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Biomedical subjects

M Dani

Publications and source records attributed to M Dani.

15 recordsLinked to original sources
PubMed2003

Novel antitenascin antibody with increased tumour localisation for Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT).

The Pretargeted Antibody-Guided RadioImmunoTherapy (PAGRIT) method is based on intravenous, sequential administration of a biotinylated antibody, avidin/streptavidin and (90)Y-labelled biotin. The hybridoma clone producing the monoclonal antitenascin antibody BC4, previously used for clinical applications, was found not suitable for further development because of the production of an additional, nonfunctional light chain. In order to solve this problem, the new cST2146 hybridoma clone was generated. The monoclonal antibody ST2146, produced by this hybridoma, having the same specificity as BC4 but lacking the nonfunctional light chain, was characterised. ST2146 was found able to bind human tenascin at an epitope strictly related, if not identical, to the antigenic epitope of BC4. It showed, compared to BC4, higher affinity and immunoreactivity and similar selectivity by immunohistochemistry. Biodistribution studies of biotinylated ST2146 and three other monoclonal antitenascin antibodies showed for ST2146 the highest and more specific tumour localisation in HT29-grafted nude mice. On the overall, ST2146 appears to be a good alternative to BC4 for further clinical development of PAGRIT.

Animals↗
PubMed2001

Designing multidrug-resistance modulators circumventing the reverse pH gradient in tumours.

Multidrug-resistant tumours often exhibit a reverse pH gradient (acid outside), as they have an acid extracellular pH (pHe) and a neutral alkaline intracellular pH (pHi). This study was designed to test the hypothesis that the ability of lipophilic drugs to mediate multidrug resistance (MDR) reversal by interacting with the membrane phospholipids may be correlated with pH in resistant tumours. The permeation properties of five MDR modulators were therefore studied at 37 degrees C by quantifying their ability to induce the leakage of Sulfan blue through unilamellar anionic liposomes, over the range pH 6.5-7.7, and in the absence of any membrane potential (pHe = pHi). The dye leakage induced by two calcium blockers (diltiazem and verapamil) and two antiparasitic agents (thioacridine derivative and mepacrine) was found to significantly increase with the pH of the medium (P < 0.001), whereas that induced by a non-ionic detergent (Triton X-100) showed almost no pH-dependent variations. This process was a cooperative one (0.8 < Hill coefficient < 8.5) and the permeation doses inducing 50% dye leakage (PD50) ranged from 1.6 to 36.0 mM. The permeation ability of the MDR modulators (log(1/PD50)) significantly increased with their octanol-buffer distributions (logD) (slope = 0.35+/-0.06; y intercept = 1.65 +/- 0.14; P < 0.0001) and significantly decreased with their net electric charge (z) (slope = -0.48+/-0.07; y intercept = 2.85+/-0.08; P < 0.0001). A highly significant multiple correlation was found to exist between the variations of log(1/PD50) with those of logD and z (dlog(1/PD50)/dlogD = 0.21 +/- 0.05; dlog(1/PD50)/dz = -0.34+/-0.07; y intercept = 2.27+/-0.17; P < 0.000001). The results provide evidence that in resistant tumours (acid pHe and neutral alkaline pHi), the MDR reversal might be enhanced by favourable drug-membrane interactions if the modulators are designed in the form of highly lipophilic (logP approximately equals 4) mono-basic drugs with a near neutral pKa (pKa approximately equals 7-8).

Antineoplastic Agents↗
PubMed2000

Thermal dependence of multidrug-resistant-modulator efficiency: a study in anionic liposomes.

This study was designed to test the hypothesis that there exists a correlation between the ability of lipophilic drugs to mediate the reversal of multidrug-resistance (MDR) by interacting with the membrane phospholipids and the metabolic level in tissues. The permeation properties of five MDR-modulators were studied by quantifying their ability to induce the leakage of Sulphan blue through unilamellar liposomes, over the temperature range 27-42 degrees C. The dye leakage induced by a non-ionic detergent (Triton X-100), two calcium blockers (diltiazem and verapamil) and two antiparasitic agents (thioacridine derivative and mepacrine) was temperature-dependent. The permeation process was a co-operative one (1.1 < Hill coefficient < 7.5) and the permeation doses inducing 50% dye leakage (PD50) were 1.5 - 14.9 mM. The permeation ability of the MDR-modulators (log(1/PD50)) decreased significantly as the net electric charge (z) increased. The passive dye leakage (deltaG < 0) was found to be an endothermic process (deltaH > 0), favoured by an increase in the membrane disorder (deltaS > 0). The apparent enthalpy factor (deltaH50) associated with 50% dye leakage increased with the net electric charge of the compound, and this energetically non-favoured event was entirely offset by the concomitant increase in the entropy factor (deltaS50). The apparent permeation enthalpy (deltaH50) and entropy (deltaS50) showed the lowest values for Triton X-100 (deltaH50 = 7.1 +/- 0.53 kJ mol(-1), deltaS50 = 76.9 +/- 1.86 Jmol(-1) K(-1)), and the highest values for mepacrine (deltaH50 = 79.5 +/- 3.80 kJmol(-1), deltaS50 = 306.7 +/- 5-97 J mol(-1) K(-1)). When the temperature was increased from 27 to 42 degrees C, the apparent Gibbs free energy (deltaG50) of the dye leakage induced by Triton X-100 decreased by less than 10% of the initial value, and that induced by mepacrine decreased by nearly 40%. The results provide evidence that in tissues with high metabolic levels and therefore high temperatures, MDR-reversal is likely to be enhanced via favourable drug-membrane interactions controlled by the electric charge of the modulators.

