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Biomedical subjects

N H Tran

Publications and source records attributed to N H Tran.

7 recordsLinked to original sources

Structural-chemical evolution within exfoliated clays.

The exfoliated (delaminated) structures of lamellar clays offer potential as precursors for the formation of various nanostructured materials. In this article, Lucentite and Laponite phyllosilicate clays, which both have empirical formulas of Na(0.33)[Mg(2.67)Li(0.33)Si4O10(OH)2] but differ in nanodimensions, have been exfoliated. Experiments were carried out for mixtures containing approximately 1 wt % phyllosilicate in a 5% aqueous solution of poly(acrylic acid) at different temperatures. X-ray diffraction and photoemission spectroscopy measurements for the solid products recovered after stirring the mixtures at 20 degrees C showed that the fully extended chains of poly(acrylic acid) were intercalated within the interlayer spaces between the silicate plates of the clays. At 85 degrees C, however, the clays were exfoliated and/or partially exfoliated. Photoemission spectroscopy also indicated that the exfoliated structures primarily consisted of silica nanoplates. 29Si nuclear magnetic resonance and oxygen K-edge near-edge X-ray absorption fine structure indicated that the surfaces of the plates were terminated by high concentrations of the silanol (-SiOH) groups, which created structural branches during intercalation. A model was developed in which intercalation and the removal of ions from the clays after the poly(acrylic acid) interactions reduced the electrostatic van der Waals forces between the plates. It was also shown that the formation of branches created a steric effect that inhibited the stacking of the plates. Together these resulted in exfoliation.

Journal Article↗

Dispersion of organically modified clays within n-alcohols.

A method for formation of polymer-clay nanocomposites involves dispersion of the nanometer silicate layers of clays into a solvent, followed by dispersion into polymers. The dispersion of layered silicates within solvents affects the structure and properties of the nanocomposites. We report the dispersion of organically modified clays, used for formation of nanocomposites with organic polymers, within a range of alcohol solvents. Experiments involved stirring a mixture containing approximately 1 wt% of alkylammonium-modified clays in n-alcohols with general molecular structure RnOH, where n represents the number of carbons of alkyl chains, varying from 2 to 8. The clays precipitated from the dispersion when RnOH solvents with n<5 were used, however, they formed gels for solvents with n5. The increased dispersion was related to the decrease of polarity and hydrogen bonding force within solvents. X-ray diffraction for the dispersed clays indicated that the interlayer spaces (1.8 nm), formed by regular stacking of the silicate layers, expanded to a maximum of 3.0 nm after treatment with RnOH with n5. The interlayer expansion was due to the intercalation of n-alcohol molecules within the interlayer spaces. It is suggested that the alkyl chains of n-alcohols remain parallel to the silicate surface in the intercalate. Preliminary experiments on the influence of these alcohol solvents on the intercalation of polyol (polyether) are also reported.

Journal Article↗

Thermal effects on reverse transcription: improvement of accuracy and processivity in cDNA synthesis.

Reverse transcription, coupled with DNA amplification, has been widely used for molecular analysis of RNAs. Reverse transcriptases are retroviral DNA polymerases that can synthesize DNA from both RNA and DNA. In general, because of the lack of 3'-->5' exonuclease activity in retroviral reverse transcriptases, the reverse transcription step is error prone. Mutations created during the reverse transcription step of cDNA synthesis or RT-PCR are delivered to the final products to be analyzed, interfering with accurate analysis of the RNAs. In addition, because reverse transcription uses RNA as a template, processive DNA synthesis by reverse transcriptase is frequently interrupted by secondary structures of the RNA templates, causing difficulties in full-length cDNA synthesis. Here, we report that an increase in reaction temperature greatly enhances both the accuracy and the processivity of reverse transcription catalyzed by murine leukemia virus (MuLV) and human immunodeficiency virus type 1 (HIV-1) reverse transcriptases.

DNA, Complementary↗

Molecular architecture of the mutagenic active site of human immunodeficiency virus type 1 reverse transcriptase: roles of the beta 8-alpha E loop in fidelity, processivity, and substrate interactions.

Human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is a putative source of the genomic hypermutation that promotes rapid evolution of HIV-1. To understand the molecular strategies that create a highly mutagenic DNA polymerase active site in HIV-1 RT, we investigated the roles of four residues in the beta 8-alpha E loop. Gln151, which interacts with the sugar of the incoming dNTP, and Lys154, which interacts with the template, yielded site-directed mutants with increased fidelity, suggesting that these residues are directly involved in the mutagenic architecture of the active site. Mutations at Gln151 and Lys154 also reduced processivity. Q151N RT showed enhanced ability to discriminate between TTP and AZT triphosphate, consistent with the observation that the Q151M mutation confers AZT resistance in vivo. Mutations at Gly152 greatly decreased RT activity; molecular modeling suggests that Gly152 is critical for the proper geometric alignment that permits base-pairing of the incoming dNTP with the template. Mutations at Trp153 reduced the expression level, and presumably the stability, of RT proteins in bacteria. These observations support the conclusion that interactions of active site residues in the beta 8-alpha E loop with incoming dNTPs and the template are determinants of the accuracy, processivity, and substrate selectivity of HIV-1 RT.

