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C Nicolini

Publications and source records attributed to C Nicolini.

At least 37 records · Page 2Linked to original sources
PubMed1996

Room-temperature single-electron junction.

The design, realization, and test performances of an electronic junction based on single-electron phenomena that works in the air at room temperature are hereby reported. The element consists of an electrochemically etched sharp tungsten stylus over whose tip a nanometer-size crystal was synthesized. Langmuir-Blodgett films of cadmium arachidate were transferred onto the stylus and exposed to a H2S atmosphere to yield CdS nanocrystals (30-50 angstrom in diameter) imbedded into an organic matrix. The stylus, biased with respect to a flat electrode, was brought to the tunnel distance from the film and a constant gap value was maintained by a piezo-electric actuator driven by a feedback circuit fed by the tunneling current. With this set-up, it is possible to measure the behavior of the current flowing through the quantum dot when a bias voltage is applied. Voltage-current characteristics measured in the system displayed single-electron trends such as a Coulomb blockade and Coulomb staircase and revealed capacitance values as small as 10(-19) F.

Journal Article↗
PubMed1996

Potentiometric alternating biosensing system for in continuum toxicity tests on cell population.

In this paper, we investigate the possibility to monitor, in continuum, changes of cellular metabolism. Examples of this feature are shown: metabolic changes due to drugs administration are evaluated as a decrease of extracellular acidification rate on CHO-K1 and HeLa cells. Parallel experiments with a standard test were performed to validate the results obtained with the biosensor.

Animals↗
PubMed1996

Effect of fixatives on calf thymocytes chromatin as analyzed by 3D high-resolution fluorescence microscopy.

Two common fixatives--glutaraldehyde and ethanol/acetic acid mixture--were studied to understand their effects on DNA distribution inside the cell nucleus. Native calf thymocytes were analyzed by using a DNA selective fluorescent dye (DAPI) and computational optical sectioning microscopy on isolated cells before and after fixation. In order to estimate quantitatively the intranuclear DNA distribution, the stained calf thymocytes images were processed by removing the out-of-focus contributions present in each optical section. Although preliminary, the results show that within individual nuclei the frequency distribution of the fluorescence intensity appears significantly and differentially altered by the two fixatives. Namely with respect to the native unfixed preparation, the ethanol/acetic acid causes the complete disappearance of the higher intensity pixels, whereas glutaraldehyde fixation can be associated with the appearance of new ones of an even higher intensity. The quantitative analysis of the processed images allowed us to reconstruct the topological distribution of DNA inside the nucleus and to correlate the data with the results obtained by differential scanning calorimetry on similar samples.

Acetates↗
PubMed1996

Langmuir-Blodgett films of photosensitive proteins.

The striking properties of monolayers and multilayers of photosensitive proteins obtained by using the Langmuir-Blodgett technique are described. The close packing of the protein molecules, which preserve most of the properties found in solution, seems to be the main cause for their thermal stability, which in some cases reached a temperature of 200 degrees C without the loss of the protein secondary structure. The review is focused on three of the most intensively studied photosensitive proteins, namely photosynthetic reaction centres, bacteriorhodopsin and bovine rhodopsin, and on their possible applications as molecular optical devices.

Bacteriorhodopsins↗
PubMed1996

Cytochrome P450scc spin state transitions in the thin solid films.

Langmuir-Blodgett films of cytochrome P450scc were prepared on the solid supports and their spectral properties were investigated. Being immobilized, hemoprotein changes its spin state from initially high to low spin. This transition is reversible since after the solubilization of hemoprotein, the spin state equilibrium is shifted towards high-spin state. Anaerobic reduction of film incorporated cytochrome P450scc by electron transfer chain (NADPH-->adrenodoxin reductase-->adrenodoxin) revealed the low rate of the reaction that coincides well with the content of the hemoprotein low-spin form. We suggest that particularly regular orientation of solid cytochrome P450scc are of crucial importance for this phenomenon.

Cholesterol↗
PubMed1995

From neural chip and engineered biomolecules to bioelectronic devices: an overview.

At the first C.E.C. Workshop, in Brussels on 28-29 November 1991, attended by over 70 leading European scientists and industrialists, bioelectronics was defined as 'the use of biological materials and biological architectures for information processing systems and new devices'. At the end of the Frankfurt Workshop, bioelectronics, specifically bio-molecular electronics, was described as 'the research and development of bio-inspired (i.e. self-assembly) inorganic and organic materials and of bio-inspired (i.e. massive parallelism) hardware architectures for the implementation of new information processing systems, sensors and actuators, and for molecular manufacturing down to the atomic scale'. The subject of this overview is to summarize some of the most significant progress in bio-molecular electronics from neural VLSI networks and bio-molecular engineering. As an example of one possible route, emphasis is placed on the results recently obtained within this laboratory.

Biofilms↗
PubMed1995

A silicon-based biosensor for real-time toxicity testing in normal versus cancer liver cells.

