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Biomedical subjects

D Bossi

Publications and source records attributed to D Bossi.

At least 19 recordsLinked to original sources
PubMed2003

A functional assessment methodology for alcohol dependent patients undergoing rehabilitative treatments.

PURPOSE: We propose a functional assessment approach for patients with alcoholic dependence of working age undergoing aerobic training. The background is the WHO indication (ICIDH-2) to use measurable 'activities' as a means to assess the individual 'participation' in social life which also implies work capacity. Defining sustainable energetic levels for the individual is an important issue for both the quantification of an effective training and the evaluation of possible improvements following training. METHODS: Fifty-six 'alcohol dependent' patients, as defined by DSM IV (Diagnostic and Statistical Manual of Mental Disorders), admitted to our Unit in a 16 month-period participated in the study. Eighteen healthy subjects served as controls (Group C). Out of all the 56 patients, 33 (Group A) underwent an aerobic training and 23 subjects (Group N) underwent the same pharmacological and psychological therapy but without aerobic training. Patients were assigned to the treatment (A) or no treatment (N) group according to a 'quasi-experimental' design (i.e. temporal selection criteria). The evaluation protocol consisted of submaximal symptom-limited tests. The tests consisted of bouts of 'basic' activities (walking, lifting, arm-work) to be performed at different intensities. We estimated the total energetic work (TW) performed in the tests by means of formulas available in the literature. The maximal energetic intensity (EI) reached during the tests was also estimated and expressed in MET (multiple of the basal metabolism). RESULTS: Significant differences in work capacity were observed between patients and healthy subjects at baseline. Group A significantly increased TW after rehabilitation, while Group N did not increment their performance at the re-test. CONCLUSIONS: The proposed approach could be useful in the functional assessment of deconditioned subjects with alcohol dependence in working age, and could monitor the changes in work capacity following training.

Adult↗
PubMed2000

A baker's yeast mutant (fil1) with a specific, partially inactivating mutation in adenylate cyclase maintains a high stress resistance during active fermentation and growth.

The initiation of fermentation in the yeast Saccharomyces cerevisiae is associated with a rapid drop in stress resistance. This is disadvantageous for several biotechnological applications, e.g. the preparation of freeze doughs. We have isolated mutants in a laboratory strain which are deficient in fermentation-induced loss of stress resistance ('fil' mutants) using a heat shock selection protocol. We show that the fil1 mutant contains a mutation in the CYR1 gene which encodes adenylate cyclase. It causes a change at position 1682 of glutamate into lysine and results in a tenfold drop in adenylate cyclase activity. The fil1 mutant displays a reduction in the glucose-induced cAMP increase, trehalase activation and loss of heat resistance. Interestingly, the fil1 mutant shows the same growth and fermentation rate as the wild type strain, as opposed to other mutants with reduced activity of the cAMP pathway. Introduction of the fil1 mutation in the vigorous Y55 strain and cultivation of the mutant under pilot scale conditions resulted in a yeast that displayed a higher freeze and drought resistance during active fermentation compared to the wild type Y55 strain. These results show that high stress resistance and high fermentation activity are compatible biological properties. Isolation of fil-type mutations appears a promising avenue for development of industrial yeast strains with improved stress resistance during active fermentation.

Adenylyl Cyclases↗
PubMed1996

Red cell physiology.

