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M Foresti

Publications and source records attributed to M Foresti.

At least 19 recordsLinked to original sources
PubMed2001

A short low-level exposure to metavanadate during a cell cycle-specific interval of time is sufficient to permanently derange the differentiative properties of Mel cells.

Mouse erythroleukemia (Mel) cells have a cell cycle-dependent high sensitivity to chemical and physical mutagens. This report shows that a 5 h exposure to 0.1 or 0.01 microg/ml metavanadate during the initial period of erythroid differentiation induction was sufficient to permanently damage the ability of treated Mel cells and their progeny to undergo erythroid differentiation, without affecting cell viability and proliferation. Conversely, a 5 h pulse of metavanadate at 1 or 10 microg/ml inhibited both differentiation and cell proliferation. The cell cycle-dependent period of mutagenesis was essential for fixation of damage in the cell genome and the progeny of the cells treated with 0.1 or 0.01 microg/ml metavanadate stably inherited an impaired capacity to differentiate. The efficiency of the DNA repair synthesis machinery during the specific period of exposure of Mel cells seemed directly involved in damage fixation. In fact, the mutagenic effects of a 0.1 microg/ml metavanadate pulse was further increased in the presence of 1 mM hydroxyurea, an inhibitor of DNA repair synthesis. In contrast, 5 microg/ml vanillin, an antimutagenic agent that stimulates repair, completely restored the capacity of progeny of cells treated with 0.1 microg/ml metavanadate to complete differentiation. Determination of [(3)H]deoxythymidine in acid-insoluble DNA indicated that incorporation was stimulated by metavanadate alone and was further increased by metavanadate plus vanillin; conversely, incorporation of thymidine was reduced in the presence of hydroxyurea. The capacity of metavanadate to permanently damage Mel cell erythroid differentiation appeared to depend on the cell cycle-related efficiency of the DNA repair systems, activated to correct the induced alteration, rather than on a specific concentration.

Animals↗
PubMed1999

Hypoxic Hypoperfusion Fails to Induce Myocardial Hibernation in Anesthetized Swine.

BACKGROUND: Congenital origin of the left coronary artery from the pulmonary artery (ALCAPA) results in chronically dysfunctional myocardium with the partial ability to recover after revascularization. We attempted to establish an ALCAPA syndrome in anesthetized pigs for 24 hours and to compare it with stunned and infarcted myocardium. METHODS AND RESULTS: In group 1 (n = 12), a bypass graft was interposed between the pulmonary artery and the left anterior descending coronary artery (LAD). Reduction of flow in the LAD with gradual increases in flow from the pulmonary artery resulted in an incremental reduction of segment shortening (8.9 +/- 5.3% at 24 hours vs 26.6 +/- 10% at baseline, P <.005). In group 3 (n = 5), 2 cycles of 10-minute LAD occlusion resulted in decreased segment shortening with slow recovery (at 24 hours 18.7 +/- 1.3% vs 24.2 +/- 4% at baseline, segment shortening with slow recovery (at 24 hours 18.7 +/- 1.3% vs 24.2 +/- 4% at baseline, P <.05). In group 3 (n = 6), 1-hour LAD occlusion reduced segment shortening at 24 hours to 4.7 +/- 5.2% (P <.005 vs baseline). Histological analysis of the LAD territory revealed severe degeneration, myolysis, and alteration of the chromatin structure in group 1 comparable to ischemic cell death in group 3, whereas control areas and the LAD area in group 2 showed only minor structural alterations. Infarct size/risk area, as measured by tetrazolium staining, was 49.8 +/- 11.2% in group 1, 9.3 +/- 8.1% in group 2 (P <.005), and 60.3 +/- 9% in group 3. CONCLUSION: Hypoxic myocardial hypoperfusion from the pulmonary artery results in myocardial necrosis in anesthetized pigs. These findings are in contrast to the concept of myocardial hibernation in the ALCAPA syndrome because in this model, hypoxic hypoperfusion failed to induce adaptation to preserve myocardial structure.

Journal Article↗
PubMed1998

Infarct Size Reduction by Ischemic Preconditioning Is a Monophasic, Short-Lived Phenomenon in Anesthetized Pigs.

