Tissue distribution and timing of appearance of polytropic envelope recombinants during infection with SL3-3 murine leukemia virus or its weakly pathogenic SL3DeltaMyb5 mutant.

A time course analysis was performed to identify the sites of formation and timing of appearance of polytropic recombinant viruses following infection of NIH/Swiss mice with the murine retrovirus SL3-3 murine leukemia virus (SL3) or with a weakly pathogenic mutant termed SL3DeltaMyb5. The results indicated that (i) polytropic recombinant viruses occur initially in the thymus of SL3-infected animals, (ii) the timing of appearance of polytropic recombinants in bone marrow is not consistent with their participation in the previously reported formation of transplantable tumor-forming cells at 3 to 4 week postinoculation, and (iii) the efficient generation of recombinant virus is correlated with efficient tumor induction.

R region sequences in the long terminal repeat of a murine retrovirus specifically increase expression of unspliced RNAs.

A stem-loop structure at the 5' end of the R region of the long terminal repeat (LTR) of the murine leukemia virus SL3 and other type C mammalian retroviruses is important for maximum levels of expression of a reporter gene under the control of the viral LTR. This element, termed the R region stem-loop (RSL), has a small effect on transcriptional initiation and no effect on RNA polymerase processivity. Its major effect is on posttranscriptional processing of LTR-driven transcripts. Here we tested whether the RSL affected the production of RNAs from a full-length SL3 genome. Mutation of the RSL in the 5' LTR of SL3 reduced the cytoplasmic levels of full-length viral transcripts but not those of spliced, env mRNA transcripts. Thus, the RSL specifically affected the cytoplasmic levels of the unspliced viral RNA. To test further whether the effect was specific for unspliced transcripts, a system was devised in which the expression of a reporter gene under the control of the viral LTR was tested in the presence or absence of an intron. Mutation of the RSL resulted in only about a twofold decline in the level of reporter gene expression when the transcripts contained an intron. However, when the intron was removed, mutation of the RSL reduced expression of the reporter gene about 10- to 60-fold in various cell lines. The secondary structure of the RSL was essential for its activity on the intronless transcript. Thus, the RSL appears to be important for the cytoplasmic accumulation of unspliced viral RNA and unspliced RNA from chimeric transcription units under the control of the viral LTR.

The secondary structure of the R region of a murine leukemia virus is important for stimulation of long terminal repeat-driven gene expression.

In addition to their role in reverse transcription, the R-region sequences of some retroviruses affect viral transcription. The first 28 nucleotides of the R region within the long terminal repeat (LTR) of the murine type C retrovirus SL3 were predicted to form a stem-loop structure. We tested whether this structure affected the transcriptional activity of the viral LTR. Mutations that altered either side of the stem and thus disrupted base pairing were generated. These decreased the level of expression of a reporter gene under the control of viral LTR sequences about 5-fold in transient expression assays and 10-fold in cells stably transformed with the LTR-reporter plasmids. We also generated a compensatory mutant in which both the ascending and descending sides of the stem were mutated such that the nucleotide sequence was different but the predicted secondary structure was maintained. Most of the activity of the wild-type SL3 element was restored in this mutant. Thus, the stem-loop structure was important for the maximum activity of the SL3 LTR. Primer extension analysis indicated that the stem-loop structure affected the levels of cytoplasmic RNA. Nuclear run-on assays indicated that deletion of the R region had a small effect on transcriptional initiation and no effect on RNA polymerase processivity. Thus, the main effect of the R-region element was on one or more steps that occurred after the template was transcribed by RNA polymerase. This finding implied that the main function of the R-region element involved RNA processing. R-region sequences of human immunodeficiency virus type 1 or mouse mammary tumor virus could not replace the SL3 element. R-region sequences from an avian reticuloendotheliosis virus partially substituted for the SL3 sequences. R-region sequences from Moloney murine leukemia virus or feline leukemia virus did function in place of the SL3 element. Thus, the R region element appears to be a general feature of the mammalian type C genus of retroviruses.

Comparison of different lasers in terms of thrombogenicity of the laser-treated vascular wall.

The thrombogenic properties of the laser-treated vascular wall are reported as quantitatively assessed on the basis of 33 experiments with peripheral canine vessel segments. Three types of surfaces, namely, 1) intact, 2) mechanically de-endothelized, and 3) postlaser, have been simultaneously exposed to platelet-enriched plasma in a sequentially organized artificial circulation system. Then the adherent platelets have been counted on the treated surfaces, and the relative thrombogenicity index has been calculated according to the equation T = (A1 - Aint)/(Ad - Aint), where Aint, Ad, A1 = the adherent platelet counts on intact, mechanically de-endothelized, and laser-treated surfaces, respectively. The following lasers have been evaluated: 1) Nd-YAG, 1,060 nm, continuous wave, 4W; 2) argon-ion, 480 and 514 nm and argon-ion, 350 nm, continuous-wave, 1 W and 400 mW [corrected], respectively; 3) excimer XeCl, 308 nm, pulsed, 30 mJ per pulse, repetition rate 10 Hz [corrected]; and also 4) the laser-heated metal probe (2 mm diameter, Trimedyne, Nd-YAG) 1,060 nm, 8 W. The thrombogenicity index values obtained were 83 +/- 7, 72 +/- 8, 57 +/- 9, 63 +/- 7, and 82 +/- 9%, respectively. The differences between these values were statistically insignificant. The data are suggestive of the essential requirement of, at least, anticoagulant therapy after laser angioplasty irrespective of the laser type.

Certain characteristics of myocardial contractility of isovolumic dog heart at randomly variable heart rhythm.

The relationship "heart rate - left ventricular pressure" was investigated in the isolated canine heart perfused with constant pressure at different preloads. Rhythmical stimulation was performed with constant stimulus interval duration and with stimulus intervals randomly changed near the average value (150-200 stimuli in series). Correlation and dispersion function analysis show that rhythm dispersion had a negative inotropic effect which was independent of the preload of the ventricle in the range of 120-180 beat/min, but this dependence occurred with low rats of stimulation. This method is proposed for the assessment of contractility under conditions of heart rate variations (physiological and pathological arrhythmias).