PubMed Health⌕ Search

Biomedical subjects

L Leoni

Publications and source records attributed to L Leoni.

At least 37 records · Page 2Linked to original sources
PubMed1993

Lectins and anti-T monoclonal antibodies-induced changes of second messengers generating enzymes in human peripheral blood mononuclear cells.

A five min. incubation of peripheral blood mononuclear cells (PBMN) with either phytohaemagglutinin (PHA) or concanavalin A (ConA) resulted in distinct subcellular redistribution patterns of phosphatidylinositol 4,5-bisphosphate phospholipase C [PtdIns(4,5)P2-PLC] and myo-inositol 1,4,5-trisphosphate monophosphatase [Ins(1,4,5)P3-monophosphatase] activities. When compared to control cells, PHA-treated PBMN cells displayed a significant increase of PtdIns(4,5)P2-PLC and Ins(1,4,5)P3-monophosphatase relative specific activities in the nuclear fraction along with an increment (D) in enzyme amount of 6.5% and 7.3%, respectively. Incubation with B66.6, an anti-CD4 monoclonal antibody (Mab) which specifically activates CD4(+)-T cells in the absence of any other stimuli, also induced changes of these activities in the nuclear fraction, thus mimicking the effect of PHA observed in helper T cell subpopulation. No changes were detected after incubation of PBMN cells with the non mitogenic anti-CD4 MAb 101-69, or with an anti-CD3 MAb which activates T cells only in the presence of a second stimulus. On the other hand, after incubation with ConA, PtdIns(4,5)P2-PLC relative specific activity was enhanced in the microsomal fraction while the Ins(1,4,5)P3-monophosphatase activity increased in both nuclear and microsomal fractions and decreased in cytosol. An increment D of 4.6% and 10.9% for PtdIns(4,5)P2-PLC and Ins(1,4,5)P3-monophosphatase, respectively, was measured in the microsomal fraction. Only after three days of incubation with a mitogenic anti-CD2 MAb Lau-2.1.2, the PtdIns(4,5)P2-PLC activity increased in the particulate fraction of PBMN similar to ConA treatment.

Antilymphocyte Serum↗
PubMed1991

Influence of DNA topology and histone tails in nucleosome organization on pBR322 DNA.

Recently, we have found that the assembly of nucleosomes reconstituted on negatively supercoiled DNA is cooperative. In the present paper the role of DNA topology and of histone tails in nucleosome assembly was explored. Reconstituted minichromosomes on relaxed DNA at different histone/DNA ratios (R) were assayed by topological analysis and electron microscopy visualization. Both methods show a linear relationship between average nucleosome number (N) and R. This suggests that in the case of relaxed DNA, cooperative internucleosomal interactions are small or absent. The influence of histone tails in nucleosome assembly was studied on minichromosomes reconstituted with trypsinized histone octamer on negatively supercoiled DNA by topological analysis. The topoisomers distribution, after trypsinization, dramatically changes, indicating that nucleosome-nucleosome interactions are remarkably decreased. These results show that, in chromatin folding, in addition to the well known role of histone H1, the interactions between histone octamer tails and DNA are also of importance.

Chromatin↗
PubMed1991

Non random positioning of reconstituted nucleosomes on polyomavirus DNA.

Nucleosome positioning on linear polyomavirus DNA was evaluated by Fourier transform analysis of data obtained by electron microscopy visualization of reconstituted nucleosomes after photoreaction with trimethylpsoralen. Results show a non random nucleosome positioning and this implies that the histone octamer discriminates among various nucleotide sequences also in the very simple model system adopted in this study. This recognition process appears rather complex because of the limited correlation between nucleosome distribution and DNA curvature, suggesting that other interactions could play a role.

DNA, Viral↗
PubMed1990

Influence of parenteral nutrition on leg nitrogen exchange in injured patients.