Anions↗
PubMed1994

Carnitine-dependent changes of metabolic fuel consumption during long-term treatment with valproic acid.

Energy metabolism was measured in children receiving long-term treatment with valproic acid. In 8 of 10 randomly selected subjects, the resting respiratory quotient was higher than in age- and sex-matched control subjects (0.91 +/- 0.01 vs 0.87 +/- 0.01; p < 0.05). A shift was observed in fuel consumption, and a significant reduction was found in the amount of fats oxidized (0.68 +/- 0.23 vs 1.18 +/- 0.18 gm.kg-1.day-1), which was accompanied by increased utilization of carbohydrates (5.31 +/- 0.79 vs 3.81 +/- 0.39 gm.kg-1.day-1) in comparison with the control subjects. The resting total energy expenditure was not affected by the treatment. The children with an altered energy consumption pattern (n = 8) received carnitine supplementation for a month; the respiratory quotient then decreased (0.87 +/- 0.02), the oxidation of fats increased (1.42 +/- 0.25), and the consumption of carbohydrates decreased (3.87 +/- 0.79), but no changes in resting energy expenditure were observed. We conclude that carnitine depletion, a known adverse effect of valproic acid administration, may result in inhibited fatty acid oxidation, leading to a shift of substrates utilized from fats to carbohydrates.

Adolescent↗
PubMed1993

Mapping quantitative trait loci (QTLs) for resistance to Gibberella zeae infection in maize.

The basic prerequisite for an efficient breeding program to improve levels of resistance to pathogens in plants is the identification of genes controlling the resistance character. If the response to pathogens is under the control of a multilocus system, the utilization of molecular markers becomes essential. Stalk and ear rot caused by Gibberella zeae is a widespread disease of corn: resistance to G. zeae is quantitatively inherited. Our experimental approach to understanding the genetic basis of resistance to Gibberella is to estimate the genetic linkage between available molecular markers and the character, measured as the amount of diseased tissue 40 days after inoculation of a suspension of Fusarium graminearum, the conidial form of G. zeae, into the first stalk internode. Sensitive and resistant parental inbreds were crossed to obtain F1 and F2 populations: the analysis of the segregation of 95 RFLP (restriction fragment length polymorphism) clones and 10 RAPD (random amplified polymorphic DNA) markers was performed on a population of 150 F2 individuals. Analysis of resistance was performed on the F3 families obtained by selfing the F2 plants. Quantitative trait loci (QTL) detection was based either on analysis of regression coefficients between family mean value and allele values in the F2 population, or by means of interval mapping, using MAPMAKER-QTL. A linkage map of maize was obtained, in which four to five genomic regions are shown to carry factors involved in the resistance to G. zeae.

Chromosome Mapping↗
PubMed1989

Nucleotide sequence of cDNA coding for saporin-6, a type-1 ribosome-inactivating protein from Saponaria officinalis.

We have isolated and sequenced partial cDNA clones that encode SO-6, a ribosome-inactivating protein from Saponaria officinalis. A cDNA library was constructed from the leaves of this plant and screened with synthetic oligonucleotide probes representing various portions of the protein. The deduced amino acid sequence shows the signal peptide and a coding region virtually accounting for the entire amino acid sequence of SO-6. The sequence reveals regions of similarity to other ribosome-inactivating proteins, especially in a region of the molecule where critical amino acid residues might participate in the active site.

Amino Acid Sequence↗
PubMed1988

A highly conserved repetitive DNA sequence, (TTAGGG)n, present at the telomeres of human chromosomes.

