[Early vs delayed radical cystectomy compared in highgrade superficial bladder tumors].

Objectives. The treatment of aggressive superficial TCC of the bladder remains controversial. In fact, although still classified as 'superficial', it has been shown that the biological characteristics of T1G3 bladder tumors are the same as those of the muscle-invasive group (T2 and above). Even with close monitoring and intensive intravesical therapy, the reported risk of muscle invasion in these patients is 53% and 1/3 die from this disease in the long-term. The aim of this study is to determine whether the timing of radical cystectomy affects the survival of patients with aggressive superficial bladder tumor. Methods. We consider 74 patients who underwent radical cystectomy between November 1994 and October 2006 before a diagnosis of T1G3 bladder tumor. These patients were divided in 2 subgroups: group A (n=27, 25 M and 2 F) who underwent immediate radical cystectomy, and group B (n=47, 40 M and 7 F) who underwent other conservative treatments before radical cystectomy. Results. The two subgroups were similar concerning age (66.29±8.37 yrs vs 66.87±8.6 yrs, respectively, p NS) and the timing of follow-up (respectively 77±45 vs 60±35 mths, p NS). Moreover, the progression-free survival was significantly higher in subgroup A (53.73±48.54 vs 31.94±35.19 mths, log-rank p<0.05) as well as the overall survival (59.73±45.37 vs 36.45±33.96 mths respectively, log-rank p<0.05). Comparing the histological examinations, the two subgroups were significantly different concerning the T stage (superficial tumors 14/27 vs 16/47, respectively, p<0.05; invasive tumors 13/27 vs 31/47, respectively, p<0.00005) and the lymphonodal dissemination (2N+/27 vs 11N+/47, respectively, p<0.0005). . Delaying radical cystectomy for aggressive superficial bladder tumors leads to a worse progression-free survival; the overall survival is likely to be due also to an early lymphonodal dissemination, which occurs extending the timing between diagnosis and radical treatment.

In fibroblasts Vegf-D expression is induced by cell-cell contact mediated by cadherin-11.

Vascular endothelial growth factors (VEGFs) are a highly conserved family of growth factors all angiogenic in vivo with mitogenic and chemotactic activity on endothelial cells. VEGFs are expressed in fibroblasts either in hypoxia or in response to growth factors. Here we report that, differently from the other members of the family, Vegf-D is induced by cell-cell contact. By in situ hybridization we demonstrated that noninteracting fibroblasts express low levels of Vegf-D mRNA, whereas contacting cells express high levels of Vegf-D transcripts. By immunostaining we observed that the surface protein cadherin-11 is localized at the opposite sites of interacting cell surfaces. Ca(2+) deprivation from the culture medium determined the loss of cadherin-11 from the cell surfaces and down-regulation of Vegf-D mRNA. Moreover, a cadherin-11 antisense RNA construct inhibited Vegf-D expression in confluent BALB/c fibroblasts, whereas in NIH 3T3 cells, which express low levels of cadherin-11, Vegf-D induction could be obtained by overexpression of cadherin-11. This suggests that cell interaction mediated by cadherin-11 induces the expression of the angiogenic factor Vegf-D in fibroblasts.

Discovery of a transient absorption edge in the X-ray spectrum of GRB 990705.

We report the discovery of a transient equivalent hydrogen column density with an absorption edge at approximately 3.8 kiloelectron volts in the spectrum of the prompt x-ray emission of gamma-ray burst (GRB) 990705. This feature can be satisfactorily modeled with a photoelectric absorption by a medium located at a redshift of approximately 0.86 and with an iron abundance of approximately 75 times the solar one. The transient behavior is attributed to the strong ionization produced in the circumburst medium by the GRB photons. The high iron abundance points to the existence of a burst environment enriched by a supernova along the line of sight. The supernova explosion is estimated to have occurred about 10 years before the burst. Our results agree with models in which GRBs originate from the collapse of very massive stars and are preceded by a supernova event.

BeppoSAX and Chandra Observations of SAX J0103.2-7209 = 2E 0101.5-7225: A New Persistent 345 Second X-Ray Pulsar in the Small Magellanic Cloud.

