Synergistic inhibition of cell-to-cell HIV-1 infection by combinations of single chain variable fragments and fusion inhibitors

Cell-to-cell spread of HIV permits ongoing viral replication in the presence of antiretroviral therapy and is suggested to be a major contributor to sexual transmission by mucosal routes. Fusion inhibitors that prevent viral entry have been developed, but their clinical applications have been limited by weak antiviral activity, short half-life, and the low genetic barrier to development of resistance. We examined the inhibitory activities of a series of single-chain variable fragments (scFvs) targeting the V3 and CD4i epitopes against both cell-free and cell-to-cell HIV infection. We found that all anti-V3 scFvs, including two newly constructed scFvs, showed broad neutralization activity against a panel of subtype B viruses compared with the corresponding IgGs. All scFvs neutralized cell-free infection by HIV-1_{JR-FL WT} and fusion inhibitor-resistant mutants. In addition, all anti-V3 scFvs and some CD4i scFvs significantly inhibited cell fusion, while their IgG counterparts did not. Furthermore, scFvs-fusion inhibitors combinations, such as C34 and SC34, showed synergistic inhibition of cell fusion by both HIV-1_{JR-FL WT} and fusion inhibitor-resistant mutants. The most prominent combinational effect was observed for 916B2 CD4i scFv with SC34. The delayed fusion kinetics of fusion inhibitor-resistant mutants partly explain their synergistic inhibition by such combinations. Our data demonstrate the advantages of using scFvs over their parent IgGs for inhibiting both cell-free and cell-to-cell infection. High synergistic inhibition of cell fusion by using scFvs-fusion inhibitors combinations suggests the possibility of intensification therapy adding this combination to current anti-HIV treatment regimens.     

Determination of peracetylated sulfoglycolipids using the azure A method

Several sulfoglycolipids, including sulfogalactosylceramide, sulfolactosylceramide, monosulfogangliotriaosylceramide, monosulfogangliotetraosylceramide, bis-sulfogangliotriaosylceramide, bis-sulfogangliotetraosylceramide, seminolipid, and lysoseminolipid, were assayed by the azure A procedure (Kean, E.L. 1968. J. Lipid Res. 9: 319-327). The color yields were different significantly among each sulfoglycolipid. The accurate determination of monosulfoglycolipids having longer carbohydrate chains than sulfolactosylceramide as well as bis-sulfoglycolipids could not be achieved because of the low color yields, turbidity, and/or formation of an additional chromogen. However, the quantitative determination of these complex sulfoglycolipids could be achieved using the azure A procedure when these compounds were assayed after peracetylation. Using the modified Kean procedure, the behavior of these sulfoglycolipids in the chloroform-methanol-water partition system was compared.     

The protective effect of astaxanthin on the ganglion cell complex in glutamate/aspartate transporter deficient mice,a model of normal tension glaucoma,analyzed by spectral domain-optical coherence tomography

Astaxanthin (AST), a natural marine carotenoid, possess a wide variety of biological functions. In particular, as a strong antioxidant, AST effectively scavenges oxygen free radicals and reduces oxidative stress. In addition, recent in vitro studies have suggested that AST attenuates glutamate-induced apoptosis and cytotoxicity. The glutamate/aspartate transporter (GLAST) deficient (GLAST^-/-) mouse is a mouse model of normal tension glaucoma (NTG) caused by both the glutamate neurotoxicity and oxidative stress in the retina. In the present study, we investigated the effects of AST on the ganglion cell complex, indicator of glaucomatous structural damage, using spectral domain-optical coherence tomography. As a result, AST significantly attenuated the thinning of ganglion cell complex in GLAST^-/- mice in comparison to an AST-free control group. Our results suggest the possibility that AST has protective effects against glutamate neurotoxicity and oxidative stress in the retina. At present, the only treatment for NTG that is available in the clinical setting is to reduce the IOP as much as possible. Thus, our results suggest that AST supplementation may be effective for some types of NTG in which glutamate neurotoxicity and oxidative stress are involved.     

Corticotrigeminal motor pathway in the rat--II. Anterio- and retrograde HRP labeling

1. Horseradish peroxidase (HRP) injection into the rat cortical jaw motor area (JMA) disclosed direct projection fibers in and around the motor trigeminal nucleus (MTN), primarily contralaterally. 2. Descending axons were found in the ventromedial half of the cerebral peduncle and MTN-projecting axons were concentrated near the descending facial nerve root. 3. HRP injection into the peduncle could label 1257 cells in the JMA, 128 cells in the taste area and 1409 cells in the neck-forelimb motor area ipsilaterally. 4. Some MTN-terminated axons could be traced from the peduncle in serial sections.