The erythrocyte, the ultimate product of mammalian erythroid maturation, appears as a highly specialized and, paradoxically, a very simplified cell. In fact it lacks a nucleus and the intracellular organelles are essentially designed to transport oxygen from the outer environment to respiring tissues through the sophisticated functional properties of intraerythrocytic hemoglobin. In this respect, since oxygen transport is such a vital process, the red cell has often been considered, in an excess of oversimplification, as merely a "biological bag' enveloping a viscous solution of concentrated hemoglobin and containing only those few enzymes which are needed to maintain the cell functionally active. However, within the cell a number of different processes are contemporaneously going on, hemoglobin acts as an oxygen and carbon dioxide transporter, glycolysis and the pentose phosphate shunt are devoted to the production of ATP and NADPH, respectively, and membrane organization provides the cell with a good deformability, allowing it to cross narrow splenic capillaries and channels without any appreciable damage for several weeks of activity. All these processes are, indeed, highly integrated and concur to define a complex scenery centered on the oxygenation-deoxygenation cycle of hemoglobin. Within this emerging scheme, hemoglobin appears to display, besides the basic function of oxygen transport, several other biological functions which are driven by the oxygen-linked conformational transition and whose relative importance, in the economy of the cell and of the organism, is not easy to qualify. Some of these aspects are described and discussed.

Cell Membrane↗
PubMed1996

Hemolytic anemias due to disorders of red cell membrane skeleton.

During the past 10 years, knowledge of the composition, function and supramolecular assembly of the red cell membrane has been greatly expanded by progress in molecular and cell biology. Detailed information on the organization of membrane cytoskeletal proteins and their molecular characterization has allowed us to correlate a number of protein abnormalities with clinical symptoms that are peculiar to hereditary hemolytic anemias (HHA). In particular, three general principles emerge that can help us to understand the pathogenetic mechanisms of HHA: (a) protein-protein and protein-lipid interactions greatly influence the correct assembly of the membrane skeleton; (b) the red blood cell (RBC) membrane skeleton mostly determines the shape (discocyte), deformability (rheologic properties) and durability (half-life and resistence to shear stress) of the erythrocytes; (c) changes in cytoskeletal composition and/or organization can produce alterations in all of the above properties, and therefore they are responsible for the onset of the hemolytic damage.

Anemia, Hemolytic↗
PubMed1991

The effect of magnesium on glycolysis of permeabilized Ehrlich ascites tumor cells.

We have previously observed that extracellular Mg2+ influences the phosphofructokinase (PFK) activity of intact Ehrlich Ascites tumour cells (EATC). In this study we have investigated the mechanism by which Mg2+ modulates this key glycolytic enzyme in EATC made permeable to the cation by either digitonin or dextran sulphate. Results showed that when Mg2+ is freely permeable to the cytosol, the in vivo PFK activity, calculated as FDP/G6P ratio, is not increased as it is in intact cells. We also observed that in permeabilized cells Mg2+ determines the increase of glucose 6 phosphate (G6P), fructose 1,6 bisphosphate (FDP) and lactate production. We hypothesize that extracellular Mg2+ regulates PFK and glycolysis in these neoplastic cells not by entering the cytosol but by a specific interaction with the plasma membrane.

Adenosine Triphosphate↗
PubMed1991

Magnesium in normal and neoplastic cell proliferation: state of the art on in vitro data.

Information about the involvement of Mg2+ in all biochemical processes that participate in cell proliferation is reviewed in order to define the role of this divalent cation in normal and pathological growth. The lack of conclusive data about cell Mg2+ homeostasis does not suggest any definitive model for its role in the control of cell proliferation. On the other hand, new important information about its absolute requirement in crucial steps of cell activation that can, beside other functions, trigger cell division, strongly support the involvement of Mg2+ in the control of cell proliferation. Studies on the growth of cells in vitro, however, while confirming the indispensible requirement for Mg2+ in extracellular media, do not completely clarify the mechanism(s) or the exact phase/point of the cell cycle where Mg2+ exerts its regulation. Furthermore, the observation that tumour cells grown in culture are influenced by external divalent cations confirms the involvement of Mg2+ in cancer as well as in normal cell proliferation. The proposed explanations (theories, hypotheses) are described and discussed.

Animals↗
PubMed1989

The effect of Mg2+ upon 6-phosphofructokinase activity in Ehrlich ascites tumor cells in vivo.