We report the results of a 1998 July BeppoSAX observation of a field in the Small Magellanic Cloud which led to the discovery of approximately 345 s pulsations in the X-ray flux of SAX J0103.2-7209. The BeppoSAX X-ray spectrum is well fitted by an absorbed power law with a photon index of approximately 1.0 plus a blackbody component with kT=0.11 keV. The unabsorbed luminosity in the 2-10 keV energy range is approximately 1.2x1036 ergs s-1. In a very recent Chandra observation, the 345 s pulsations are also detected. The available period measurements provide a constant period derivative of -1.7 s yr-1 over the last 3 years, making SAX J0103.2-7209 one of the most rapidly spinning up X-ray pulsars known. The BeppoSAX position (30&arcsec; uncertainty radius) is consistent with that of the Einstein source 2E 0101.5-7225 and the ROSAT source RX J0103.2-7209. This source was detected at a luminosity level of a few times 1035-1036 ergs s-1 in all data sets of past X-ray missions since 1979. The ROSAT HRI and Chandra positions are consistent with that of a mV=14.8 Be spectral-type star already proposed as the likely optical counterpart of 2E 0101.5-7225. We briefly report and discuss photometric and spectroscopic data carried out at the ESO telescopes 2 days before the BeppoSAX observation. We conclude that SAX J0103.2-7209 and 2E 0101.5-7225 are the same source: a relatively young and persistent X-ray pulsar in the SMC.

Protein kinase CK2alpha' is induced by serum as a delayed early gene and cooperates with Ha-ras in fibroblast transformation.

Protein kinase CK2 is an ubiquitous and pleiotropic Ser/Thr protein kinase composed of two catalytic (alpha and/or alpha') and two noncatalytic (beta) subunits forming a heterotetrameric holoenzyme involved in cell growth and differentiation. Here we report the identification, cloning, and oncogenic activity of the murine CK2alpha' subunit. Serum treatment of quiescent mouse fibroblasts induces CK2alpha' mRNA expression, which peaks at 4 h. The kinetics of CK2alpha' expression correlate with increased kinase activity toward a specific CK2 holoenzyme peptide substrate. The ectopic expression of CK2alpha' (or CK2alpha) cooperates with Ha-ras in foci formation of rat primary embryo fibroblasts. Moreover, we observed that BALB/c 3T3 fibroblasts transformed with Ha-ras and CK2alpha' show a faster growth rate than cells transformed with Ha-ras alone. In these cells the higher growth rate correlates with an increase in calmodulin phosphorylation, a protein substrate specifically affected by isolated CK2 catalytic subunits but not by CK2 holoenzyme, suggesting that unbalanced expression of a CK2 catalytic subunit synergizes with Ha-ras in cell transformation.

Embryonic expression pattern of the murine figf gene, a growth factor belonging to platelet-derived growth factor/vascular endothelial growth factor family.

Morphogenesis, growth and differentiation of tissues and organs require cell interactions mediated by signal molecules, their receptors and transcriptional control systems. c-fos-induced growth factor (figf) is a new secreted member of the platelet-derived growth factor/vascular endothelial growth factor (PDGF/VEGF) family with mitogenic activity on fibroblasts. Here we studied figf expression during murine embryonic development. figf expression was detected with a dynamic pattern in several body structures and organs such as limb buds, acoustic ganglion, teeth, heart, anterior pituitary as well as lung and kidney mesenchyme, liver, derma, and periosteum of the vertebral column.

Human FIGF: cloning, gene structure, and mapping to chromosome Xp22.1 between the PIGA and the GRPR genes.

We report the identification, structural characterization, and mapping of the human FIGF gene. FIGF is the human homologue of mouse figf (c-fos-induced growth factor), a new member of the platelet-derived growth factor/vascular endothelial growth factor (PDGF/VEGF) family. It codes for a secreted factor with mitogenic and morphogenic activity on fibroblast cells. The predicted amino acid sequence of FIGF is 84% identical to that of the mouse protein, and it is highly conserved (up to 40%) in the dimerization domain with respect to the VEGF members of the family. The 2.5-kb mRNA of FIGF was detected in adult lung and heart tissues. The gene spans about 50 kb and is organized into seven exons and six introns. The FIGF promoter contains an optimal AP-1-binding site and lacks a canonical TATA box. Fluorescence in situ hybridization mapped FIGF to chromosomal region Xp22.1. The subsequent identification of YAC positive clones from this region allowed us to refine the map and localize FIGF centromeric to the phosphatidylinositol glycan complementation class A (PIGA) gene and telomeric to the gastrin-releasing peptide receptor (GRPR) gene. FIGF and PIGA genes lie next to each other in a head-to-tail orientation, with the FIGF polyadenylation signal about 12 kb from the PIGA transcriptional start site.

Identification of a c-fos-induced gene that is related to the platelet-derived growth factor/vascular endothelial growth factor family.

Using a mRNA differential screening of fibroblasts differing for the expression of c-fos we isolated a c-fos-induced growth factor (FIGF). The deduced protein sequence predicts that the cDNA codes for a new member of the platelet-derived growth factor/vascular endothelial growth factor (PDGF/VEGF) family. Northern blot analysis shows that FIGF expression is strongly reduced in c-fos-deficient cells. Transfection of exogenous c-fos driven by a constitutive promoter restores the FIGF expression in these cells. In contrast, both PDGF and VEGF expression is unaffected by c-fos. FIGF is a secreted dimeric protein able to stimulate mitogenic activity in fibroblasts. FIGF overexpression induces morphological alterations in fibroblasts. The cells acquire a spindle-shaped morphology, become more refractive, disorganized, and detach from the plate. These results imply that FIGF is a downstream growth and morphogenic effector of c-fos. These results also suggest that the expression of FIGF in response to c-fos activation induces specific differentiation patterns and its aberrant activation contributes to the malignant phenotype of tumors.

