Illness and Diseases

Generation of ds DNA template from synthetic oligos. Part 2

The use of fully synthetic oligonucleotides as positive control material has been described recently. However, there were limitations as to the length of the oligonucleotide, which was generally unable to reach more than 130 bases. In many situations, for instance in the case of drug resistance mutations, nucleotide substitutions are often scattered within the target […]

September 21, 2011 – 11:08 pm | By Dan Frost | Posted in Other | Comments (0)

Synthetic long oligonucleotides. Part 2

Significant advances in molecular biology and human genomic research has paved the way for a host of new genetic diagnostic tests, including gene sequencing, detection, identification and genotyping of organisms using real time polymerase chain reaction (PCR) or other amplification techniques such as multiplex PCR, reverse line blot hybridization (RLB), Ligase chain reaction (LCR), Rolling […]

September 15, 2011 – 4:48 pm | By Dan Frost | Posted in Other | Comments (0)

Synthetic long oligonucleotides

Background Positive controls are an integral component of any sensitive molecular diagnostic tool, but this can be affected, if several mutations are being screened in a scenario of a pandemic or newly emerging disease where it can be difficult to acquire all the necessary positive controls from the host. This work describes the development of […]

September 8, 2011 – 5:16 am | By Dan Frost | Posted in Other | Comments (0)

Non-structural protein 1 of avian influenza A viruses differentially inhibit NF-kappaB promoter activation

Background Influenza virus infection activates NF-κB and is a general prerequisite for a productive influenza virus infection. On the other hand, non-structural protein 1 (NS1) suppresses this viral activated NF-κB, presumably to prevent expression of NF-κB mediated anti-viral response. NS1 proteins of influenza A viruses are divided into two groups, known as allele A and […]

August 23, 2011 – 3:11 pm | By Dan Frost | Posted in Other | Comments (0)

Entry in the absence of membrane fusion. Part 2

Additional evidence supporting alphavirus entry via a pore at the plasma membrane was obtained using freeze-fracture (Kononchik, Vancini and Brown, Virology, in press). In these experiments, high MOI of SVHR was adsorbed onto mosquito cells for half an hour after which the virus was cross-linked to cell surface proteins with gluteraldehyde. These samples were then […]

August 14, 2011 – 11:02 pm | By Dan Frost | Posted in Diseases | Comments (0)

A measure of successful virus entry

There are numerous techniques that can be used to examine the interactions between the virus and the cell. These techniques can generally fall into two categories: direct and indirect observation. Direct observation uses familiar techniques such as thin-section microscopy, cryo-electron microscopy, tomography, and other less familiar techniques like freeze-fracture immunolabeling to examine virus-cell interactions. When […]

July 27, 2011 – 10:51 pm | By Dan Frost | Posted in Diseases | Comments (0)

Adsorption and receptor recognition. Part 2

As arboviruses, alphaviruses infect insect and vertebrate hosts. Since alphaviruses need to infect cells which provide widely divergent biochemical and genetic environments, it is likely that they either use a ubiquitous receptor, or are able to use multiple proteins as a receptor. The receptor/s has not been identified. Many proteins and polysaccharides have been implicated […]

July 23, 2011 – 5:32 pm | By Dan Frost | Posted in Diseases | Comments (0)

Adsorption and receptor recognition

A second caveat which seriously affects the quality of the data is that the entity to be studied not be distorted by the ability of proteins to be manipulated and expressed in some form in E. coli. There are currently crystal structures of an E1-E2 fusion protein (PDB codes 3MUU), chikungunya glycoproteins (PDB codes: 3N40;3N41;3N42;3N43;3N44;2XFB;2XFC) […]

July 19, 2011 – 9:49 pm | By Dan Frost | Posted in Other | Comments (0)

Alphavirus Structure. Part 2

The current molecular model for how the virus transfers its RNA into the cell is hypothesized to be by low pH mediated membrane fusion, after endocytosis of the virus attached to its receptor. E2 contains the receptor binding sequence while E1 is known to contain the properties necessary for membrane fusion. It is predicted that […]

July 15, 2011 – 6:54 pm | By Dan Frost | Posted in Diseases | Comments (0)

Alphavirus Structure

Alphaviruses are small 70 nm viruses that have 240 copies each of three structural proteins, E1, E2 and capsid (C) assembled in a 1:1:1 stoichiometry. These three proteins create two nested T = 4 icosahedral shells that sandwich a host derived lipid bilayer. The outer protein shell is composed of E1 and E2 heterodimers that […]

July 5, 2011 – 7:21 pm | By Dan Frost | Posted in Diseases | Comments (0)