Recurrent erosion after photorefractive keratectomy

Recurrent erosion of the corneal epithelium has been treated successfully by phototherapeutic keratectomy when convention therapy has failed. We present a case of recurrent corneal erosion occurring 1 year after photorefractive keratectomy in a 55-year-old myopic woman. No corneal abnormalities were previously identified in either the affected or the fellow eye. This case demonstrates the potential for ineffective healing of the corneal epithelial basement membrane complex after excimer laser photorefractive keratectomy.     

Frequent clones of p53-mutated keratinocytes in normal human skin

The multiple genetic hit model of cancer predicts that normal individuals should have stable populations of cancer-prone, but noncancerous, mutant cells awaiting further genetic hits. We report that whole-mount preparations of human skin contain clonal patches of p53-mutated keratinocytes, arising from the dermal-epidermal junction and from hair follicles. These clones, 60-3000 cells in size, are present at frequencies exceeding 40 cells per cm2 and together involve as much as 4% of the epidermis. In sun-exposed skin, clones are both more frequent and larger than in sun-shielded skin. We conclude that, in addition to being a tumorigenic mutagen, sunlight acts as a tumor promoter by favoring the clonal expansion of p53-mutated cells. These combined actions of sunlight result in normal individuals carrying a substantial burden of keratinocytes predisposed to cancer.     

Apoptosis as a cause of death in measles virus-infected cells

To determine the mechanism of measles virus-induced cell death, we studied the infection of Vero cells and monocytic cell lines with wild-type (Chicago-1) and vaccine (Edmonston) strains of measles virus. DNA fragmentation indicative of apoptosis was apparent by flow cytometry, agarose gel electrophoresis, and electron microscopy. Within syncytia, DNA strand breaks were demonstrated by end labeling with terminal transferase and then by visualization.     

Angiotensin II regulates the expression of plasminogen activator inhibitor-1 in cultured endothelial cells. A potential link between the renin-angiotensin system and thrombosis

Plasminogen activator-inhibitor C-1 (PAI-1) plays a critical role in the regulation of fibrinolysis, serving as the primary inhibitor of tissue-type plasminogen activator. Elevated levels of PAI-1 are a risk factor for recurrent myocardial infarction, and locally increased PAI-1 expression has been described in atherosclerotic human arteries. Recent studies have shown that the administration of angiotensin converting enzyme inhibitors reduces the risk of recurrent myocardial infarction in selected patients. Since angiotensin II (Ang II) has been reported to induce PAI-1 production in cultured astrocytes, we have hypothesized that one mechanism that may contribute to the beneficial effect of angiotensin converting enzyme inhibitors is an effect on fibrinolytic balance. In the present study, we examined the interaction of Ang II with cultured bovine aortic endothelial cells (BAECs) and the effects of this peptide on the production of PAI-1. 125I-Ang II was found to bind to BAECs in a saturable and specific manner, with an apparent Kd of 1.4 nM and Bmax of 74 fmol per mg of protein. Exposure of BAECs to Ang II induced dose-dependent increases in PAI-1 antigen in the media and in PAI-1 mRNA levels. Induction of PAI-1 mRNA expression by Ang II was not inhibited by pretreating BAECs with either Dup 753 or [Sar1, Ile8]-Ang II, agents that are known to compete effectively for binding to the two major angiotensin receptor subtypes. These data indicate that Ang II regulates the expression of PAI-1 in cultured endothelial cells and that this response is mediated via a pharmacologically distinct form of the angiotensin receptor.     

Assessment of bog-body tissue preservation by pyrolysis-gas chromatography/mass spectrometry

Flash pyrolysis-gas chromatography/mass spectrometry (py-GC/MS) was used to assess the quality and mechanism of protein preservation in the tissue of Iron Age bog bodies from Lindow, UK, and south-eastern Drenthe, The Netherlands. Abundant pyrolysis products of the fresh skin tissue, including 2,5-diketopiperazines of Pro-Gly, Pro-Ala, Pro-Val, Pro-Pro and Hyp, were readily assigned to specific amino acid or dipeptide moieties. Comparison of the pyrolysates of the bog-body tissues with that of modern samples revealed qualitative similarities suggesting good preservation of the collagen and non-collagenous proteins in the ancient tissues. Examination of the pyrolysates of samples of fresh calf skin, which had been treated with various vegetable tanning agents, clearly revealed markers of non-hydrolysable tannins including 1,2-benzenediol, 1,3-benzenediol and 1,2,3-benzenetriol, although chromatographic quality inevitably diminished with increasing functionalization of the compounds. Such markers were not detected in the pyrolysates of the bog-body tissues. Instead 4-isopropenylphenol, a characteristic pyrolysis product of Sphagnum moss, was detected in both solvent-extracted and base-treated samples of tissue. The presence of 4-isopropenylphenol in the pyrolysates of the bog-body tissues provides evidence that their preservation involves reactions of amino acids with sphagnum acid, and possibly other agents derived from the peat. The study constitutes the first chemical characterization of the pyrolysis products of modern and ancient collagen.     

Sensory Evaluation and Acceptability of Cookies Enriched with Sweet Lupine Flour (Lupinus albus cv Multolupa)

Cookies enriched with 0, 5, 10, 15, 20, and 25% full-fat sweet lupine flour (FFSL) were evaluated by a sensory panel using the rank of preference and paired comparison tests. Cookies with 0, 5, and 10% FFSL were preferred while those containing 20 and 25% FFSL were rejected (p≤0.01). Studies conducted with school children showed similar acceptability for 0 and 10% FFSL-containing cookies which was different (p = 0.05) from those containing 20% FFSL. Fortification of the basic formula with 10% FFSL was recommended on the basis of acceptability.     

Thrombolytic and endovascular treatment of peripartum iliac vein thrombosis: a case report

This case report details the multidisciplinary treatment of peripartum left iliac vein thrombosis using percutaneous catheter-directed urokinase thrombolysis and balloon thromboplasty. Enhanced chances for long-term patency and the normalization of venous function make these minimally invasive procedures accepted options for the treatment of iliofemoral deep venous thrombosis in selected peripartum patients.     

Preparation of compound-specific and group-specific antibodies to 7-methylguanine and related 7-alkylguanines and their use in immunoaffinity purification

The preparation and characteristics of compound-specific and group-specific antibodies against 7-alkylguanines (7-alkGua) are described. A compound-specific antibody against 7-methylguanine was prepared using a hapten bound to carrier protein through the N2 position. In a competitive enzyme-linked immunosorbent assay (ELISA) 7-methylguanine (7-MeGua) showed 50% inhibition (I50%) at 10 pmol/well at room temperature, but the inhibition was found to be 40 times better at 4 degrees C (I50% at 250 fmol/well). When the antibody was bound to protein A-Sepharose CL4B 7-MeGua was retained in immunoaffinity columns. A group-specific antibody to 7-alkGua was prepared using 7-(2-carboxyethyl)guanine (7-CEGua) bound to carrier protein via the carboxyl group. In a competitive ELISA, this antibody cross-reacted well with 7-CEGua, 7-ethylguanine (7-EtGua), 7-(2-hydroxyethyl)guanine (7-HOEtGua) and 7-(2',3'-dihydroxy)-propylguanine (7-DHPGua) and some inhibition was seen with 7-MeGua. Immunoaffinity columns prepared from this antibody retained a number of 7-alkGua of diverse structure. 7-EtGua in calf thymus DNA treated with diethylsulphate and ethylnitrosourea was isolated by immunoaffinity purification and quantified by HPLC-fluorescence. These results illustrate the potential of immunoaffinity purification for both individual DNA adducts and groups of adducts.