Cloning, characterization, and modeling of a monoclonal anti-human transferrin antibody that competes with the transferrin receptor.

In this report we describe the isolation and characterization of a monoclonal antibody against human serum transferrin (Tf) and the cloning and sequencing of its cDNA. The antibody competes with the transferrin receptor (TR) for binding to human Tf and is therefore expected to bind at or very close to a region of interaction between Tf and its receptor. From the deduced amino acid sequence, we constructed a 3-dimensional model of the variable domains of the antibody based on the canonical structure model for the hypervariable loops. The proposed structure of the antibody is a first step toward a more detailed characterization of the antibody-Tf complex and possibly toward a better understanding of the Tf interaction with its receptor. The model might prove useful in guiding site-directed mutagenesis studies, simplifying the experimental elucidation of the antibody structure, and in the use of automatic procedures to dock the interacting molecules as soon as structural information about the structure of the human Tf molecule will be available.

Was it infarction or haemorrhage? A clinical diagnosis by means of the Allen score.

Since the clinical distinction between haemorrhagic and ischaemic stroke cannot be achieved with a simple clinical evaluation, and it is virtually impossible to submit all stroke patients to CT, a weighted clinical score may offer some advantages to physicians who are involved in stroke management. The Allen score (also referred to as the Guy's Hospital score), a validated clinical score, has been tested in two different clinical settings, comprising 289 patients. When only the values under 4 and those over 24 are taken into account (i.e. greater than 90% probability of ischaemia and haemorrhage), the global accuracy of the score is 97%, and the diagnostic gain (given a pretest probability for haemorrhage of 11% and a likelihood ratio of 194) is 85%. Therefore, we conclude that this simple clinical method can be used for epidemiological studies of stroke incidence and outcome, as well as for a first bedside screening to decide which patients should have priority for CT.

Intra- and inter-observer variability of basal flow velocity and vascular reactivity measurements using transcranial Doppler sonography.

The reliability of blood flow velocity measurements performed using transcranial Doppler sonography was investigated with regard to the intra- and inter-observer variability. We compared the flow velocity values recorded in the intracranial vessels by two experienced investigators both under rest conditions and during hypercapnia induced by an apnea test. The results showed how flow velocity values measured at the level of the middle cerebral artery, anterior cerebral artery, ophthalmic artery and basilar artery were not significantly influenced by intra- or inter-examiner variability. The flow velocity increases recorded from the middle cerebral artery during the activation test proved to be reproducible when the measurements were repeated by the same examiner, but more variable when detected by different investigators. On the basis of the available data, the clinical relevance of transcranial Doppler results must take into account the degree of reliability of its measurements.

Carrier-Mediated Uptake of Abscisic Acid by Suspension-Cultured Amaranthus tricolor Cells.

Abscisic acid (ABA) uptake by Amaranthus tricolor cell suspensions was found to include both a nonsaturable component and a saturable part with K(m) of 3.74 +/- 0.43 micromolar and an apparent V(max) of 1.5 +/- 0.12 nanomoles per gram per minute. These kinetic parameters as well as the uptake by intact cells at 0 degrees C or by frozen and thawed cells, are consistent with operation of a saturable carrier. This carrier-mediated ABA uptake was partially energized by DeltapH: it increased as the external pH was lowered to pH 4.0; it decreased after the lowering of the DeltapH by the proton ionophore carbonylcyanide-m-chlorophenylhydrazone or after the altering of metabolically maintained pH gradient by metabolic inhibitors (KCN, oligomycin). The carrier is specific for ABA among the plant growth regulators tested, is unaffected by (RS)-trans-ABA and was inhibited by (S)-ABA, (R)-ABA, and also by the ABA analog LAB 173711.

[An in vivo study of phytochrome in seeds of Pinus nigra Arn by differential spectrophotometry]. / Etude du phytochrome des graines de Pinus nigra Arn par spectrophotométrie bichromatique in vivo.

Phytochrome photoconversions Pr→Pfr and Pfr→Pr can be measured by differential spectrophotometry in "dry" seeds (6% water content) of Pinus nigra Arn. A red light irradiation given before imbibition induces germination when the seeds are subsequently wetted and kept in darkness.In continuous darkness the phytochrome content shows a drastic increase at the beginning of moistening.The detectable pigment is entirely in the Pr form. The normal Pfr→Pr dark reversion is observed. Pfr destruction does not take place.