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Biomedical subjects

M Cinelli

Publications and source records attributed to M Cinelli.

At least 19 recordsLinked to original sources
PubMed2006

Systemic sclerosis fibroblasts inhibit in vitro angiogenesis by MMP-12-dependent cleavage of the endothelial cell urokinase receptor.

Failure of endothelial cells to develop new vessels in response to hypoxia is a distinctive feature of systemic sclerosis (SSc) in the avascular phase. We have previously shown that SSc endothelial cells over-express matrix metalloproteinase-12 (MMP-12), which blocks angiogenesis by cleavage of the endothelial urokinase-type plasminogen activator receptor (uPAR). In the present study, we have investigated whether over-expression of MMP-12 and of angiostatic factors, or hypo-expression of angiogenic factors by SSc fibroblasts, contributes to impaired angiogenesis in SSc. Dermal fibroblasts were isolated from healthy subjects (N-Fb) and patients with diffuse SSc (SSc-Fb). Angiogenesis of target normal human microvascular endothelial cells (H-MVECs) was assayed by Matrigel invasion, cell proliferation, and capillary morphogenesis. uPAR cleavage and MMP-12 activity were evaluated by western blotting. We show that the over-expression of MMP-12 by SSc-Fb determines uPAR cleavage in H-MVECs. Conditioned medium from SSc-Fb impaired H-MVEC proliferation, invasion, and capillary morphogenesis. Anti-MMP-12 antibodies restored such impairment. Altered expression of angiostatic/angiogenic factors, including transforming growth factor beta1, did not account for SSc-Fb-dependent impairment of angiogenesis. The over-expression of MMP-12 by both SSc-Fb and SSc endothelial cells indicates that MMP-12 over-production may have a critical pathogenic role in SSc-associated vascular alterations.

Blotting, Western↗
PubMed2005

Raloxifene reduces urokinase-type plasminogen activator-dependent proliferation of synoviocytes from patients with rheumatoid arthritis.

Extracellular fibrinolysis, controlled by the membrane-bound fibrinolytic system, is involved in cartilage damage and rheumatoid arthritis (RA) synovitis. Estrogen status and metabolism seem to be impaired in RA, and synoviocytes show receptors for estrogens. Our aims in this study were to evaluate in healthy and RA synoviocytes the effects of Raloxifene (RAL), a selective estrogen receptor modulator (SERM), on: proliferation; the components of the fibrinolytic system; and chemoinvasion. The effects of RAL were studied in vitro on synoviocytes from four RA patients and four controls. Proliferation was evaluated as cell number increase, and synoviocytes were treated with 0.5 microM and 1 microM RAL with and without urokinase-plasminogen activator (u-PA) and anti-u-PA/anti-u-PA receptor (u-PAR) antibodies. Fibrinolytic system components (u-PA, u-PAR and plasminogen activator inhibitor (PAI)-1) were assayed by ELISA with cells treated with 0.5 microM and 1 microM RAL for 48 h. u-PA activity was evaluated by zymography and a direct fibrinolytic assay. U-PAR/cell and its saturation were studied by radioiodination of u-PA and a u-PA binding assay. Chemoinvasion was measured using the Boyden chamber invasion assay. u-PA induced proliferation of RA synoviocytes was blocked by RAL (p < 0.05) and antagonized by antibodies alone. The inhibitory effect of RAL was not additive with u-PA/u-PAR antagonism. RA synoviocytes treated with RAL showed, compared to basal, higher levels of PAI-1 (10.75 +/- 0.26 versus 5.5 +/- 0.1 microg/10(6) cells, respectively; p < 0.01), lower levels of u-PA (1.04 +/- 0.05 versus 3.1 +/- 0.4 ng/10(6) cells, respectively; p < 0.001), and lower levels of u-PAR (11.28 +/- 0.22 versus 23.6 +/- 0.1 ng/10(6) cells, respectively; p < 0.001). RAL also significantly inhibited u-PA-induced migration. Similar effects were also shown, at least partially, in controls. RAL exerts anti-proliferative and anti-invasive effects on synoviocytes, mainly modulating u-PAR and, to a lesser extent, u-PA and PAI-1 levels, and inhibiting cell migration and proliferation.

