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L Leoni

Publications and source records attributed to L Leoni.

52 records · Page 3Linked to original sources

Folding of single-stranded DNA on the histone octamer.

A complex between the single-stranded DNA of the bacteriophage M13 and the histone octamer was analyzed by electron microscopy, low-angle X-ray diffraction and nuclease analysis. The morphology and the diffraction pattern of the complex strongly resemble those of the nucleosome. These results, as well as the finding of a protected DNA fragment about 100 nucleotides long following single-stranded DNA specific nuclease digestion, indicate that 'a nucleosome-like' complex can be formed between single-stranded DNA and the histone octamer. Competition experiments suggest that under physiological conditions the histone octamer is transferred from single- to double-stranded DNA.

Chromatin↗

Parenteral nutrition of injured patients: Effect of manipulation of aminoacid infusion (increasing branched chain while decreasing aromatic and sulphurated aminoacids).

Sixteen critically ill injured patients received parenteral nutrition providing nitrogen (0.34 g kg(-1) day(-1)) and glucose (32 kcal kg(-1) day(-1)) for 5 days. They were randomly divided into two groups with respect to aminoacid supply: an essential aminoacid solution vs the same solution enriched in branched chain amino acid (BCAA) content and decreased in phenylalanine and methionine content (mean BCAA intake, 0.55 vs 0.69 g kg(-1) day(-1)). Basal values of nitrogen metabolism without treatment showed no difference between the two groups. Nitrogen losses and 3 methylhistidine (3-MEH) excretion were elevated; the plasma aminoacid pattern was altered by the trauma and except for phenylalanine, aspartate and glutamate, plasma aminoacid concentrations were decreased below normal values. Net muscular aminoacid output was demonstrated by femoral arterio-venous (av) differences that were all negative except for glutamate and citrulline TPN with both solutions improved the nitrogen balance and reduced the negative aminoacid balance across the leg. Adjusting a TPN regimen to increase the BCAA content without altering the total nitrogen infused, had no effect on overall nitrogen balance, but exerted a beneficial effect on body protein catabolism, as assessed by the urinary 3-MEH excretion rate, and a transient improvement in the aminoacid balance across the leg at the peak of the infusion. The short-lived effect of BCAA suggest a metabolic effect of these aminoacids which deserves further study.

Journal Article↗

The main determinants of nitrogen balance during total parenteral nutrition in critically ill injured patients.

Factors influencing nitrogen balance during total parenteral nutrition have been investigated in 34 critically ill injured patients studied during the first 6 days after trauma. Basal nitrogen balance was severely negative (-0.26 +/- 0.12 (SD) g X kg-1), but improved consistently during treatment. Nitrogen intake proved to be the major determinant of a positive, or less negative, nitrogen balance, only secondarily followed by total energy intake corrected to predicted basal energy expenditure, according to multiple regression analysis. The amount of non-protein calories and the non-protein calorie to nitrogen ratio appeared to have little significance on nitrogen balance, when corrected for the two former variables.

Adolescent↗

Nucleosome "phasing" and cruciform structures in circular supercoiled pBR322 DNA.

Cruciform structures have been detected in pBR322 supercoiled DNA, both in its naked state and when complexed with histone octamer, using S1 endonuclease cleavage and EcoRI restriction. An inspection of the DNA sequence shows that the S1-hypersensitive sites are very near to AT-rich regions of pBR322 genome. A nucleosome "phasing" in these regions, as found on AT-rich regions of SV40 DNA (15), has been shown by restriction enzymes analysis. On the basis of these results it can be proposed that cruciform structures protrude on the nucleosome surface. This model explains the reason why these structures, which need high superhelical density, can exist in supercoiled DNA partially relaxed by nucleosome formation.

Base Sequence↗

Interactions of the histone octamer with single-stranded DNA. Sedimentation analysis and low-angle X-ray diffraction.

A complex between 140-160 nucleotide single-stranded DNA and the octamer of histones was formed and analyzed by electron microscopy and X-ray low angle diffraction. The morphology of the complex is very similar to that of the nucleosome; the diffraction pattern appears less defined than for chromatin showing broader maxima in the same positions. These results strongly suggest that this particle has a geometry very similar to that of the fundamental subunit of chromatin. The possibility of artifacts due to renaturation reaction promoted by histones is ruled out by the analysis of the complex with S1 nuclease and by the formation of a 'nucleosome like' particle using poly(dT) instead of DNA. Association of the histone octamer with either the 140-160 nucleotide single-stranded DNA or the 140-160 bp double-stranded DNA was evaluated at different histone/DNA input ratios. In both cases, the formation of the complex appears to be regulated by comparable association constants, and in both cases the trend of the complexation reaction in function of the temperature is almost the same. These results suggest that an alternative binding of the histone octamer to double-stranded or to single-stranded DNA requires low energy charge and may be involved in the processes of replication and transcription of the 'active chromatin'.

Animals↗

Heterogeneity of mitochondrial DNA from Saccharomyces cerevisiae and genetic information for tRNA.

Mitochondrial DNA from wild-type Saccharomyces cerevisiae and from an "extreme" petite mutant were analyzed by hybridization of several tRNAs on DNA fragments of different buoyant density, obtained by sonication and fractionation on a CsCl gradient. The hybridization patterns show that the genes for tRNAser, tRNAphe, tRNAhis, tRNAval, tRNAileu are present on wild-type mitochondrial DNA, while only genes for tRNAser and tRNAhis are present on petite mitochondrial DNA; moreover the hybridization patterns indicate that these genes are not clustered and suggest that more than one gene might exist for tRNAser and tRNAhis.

Centrifugation, Density Gradient↗

Influence of total parenteral nutrition on protein metabolism following acute injury: assessment by urinary 3-methylhistidine excretion and nitrogen balance.

The use of total parenteral nutrition after acute injury, either surgical or accidental, is widely accepted for its important benefits, although it is not yet completely understood whether a reduction of body protein catabolism can be effectively achieved. We applied total parenteral nutrition to 14 critically ill patients after either trauma or major surgery. Their daily nitrogen balance, urinary 3-methylhistidine and creatinine excretion, and molar 3-methylhistidine/creatinine ratio, during initial 24-hr fasting period, were respectively -0.19 +/- 0.01 (SEM) g kg-1, 5.46 +/- 0.47 mumol kg-1, 27 +/- 4 mg kg-1, and 0.030 +/- 0.005. Daily nonprotein calorie intake of 31.11 +/- 0.58 kcal kg-1, as glucose, and administration of nitrogen 0.350 +/- 0.004 g kg-1, as 10% crystalline L-aminoacids solution, and insulin 1 IU every 5.03 +/- 0.14 g of glucose, resulted in progressive decline of urinary 3-methylhistidine (4.21 +/- 0.43 mumol kg-1, p less than 0.001), creatinine (22 +/- 2 mg kg-1, NS), and their molar ratio (0.022 +/- 0.002, NS). Mean nitrogen balance resulted in 0.032 +/- 0.008 g kg-1. Since urinary 3-methylhistidine role as a marker of protein catabolism is well established, its decrease under total parenteral nutrition together with greatly improved nitrogen balance, demonstrates that our treatment can effectively quench protein catabolism, meanwhile enhancing protein synthesis.

Adult↗