genomic extraction kit (QIAamp DNA Mini Kit, Qiagen). Construction of the library and sequencing were performed by the Biochemistry Department, Medicine Faculty, Chulalongkorn University, Thailand. The library was prepared using a NEBNext Ultra DNA Library Prep Kit for Illumina. Whole genome sequencing and assembly were analyzed using a HiSeq 2000 Illumina system (New England Biolabs). PCR and DNA sequencing of the RD16 region (Rv3405c) Genomic DNA from the working seed BCG (lot No. FB03012) was extracted using a genomic extraction kit (QIAamp DNA Mini Kit, Qiagen
Wichit Thaveekarn, Sunchai Payungporn, Narumol Pakmanee, Sunutcha Suntrarachun, Suchitra Khunsap and Suthidee Petsong
Methee Sriprapun, Chalinee Laosakul, Sunisa Krajiw, Kesinee Arunyingmongkol, Padet Siriyasatien and Wanla Kulwichit
ranging from asymptomatic infection, dengue fever and, dengue hemorrhagic fever (DHF) to dengue shock syndrome [ 2 ]. Previous studies by our group and by others have shown that, during acute infection, DENV genome is detectable in blood, saliva, and urine samples [ 3 - 6 ]. In addition, concurrent infections with multiple serotypes can occur in both infected mosquitoes and humans [ 7 - 10 ]. Here, we present a case of dual DENV infections with DENV2 and DENV4 serotypes in the urine at a single time point. The ‘minor’ serotype disappeared in subsequent specimens
Wasin Laohavinij and Apiwat Mutirangura
, including Student t , Pearson χ 2 , and Benjamini false discovery rate (FDR) tests, to identify the genes and the molecular pathways associated with the aging process and various types of exercise. The official names of significant genes and their molecular functions, and their molecular and cellular phenotypes, were evaluated by GeneCards, a human gene database maintained by the Crown Human Genome Center at the Weizmann Institute of Science ( http://www.genecards.org/ ). Methods Overall methodical framework is shown in Figure 1 . Figure 1 The overall
Haifeng Wang, Delin Yang, Jiansong Wang and Hongyi Xu
Background: Metastasis of tumor implantation includes a series of processes from detachment from the primary tumor to formation of the implanted metastase. Tumor cells survival in urine is a necessary condition for metastasis. Adaptation to urine is essential for this.
Objective: Establish a urine-resistant cell sub-strain of human bladder cancer cell line (ET cell lines), and study different characteristics compared to parent cells.
Methods: EJ cell lines were cultured in nutrient medium. Urine-resistance cell sub-strain (EJ-U) was harvested after prolonged culture by gradually increasing the concentration of urine. Gen chip was used to detect the genome series of EJ and EJ-U and to analyze the difference of gene expression.
Results: EJ-U in urine had a higher survival rate after 24 hours in urine compared with EJ. The EJ-U had almost the same growth velocity with EJ, and they had the analogous growth curves. The time-duration for EJ-U to survive was longer than EJ in urine. In gene ontology analysis, 272 significant different genes were found.
Conclusion: EJ-U cell sub-strain was more adaptable than its parent cell lines EJ. The different genes may explain the reason why bladder cancer cells could survive for a long time in urine.
Mohammad A. Farraj, Gabi M. Abusada, Abed Alraoof M. Saleem, Ayyub Y. Joaidi, Raba M. Radad, Hiba N. Atrash, Israr N. Sabri and Tamer A. Essawi
Background: Neisseria gonorrohoeae is an exclusive human pathogen that primarily infects the urogenital epithelia. Infections caused by N. gonorrhoeae are considered the second major cause of sexually transmitted disease after Chlamydiae worldwide. Although the urethra and the uterine cervix serve as the initial sites for gonococcal infections in men and women, infection of the conjunctiva, pharynx, tendons, joints, as well as rectal mucosa are also reported. Objectives: The objectives of this study were to introduce molecular techniques such as polymerase chain reaction (PCR) to detect N. gonorrhoeae directly from endocervical swabs. In addition, it provides a picture of Neisseria gonorrohea infection among a sample of Palestinian women in West Bank. Methods: Two hundred and thirteen endocervical swabs were collected from sexually active married women with endocervical abnormalities attending healthcare clinics. DNA was extracted directly from the swabs and PCR was performed using specific primers targeting the orf1 region of the genome. Results: The results obtained indicated that the percentage of positive cases of N. gonorrhoeae among the women tested was 1.40%. Conclusion: Implementing guidelines for comprehensive screening of men and women with more sensitive tests may improve detection and management of sexually transmitted infections.
