Biomedical subjects

L T Do

Publications and source records attributed to L T Do.

4 recordsLinked to original sources

PubMed1998

In vivo tear-film thickness determination and implications for tear-film stability.

PURPOSE: Previous measurements of tear-film thickness in vivo are limited and cannot be easily applied in a clinical setting. A novel technique to measure tear-film thickness indirectly is introduced here, requiring only a slit lamp, video camera, and computer. A recent fluid mechanical theory relates tear-film thickness h, to the tear meniscus radius R, tear surface tension or, tear viscosity mu, and upper lid velocity U. This theory yields the result that h/R = 2.12 (microU/sigma)2/3. All parameters except h/R are taken as known physical constants, and R was measured for each subject, allowing the above equation to establish h. Tear-film breakup was also evaluated and correlated with tear-film thickness. METHODS: A clinical study was performed in which aqueous tear-film thickness was determined for 45 subjects, including 24 non-lens subjects, 15 hydrogel contact lens wearers, and 6 RGP lens wearers. R was measured by instilling fluorescein dye in the form of an eyedrop and videotaping the tear meniscus in profile. Tear-film breakup was videotaped through the ocular port of the slit lamp and evaluated based on a severity scale. RESULTS: Aqueous tear-film measurements are in the same range as literature values, with most measured values falling between 6 and 12 microm. Average tear-film thicknesses for non-lens, hydrogel, and RGP subjects are 10.4, 6.5, and 5.8 microm, respectively. Tear-film breakup is most severe in subjects with thin tear films, especially in contact-lens wearers. CONCLUSIONS: Tear-film thickness is an important parameter that varies among individuals. These variations correlate with differences in tear-film stability.

Adult↗

PubMed1998

A novel sensitive and selective bioassay for human type I interferons.

We describe a novel MxA gene-induction assay for type I interferons (IFN-alpha and IFN-beta) based on the specific induction of the MxA gene in cultured human cells. Accumulated intracellular MxA protein is determined by immunologic measurement by a rapid method using commercially available materials. IFN activity can be measured accurately over a concentration range of 0.1-30 IU/ml. In contrast, type II IFN and other cytokines are not significantly detected. The MxA-induction assay has advantages in terms of specificity, reliability, and sensitivity over other methods for assaying type I IFN. It has also been adapted and validated for measuring the titers of anti-IFN-beta neutralizing antibodies in human sera.

Antiviral Agents↗

PubMed1998

A novel bioassay for the determination of neutralizing antibodies to IFN-beta1b.

We have adapted the new MxA gene-induction bioassay to measure neutralizing antibodies to interferon-beta1b (IFN-beta1b, the active ingredient in Betaseron) in sera from patients treated with Betaseron. This antibody assay has been validated to quantify neutralizing titers of 1:20 and above, with a precision of +/- 0.20 in log10. We have used this MxA gene-induction antibody assay to reinvestigate serum samples from multiple sclerosis (MS) patients treated with Betaseron. The titers measured were closely comparable to those obtained in antiviral assays. Data obtained by both methods show that neutralizing antibodies may appear and subsequently disappear over time in the sera of some patients treated with Betaseron. Sera from some patients contain binding antibodies to IFN-beta1b. It was shown that binding antibody titers do not correlate quantitatively or qualitatively with neutralizing antibody titers, and indeed, a number of patients develop high levels of binding antibodies but never form measurable levels of neutralizing antibodies.

Antigen-Antibody Reactions↗

PubMed1996

Site-directed mutagenesis of the subunit PsaC establishes a surface-exposed domain interacting with the photosystem I core binding site.

We have postulated that the orientation of PsaC on the photosystem I core involves electrostatic interactions between charged residues on the core binding site and the subunit [Rodday, S. M., Jun, S.-S., & Biggins, J. (1993) Photosynth. Res. 36, 1-9]. We, therefore, changed eight acidic residues on PsaC to arginine and examined the efficiency of the mutant subunits in the reconstitution of P700-Fx cores in vitro. Reconstitution of the cores by the mutant subunits was determined by analysis of the kinetics of recombination reactions between P700+ and reduced acceptors as measured optically. Restoration of complete forward electron transfer, indicative of efficient subunit binding, was estimated from the ca. 30 ms decay component in the flash transients. Slightly reduced levels of reconstitution were observed for the mutants D24R, E46R/D47R. D61R, and E72R. In contrast, mutants D9R, E27R, and D32R showed significantly lower efficiencies. The presence of the iron-sulfur centers, FA and FB, in these three mutant subunits was confirmed by low-temperature EPR spectroscopy indicating that the polypeptides had folded correctly. We conclude that the introduction of positively charged side chains at positions 9, 27, and 32 seriously disrupts PsaC binding. However, when the wild-type acidic residues in these positions were changed to alanine, only mutant D9A showed a reduced level of reconstitution, suggesting that this aspartate is the most important residue participating in the electrostatic interaction with the core. The results are discussed in relation to the photosystem I crystal structure and support an orientation of PsaC on the core such that center FB is proximal to Fx.

Alanine↗