THE OSTIA VENAE HEPATICAE AND THE RETHROHEPATIC ...

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NEUTRALIZING MONOCLONAL ANTIBODY TO BURKHOLDERIA PSEUDOMALLEI CYTOLETHAL TOXIN Pattama Ekpo1, Supinya Pongsunk2 and Takayuki Ezaki3 1Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2Department of Microbiology, Faculty of Medicine, Srinakharinwirot University, Bangkok, Thailand and 3Department of Microbiology, Gifu University School of Medicine, Gifu, Japan. Little is known about the virulence factors associated with infection by Burkholderia pseudomallei, the causative organism of melioidosis. The toxins are ones of the virulence factors that might be responsible for the rapid onset of the melioidosis and dramatic course of septicemic melioidosis. In this study, a cytolethal toxic activity of the crude culture filtrate (CCF) of B. pseudomallei to cell culture, THP-1 cell; human monocyte cell line, was studied. The highest activity of the cytolethal toxin was in the CCF fraction with MW ranging from >30,000-100,000. The evidence from a laser scanning confocal microscope that used for examining the THP-1 cells at time intervals after adding the toxin showed that there were many pores on the cells’ surface before lysis. Therefore, the mechanism of the toxin might generate holes on the cells’ membrane. We produced a monoclonal antibody (MAb) by using the CCF as immunogen. The MAb was specific to the 30-kDa protein of B. pseudomallei. The MAb can neutralize the cytolethal toxin in the CCF and the CCF fraction (MW>30,000-100,000). The purified cytolethal toxin will be further studied for its role in pathogenesis. It might be a candidate for one of the factors involved in the rapid onset and dramatic course of septicemic melioidosis. CLONING AND CHARACTERIZATION OF A NONHEMOLYTIC PHOSPHOLIPASE C GENE FROM BURKHOLDERIA PSEUDOMALLEI Sunee Korbsrisate, 2* Nuttiga Suwanasai, 1 Amornurt Leelaporn, 2 Takayuki Ezaki, 3 Yoshiaki Kawamura, 3 and Suttipant Sarasombath 3 1 Department of Immunology and 2 Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University Bangkok 10700, Thailand, and 3 Department of Microbiology, Gifu University School of Medicine, Gifu 500, Japan3 We cloned and characterized a phosphatidylcholine hydrolyzing phospholipase C (PC- PLC) gene from Burkholderia pseudomallei. DNA sequence analysis of the gene indicated an open reading frame coding for 700 amino acids with a 34-amino-acid signal peptide. When cleaved, this yields a secreted 73-kDa mature protein. The deduced amino acid sequence exhibited 48% similarity to that of a nonhemolytic PLC from Pseudomonas aeruginosa. The expressed PC-PLC was heat stable, nonhemolytic for sheep erythrocytes, and active between pH2 and 8. Western blot analysis with sera from melioidosis patients indicated that they produced immunoglobulin M antibodies against this PC-PLC protein. (276) 207 (277)
208 Faculty of Medicine Siriraj Hospital CHARACTERIZATION OF A PHASE 1-D EPITOPE ON SALMONELLA TYPHI FLAGELLIN AND ITS ROLE IN THE SERODIAGNOSIS OF TYPHOID FEVER Sunee Korbsrisate 1 , Ariya Thanomsakyuth 1 , Napatawn Banchuin 1 , Stan McKay 2 , Moazzem Hossain 3 and Suttipant Sarasombath 1 1 Department of Immunology, Faculty of Medicine Siriraj Hospital,Mahidol University, 2 World Vision International cambodia, Phnom-Penh, Kingdom of Cambodia, 3 Sir Salimullah Medical College, Dhaka, Bangladesh. A monoclonal antibody (Mab) directed against Samonella typhi 52 kDa flagellin protein has been previously produced by our group. In this study, we have demonstrated that the epitope specific to the Mab is unique to phase 1-d. To map the epitope, plasmids encoding different regions of S. typhi flagellin gene were constructed. Analysis of protein produced from each recombinant plasmid indicated that the epitope specific to the Mab resided within amino acids 171-303 (region IV) of S. typhi flagellin protein. The recombinant region IV flagellin was used to develop an ELISA for the detection of IgM antibody to S. typhi in serum. In the hemoculture-positive typhoid group, the developed ELISA was positive in 77 of 92 cases. In patients with non-typhoidal Salmonella, gram-positive and gram-negative bacteria or dengue virus, the ELISA was negative in all 78 cases. Two from 116 healthy control subjects had positive reactions with the assay. The calculated sensitivity, specificity, positive and negative predictive values of the test were 83.7%, 99.0%, 97.5% and 92.8%, respectively. With such high validity together with the requirement of only a single serum specimen and one day for performing the test, the developed ELISA should become a