BACKGROUND: Controversy exists concerning the duration of infarct size reduction with ischemic preconditioning in different species. In the present study, we (a) evaluated the time course of protection with preconditioning and (b) sought to determine whether late protection (the "second window") after 24 hours is manifest in the open-chest pig model. METHODS AND RESULTS: Six groups of pentobarbital-anesthetized pigs underwent 1 hour of left anterior descending coronary artery occlusion and 2 hours of reperfusion. Group 1 served as control, and pigs in group 2 received two 10-minute episodes of preconditioning ischemia followed by 30 minutes of reperfusion before the sustained 1-hour occlusion. In groups 3-6, the period of intervening reperfusion between the preconditioning stimulus and the index ischemia was extended to 60, 90, and 300 minutes and 24 hours, respectively. The area at risk was determined by fluorescein dye injection, and infarct size was measured by incubation in p-nitrobluetetrazolium and expressed as percent of the risk area. Infarct size in preconditioned pigs (group 2) was significantly reduced compared with controls (25.6 +/- 3.9% v 71.3 +/- 5.9%, P <.001). Extension of the intervening reperfusion to 60, 90, and 300 minutes and 24 hours resulted in infarct sizes of 64.5 +/- 5.5%, 67.2 +/- 8%, 62.6 +/- 6.1%, and 75.3 +/- 7%, respectively (P = NS v control). CONCLUSIONS: The infarct size-limiting effects of ischemic preconditioning last less than 1 hour in the pig model. Moreover, in contrast to other species, a late protection at 24 hours after the preconditioning stimulus was not detected. These results indicate that precondition-induced reduction of infarct size is monophasic in anesthetized pigs.

Journal Article↗
PubMed1997

Two periods of sensitivity to mutagens in induced Mel cells with different outcomes.

Mouse erythroleukemia (Mel) cells are particularly sensitive to mutagenic agents between 18 and 24 h from the start of induction (Foresti, M.L. Gaudio, G. Geraci and P. Manduca (1986) Inhibition of dimethyl sulfoxide induced erythropoietic differentiation of murine erythroleukemia cells in culture. Cancer Res., 46, 6260-6263). We show here the occurrence of another period of sensitivity during the initial 5 h after the addition of the inducer dimethyl sulfoxide (DMSO) to the culture medium. The sensitivity to the mutagenic action of a sublethal 3-s pulse of UV light (13.5 J/m2) was monitored on the progeny of the irradiated cells at day 5 after the start of induction. The effects were analysed on functions strictly linked to the final expression of the differentiated phenotype: hemoglobin concentration, percent cells producing hemoglobin (%B+), activity of delta-amino levulinic acid dehydrase (ALA-DH) and presence of globins. Each function appeared differently and selectively affected in the progeny of the cells depending on the exact time of irradiation during the period of sensitivity Specifically, cells irradiated at hour 3 after induction show both hemoglobin concentration and ALA-DH activity values increased by a factor 3 over controls. Cells irradiated at hour 5 show an almost complete halt in cell induction and the other tested functions show minimal values. Cells are nearly insensitive to irradiation at later times, until hour 20, after which a second period of sensitivity with peak value at hour 22 occurs at which time hemoglobin concentration in the progeny of irradiated cells is increased by a factor 3 over controls, ALA-DH activity is increased by a factor 15 while percent B+ value is at its minimum. The differential effects of UV irradiation on Mel cell functions in the first and in the second period of sensitivity to mutagens confirm the hypothesis that the consequences of a mutational event are strictly dependent on the functional state of the cell. The 1-5 h period of sensitivity in which Mel cells fix the effects of the mutagen in their genome corresponds to increased thymidine incorporation not correlated with cell duplication.

Animals↗
PubMed1993

Inhibition of erythroid differentiation in MEL cells by UV irradiation. Cell cycle and DNA repair activity.

Irradiation with a 3-s pulse of 254 nm UV light has been used to study sensitivity to mutagenic agents of mouse erythroleukemia (MEL) cell cultures in correlation with the cell cycle. A dose of UV irradiation was chosen that had no consequences for cell viability and growth. For this reason phenotypic effects were monitored on the progeny of all cells of the irradiated cultures by scoring those unable to undergo erythroid differentiation upon induction with dimethyl sulfoxide. The very short period of irradiation made it possible to show that MEL cells, synchronized by two sequential blocks of deoxythymidine and one of hydroxyurea (HU), are sensitive to UV irradiation only in a very short period of time at about 60 min after release from HU block. Determinations of deoxythymidine incorporation into DNA show that this time period corresponds only marginally to the initial part of the S phase during which irradiation has no consequences for cell properties. Cells are not sensitive to UV irradiation in G1 and in G2/M unless, immediately after irradiation and for the following 2 h, cultures are treated with 1 mM HU to interfere with DNA repair. Alkaline sucrose gradient analyses show at all tested times that irradiation leads to fragmentation of cell DNA. The data suggest that an immediate increase of deoxythymidine incorporation into DNA following irradiation is not necessary for the efficient repair of damaged DNA. In fact, the percent of cells expressing the erythroid phenotype is normal in the progeny of cells irradiated in G2/M, when TdR incorporation is at a minimum. Repair activities appear then to be mechanistically divided into two phases, (1) recognition labeling of the altered sites and (2) reconstitution of the DNA sequences. The first activity appears to be operative at all phases of the cycle, the second activity is little or not operative in G2/M, possibly delayed to the following G1 period.