Arthritis, Rheumatoid↗
PubMed2005

Deflazacort modulates the fibrinolytic pattern and reduces uPA-dependent chemioinvasion and proliferation in rheumatoid arthritis synoviocytes.

OBJECTIVE: Extracellular fibrinolysis, controlled by the cell-associated fibrinolytic system (urokinase plasminogen activator, uPA; uPA receptor, uPAR; plasminogen activator inhibitor type-1, PAI-1), is involved in cartilage damage generation and in rheumatoid arthritis (RA) synovitis. Since steroids reduce the rate of radiological progression of RA, we planned to evaluate in healthy and RA synoviocytes the effects of the steroid deflazacort on uPA, uPAR and PAI-1 expression, and subsequent phenotypic modifications in terms of uPA/uPAR-dependent invasion and proliferation. METHODS: uPA, uPAR and PAI-1 levels were studied by ELISA, RT-PCR (uPAR) and zymography (uPA) in synoviocytes from four RA patients and four healthy controls. Chemoinvasion was assessed by the Boyden chamber invasion assay, using Matrigel as the invasion substrate. Proliferation was evaluated by cell counting. Both invasion and proliferation were measured upon treatment with deflazacort 5 muM with or without parallel stimulation with uPA 500 ng/ml or in the presence of monoclonal anti-uPA and anti-uPAR antibodies. RESULTS: Invasion and proliferation of RA synoviocytes require a proper functional balance of the fibrinolytic system. Both deflazacort and monoclonal antibodies against uPA and uPAR reduced expression and activity of the system, thus inhibiting invasion and proliferation. In RA synoviocytes, deflazacort induced higher PAI-1 and lower uPA and uPAR levels, as well as a decrease in uPA enzymatic activity. The levels of uPAR mRNA were concomitantly reduced, as was uPA-induced chemoinvasion. All these effects were also shown in controls, though to a lesser extent. CONCLUSIONS: Deflazacort might control RA synovial proliferation and invasion by differential modulation of single members of the fibrinolytic system.

Adult↗
PubMed2001

Prospective comparison of arm veins and greater saphenous veins as infrageniculate bypass grafts.

OBJECTIVES: to compare arm and saphenous veins for infrageniculate bypass grafting. DESIGN: prospective non-randomised study. MATERIALS: two hundred patients, of which 197 had ischaemic tissue loss or rest pain. METHODS: two hundred and eleven infrageniculate vein bypass procedures using 176 greater saphenous veins and 35 arm veins. RESULTS: the cumulative primary graft patency rate at 1-month and 2 years was 80% and 61% for saphenous vein and 89% and 42% for arm vein. The corresponding rates for secondary patency were 84.5% and 68%, and 91% and 57%, respectively. These results corresponded to a relative risk of secondary failure of 1.53 (95% CI 0.71, 3.31) for arm vein grafts. In subgroup analyses, this estimate was 0.93 and 2.1 for primary vs secondary bypasses and 0.38 and 2.06 for single-vein vs spliced-vein bypasses. Among arm veins, cephalic vein grafts performed better than basilic vein grafts. Early mortality was 14% for arm vein and 10% for saphenous vein. CONCLUSION: in the setting of infrageniculate bypass grafting, arm vein grafts are not equivalent to greater saphenous vein grafts, but contribute importantly to a policy of using autologous veins. The possibility of equivalence remains for the arm vein graft that uses a cephalic vein or is a primary procedure.

Aged↗
PubMed2000

Bypass to the genicular arteries for revascularisation of the lower limb.