The effect of Mg2+ addition to intact Ehrlich ascites tumor cells (EATC) has been investigated. A decrease of glucose 6-phosphate (G6P) content and an increase of fructose 1,6-diphosphate (FDP) content are detected in glucose utilizing EATC incubated with increasing Mg2+ concentrations (from 0 to 5.0 mM). The strong enhancement of FDP/G6P ratio is taken as evidence for in vivo stimulation of phosphofructokinase 1 (PFK) (ATP:D-fructose-6-phosphate 1-phosphotransferase; EC 2.7.1.11). A similar effect can be observed when glucose is replaced by fructose as the glycolytic substrate. Stimulation of PFK is paralleled by substantial depletion of ATP. Cytochalasin B prevents the observed phenomena. Cell total Mg increases by about 15% when EATC are incubated with 5 mM Mg2+. The overall data show that extracellular Mg2+ may modulate glycolytic flux in EATC in vivo. Implications and significance of these phenomena in the regulation of cancer cell metabolic features are discussed.

3-O-Methylglucose↗
PubMed1988

Impairment of microsomal calcium sequestration activity upon superoxide dismutase depletion in rat liver.

We have studied the Ca2+-accumulation activity of microsomal vesicles isolated from the liver of rats held for from 2 to 8 weeks on a copper-deficient diet. With this treatment that deeply modifies fatty acid composition, microsomal membranes show progressively lower Ca2+ sequestration. The activity can be fully restored upon physiological copper supply to the depleted animals. The determination of kinetic parameters of microsomal Ca2+ uptake shows that copper deficiency affects mainly the apparent velocity, leaving unaffected the apparent affinity of the pump for Ca2+. Many similarities were found between this model and the Morris hepatomas with different growth rate. The data support the hypothesis that the oxidative stress imposed on the cell by the loss of superoxide dismutase can influence many cell features, with different implications in the regulation of several biological and biochemical functions.

Animals↗
PubMed1987

Further observations on the effect of calcium ionophores on ascites tumor cells.

The effect of the Ca2+ ionophore ionomycin on neoplastic thymocytes in comparison to its effect on normal thymus cells was studied. Ionomycin increases intracellular Ca2+ in normal lymphocytes but fails to increase Ca2+ in neoplastic thymocytes. In these cells the ionophore causes a transient increase in cytosolic free Ca2+. The lack of effect of ionomycin reproduces that of A23187, but it does not depend on reduced availability of intracellular Mg2+ to exchange with Ca2+; it appears to depend on the strong activity of the plasma membrane Ca2+-extruding pump that counteracts ionomycin permeabilization and that can be partly inhibited by the calmodulin inhibitor R24571 (calmidazolium). Neoplastic thymocytes show a high content of magnesium, the intracellular binding of which is efficiently regulated by endogenous ATP. The data show also an interesting correlation between the regulation of energy metabolism (aerobic glycolysis) and cation homeostasis in the neoplastic cells studied.

Animals↗
PubMed1985

Involvement of the Ca2+/phospholipid-dependent protein kinase in the G1 transit of T51B rat liver epithelial cells.

The G0----G1 and G1----S transitions (but not the intervening events) in the G1 phase of T51B rat liver epithelial cells in serum-stimulated confluent cultures required a high concentration of extracellular Ca2+ and were accompanied or immediately preceded by increases in the amount of EDTA-extractable protein kinase C, a Ca2+/phospholipid-dependent enzyme. Involvement of this Ca2+-dependent enzyme in the two Ca2+-dependent transitions was further indicated by the facts that 12-O-tetradecanoyl phorbol-13-acetate (TPA), a compound that stimulated protein kinase C from T51B cells even in the absence of Ca2+, enabled these cells to transit G1 in Ca2+-deficient medium, while a TPA analogue (4 alpha-phorbol-12, 13-didecanoate (4 alpha-PDD) that did not stimulate the enzyme in cell-free preparations did not promote G0----G1 or G1----S transit in Ca2+-deficient medium.

