The use of objective measures of asthma severity in primary care: a report from ASPN

This study compared the use of 2 1/2-hour multimedia workshop with distribution of an algorithm on the ability of fourth-year medical students to present a stop-smoking plan to a simulated patient. Results showed that students who participated in the workshop performed statistically significantly better on the skill areas of providing information, eliciting and responding to feeling and on content areas of past experience with quitting, resources available for change and negotiating a plan. There were no significant differences in the skill area of eliciting information and the content areas of motivation to stop smoking, factors that inhibit change and problems affecting the plan. Neither of the groups performed very well. The highest number of available points obtained by both groups was in eliciting information (53% in the algorithm group and 64% in the formal training group); however, most of the values were in the range of 10%-25% of possible points. Suggested reasons for the low values may be due to the specific items rated, the teaching methods or the time needed to assimilate new skills.     

Activated platelet aggregates in thrombotic thromboctyopenic purpura: decrease with plasma infusions and normalization in remission

Circulating activated platelet aggregates (aPA) were assayed by flow cytometry employing mAb alpha-CD62p in eight patients with thrombotic thrombocytopenic purpura (TTP). Elevation of aPA was observed in all patients in active stages of TTP; aPA normalized in remission. Plasma infusions with plasmapheresis decreased aPA in responding patients. The rise and fall of aPA preceded relapses and improvements, respectively. These changes were seen prior to the traditional indicators, LDH, haematocrit, and platelet count. Incubation of plasma from TTP patients with normal whole blood induced formation of aPA; this effect was significantly greater than that of plasmas from ITP patient controls (P < 0.01), suggesting the presence of an aPA-promoting factor in TTP plasma. Parallel experiments using a platelet aggregometer failed to detect effect of TTP plasma on normal blood. In summary, aPA appear to be a marker of disease activity, rising with relapse, falling with plasma therapy, and normalizing in remission. The flow cytometric assay of aPA is more sensitive than aggregometry in detecting the putative aPA-promoting factor in TTP.     

Induction of interleukin-1 and glucocorticoid hormones by HIV promotes viral replication and links human chromosome 2 to AIDS pathogenesis: genetic mechanisms and therapeutic implications

Human immunodeficiency virus may regulate its replication by stimulating the synthesis of interleukin-1. Interleukin-1, in turn, has the ability to stimulate the human immunodeficiency virus enhancer region. The human genes responsible for interleukin-1 and interleukin-1 receptor antagonist synthesis are located on the long arm of chromosome 2. Coincidentally, the trans-activation responsive ribonucleic acid element in the R region of the long terminal repeat of human immunodeficiency virus-1 has been found to interact directly with a factor present on the long arm of chromosome 2 to facilitate transactivation by the human immunodeficiency virus Tat protein. The human CD26 gene is also located on the long arm of chromosome 2. CD26 is a lymphocyte cell surface antigen that is stimulated by interleukin-1 and serves with CD4 as a coreceptor that interacts with the V3 loop in gp120 of human immunodeficiency virus. The human immunodeficiency virus-induced interleukin-1 excess, thus, serves human immunodeficiency virus by enhancing replication, and by increasing human immunodeficiency virus infectivity via activation of CD26. IL-1 also adversely affects acquired immune deficiency syndrome-related Kaposi's sarcoma. Several genetic treatments for human immunodeficiency virus infection are proposed.     

Solution structure of the transforming growth factor beta-binding protein-like module, a domain associated with matrix fibrils

Here we describe the high resolution nuclear magnetic resonance (NMR) structure of a transforming growth factor beta (TGF-beta)-binding protein-like (TB) domain, which comes from human fibrillin-1, the protein defective in the Marfan syndrome (MFS). This domain is found in fibrillins and latent TGF-beta-binding proteins (LTBPs) which are localized to fibrillar structures in the extracellular matrix. The TB domain manifests a novel fold which is globular and comprises six antiparallel beta-strands and two alpha-helices. An unusual cysteine triplet conserved in the sequences of TB domains is localized to the hydrophobic core, at the C-terminus of an alpha-helix. The structure is stabilized by four disulfide bonds which pair in a 1-3, 2-6, 4-7, 5-8 pattern, two of which are solvent exposed. Analyses of MFS-causing mutations and the fibrillin-1 cell-binding RGD site provide the first clues to the surface specificity of TB domain interactions. Modelling of a homologous TB domain from LTBP-1 (residues 1018-1080) suggests that hydrophobic contacts may play a role in its interaction with the TGF-beta1 latency-associated peptide.     