OBJECTIVE: To describe an initial experience with infrainguinal bypass grafts inserted distally in a genicular artery. DESIGN: Retrospective case series study. SUBJECTS AND METHODS: Eleven patients with Grade III chronic limb ischaemia in whom arteriography showed femoropopliteal occlusive disease and at least one genicular branch suitable for receiving a bypass. Bypass grafts were done to the descending genicular artery (n=4) or the medial sural artery (n=6) using segments of autologous veins; one bypass was not completed. RESULTS: Primary graft patency and foot salvage rates were 73% at 1 month and 24 months of follow-up. Patient survival rate was 100% and 90%, respectively. Major amputation was required in two of three patients following early graft failure. Of the eight patients who had a patent graft, the Doppler ankle-brachial systolic pressure index showed no change in one patient, an increase of 0.13-0.66 in six patients, and was not measured in one patient. The former patient underwent a below-knee amputation whereas the other seven patients showed complete healing of their skin ulcers and sites of minor amputation. CONCLUSION: The genicular bypass is a useful alternative that may extend the limits of infrainguinal arterial reconstruction with autologous tissue and the potential for long-term patient benefit.

Aged↗
PubMed1998

Chronic treatment of human fibroblasts cultures with diacylglycerol induces down-regulation of p53 functional activity.

We found that many spontaneous human tumors exhibit increased levels of endocellular diacylglycerol (DAG) which is synthesized de novo as a byproduct of glycolysis. It has been shown that DAG mimics phorbol esters as a full tumor promoter in mouse skin carcinogenesis. A short term DAG treatment activates protein kinase C (PKC), while a long term "chronic" treatment down-regulates PKC. We show here that chronic treatment of human fibroblast with DAG induces p53 down-regulation and inhibition of p53 functional activity, and protection from UV-induced apoptosis. As PKC phosphorylation is necessary for p53 functional activity, we propose that chronic DAG treatment mimics the same event occurring in vivo for the effect of glycolysis in tumor progression.

Animals↗
PubMed1998

Redundant down-regulation pathways for p53.

The aim of this review is to underline the redundancy of down-regulation pathways for p53, at the light of the two more important degradative systems: calpains and ubiquitin-dependent pathways. The MDM2 feed-back loop is also illustrated, as well as the phosphorylative/dephosphorylative regulation of the latent and active p53 isoforms. The mechanisms prolonging p53 half life, following irradiation, are also discussed.

Animals↗
PubMed1998

Acetylation and phosphorylation of the carboxy-terminal domain of p53: regulative significance.

Three fundamental domains are conventionally distinguished on the p53 molecule: an NH2 domain involved in transcription, a central core domain involved in specific DNA binding to the consensus sequences, and a carboxy-terminal domain of about 30 amino acids working as a basic regulatory domain, exhibiting aspecific DNA binding, tetramerization, and nuclearization. The presence of an unmodified carboxy-terminus does not allow the specific transactivation transcriptional function of the p53 protein. Therefore, for the activation of the protein function the carboxy-terminus must be modified. In the present commentary we discuss the role of two covalent modification events occurring at the carboxy-terminus, namely phosphorylation and acetylation, as well as the specific role of these events in the functional regulation of p53 molecule.

Acetylation↗
PubMed1997

Complex interplay among apoptosis factors: RB, p53, E2F, TGF-beta, cell cycle inhibitors and the bcl2 gene family.

Apoptosis is a fundamental cell program as important as growth, differentiation and quiescence. It regulates tissue development, homeostasis and it is a basic defence against cancer. The cell can undertake multiple apoptotic pathways, where different elements are involved. In this report, we would like to stress particularly that the p53/RB pathway and its complex, interplay with the bcl2 gene family, where paramount elements of apoptosis regulation are operating. It is generally believed that bcl2 blocks apoptosis at the level of the activation of ICE-(Interleukin Converting Enzymes)-like proteases [1,2]. The interconnection between apoptosis and cell cycle is very important and complex and we will start the story from this very up-to-date point of view.

Animals↗
PubMed1995

Phorbol esters attenuate the expression of p53 in cells treated with doxorubicin and protect TS-P53/K562 from apoptosis.

