The PML/RARalpha fusion protein inhibits tumor necrosis factor-alpha-induced apoptosis in U937 cells and acute promyelocytic leukemia blasts

We investigated the effect of the acute promyelocytic leukemia (APL) specific PML/RARalpha fusion protein on the sensitivity to TNF-alpha-mediated apoptosis. The U937 leukemia cell line was transduced with PML/RARalpha cDNA. PML/RARalpha expression caused a markedly reduced sensitivity to TNF-alpha, even if apoptosis was triggered by agonistic antibodies to TNF-alpha receptors I and II (TNF-alphaRI, II). PML/RARalpha induced a 10-20-fold decrease of the TNF-alpha-binding capacity via downmodulation of both TNF-alphaRI and TNF-alphaRII: this may mediate at least in part the reduced sensitivity to TNF-alpha. Furthermore, the fusion protein did not modify Fas expression (CD95) or sensitivity to Fas-mediated apoptosis. The pathophysiological significance of these findings is supported by two series of observations. (a) Fresh APL blasts exhibit no TNF-alpha binding and are resistant to TNF-alpha-mediated apoptosis. Conversely, normal myeloblasts-promyelocytes show marked TNF-alphaR expression and are moderately sensitive to TNF-alpha-mediated cytotoxicity. Similarly, blasts from other types of acute myeloid leukemia (AML M1, M2, and M4 FAB types) show an elevated TNF-alpha binding. (b) The NB4 APL cell line, which is PML/RARalpha+, shows low TNF-alphaR expression capacity and is resistant to TNF-alpha-triggered apoptosis; conversely a PML/RARalpha- NB4 subclone (NB4.306) exhibits detectable TNF-alpha-binding capacity and is sensitive to TNF-alpha-mediated cytotoxicity. These studies indicate that the PML/RARalpha fusion protein protects against TNF-alpha-induced apoptosis, at least in part via downmodulation of TNF-alphaRI/II: this phenomenon may play a significant role in APL, which is characterized by prolonged survival of leukemic blasts.     

Detection of p53 point mutations by double-gradient, denaturing gradient gel electrophoresis

Genetic instability is a typical feature of tumor cells. This evidence has stimulated the development of rapid methods for detection of gene mutations. A new, improved protocol for denaturing gradient gel electrophoresis (DGGE), to screen for point mutations in genomic DNA, is reported: double gradient (DG) DGGE. In this technique, to the primary, denaturing gradient (typically 30-80% or 40-80% urea/formamide) a secondary gradient, colinear with the first, is superimposed: a porosity gradient (typically 6.5-12% polyacrylamide). The secondary gradient acts by recompacting smeared and diffuse bands of heteroduplexes, which are often indistinguishable from background fluorescence, and by augmenting the resolution between closely spaced homoduplex zones. This allows proper densitometric quantitation of the ratio of the two homoduplex bands. The reliability of this technique has been documented by detection of a number of mutations in exons 6 and 8 of the p53 gene which had escaped revelation by single-strand conformational polymorphism (SSCP) analysis. Additionally, the precise assessment of ratio of the doublet of homoduplex bands has allowed quantitation of the extent of p53 mutation in a mixed cell population extracted from a tumor specimen.     

Vero cytotoxin-producing Escherichia coli O157 outbreaks in England and Wales, 1995: phenotypic methods and genotypic subtyping

Vero cytotoxin-producing Escherichia coli O157 belonging to four phage types (PTs) caused 11 outbreaks of infection in England and Wales in 1995. Outbreak strains of different PTs were distinguishable by DNA-based methods. Pulsed-field gel electrophoresis best discriminated among strains belonging to the same PT, distinguishing six of the seven PT2 outbreak strains and both PT49 outbreak strains.     

Heparin coating of extracorporeal circuits inhibits contact activation during cardiac operations

OBJECTIVE: The choice of a valve substitute remains a challenge in young patients, with numerous reports of early degeneration and calcification of biological valves in this age group. Therefore an assessment of the long-term results after mechanical aortic valve replacement in children was initiated. METHODS: A retrospective study was conducted in 54 consecutive patients aged 1.1 to 17 years (mean 12.8 +/- 4 years) operated on between 1975 and 1993. Aetiology was congenital in 34 patients, rheumatic in 13, infectious in 5, and dystrophic in 2. Concomitant surgery included mitral valve replacement (10), aortic annulus enlargement (9), correction of truncus arteriosus (7), Bentall operation (2), coarctation repair (2), tricuspid valvuloplasty (2), correction of double outlet right ventricle (1), and replacement of a right ventricle to pulmonary artery conduit (1). A Bjork-Shiley valve was implanted in 14 patients, and a St Jude Medical valve in 40. All patients were given Warfarin with a monthly INR control. Follow-up was completed through questionnaires mailed to referring physicians and direct clinical examination. RESULTS: Overall early mortality was 13% (7 cases), and 6% (2 cases) in the 32 patients operated on after 1984. Follow-up was complete in 45 survivors (2 lost to follow-up), with a total follow-up of 261 patient-years. There were 6 late deaths, 4 being cardiac and due to persistent LV dysfunction, and 2 valve-related, due respectively to major gastro-intestinal bleeding and massive thromboembolism. Linearized rates of valve thrombosis and anticoagulant-related hemorrhage were both 0.3% per patient-year. Actuarial survival rate was respectively 84.5% at 5 years and 70.2% at 10 years. Reoperation was necessary in 3 patients for recurrent LV outflow tract obstruction. One patient with severe LV dysfunction is awaiting a heart transplant. CONCLUSION: We conclude that the longterm outcome after mechanical aortic valve replacement in children and adolescents is satisfactory and comparable to currently available reports on biological substitutes. The mandatory anticoagulant therapy is well tolerated in this age group.     