Body N balance, 3-methylhistidine (MEH) excretion, amino acid (AA) plasma concentration, and fluxes across the leg were investigated both during fasting and during parenteral nutrition of injured patients in order to better understand protein-sparing mechanisms induced by metabolic support in the whole body and in skeletal muscle. Patients were randomized to receive 15 or 30 kcal/kg.day coupled with 0.30 g of N either with standard or branch-chain (BC)-enriched AA solutions. During fasting, patients were highly catabolic (N balance -14.7 +/- 1.2 g N/m2.day, MEH excretion 422 +/- 25 mumol/m2.day) and showed a high efflux of AA N from the leg (5.08 +/- 2.1 g N/m2.day) without difference between the groups. During treatment, body N balance (-5.55 +/- 0.88, p less than .001) and MEH excretion (284 +/- 20, p less than .001) were significantly reduced without difference among the groups; also, AA N leg efflux (2.64 +/- 0.47, p less than .001) was reduced. Moreover, considering the effect of calorie load, patients receiving 30 kcal/kg.day showed a lower efflux of total AA N and of some AA considered as markers of muscle protein catabolism, such as phe, lys, met, and glu. The main difference between solutions was in the efflux of BCAA; particularly, val and leu efflux was turned into uptake in the BCAA group. No significant difference among the groups was found in N balance and MEH excretion during treatment. In brief, muscle catabolism was reduced in an amount dependent on glucose and insulin load, but it was not influenced by BCAA supply. Whole body net protein catabolism was reduced through different mechanisms, either an increased visceral N retention or a decreased muscle N loss. However, muscle N loss was never abolished even in the high calorie groups.

Adolescent↗
PubMed1989

Cooperativity in nucleosomes assembly on supercoiled pBR322 DNA.

Many studies have shown that in reconstituted chromatin model systems, containing only purified DNA and histone octamer, nucleosomes can adopt well defined locations with respect to DNA nucleotide sequence. Recently, nucleosome-nucleosome interactions were suggested as one of the factors underlying preferential nucleosomes positioning. In the present paper this aspect has been studied by topological analysis and electron microscopy visualization of minichromosomes reconstituted at different histone/DNA ratios. Both methods suggest that cooperativity plays a role in nucleosomes formation. A linear cooperative model in which nucleosomes are formed on discrete sites with cooperative interactions occurring only between nearest neighbours allows to calculate the cooperative constant. The reported results show that basic interactions, which are of relevance in the process of chromatin folding, are present also in very simple model system.

Animals↗
PubMed1989

Specific interactions between DNA left-handed supercoils and actinomycin D.

The interactions between the natural cyclopentapeptide antibiotic actinomycin D (ACT) and circular pBR322 DNA have been studied by freezing the topological state of the DNA in the complex by topoisomerase I reaction. Both supercoiled and relaxed DNAs, in the complexes at low antibiotic/DNA base-pair ratios, showed a dramatic decrease in linking number that cannot be explained by taking into account only the generally accepted unwinding of 28 degrees for each ACT molecule bound. Recent results derived from the crystallographic analysis of the complex between GpC and ACT suggest that ACT could mediate non-covalent cross-links between distant sections of DNA. Bridges between ACT and different sections of the pBR322 double helix could also explain our results. Two-dimensional gel electrophoresis of ACT-relaxed pBR322 DNA complexes reveals that all supercoils induced by ACT are negative. Two models of the complexes which correspond to the stabilization of DNA crossing by one or two molecules of ACT are proposed. In both cases the ability of ACT to stabilize only DNA left-handed supercoils is derived from the chirality of ACT, when it interacts with DNA.

Animals↗
PubMed1989

[Effects of various caloric intakes on nitrogen metabolism of the body, the muscles and the viscera during total parenteral nutrition with aminoacidic solution enriched with branched-chain amino acids].

Nitrogen metabolism was studied in ten injured/septic patients by means of a two compartment model, differentiating muscle from non muscle (central) tissue. During fasting muscle tissue loses a consistent amount of aminoacids which is retained in part by central tissue, giving rise to a positive nitrogen balance, whilst three methyl histidine and body nitrogen output are elevated. Total parenteral nutrition (glucose 15 kcal kg-1 day-1, nitrogen 0.30 g kg-1 die-1) improved body nitrogen balance and three methyl histidine excretion, but did not affect significantly either muscle or central nitrogen balance. Increasing caloric support to 30 kcal kg-1 day-1 did not showed any further effect on body nitrogen balance and three methyl histidine, while it improved significantly vs basal muscle nitrogen balance, but did not affect central nitrogen balance.

Amino Acids, Branched-Chain↗
PubMed1988

Kinetic persistence of cruciform structures in reconstituted minichromosomes.

