Atopic dermatitis. How to incorporate advances in management.

Atopic dermatitis is a common, genetically determined skin disease that most likely results from an immunologic imbalance. In the majority of patients, conservative topical steroid treatment along with patient education suffices.

In vivo resistance to a human immunodeficiency virus type 1 proteinase inhibitor: mutations, kinetics, and frequencies.

Resistance to saquinavir (Ro 31-8959), an inhibitor of human immunodeficiency virus type I proteinase, was studied in peripheral blood mononuclear cell-derived proviral DNA from patients undergoing prolonged treatment. A Leu90-->Met exchange was the predominant resistance mutation in vivo; Gly48-->Val or doubly mutant virus was rarely observed. After 8-12 months of treatment with saquinavir alone (600 mg, 3 times/day) or in combination with zidovudine (200 mg, 3 times/day), approximately 45% of all patients carried provirus with mutant proteinase; the incidence was lower (22%) in patients treated with a combination of saquinavir, zidovudine, and dideoxycytidine. There was a good relationship between genotypic analysis of saquinavir resistance and data from virus assays, confirming that Leu90-->Met and Gly48-->Val are the essential exchanges in the proteinase that determine loss of sensitivity to this inhibitor. Absence of genotypic resistance correlated with a sustained decrease in plasma viral RNA. There was a positive correlation between a Met90 mutation and some residues at natural polymorphic sites (positions 10, 36, 63, and 71).

Isolated in-vitro perfusion of pig hearts obtained from the abattoir: an alternative to animal experiments?

Isolated pig hearts (German farm pigs) were characterized after global in-vivo ischaemia as a potential alternative to in-vivo animal studies. Hearts were harvested from adult farm swine at the abattoir 10.3 +/- 2.1 min after incision of the carotid artery. They were immediately perfused and thereafter stored in ice-cold cardioplegic (St Thomas's) solution. After 38 +/- 3 min, retrograde perfusion was started with oxygenated pig blood (37 degrees C; 5000 U Heparin.l-1; pH 7.38 +/- 0.1; 11 mmol glucose.l-1) at a flow rate of 85 ml.min-1 100 g-1 wet weight (gww-1) for 30 min (n = 10). Additionally, shortly after obtaining the hearts, ATP and CP content were measured by enzymatic tests in 10 pigs at the beginning and after 15 and 30 min of reperfusion. Heart rate was 90 +/- 14 min-1 with little variation during 30 min. Perfusion pressure increased from 89 +/- 17 mmHg to 100 +/- 17 mmHg (NS). Wet weight rose from 488 +/- 33 to 548 +/- 45 g (P < 0.002). CK increased from 2180 +/- 558 to 5900 +/- 1018 U.l-1 (P < 0.001). Calcium in the perfusate decreased from 2.45 +/- 0.15 to 2.2 +/- 0.25 mmol.l-1 and magnesium increased from 0.85 +/- 0.2 to 1.79 +/- 0.35 mmol.l-1 (both P < 0.001). The transmural ATP and CP content was 2.8 +/- 0.48 and 5.08 +/- 0.88 mumol.gww-1.ATP fell moderately during reperfusion to 2.6 +/- 0.35 mumol (NS) and CP rose to 6.0 +/- 1.2 mumol (P < 0.04).(ABSTRACT TRUNCATED AT 250 WORDS)

Conserved TAAAT motif in vaccinia virus late promoters: overlapping TATA box and site of transcription initiation.

We have characterized the vaccinia virus 11-kd late promoter through 5' and 3' deletions and site-directed mutagenesis. The promoter function appears to be contained within an approximately 30-bp fragment, which after translocation is able to direct RNA synthesis late in infection at a reduced level. We demonstrate that a TAAAT sequence in the proximal part of the promoter is essential for its function. This cis-acting element is highly conserved within vaccinia virus late promoters and overlaps the site of transcription initiation. Deletions or mutations within this conserved element completely inactivate the promoter. The evidence indicates that the TAAAT motif functions as a TATA box. The region immediately upstream of the TAAAT motif determines the promoter strength.

Mapping of a gene coding for a major late structural polypeptide on the vaccinia virus genome.

Cell-free translation of total RNA isolated from vaccinia virus-infected cells late in infection results in a complex mixture of polypeptides. A monospecific antibody directed against one of the major structural proteins of the virus particle immunoprecipitated a single polypeptide with a molecular weight of 11,000 (11K) from this mixture. Immunoprecipitation was therefore used to identify the structural polypeptide among the in vitro translation products of RNA purified by hybridization selection to restriction fragments of the vaccinia virus genome. This allowed us to map the mRNA coding for the 11K polypeptide to the extreme left-hand end of the HindIII E fragment. Detailed transcriptional mapping of this region of the genome by nuclease S1 analysis revealed the presence of a late RNA transcribed from the rightward-reading strand. Its 5' end mapped at ca. 130 base pairs to the left of the HindIII site at the junction between the HindIII F and E fragments. The map position of this RNA coincided precisely with the map position of the late message coding for the 11K polypeptide.