological limitations of myocardial perfusion SPECT. To overcome these limitations, algorithms for attenuation, scatter correction and resolution recovery (ASCRR) is being developed, while quantitative myocardial SPECT has also become available. In this study, we investigated the efficacy of an ASCRR-corrected quantitative myocardial SPECT method for the diagnosis of coronary artery disease (CAD). Materials and Methods: Seventy-five patients (M:F=51:24, $61.0{\pm}8.9$ years old) suspected of CAD who underwent coronary angiography (CAG) within $7{\pm}12$ days of SPECT(Group-I) and 20 subjects (M:F=10:10, age $40.6{\pm}9.4$) with a low likelihood of coronary artery disease (Group-II) were enrolled. Tl-201 rest/ dipyridamole-stress Tc-99m-MIBI gated myocardial SPECT was performed. ASCRR correction was peformed using a Gd-153 line source and automatic software (Vantage-Pro; ADAC Labs, USA). Using a 20-segment model, segmental perfusion was automatically quantified on both the ASCRR-corrected and uncorrected images using an automatic quantifying software (AutoQUANT; ADAC Labs.). Using these quantified values, CAD was diagnosed in each of the 3 coronary arterial territories. The diagnostic performance of ASCRR-corrected SPECT was compared with that of non-corrected SPECT. Results: Among the 75 patients of Group-I, 9 patients had normal CAG while the remaining 66 patients had 155 arterial lesions; 61 left anterior descending (LAD), 48 left circumflex (LCX) and 46 right coronary (RCA) arterial lesions. For the LAD and LCX lesions, there was no significant difference in diagnostic performance. In Group-II patients, the overall normalcy rate improved but this improvement was not statistically significant (p=0.07). However, for RCA lesions, specificity improved significantly but sensitivity worsened significantly with ASCRR correction (both p<0.05). Overall accuracy was the same. Conclusion: The ASCRR correction did not improve diagnostic performance significantly although the diagnostic specificity for RCA lesions improved on quantitative myocardial SPECT. The clinical application of the ASC-RR correction requires more discretion regarding cost and efficacy.

Relationship between Helicobacter pylori infection and iron-deficiency anemia in infants and children (영유아에서 Helicobacter pylori 감염과 철결핍성 빈혈과의 관계 연구)

  • Son, Meong Hi;Yeom, Jung Suk;Park, Ji Suk;Park, Eun Sil;Seo, Ji Hyun;Lim, Jae Young;Park, Chan Hoo;Woo, Hyang Ok;Youn, Hee Shang
    • Clinical and Experimental Pediatrics
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    • v.52 no.5
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    • pp.544-548
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    • 2009
  • Purpose : To elucidate a potential association between Helicobacter pylori (HP) infection and iron-deficiency anemia (IDA) in infants and children in terms of the other factors related to iron utilization and storage although the association of ferritin was previously studied. Methods : We evaluated 135 infants (aged 6-24 months) admitted at Gyeongsang National University Hospital from 2000 to 2006. Western blot assays using the HP CagA antigen (120 kD) were conducted to identify infections. The concentrations of six parameters were measured: hemoglobin (Hb), serum ferritin, soluble serum transferrin receptors, interleukin-6, prohepcidin, and C-reactive protein. In addition, the infants were classified into IDA, anemia from inflammation (AI), unclassified anemia (UCA), and normal groups on the basis of Hb and ferritin concentrations. Results : In the IDA group (n=20), seven infants were infected with HP, with the other infants showing no evidence of infection. The mean Hb levels in the IDA group were significantly lower in HP-infected infants than those uninfected (7.1 vs. 8.2 g/dL, respectively); the mean ferritin levels were also significantly lower in the infected infants (3.2 vs. $6.8{\mu}g/L$). The other four parameters did not differ significantly among the IDA infants. No correlations were found between the six parameters and HP infection status in the other groups. Conclusion : There were no significant differences in the HP infection rates among the study groups. However, in the IDA group, the HP-infected infants had significantly lower serum ferritin and Hb levels than the HP-negative infants (P<0.05).

Coronary Artery Bypass Graft (CABG) for Intracavitary Coronary Artery Disease - A case report- (심강내관상동맥(Intracavitary Coronary Artery)질환에 대한 관상동맥우회술 -수술치험 1예-)

  • Kim Su-Wan;Sung Kiick;Park Pyo Won;Jun Tae-Gook;Park Kay-Hyun;Lee Young Tak
    • Journal of Chest Surgery
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    • v.38 no.7 s.252
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    • pp.504-506
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    • 2005
  • Intracavitary coronary artery is variant anomalous entrance to right ventricular cavity of left anterior descending artery. Since the disease is extremely rare, there has not been any report of it in Korea and is only found in $0.2\~0.3\%$ of all CABG patients. It is very difficult to be diagnosed by preoperative coronary arteriography (CAG) and secure suture is needed for right ventriculotomy after CABG due to bleeding from right ventricle. Horizontal mattress suture with pledget has been recommended but, it could compress the myocardium surrounding ventriculotomy and result in disturbed flow of left anterior descending artery branch and perforating artery. So we used simple interrupted suture and the patient was recovered as other CABG patients without complications.