The induction of p53 by doxorubicin in normal human fibroblasts was completely reverted by TPA, a phorbol ester. A ts-p53 mutant protein which is ineffective at 37 degrees C, but behaves in a wild-type fashion at 32 degrees C, was overexpressed in the p53-null human leukemia cell line K562. Wild-type-p53 overexpression induced apoptosis, whereas TPA protected K562 cells from this phenomena. By analogy with the observed human fibroblasts, TPA was found to decrease p53 amount. The TPA-dependent down-regulation of p53 could explain the chromosomal gross alterations typical of cells subjected to a chronic TPA treatment, alterations also found in cells defective for p53 function.

Apoptosis↗
PubMed1995

Dominant oncogenes, tumor suppressors, and radiosensitivity.

A variety of conflicting results appeared in the literature concerning the effect of dominant oncogenes on the sensitivity to irradiation and to anticancer agents in a number of cell lines of human and animal origin. In this report we provide evidence supporting the hypothesis that the tumor suppressor gene p53 and the apoptosis suppressor gene bcl2 modulate the effect of dominant oncogenes and that the effect of dominant oncogenes on resistance or sensitivity is dependent on the balance between the expression of p53 and bcl2.

Apoptosis↗
PubMed1994

Bcl-2 overexpression abolishes early calcium waving preceding apoptosis in NIH-3T3 murine fibroblasts.

Overexpression of the apoptosis protection gene bcl-2 abolished the earliest response to serum withdrawal in NIH-3T3 murine fibroblasts, i.e., the abrupt cytoplasmic free calcium drop. This phenomenon, also observed in a myeloid cell line, led us to propose this ionic waving as an early apoptotic signal. Its abolition by bcl-2 overexpression suggests that this gene plays a role also on early events "priming" apoptosis.

3T3 Cells↗
PubMed1994

Apoptosis and the cell cycle.

This brief review examines the strict relationships between cell apoptosis and G1 cyclins. It has been shown that the basic role of G1 cyclins is in regulating G1 progression and G1/S transition (the critical cycle point for cell program decisions, including apoptosis) a fatal program for cells unable to bypass G1/S checkpoint 1. Notably, both of the two giant regulators of checkpoint 1 (i.e., p105RB [retinoblastoma oncosuppressor-encoded protein] and p53 dependent WAF1/CIP1) are influenced by or influence G1 cyclins: cyclin E/cdk2 kinase complexes hyperphosphorylate p105RB, induce E2F release, and free G1 exit. On the other hand, p21-WAF1/CIP1 is an inhibitor of cyclin-dependent kinases blocking cells at G1/S. Thus, G1 cyclin activity appears as a conditio sine qua non for G1 exit and apoptosis escape.

Animals↗
PubMed1993

Apoptosis induction in 32D cells by IL-3 withdrawal is preceded by a drop in the intracellular calcium level.

The 32D murine myeloid cell line is dependent on interleukin-3 (IL-3) for growth in vitro. We show here that IL-3 induces a slow increase in endocellular calcium, which is sizeable two hours after the addition. Withdrawal of the cytokine induces apoptosis, whose classic late events are evident after 16/18 hours, and are preceded by a calcium drop during the first 2/3 hours after IL-3 subtraction. Calcium drop is here proposed as a trigger of the apoptotic process, in agreement with other recently reported findings.

Animals↗
PubMed1986

Viminol R2 analgesic activity in patients with postoperative pain: comparison with pentazocine.

The analgesic activity of Viminol R2 was compared to that of Pentazocine and placebo in a double-blind, "between subjects" study involving 42 patients with postoperative pain randomly assigned to 3 groups of 14 patients each and treated with a single intravenous dose of Viminol R2 10 mg, Pentazocine 30 mg or placebo. Pain relief scores were recorded over a 2 h period after treatment. The analgesic activity of Viminol R2 and Pentazocine was significantly greater than that of placebo both in terms of time-course of action and frequency of classes of satisfactory analgesia. No statistically significant differences were found between Viminol R2 and Pentazocine. No side effects were observed in any patients treated with Viminol R2.

Adult↗