valuable diagnostic test for typhoid fever. DETECTION OF IgM ANTIBODY AGAINST REGION IV FLAGELLIN OF SALMONELLA PARATYPHI A Sunee Korbsrisate 1 , Suttipant Sarasombath 1 , Nuttaya Praaporn 1 , Pattara Iamkamala 2 , Moazzem Hossain 3 and Stan Mckay 4 1 Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Department of Medicine, Lardprao General Hospital, Bangkok, Thailand, 3 Sir Salimullah Medical College, Dhaka, Bangladesh, 4 World Vision International Cambodia, Phnom-Penh, Kingdom of Cambodia. Salmonella paratyphi A is a pathogenic bacterium that causes paratyphoid fever. The current laboratory diagnostic techniques are unsatisfactory. To improve diagnosis, a plasmid (pSK-8E) encoding phase 1 flagellin gene nucleotide position 452-890 from S. paratyphi A has been constructed. The recombinant protein expressed from the plasmid has been used to develop an indirect ELISA for IgM antibody detection. Sera from patients with hemoculture positive for S. paratyphi A, S. typhi, other gram-positive and gram-negative bacteria, and dengue hemorrhagic fever as well as from healthy control subjects were tested. Sensitivity, specificity, positive and negative predictive values of the test were 56.9%, 98.8% 90.6% and 92.1%, respectively. Since the sensitivity was low, the explanation for this result was investigated. It was found that the sensitivity of the test could be increased to 83.3% if the sera were obtained 9-12 days after onset of fever. The sera obtained earlier or later gave only 33.3% and 66.6% sensitivity, respectively. This result suggests that the IgM antibody detection assay which we have developed is a valuable tool for diagnosis of S. paratyphi A infection when the serum samples are taken at the appropriate time. (278) (279)
Page 1 and 2: THE OSTIA VENAE HEPATICAE AND THE R
Page 3 and 4: WHEN SHOULD WE ORDER PREOPERATIVE C
Page 5 and 6: toneal lidocaine instillation (Grou
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Page 17 and 18: IMPROVED DETECTION OF RADIOFREQUENC
Page 19 and 20: THE PREVALENCE OF CHLAMYDIA PNEUMON
Page 21 and 22: THE PREVALENCE OF CHLAMYDIA PNEUMON
Page 23 and 24: COMPARATIVE TREATMENT OF ORAL CANDI
Page 25 and 26: DEEP FUNGAL AND HIGHER BACTERIAL SK
Page 27 and 28: CLINICAL AND MICROBIOLOGIC FINDINGS
Page 29 and 30: LASER HAIR REMOVAL AFFECTS SEBACEOU
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Page 37 and 38: 202 Faculty of Medicine Siriraj Hos
Page 39 and 40: 204 PHOTOTESTING IN ORIENTAL PATIEN
Page 41: 206 specimens, respectively. The sp
Page 47 and 48: 212 HEMOGLOBINOPATHIES : A COMPARAT
Page 49 and 50: 214 tomized cases were 137.99³29.5
Page 57 and 58: 222 Characterization of mutations o
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Page 71 and 72: 236 RISK OF OMENTAL METASTASIS IN E
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Page 81 and 82: 246 THE MANAGEMENT OF SACROCOCCYGEA
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Page 87 and 88: 252 V. Rojvanit AVOIDNESS AND TREAT
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258 Faculty of Medicine Siriraj Hos
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260 INTESTINAL PARASITIC INFECTIONS
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272 The 1,020 gm., 19.5 x 18.5 x 5.
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276 Objective : To measure the rela
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280 formula ketogenic diet was used
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282 INTERMEDIATE TERM FOLLOW-UP ON
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292 NEONATAL SCREENING FOR CONGENIT
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294 SENSITIVE AND SIMPLE HPLC ASSAY
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296 (EIA) and the isoform of COX pr
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298 maintenance of local homeostasi
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300 Faculty of Medicinr Siriraj Hos
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314 NORMAL APPEARANCES OF TECHNETIU
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316 that will deliver 2 Gy to the b
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322 THE RELATIONSHIP BETWEEN MYOFAS
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328 Result.At the 5-month postopera
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Mahidol University Annual Research
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336 individuals with SAO and in the
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342 EVALUATION OF MONOCLONAL ANTIBO
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