Protein-disulfide isomerase-mediated reduction of the A subunit of cholera toxin in a human intestinal cell line

A key step in the action of cholera toxin (CT) is the reduction of its A subunit to the A1 peptide. The latter is an ADP-ribosyltransferase, which activates the alpha-subunit of the stimulatory G protein of adenylyl cyclase. In this study, the enzymatic reduction of membrane-bound CT in CaCo-2 human intestinal epithelial cells was characterized. Whereas diphtheria toxin was found to be reduced by a cell surface population of protein-disulfide isomerase (PDI) and its cytotoxicity was inhibited by p-chloromercuribenzenesulfonic acid, bacitracin, or anti-PDI antibodies, these inhibitors had no effect on CT reduction or activity in intact cells. In contrast, the reduction of CT in vitro by either postnuclear supernatants (PNS) or microsomal membranes in the presence of Triton X-100 was significantly inhibited by p-chloromercuribenzenesulfonic acid and bacitracin. Anti-PDI monoclonal antibodies likewise inhibited the in vitro reduction of CT and also were effective in depleting reductase activity from PNS. Since inhibition and depletion were not observed in the absence of detergent, these results suggested that the reductase activity was a soluble component localized to the lumen of microsomal vesicles and correlated with the presence of protein-disulfide isomerase. This was further confirmed by showing a corresponding depletion of reductase activity and PDI in alkali-treated microsomes. This activity was restored when purified bovine PDI was added back to alkali-treated microsomes in a redox buffer that reflected conditions found in the lumen of the endoplasmic reticulum (ER). When the CT-related reductase activity was assayed in subcellular fractions of PNS-derived membranes isolated on a 9-30% Iodixanol gradient, the activity, as measured by CT-A1 peptide formation localized to those fractions containing PDI. Likewise CT-A1 peptide formed in intact cells co-localized to those membrane fractions containing the majority of cellular PDI. Furthermore, the banding density corresponded to a region of the gradient containing ER-derived membranes. These results indicated that CT was a substrate for PDI-catalyzed reduction in intact cells and supported the hypothesis that CT reduction and activation occurs in the ER.     

Further observations on histological changes at the ureteroileal junction in ileal conduits

Seventy-two ureteroileal anastomoses taken from ileal conduits removed from 62 patients were examined histologically to characterize the range of mucosal and stromal changes at these sites. All 72 demonstrated variable amounts of subepithelial chronic inflammation and fibrosis. Other histological features included: cystic spaces lined by transitional epithelium (N = 29; 40%; average diameter 1.2 mm); cystic spaces lined by mixed intestinal/transitional epithelium (N = 5; 7%; average diameter 0.77 mm); and cystically dilated intestinal glands (N = 21; 29%; average diameter 0.24 mm). The latter were associated with overgrowth by transitional epithelium, which had prevented mucus drainage. Twenty-one (29%) had mucus pools with no epithelial lining (average diameter 1.2 mm), and polypoidal protrusions into the lumen of the anastomosis were found containing mucus pools (N = 4; 6%; average diameter 1.4 mm), transitional-lined cysts (N = 5; 7%; average diameter 2.2 mm), and mixed intestinal/transitional-lined cysts (N = 2; 3%; average diameter 2.5 mm). Focal rupture of dilated intestinal glands with interstitial pooling of mucus was not uncommon, and marked dystrophic calcification was found in 1 case within a large collection of extracellular mucus. This series confirms that inflammation, fibrosis, and glandular overgrowth by transitional epithelium are common occurrences at ureteroileal anastomosis sites. Subsequent gland rupture may result in sizable accumulations of interstitial mucus, and rarely in marked dystrophic calcification.