Amino Acid Substitution↗

New human immunodeficiency virus, type 1 reverse transcriptase (HIV-1 RT) mutants with increased fidelity of DNA synthesis. Accuracy, template binding, and processivity.

Infidelity of DNA synthesis by human immunodeficiency virus, type 1 reverse transcriptase (HIV-1 RT) is a presumptive determinant of HIV-1 hypervariability and is incompletely understood at the mechanistic and structural levels. Amino acid substitution at only three residues, including Asp-76 (Kim, B., Hathaway, T. R., and Loeb, L. A. (1996) Biochemistry 37, 5831-5839), is known to increase fidelity. We report here that substitution at Arg-78 can also increase accuracy. Mutant R78A RT showed reduced primer extension in misincorporation assays lacking a complementary dNTP and exhibited a 9-fold decrease in mutation frequency in the M13mp2 lacZ forward mutation assay. Previous structural studies indicate that Arg-78 and Asp-76 lie in a region that interacts with template nucleotides. Interestingly, R78A RT exhibited 6- to 8-fold decreases in binding affinity (K(d)) for RNA and DNA templates relative to wild type RT. In contrast, D76V RT, which also increases fidelity (Kim et al., 1996), showed a 6- to 7-fold increased affinity. The processivity of R78A RT on both RNA and DNA templates was substantially reduced relative to wild type RT, whereas the processivity of D76V RT was increased. We discuss relationships of fidelity, template binding, and processivity in these and other HIV RT mutants.

Amino Acid Substitution↗

Lung pressures and gas transport during high-frequency airway and chest wall oscillation.

The major goal of this study was to compare gas exchange, tidal volume (VT), and dynamic lung pressures resulting from high-frequency airway oscillation (HFAO) with the corresponding effects in high-frequency chest wall oscillation (HFCWO). Eight anesthetized paralyzed dogs were maintained eucapnic with HFAO and HFCWO at frequencies ranging from 1 to 16 Hz in the former and 0.5 to 8 Hz in the latter. Tracheal (delta Ptr) and esophageal (delta Pes) pressure swings, VT, and arterial blood gases were measured in addition to respiratory impedance and static pressure-volume curves. Mean positive pressure (25-30 cmH2O) in the chest cuff associated with HFCWO generation decreased lung volume by approximately 200 ml and increased pulmonary impedance significantly. Aside from this decrease in functional residual capacity (FRC), no change in lung volume occurred as a result of dynamic factors during the course of HFCWO application. With HFAO, a small degree of hyperinflation occurred only at 16 Hz. Arterial PO2 decreased by 5 Torr on average during HFCWO. VT decreased with increasing frequency in both cases, but VT during HFCWO was smaller over the range of frequencies compared with HFAO. delta Pes and delta Ptr between 1 and 8 Hz were lower than the corresponding pressure swings obtained with conventional mechanical ventilation (CMV) applied at 0.25 Hz. delta Pes was minimized at 1 Hz during HFCWO; however, delta Ptr decreased continuously with decreasing frequency and, below 2 Hz, became progressively smaller than the corresponding values obtained with HFAO and CMV.

Animals↗

Single dose artemisinin-mefloquine versus mefloquine alone for uncomplicated falciparum malaria.

The efficacy of the combination of a single oral dose of 500 mg artemisinin with a single 500 mg oral dose of mefloquine (AM) in the treatment of uncomplicated falciparum malaria was compared to mefloquine therapy alone (M) in a double-'blind' randomized study in an endemic area in the south of Viet Nam where single low dose treatment was employed and where mefloquine had been recently introduced. 231 patients, 117 AM and 114 M, were studied. Failure of therapy occurred in 1 AM patient and in 3 M patients. The radical cure rate was 84% for the AM regimen and 65% for the M regimen (P = 0.002). Recrudescence (including an unknown percentage of reinfections) occurred in 15% of AM patients and in 30% of M patients (P = 0.01). The mean parasite clearance time was 40 h (SD = 16) for AM and 60 h (SD = 27) for the M regimen (P = 0.0001). No effect of artemisinin was noted on gametocytes present on admission, but new gametocytes developed less frequently in the AM group. The addition of a single dose of 500 mg artemisinin to 500 mg mefloquine increased the efficacy and reduced the rate of recrudescence, but this regimen was not adequate and, for short course regimens, more doses of artemisinin as well as higher, doses of mefloquine should be studied.

Adolescent↗