Sunutcha Suntrarachun, Surasak Akesowan and Thaweesak Tirawatnapong
Background: Equine infectious anemia virus (EIAV) is a lentivirus with an almost worldwide distribution, infecting equids. It causes a persistent infection that is characterized by recurring episodes of fever, anemia, and thrombocytopenia. Most of the horses may control EIAV replication within a year, remaining persistently infected without clinical signs of disease. Objective: Detect EIA nucleic acid from peripheral blood of asymptomatic horses using nested PCR. Materials and method: We used nested PCR, amplifying P26 gag gene of EIAV, for direct detection of viral RNA in plasma and proviral DNA from PBMC in asymptomatic carrier horses in comparison with the Coggins test. EIA nucleic acid was prepared from 20 seropositive and five EIAV seronegative horses. Amplification of 246 bp expected size fragments was obtained using two different sets of primers targeting the P26 gag gene. Results: Among 20 seropositive horses, nine samples were positive for RNA and DNA. The five samples were positive for DNA but not for RNA, which indicates that the virus integrated into the host cell genome with a low level of viral replication. However, six samples were negative for both DNA and RNA. False negative could result due to primer failure caused by gag sequences variation among strains circulating in Thailand when compared with various strains from other parts of the world. EIAV antigens may also be prepared from cell cultures contaminated with other retroviruses thus causing false positives with the Coggins test. Conclusion: Nested PCR can be a useful tool for detecting the presence of EIAV in asymptomatic carrier horses. This may be especially true during the acute stage of the disease where the viremia levels are usually at the highest levels before detectable antibodies appear.
Chatchawit Aporntewan and Apiwat Mutirangura
millions of expression profiles-database and tools update. Nucleic Acids Res. 2007; 35(Database issue):D760-5. 5. Barrett T, Suzek TO, Troup DB, Wilhite SE, Ngau WC, Ledoux P, et al. NCBI GEO: mining millions of expression profiles-database and tools. Nucleic Acids Res. 2005; 33(Database issue):D562-6. 6. Butte AJ. Translational bioinformatics applications in genome medicine. Genome Med. 2009; 1:64. 7. Pyeon D, Newton MA, Lambert PF, den Boon JA, Sengupta S, Marsit CJ, et al. Fundamental differences in cell cycle
Uraiwan Kositanont, Patcharee Prasajak, Suwanna Trakulsomboon, Noppadol Sangjun and Duangporn Phulsuksombati
medium of bovine albumin and polysorbate 80. Am J Vet Res. 1965; 26:45-51. 16. Galloway RL, Levett PN. Evaluation of a modified pulsed-field gel electrophoresis approach for the identification of Leptospira serovars. Am J Trop Med Hyg. 2008; 78:628-32. 17. Nascimento AL, Verjovski-Almeida S, Van Sluys MA, Monteiro-Vitorello CB, Camargo LE, Digiampietri LA, et al. Genome features of Leptospira interrogans serovar Copenhageni. Braz J Med Biol Res. 2004; 37: 459-77. 18. Tamai T, Sada E, Kobayashi Y. Restriction endonuclease
Human genome variability occurs by chance over several generations. When genetic variation occurs and disrupts the function of a particular gene, it is called mutation. When genetic variation occurs, but does not affect any particular gene function, it is called polymorphism. Despite the lack of functional effect, gene polymorphisms are known to be associated with certain disease conditions. The association can occur because the particular polymorphism is linked to a not-yet-known disease gene mutation. It is well known that the incidence of many diseases
Chotika Suyarnsestakorn, Thatchawan Thanasupawat, Kantima Leelahavanichkul, J. Silvio Gutkind and Apiwat Mutirangura
:4839-44. 11. Holmes SE, O’Hearn EE, McInnis MG, Gorelick- Feldman DA, Kleiderlein JJ, Callahan C, et al. Expansion of a novel CAG trinucleotide repeat in the 5' region of PPP2R2B is associated with SCA12. Nat Genet.1999; 23:391-2. 12. Holmes SE, O’Hearn E, Margolis RL. Why is SCA12 different from other SCAs? Cytogenet Genome Res. 2003; 100:189-97. 13. Chun HH, Gatti RA. Ataxia-telangiectasia, an evolving phenotype. DNA Repair (Amst). 2004; 3: 1187-96. 14. Strack S, Zaucha JA, Ebner FF, Colbran RJ, Wadzinski BE. Brain protein