Cruciforms persist in reconstituted minichromosomes, as revealed by cleavage with specific nucleases and hybridization with synthetic oligonucleotides. Relaxation by topoisomerase I suggests that cruciforms are located mainly on internucleosomal DNA and that their persistence on minichromosomes may be due to kinetic effects. The analysis of the kinetic behaviour of cruciforms in minichromosomes shows a definite velocity of reabsorption with respect to stable cruciforms in supercoiled naked DNA. An explanation based on suppression of the untwisting of linker DNA due to adjacent nucleosomes is proposed.

Chromosomes↗
PubMed1988

Preferential positioning of nucleosomes on pBR322 as evaluated via Fourier transform of data from electron microscopy.

Nucleosome positioning on pBR322 DNA has been evaluated by electron microscopy visualization, after psoralen cross-linking. The distribution function of nucleosomes on pBR322 DNA has been calculated analyzing the data of the electron microscopy via Fourier transform. This function shows definite maxima, which indicate differential interactions of the histone octamer to different DNA sequences.

Binding Sites↗
PubMed1988

Essential and non-essential amino acid requirement in injured patients receiving total parenteral nutrition.

The metabolic derangements of injury are known to influence nitrogen (N) requirements whilst less is known about individual amino acid (AA) requirements. This study was designed to investigate prospectively N vs AA requirement in 36 injured patients treated with total parenteral nutrition (TPN). The non-protein caloric input was 30 kcal kg-1 day-1 and three AA solutions were assessed containing the same AAs but in different proportion. Overall N intake was set at 0.35 g N kg-1 day-1 for solution A and B and 0.24 g N kg-1 day-1 for solution C. Solution B was similar to A, both being enriched in branched chain AAs (BCAA: 0.69 g kg-1 day-1 in B compared with 0.55 g kg-1 day-1 in A) while decreased in aromatic and sulphurated forms (1.75 times the normal need). Solution C was designed to maintain a daily input of BCAA similar to A (0.52 g kg-1 day-1) but with the supply of aromatic and sulphurated AA between solutions A and B, the supply of other AAs (lysine, theonine, histidine, arginine, glycine) being dependent on the selected N intake. For all the essential AAs the supply was always greater than normal allowances. Increasing BCAA over 0.55 g kg-1 day-1 did not improve N balance when N intake was 0.35 g kg-1 day-1, whilst nutrition with solution C was unable to maintain N balance. Moreover we found indirect evidence that this N intake, 0.52 g kg-1 day-1 was more sparing than 0.37 g kg-1 day-1 of BCAA.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1986

Persistence of cruciform structure and preferential location of nucleosomes on some regions of pBR322 and ColE 1 DNAs.

Inverted repeats of pBR322 and ColE 1 DNAs have been analyzed for the presence of cruciform structures upon formation of nucleosomes, using S1, P1 and restriction enzyme analysis. In both cases the fraction of molecules showing nuclease-sensitive sites is unaffected by the DNA relaxation, owing to the formation of nucleosomes. A kinetic mechanism, based on the freezing of cruciform structures on the nucleosome surface or nearby, is proposed. This hypothesis is supported by a preferential location of nucleosomes at the DNA sequences containing the nuclease-sensitive sites, as indicated by restriction enzyme analysis and electron microscopy visualization after psoralen cross-linking.

DNA Restriction Enzymes↗
PubMed1986

Main nitrogen balance determinants in malnourished patients.

Factors influencing nitrogen balance during total parenteral nutrition have been investigated in 38 malnourished patients studied for a cumulative period of 280 days. According to multiple regression analysis, nitrogen intake (0.213 +/- 0.004 g kg-1 day-1, mean +/- SD) proved to be the major determinant of a positive nitrogen balance (0.018 +/- 0.004 g kg-1 day-1), followed by non-protein energy intake (43.3 +/- 0.5 kcal kg-1 day-1). Total calorie intake to predicted basal energy expenditure and non protein calorie to nitrogen ratios appeared to have little significance on nitrogen balance, when corrected for the two former variables.

Female↗
PubMed1986

Parenteral nutrition of injured patients.

Injured patients treated with fructose 1,6-diphosphate (1 millimole of phosphate per kilogram per day) together with parenteral nutrition had a better nitrogen balance than patients treated with isocaloric nutrition and an inorganic source of phosphate. Excretion of 3-methylhystidine was similar while tyrosine and alanine output from the extremities was lower in the group of patients given fructose 1,6-diphosphate. The data indicates that the protein sparing action of fructose 1,6-diphosphate is exerted through an increased protein synthesis.

Abdominal Injuries↗