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The effect of nicotinamide on poly(A)-containing messenger RNA from lungs of inbred mice.
A sensitive method for the isolation of poly(A)+ messenger RNA from the lungs of mice was developed. The poly(A)+ messenger RNA from inbred mice in the C3H and DBA strains exhibited identical molecular weights, sedimentation rates, and densities in linear sucrose gradients. The poly(A)+ messenger RNA isolated from the lungs of inbred C3H mice was quantitatively converted into cDNA by the reverse transcriptase. Deletion of the 3′ terminal 84 nucleotides from the poly(A)+ messenger RNA resulted in a loss of almost 90% of the specific activity. nicotinamide (1 mg/100 g body weight) per se had no effect on the specific activity of the messenger RNA. However, in the presence of nicotinamide, a qualitative and quantitative increase of the specific activity was found. When the nicotinamide-treated poly(A)+ messenger RNA from the lungs of C3H mice was injected into C3H mice, a significant increase in the number of the antibody plaque forming cells was found after the third immunization.Polymerase chain reaction to determine HIV type and drug resistance genotype in Thai patients with acquired immune deficiency syndrome.
To develop a simple and sensitive method for the determination of HIV type and drug resistance genotype in Thai patients with AIDS. Twenty-five peripheral blood samples from Thai patients with AIDS were evaluated by PCR for HIV-1 type determination using serotypic probes and for drug resistance genotype using the reverse transcriptase (RT) primers. PCR results were confirmed by PstI restriction analysis. Seventeen (68%) of the patients had viral isolates classified as subtype A or subtype B and three of these 17 patients had multiple genotypes A or B. Ten patients (40%) had unique genotypes identified by the serotypic probes. One patient had genotype A or B and two other patients had only genotype B. All three patients had previously received the reverse transcriptase inhibitors didanosine or stavudine. It was demonstrated that the differences in sensitivity of different probes for genotyping and the detection of minor variants can be minimized by careful experimental design. This method is a useful tool for determining HIV type and resistance genotype in Thai patients with AIDS.Q:
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