Animals↗
PubMed1992

Selective gene mutation in MEL cells.

MEL cells, undergoing erythroid differentiation and parasynchronized by dimethyl sulfoxide (DMSO) induction, were irradiated with a 3-s pulse of UV light at sublethal dose. A large number of clones deficient in different gene functions are found in the progeny of the treated cells, if the pulse irradiation is performed 18-24 h from the start of DMSO induction. Kinetics of thymidine incorporation into DNA show that the period of sensitivity corresponds to the S phase. The results show that the activities of the tested genes are differently affected depending on the exact time of cell irradiation. Maximum percent inhibition of cells not expressing glucose-6-phosphate dehydrogenase (G-6-PD) (70%) is produced by irradiating at 20 h from the start of DMSO induction; 6-phosphogluconate dehydrogenase (6-PGD) (55%), and hypoxanthine (guanine) phosphoribosyltransferase (HPRT) (33%), at 21 h; hemoglobin (50%), at 22 h. The time difference in the sensitivity to UV light is highly reproducible and has been exploited to isolate, with high efficiency, cellular clones deficient in any one of the tested functions. Determinations of enzymatic activities on cell lysates show that the expression of tested genes is actually altered in cells that, on the basis of cytochemical tests, appear unaffected by UV irradiation. While the production of mutant clones is observed only during the S phase of the cell cycle, immediate statistical damage of the cellular DNA is produced at all times of irradiation. This finding excludes that the two types of phenotypic alterations, blocked or altered gene expression, both propagated in the progeny of the cells as clonal properties, may derive from a preferential alteration of those functions during the S phase.

Cell Differentiation↗
PubMed1991

[Primary empty sella. Incidence in 500 asymptomatic subjects examined with magnetic resonance].

In 500 consecutive patients, aged 11-82 years, who underwent MR imaging of the brain for a variety of conditions not related to pathologic processes of the sellar or juxtasellar regions, the authors detected primary totally empty sella in 28/248 males (11.3%) and in 34/252 females (13.5%). Primary partially empty sella was found in 40/248 males (16.1%) and in 38/252 females (15.1%). A progressive increase in the incidence of the signs of primary empty sella was observed with aging, the increase being more conspicuous in the 5th decade of life in females (37.5%) and in the 6th decade in males (40%). On the whole, signs of primary empty sella were detected in 140/500 cases (38%)--namely, in 9.6% of the subjects under 40 and in 39.9% of those above 40. The incidence of primary empty sella, unrelated to any other clinical condition, seems to support its inclusion into paraphysiologic variants. On the basis of the data reported in literature, the authors consider the factors possibly playing a role in this condition. They seem to be: insufficiency or absence of diaphragma sellae, CSF pressure, and pituitary involution related to aging.

Adolescent↗
PubMed1990

Inhibition of MEL cells' capacity to undergo erythroid differentiation by chemicals added during induction.

Erythroid differentiation of murine erythroleukemia (MEL) cells, as induced by dimethyl sulfoxide, can be suppressed by chemicals at very low concentrations, not affecting cell viability and proliferation, if present in the culture medium between 18 and 24 h after addition of the inducer. The effect is apparent on the progeny of the treated cells and is determined, between day 3 and 5 following DMSO induction, as percent value of cells expressing the erythroid phenotype. Cultures showing decreased values are no longer terminal and a large number of clones, incapable of expressing the erythroid phenotype, can be isolated from them. In contrast, induced cultures are terminal if the added chemicals do not decrease the expression of the erythroid phenotype. Incorporation of thymidine into induced cultures reveals that maximal sensitivity of MEL cells to chemicals coincides with DNA duplication. In all affected cells, the inhibition to undergo erythroid differentiation is transmitted from one cell generation to the next.

Cell Differentiation↗
PubMed1986

Fluorescence polarization spectroscopy and time-resolved fluorescence kinetics of native cancerous and normal rat kidney tissues.

Steady state fluorescence polarization spectra and time-resolved emission decay kinetics have been measured in vitro from malignant and normal rat kidney tissue. The degrees of polarization and emission lifetimes from the cancerous and normal systems are different. The spectroscopic differences are attributed to environmental transformations local to the native flavin and porphyrin fluorophors' binding sites.

Animals↗
PubMed1986

Inhibition of dimethyl sulfoxide induced erythropoietic differentiation of murine erythroleukemia cells in culture.

The dimethyl sulfoxide induced erythropoietic differentiation of murine erythroleukemia cells, as determined by scoring benzidine positive cells, is inhibited by mitomycin C at concentrations that have no effect on cell proliferation. The inhibition occurs only when cells are treated with mitomycin C during induction and has a limit value of about 50%, independent of mitomycin C concentration. This limit value does not depend on cell heterogeneity since genetically homogeneous subclones, derived from DS19 clone, show levels of mitomycin C inhibition between 16 and 50%. Treatment with mitomycin C at different times after dimethyl sulfoxide addition shows that cell sensitivity to inhibition is not homogeneous during the induction period; it is maximal between 18 and 24 h from the start of induction and is observed with a concentration of mitomycin C as low as 25 fM. The inhibition of the benzidine positive phenotypic expression appears irreversible since this effect is observed on cells even several generations after those which were actually treated.