Can we rely on self-reported smoking habits?

The reliability of self-reporting on smoking habits has been evaluated by comparing the reported smoking habits with the concentration of serum thiocyanate, which is higher in smokers than in non-smokers and increases with increasing cigarette consumption. When a smoker stops smoking, the level of serum thiocyanate decreases and falls to the level observed in non-smokers after about one month. When the questions asked about smoking were neutral, the reported smoking habits were generally reliable. However, in cases where the questions asked and the interview situation could be regarded as unpleasant, the reported smoking habits were not always correct. Some smokers underreported the number of cigarettes they smoked on a daily basis and some of them denied smoking altogether.     

Pulse oximetry in the diagnosis of non-critical peripheral vascular insufficiency

Pulse oximetry was used to detect return of pulsatile flow in 27 subjects during reactive hyperaemia following 3 min of total limb ischaemia induced by above knee tourniquet occlusion. Fourteen patients with exercise induced leg pain had 18 symptomatic limbs tested. Thirteen controls had 25 limbs tested. Return of pulsatile flow during reactive hyperaemia occurred within 20 s of tourniquet release in the 25 control limbs which was then regarded as normal. The mean time for return of pulsatile flow in 18 symptomatic limbs was 53 +/- 37 s (P < 0.05 versus controls). Three limbs had a normal value, two of which did not have peripheral vascular disease. Pulse oximetry correctly identified all 25 asymptomatic limbs and 15 of 16 patients with claudication secondary to peripheral vascular disease (PVD). This modification of the reactive hyperaemia test using the pulse oximeter is simple and quick to perform. It has potential as a non-invasive screening test for PVD, suitable for outpatient assessment.     

Adult age differences in the inhibition of return of visual attention.

Responses to targets are slower when they appear at a location to which attention has previously been directed than when they appear at other locations. This inhibition of return (IOR) effect is subserved by posterior brain attentional systems. In 4 experiments the IOR effect in elderly adults was found to be at least as large as in young adults for both discrimination tasks and for detection tasks. The time course and the spread of inhibition within the visual field were also equivalent in the 2 age groups. Additive factors logic was then used to test the hypothesis that the Stroop and IOR effects are due to a common mechanism, a failure to suppress attention. This hypothesis was not confirmed. The results of the 6 experiments are consistent with the hypothesis that there are changes in posterior brain systems responsible for selective attention to a location, contrary to prior claims. They cannot be explained by a general slowing of processing in old age. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Bovine internal mammary artery as a conduit for coronary revascularization: long-term results

Preterm infants may become apneic during immediate postoperative period. We studied prospectively postoperative apneic attack in 167 preterm infants after inguinal herniorrhaphy with nitrous oxide-halothane anesthesia. Their mean gestational age was 30 +/- 3 weeks. The mean postnatal age at operation was 14 weeks. The post-conceptual age varied between 36 and 56 weeks. The mean birth weight was 1351 +/- 395 kg. Although many of them had a risk factor of postoperative apneic attack, i.e.a young post-conceptual age (less than 41 weeks), a light weight at operation (below 3000g), only one infant who had received emergency operation had an episode of apneic attack up to 2 postoperative days. For preventing postoperative apneic attack in preterm infants after inguinal hernia, we recommend the use of halothane anesthesia and the attention until a complete awakening.     

Inhibition of urokinase synthesis and cell surface binding alters the motile behavior of embryonic endocardial-derived mesenchymal cells in vitro

The expression of the serine protease urokinase is elevated during the epithelial-mesenchymal transformation of the endocardium in the developing avian heart. Elevated urokinase expression is associated with the migrating mesenchymal cells of the atrioventricular canal and bulbotruncus and not the myocardium. Treatment of isolated endocardial-derived mesenchymal cells with phosphatidyinositol-specific phospholipase C released urokinase and its receptor from the cell surface and caused significant alterations in cell morphology and motility. Likewise inhibition of urokinase synthesis by treatment of cells with antisense oligonucleotides also inhibited the migration and motility of the endocardial-derived cells. These results suggest an important role for this enzyme in cell-matrix interactions and cell migration during development.     

Projective reconstruction and invariants from multiple images

This correspondence investigates projective reconstruction of geometric configurations seen in two or more perspective views, and the computation of projective invariants of these configurations from their images. A basic tool in this investigation is the fundamental matrix that describes the epipolar correspondence between image pairs. It is proven that once the epipolar geometry is known, the configurations of many geometric structures (for instance sets of points or lines) are determined up to a collineation of projective 3-space 𝒫³ by their projection in two independent images. This theorem is the key to a method for the computation of invariants of the geometry. Invariants of six points in 𝒫³ and of four lines in 𝒫³ are defined and discussed. An example with real images shows that they are effective in distinguishing different geometrical configurations. Since the fundamental matrix is a basic tool in the computation of these invariants, new methods of computing the fundamental matrix from seven-point correspondences in two images or six-point correspondences in three images are given