new CO2 incubators

not easy to lift that freaking heavy thing over the new one on the new black frame. both CO2 now for 37C. One for infected and the other clean cells. Hohohho...one step nearer to CO2 poisoning...5% + 5% = 10%..hiak...hiak...hiak...

stupid cat

somebody's cat trying to get into my house by going through the window. Grrrrh...
It managed to get in last week and sat on my couch, dropped lots of fur around.... which reminds me, i need to clean my windows soon.

bang!

the last week see accident at AYE



then yesterday also see got bang around



forgot to take down car number...

my name!!!

J Immunol. 2007 Mar 1;178(5):2699-705.

Immunization of Flavivirus West Nile Recombinant Envelope Domain III Protein Induced Specific Immune Response and Protection against West Nile Virus Infection.

Chu JH, Chiang CC, Ng ML.

Flavivirology Laboratory, Department of Microbiology, National University of Singapore, Singapore.

The domain III of the West Nile virus (WNV) envelope glycoprotein (E) was shown to serve as virus attachment domain to the cellular receptor, and neutralizing Abs have been mapped to this specific domain. In this study, domain III of the WNV E protein (WNV E DIII) was expressed as a recombinant protein and its potential as a subunit vaccine candidate was evaluated in BALB/C mice. Immunization of WNV E DIII protein with oligodeoxynucleotides (CpG-DNA) adjuvant by i.p. injection was conducted over a period of 3 wk. The immunized mice generated high titer of WNV-neutralizing Abs. Murine Ab against WNV E DIII protein was also capable of neutralizing Japanese encephalitis virus. The IgG isotypes generated were predominantly IgG2a in the murine sera against the recombinant protein. Splenocyte cultures from the mice coadministrated with WNV E DIII protein and CpG secreted large amounts of IFN-gamma and IL-2 and showed proliferation of T cells in the presence of WNV E DIII protein. Overall, this study highlighted that recombinant WNV E DIII protein delivered in combination with CpG adjuvant to mice generated a Th1 immune response type against WNV and can serve as a potential vaccine to prevent WNV infection.

PMID: 17312111 [PubMed - in process]

paper

Antiviral Res. 2006 Dec;72(3):216-23. Epub 2006 Jul 7.

Expression of vector-based small interfering RNA against West Nile virus effectively inhibits virus replication.

• Ong SP,
• Choo BG,
• Chu JJ,
• Ng ML.

Flavivirology Laboratory, Department of Microbiology, 5 Science Drive 2, National University of Singapore, 117597 Singapore, Singapore.

RNA interference is one of the effective emerging anti-viral strategies to inhibit virus infection in cells. In this study, a small interfering RNA expressing vector (pSilencer-NS5) targeting the NS5 gene of West Nile virus (WNV) was employed to target and destroy WNV transcripts. Real-time PCR revealed drastic reduction in WNV RNA transcripts in pSilencer-NS5-transfected Vero cells. The virus infectious titre was also significantly reduced by 90% as determined by plaque assays. The resulting decrease in virus replication was shown to be specific since both scrambled and nucleotide(s) mismatch siRNA against WNV NS5 gene did not have any effect on WNV productive yields. Furthermore, Western immunoblot analysis on the expression of viral NS5 and envelope (E) proteins showed significant down-regulation on the expression of viral NS5 and envelope (E) proteins in virus-infected cells that were pre-transfected with pSilencer-NS5. These data clearly supported the notion that the expression of vector-based siRNA against WNV NS5 gene is able to exert its silencing effect on WNV-infected cells without inducing cytotoxicity, hence holding promise in therapeutic treatment of this important emerging infectious disease.

PMID: 16870272 [PubMed - indexed for MEDLINE]

Dept open house

The department had an open house for year 1 and 2 students to check out the PI's in 'da house so more students will be attracted to come in. We were supposed to have a lab poster and so I gathered some pictures and printed an A3 poster out and stuck it to the board. In the end, only 3 labs had posters and no one asked us what our lab was about. We just talked to each other and ate free food. One Saturday wasted. There was a talk session where different PI's gave outlines of thier work (mine didnt) and then lunch and then more talk and then supposedly the poster session. About 50 students turned up; most came in time for lunch.

My open house

Last last week, the lab went to my place for an open house. We had a $200 Pen Cai, a barrel of KFC and some extra food from the EM unit that was having some seminar. Here are the pictures. Oh yes, Ipoh boy turns another year more mature.








Prof got a CNY hamper just two days before that contained amongst other things, that yellow dome shape cake. We thought it was a gigantic pineapple tart but it turned out to be a pumpkin filled thingy. The chocolate brownie brought us as always to cloud9.5.
Nuff said.
Happy V day everyone. Will be spending mine with the female mice at the animal house.

microbio open house

HAving an open house this Saturday and I made a poster of our lab people. I know that most people after seeing it will want to kill me. But here it is anyway.

last year's christmas lunch

We went to Oriental for lunch a few days after Christmas last year. Yeah, I know, its kinda late to post this but since I have free time today (read: all my cell culture stock contaminated) I got some time to expand.


Melt the world cafe. We were early (11:45am,) and doors only open at 12 and they have not set our table yet. So got to wait.


The table


The waiter's hand


The ambiance


Most importantly, THE FOOD!!!














The gluttons.


The glutton^n


The reflection on the ceiling


The group


The vain two.


The end.

sorry

Hi faithful readers. Sorry for the loooong absence. I have been terribly lazy/busy/sick/etc. and did not post.

Today Ad came back to lab after his holiday to some northern hemisphere region and brought back food!




This one good. Rum




This one not so good. Whisky


This one everybody liked


That person cannot have fourth rounds.





Yeah, thats my new home. And thats why these few days/weeks I did not post. Was busy with the new house reno and S&P stuff. When its done, everybody's invited! (international transport not provided).

guess

Guess what this is!

microbes and infection corrected proof

Original article

The envelope glycoprotein domain III of dengue virus serotypes 1 and 2 inhibit virus entry

J.F.L. Chin^a, J.J.H. Chu^a and M.L. Ng<sup>, a,

a</sup>Flavivirology Laboratory, Department of Microbiology, 5 Science Drive 2, National University of Singapore, Singapore 117597, Singapore

Received 28 July 2006; accepted 30 September 2006. Available online 6 December 2006.

Abstract

Dengue virus (DV) is a flavivirus and its urban transmission is maintained largely by its mosquito vectors and vertebrate host, often human. In this study, investigation was carried out on the involvement of domain III of the envelope (E) glycosylated protein of dengue virus serotypes 1 and 2 (DV-1 and DV-2 DIII) in binding to host cell surfaces, thus mediating virus entry. Domain III protein of flavivirus can also serve as an attractive target in inhibiting virus entry. The respective DV DIII proteins were expressed as soluble recombinant fusion proteins before purification through enzymatic cleavage and affinity purification. The purified recombinant DV-1 and DV-2 DIII proteins both demonstrated the ability to inhibit the entry of DV-1 and DV-2 into HepG2 cells and C6/36 mosquito cells. As such, the DV DIII protein is indeed important for the interaction with cellular receptors in both human and mosquito cells. In addition, this protein induced antibodies that completely neutralized homologous dengue serotypes although not with the same efficiency among the heterologous serotypes. This observation may be of importance when formulating a generic vaccine that is effective against all dengue virus serotypes.

Keywords: Flavivirus; Virus entry; Vaccine; Neutralizing antibodies

Thai junk food

Ed's 'friend' went to Thailand and got back this for the lab. The banana chips still my fav.