Animals↗
PubMed1985

The influence of extracellular calcium on the distribution of protein kinase C in non-neoplastic and neoplastic rat liver cells.

Non-neoplastic T51B rat liver epithelial cells cannot proliferate in Ca2+-deficient medium. This proliferative inhibition in Ca2+-deficient medium is accompanied by a large reduction in the amount of cellular EDTA-extractable Ca2+/phospholipid-dependent protein kinase (protein kinase C) activity. By contrast, tumorigenic epithelial cells from several Morris hepatomas proliferate in Ca2+-deficient medium and either maintain or greatly increase their content of EDTA-extractable protein kinase C.

Animals↗
PubMed1982

Calcium permeability of Ehrlich ascites tumour cell plasma membrane in vivo.

Passive Ca2+ entry into Ehrlich ascites tumour cells has been investigated. Passive equilibrium of Ca2+ takes place in ascites tumour cells only under conditions of exhaustive energy depletion. The specific Ca2+ ionophore A23187 does not affect Ca2+ entry into ascites tumour cells under active metabolic conditions, but it increases the rate of Ca2+ equilibration in ascites tumour cells in the early stages of energy depletion. The results of the present experiments lead to the conclusion that in ascites tumour cell plasma membrane Ca2+ permeability is not a limiting step in the regulation of intracellular calcium content, while the energy-dependent Ca2+ extrusion is the main mechanism that prevents uncontrolled intracellular Ca2+ increase. The results taken together support the hypothesis that increased Ca2+ influx into the cell, caused by plasma membrane alteration, is responsible for permanently elevated mitotic activity and for deranged metabolic behavior of these neoplastic cells.

Anaerobiosis↗
PubMed1981

Lack of effect of the Ca2+ ionophore A23187 on tumour cells.

The Ca2+ ionophore A23187 increases intracellular calcium content in normal thymic cells, while it is without effect on the corresponding neoplastic cell (Ascites thymoma) and on Ehrlich ascites tumour cells. The A23187-induced total cell calcium increase in normal thymocytes takes place both in control and energy-depleted cells, while it is lacking in neoplastic cells. In addition the ionophore stimulates aerobic glycolysis of normal thymocytes, whereas it is ineffective on neoplastic cells. The study of intracellular calcium exchange properties reveals that in normal cells the ionophore A23187 provokes a 60% increase of the exchangeable pool together with a more significant, 4-fold enlargement of the unexchangeable pool. These effects are lacking in cancer cells. The data give rise to interesting considerations concerning the regulation and compartmentalization of calcium in neoplastic cells. The results will be also discussed in relation to the models that predict altered cell calcium metabolism as a cause of cancer cell high aerobic glycolysis and uncontrolled growth.

Animals↗
PubMed1981

Further observations on calcium and other divalent cations metabolism in intact Ehrlich ascites tumour cells.

The metabolism of calcium has been investigated in the Ehrlich Ascites Tumour Cells (ATC). ATC extrude Ca2+ actively by an energy-dependent mechanism, supported by both respiration and glycolysis. Extrusion takes place even against a very steep concentration gradient (10 mM Ca2+). Cell calcium content is decreased by monovalent cations (Na+,K+ and Li+), which act independently from their metabolic effects. La3+ inhibits ATC Ca2+ extrusion whereas Ruthenium Red slightly decreases cell calcium content. The antibiotic ionophore A 23187 strongly increases ATC Ca2+ level. the metabolism of other divalent cations (Mg2+, Sr2+ and Mn2+) has been studied. Mg2+ does not show appreciable changes in the various metabolic conditions tested, while Mn2+ and Sr2+ behave quite differently from Ca2+, suggesting a different distribution of these cations in ATC. The experimental findings indicate that Ehrlich Ascites Tumour Cells regulate their calcium content by mechanisms related to plasma membranes while the size and activity of mitochondrial compartment is of minor importance.

Animals↗