Animals↗
PubMed1985

[Fibroadenolipoma of the breast].

Three cases of fibroadenolipoma (adenolipoma, hamartoma) of the breast are reported out of sixteen thousand mammographies. Their typical radiological findings are described.

Adenofibroma↗
PubMed1983

Influence of amphotericin B on leucine uptake in 3T3 cells.

By studying the effect of leucine competitors we found that activation of the specific leucine-transport system underlies the enhancement of leucine uptake in mouse 3T3 fibroblast cells induced by sublethal doses of Amphotericin B (synergic effect). The relation of the antibiotic activity and the alteration of the membrane cholesterol interaction with lipids is discussed.

Amino Acids↗
PubMed1979

Threonine deaminase: autogenous regulator of the ilv genes in Escherichia coli K-12.

In this paper we analyze the effect of mutations in three genes, ilvO, ilvA and rho, on the expression of the ilvEJGDA gene cluster of Escherichia coli K-12. The ilvO603 mutation causes a cis-dominant derepression of the ilvEJGDA genes. In particular, the ilvG gene, not expressed in the wild type, becomes expressed in the ilvO603 strain. We have introduced ilvA mutations (ilvA454 or ilvA628) in the ilvO603 strain and we show that ilvG expression requires the presence in cis of both an ilvO603 mutation and of an ilvA+ allele. The ilvG gene is not expressed when in trans is present an ilvO+, ilvA+ genotype. However, it is expressed when the chromosome in trans is ilvO603, ilvA+ (ilvG-). We suggest that ilvO603 is part of ilvA, the structural gene for threonine deaminase, and that threonine deaminase from the ilvO603 mutant binds the ilvO603 site and not the ilvO+ site. Therefore, the ilvA gene product would be a cis-acting protein. Mutations in the rho gene cause derepression of the ilvEJGDA gene cluster without a concomitant expression of the ilvG gene. We show that introduction of either a rho-218 or a rho-115 mutation into the ilvO603, ilvA454 double mutant causes expression of ilvG. We therefore suggest that the ilvA gene product, threonine deaminase, is involved in termination of transcription as an antagonist of the rho gene product. Introduction of ilvA454 into an ilvO603 strain causes also a decrease in expression of the ilvE, ilvJ and ilvD genes. This effect is maximum in the case of the ilvD gene and we studied it in detail in isogenic strains containing also the rho-218 mutation.

Escherichia coli↗
PubMed1977

Endopolygalacturonase from Rhizoctonia fragariae. Purification and characterization of two isoenzymes.

An electrophoretically homogeneous preparation of endo-polygalacturonase (poly(1,4-alpha-D-galacturonide)glycanohydrolase, EC 3.2.1.15) from culture filtrates of Rhizoctonia fragariae, a pathogenic agent in strawberry plants, was resolved into two isoenzymes when subjected to isoelectrofocusing in a narrow pH range. The isoelectric points of the two isoenzymes were 6.76 +/- 0.03 and 7.08 +/- 0.05. The two polygalacturonases exhibited similar substrate specificity, pH optimum and pattern of degradation of sodium polypectate. The two enzymes consisted of a single polypeptide chain which had an apparent molecular weight of 36 000 as determined by gel filtration on Sephadex G-100.

Glycoside Hydrolases↗
PubMedDate not supplied

Effect of bacitracin on erythroid differentiation of MEL cells.

Bacitracin, an antibiotic widely utilized in clinical and veterinary use, was tested on murine erythroleukemia (MEL) cells. Tests were performed to evaluate the capacity of the drug to interfere with erythroid differentiation. Cells were exposed to a single treatment in S phase at sublethal doses of bacitracin. Two responses were found depending on the drug concentration. At higher concentrations (25 micrograms/ml and 250 ng/ml) a reduction in number of differentiating cells was observed but the kinetics of the process remained unchanged. At lower concentrations (from 2.5 ng/ml to 2.5 fg/ml) a dramatic alteration of the dynamic of differentiation was found. These two responses are related to different activities of the DNA repair mechanisms. Higher doses of bacitracin stimulate repair while lower concentrations are not able to active repair, as demonstrated by tests with hydroxyurea. The bacitracin-induced damage can be considered a stable genetic and/or epigenetic alteration, as demonstrated by the high frequency of mutant clones isolated from low-dose treated cells. The suitability of MEL cells system in evaluating genotoxicity of drugs for veterinary use is underlined.

Animals↗