A highly conserved repetitive DNA sequence, (TTAGGG)n, has been isolated from a human recombinant repetitive DNA library. Quantitative hybridization to chromosomes sorted by flow cytometry indicates that comparable amounts of this sequence are present on each human chromosome. Both fluorescent in situ hybridization and BAL-31 nuclease digestion experiments reveal major clusters of this sequence at the telomeres of all human chromosomes. The evolutionary conservation of this DNA sequence, its terminal chromosomal location in a variety of higher eukaryotes (regardless of chromosome number or chromosome length), and its similarity to functional telomeres isolated from lower eukaryotes suggest that this sequence is a functional human telomere.

Base Sequence↗
PubMed1987

Effect of chronic valproic acid treatment on plasma and urine carnitine levels in children: decreased urinary excretion.

Plasma levels and urinary carnitine excretion rates were determined in children treated with valproic acid (n = 11) and in age and sex matched controls (n = 11). Urine was collected throughout two consecutive 24 h periods in both groups, and blood samples were taken on the first day of collection after an overnight fast. The plasma level of total and free carnitine was significantly lower in the treated group (24.3 +/- 2.2 vs 34.9 +/- 2.4 and 16.8 +/- 1.8 vs 26.5 +/- 2.1 nanomol/ml; values are means +/- SEM), while there was no significant alteration in the acylcarnitine fraction. In the treated group of children a significant reduction was found in the plasma beta-hydroxybutyrate level indicating a limited fatty acid utilization (23.2 +/- 2.5 vs 81.9 +/- 7.8 nanomol/ml). Urinary total and free carnitine decreased from 286.4 +/- 57.8 to 120.8 +/- 18.2 and from 154.3 +/- 33.6 to 21.2 +/- 5.8 mumol/day, respectively; the acyled fraction was not significantly reduced. In one child, urinary carnitine excretion was followed during the first ten days of treatment. After the 2nd day a decrease of the total and free fraction was observed, confirming previous data obtained during chronic VPA treatment. It has been concluded that the decreased plasma carnitine associated with chronic VPA treatment is not a result of an increased excretion rate, but more likely the consequence of a relatively insufficient endogenous carnitine synthesis. The decreased plasma BOB level probably due to limited fatty acid utilization might also be a metabolic consequence of depressed carnitine concentration.

Adolescent↗
PubMed1985

Plasmid pFCE4: a new system of Escherichia coli expression-modification vectors.

Two versatile expression-modification vectors were obtained by inserting the origin of replication (ori) of phage f1 into the expression vector pOTS. The resulting plasmids produce large amounts of coding or noncoding ssDNA (depending on ori orientation in pFCE4+ and pFCE4-) and excrete it into the medium as virus-like particles following infection with phage f1. These features make them suitable for dideoxy chain termination sequencing, oligonucleotide directed mutagenesis and gene expression without further manipulations. The human IFN alpha-2 gene, lacking the codon for the first amino acid, cysteine, was efficiently expressed by these vectors.

Base Sequence↗
PubMed1982

Calorimetric study of the binding reaction of concanavalin A with immunoglublins.

The thermal effects associated with the binding reaction of concanavalin A and immunoglobulins have been measured by calorimetry. IgG solutions do not generate heat on mixing with concanavalin A, confirming the low reactivity of IgG for the lectin molecule. IgM solutions, on the other hand, show a substantial enthalpic contribution of delta H = -24 +/- 1kJ/site for the binding reaction between the polysaccharide chains of human IgM macroglobulin and concanavalin. A stoichiometry is 10 monovalent concanavalin A molecules per intact macroglobulin and two bivalent concanavalin A molecules per two sites on the heavy chain of the reduced macroglobulin subunit.

Calorimetry↗
PubMed1981

Calorimetric study of concanavalin A binding to saccharides.

The binding of concanavalin A in the dimer form to various saccharides has been studied by calorimetry, and estimates of the binding enthalpy and binding constants have been calculated. Methyl alpha-D-mannoside and methyl alpha-D-glucoside have a -- delta H0 of 21.5 and 11.5 kJ/mol, respectively, at both pH 4 and 4.5. The p-nitrophenyl derivatives react with enthalpic values of 15.6 and 14.6 kJ/mol. The galactosepyranosides show no heat effects during mixing with the protein solutions. The apparent binding enthalpies calculated from the variations of the equilibrium constants with temperature are in good agreement with the values measured experimentally. The two binding sites of the dimer form of concanavalin A are equal and independent, and the low enthalpies obtained do not justify a large conformational change during the reaction. The binding reaction has also been estimated for other sugars normally contained in glycoproteins.

Binding Sites↗