Characterization of binding specificity using GST-conjugated mutant huntingtin epitopes in surface plasmon resonance (SPR)

  • Cho, Hang-Hee;Kim, Tae Hoon;Kim, Hong-Duck;Cho, Jae-Hyeon
    • Korean Journal of Veterinary Service
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    • v.44 no.4
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    • pp.185-194
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    • 2021
  • Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

Clinical Significance of Reverse Redistribution Phenomenon on Delayed Tc-99m Tetrofosmin Myocardial Perfusion Imaging in Patients with Acute Myocardial Infarction (급성 심근경색 환자의 Tc-99m Tetrofosmin 심근 관류 지연영상에서 관찰되는 역재분포 현상의 임상적 의의)

  • Park, Soon-Ah;Kim, Dae-Weung;Kim, Chang-Guhn;Jeong, Jin-Won;Kim, Nam-Ho;Yun, Kyeong-Ho
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.2
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    • pp.112-119
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    • 2009
  • Purpose: This study was performed to investigate the clinical significance of reverse redistribution(RR) phenomenon detected on delayed Tc-99m tetrofosmin myocardial single photon emission computed tomography(SPEG) in patients with acute myocardial infarction after revascularization. Materials and Methods: A Tc-99m tetrofrosmin myocardial SPECT was performed in 67 consecutive patients after revascularization for acute myocardial infarction. Myocardial SPECT imaging was performed for early imaging at 40 min and for delayed imaging at 180 min after reinjection at myocardial stress. Regional myocardial uptakes were scored by 4-point scoring in the left ventricular wall divided into 17 segments. Reverse redistribution was defined as an increase of more than 2 point in the activity score on the delayed image. Follow-up myocardial SPECT and coronary angiography(CAG) were performed 9 months later. Results: On myocardial SPECT performed following revascularization, RR was observed in 100 of all 319 segments(31%) and in 43 patients(64%). The abnormalities of perfusion and regional wall motion were more severe in the patients with RR compared to those without RR(p<0.05). On follow-up myocardial SPECT, the myocardial perfusion, regional wall motion, and myocardial thickness were significantly improved in the patients with RR(p<0.05) however, these changes were not significant in those without RR. There was no significant difference between the patients with RR and those without RR in the occurrence of restenosis on CAG. Conclusions: In patients with acute myocardial infarction, the regions showing the RR phenomenon on delayed Tc-99m tetrofosmin SPECT may reflect viable myocardium and indicate recovery of salvaged myocardium.

Genomic Diversity of Helicobacter pylori

  • Lee, Woo-Kon;Choi, Sang-Haeng;Park, Seong-Gyu;Choi, Yeo-Jeong;Choe, Mi-Young;Park, Jeong-Won;Jung, Sun-Ae;Byun, Eun-Young;Song, Jae-Young;Jung, Tae-Sung;Lee, Byung-Sang;Baik, Seung-Chul;Cho, Myung-Je
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.519-532
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    • 1999
  • Helicobacter pylori is a causative agent of type B gastritis and plays a central role in the pathogenesis of gastroduodenal ulcer and gastric cancer. To elucidate the host-parasite relationship of the H. pylori infection on the basis of molecular biology, we tried to evaluate the genomic diversity of H. pylori. An ordered overlapping bacterial artificial chromosome (BAC) library of a Korean isolate, H. pylori 51 was constructed to set up a genomic map. A circular physical map was constructed by aligning ApaI, NotI and SfiI-digested chromosomal DNA. When the physical map of H. pylori 51 was compared to that of unrelated strain, H. pylori 26695, completely different restriction patterns were shown. Fifteen known genes were mapped on the chromosome of H. pylori 51 and the genetic map was compared with those of strain 26695 and J99, of which the entire genomic sequences were reported. There were some variability in the gene location as well as gene order among three strains. For further analysis on the genomic diversity of H. pylori, when comparing the genomic structure of 150 H. pylori Korean isolates with one another, genomic macrodiversity of H. pylori was characterized by several features: whether or not susceptible to restriction digestion of the chromsome, variation in chromosomal restriction fingerprint and/or high frequency of gene rearrangement. We also examined the extent of allelic variation in nucleotide or deduced amino acid sequences at the individual gene level. fucT, cagA and vacA were confirmed to carry regions of high variation in nucleotide sequence among strains. The plasticity zone and strain-specific genes of H. pylori 51 were analyzed and compared with the former two genomic sequences. It should be noted that the H. pylori 51-specific sequences were dispersed on the chromosome, not congregated in the plasticity zone unlike J99- or 26695-specific genes, suggesting the high frequency of gene rearrangement in H. pylori genome. The genome of H. pylori 51 shows differences in the overall genomic organization, gene order, and even in the nucleotide sequences among the H. pylori strains, which are far greater than the differences reported on the genomic diversity of H. pylori.

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