The potentiometric alternating biosensor (PAB) system has been utilized to monitor the effects on normal versus cancer cells of Mitoxantrone, a potent inhibitor of nucleic acid synthesis commonly utilized in the clinical treatment of hepatocarcinomas. In this first series of toxicity tests, we have utilized a primary culture of rat hepatocytes and a stabilized line named HepG2 from a human hepatoma. Primary cultures of hepatocytes represent a unique tool in toxicology/pharmacology as shown in previous works. A suitable microvolume flow chamber has been designed and produced. The chamber is equipped with inlet and outlet circuits and with a fixed transducer (Si/SiO2/Si3N4 chip) facing a cover slip on which cells grow. The PAB system is here utilized only in the pH-sensitive configuration with a single measuring spot, warranting an accurate determination of the change in the extracellular acidifcation rate resulting from drug administration. We have also observed the changes in the acidification rate of 3T6 mouse fibroblasts, induced by a treatment with Ara-C, a well-known DNA antimetabolite. New insights can be obtained from these studies into the drug response of normal versus cancer cells and into the feasibility of real-time PAB monitoring of drug toxicity and efficacy, towards effective human cancer treatment. The effect of increasing drug concentrations on cellular metabolism is here compared with the results obtained from conventional tests (optical microscopy, Neutral Red and Trypan Blue assays).

Animals↗
PubMed1995

Characterization of Langmuir-Blodgett films of rhodopsin: thermal stability studies.

Two-dimensional close packing of purified bovine rhodopsin, made by the Langmuir-Blodgett technique, was characterized by small angle x-ray scattering and nanogravimetric measurements. The area occupied by a molecule of rhodopsin in the film was approximately 1100 Angstrum2 and the periodicity of the layers resulted in 59 Angstrum. The circular dichroism measurements showed that bleached rhodopsin in Langmuir-Blodgett film had high thermal stability, in fact, reaching a temperature of 150 degrees C without a loss of the secondary structure. Moreover, when the film was made up in the dark, rhodopsin maintained its stability up to at least 200 degrees C and its characteristic absorbance peak at 500 nm up to about 90 degrees C.

Animals↗
PubMed1995

Polarized light scattering: a biophysical method for studying bacterial cells.

We outline a method which uses differentially polarized light scattering to study the properties of bacterial cell suspensions, i.e., spores of Bacillus Subtilis. The identification of bacterial cells of different strains, with and without plasmid insertion, was performed by means of differential polarization light scattering (DPLS). The samples displayed distinct angular behaviors for the S14 and S34 normalized scattering parameters of the Mueller matrix depending on the strain and on a plasmid insertion in the chromosomal unit. These experiments, performed blindly, point out the possibility of achieving real time identification of micro-organisms by DPLS spectrometry.

Bacillus subtilis↗
PubMed1994

Ethidium bromide intercalation and chromatin structure: a spectropolarimetric analysis.

Native chromatin has been characterized at different ionic strengths, according to different methods of preparation, by means of circular dichroism and ethidium bromide intercalation. For a more precise interpretation of these results, a series of analogous experiments on phagic DNA, as a function of its circularization, and on mononucleosomes have been performed. Results are discussed here in terms of high order chromatin structure, DNA supercoiling, and topological constraints.

Animals↗
PubMed1993

Thermal stability of protein secondary structure in Langmuir-Blodgett films.

The temperature dependence of the secondary structure of photosynthetic reaction centres from Rhodobacter sphaeroides in solution and in Langmuir-Blodgett film was studied by circular dichroism. It was shown that the secondary structure of the protein was not affected in Langmuir-Blodgett films by heating up to 200 degrees C, while in solution it was completely lost at 55 degrees C. Molecular order rather than decreased hydration degree was held responsible.

Circular Dichroism↗
PubMed1993

DNA internal motions within nucleosomes during the cell cycle and as a function of ionic strength.

We have used 31P NMR spectra to show that DNA internal motions are greatly hindered within oligonucleosomes. The fluctuations seem to be a function of both the cell cycle and the number of nucleosomes interlinked. Namely, the resonance areas, directly related to unbound phosphate, are consistently smaller in M-phase than in S-phase; at the same time, the resonance line width, inversely related to base plane, deoxyribose, and phosphate internal motions, is consistently larger in mononucleosomes than in oligonucleosomes. In all cases, the removal of chromosomal proteins, by a progressive increase of ionic strength up to 2 M NaCl, increases the internal motion, as monitored by a decrease in line width toward that of free DNA. While for both oligo- and mononucleosomes in S-phase the decrease in line width is strictly correlated to a sharp increase in resonance area, in M-phase it is not, with the 31P resonance area rather low even at 2.0 M NaCl extraction. Similarly, while S-phase 31P line widths steadily grow from mono- to oligonucleosomes, in M-phase they do not. Moreover, the increase of the ionic strength to 0.6 M NaCl, as compared to 0.35, 1.2, and 2 M NaCl, displays significant variations on 31P line width and resonance area, independent of the cell cycle phase and the number of nucleosomes interlinked. These observations agree with earlier suggestions on the differential role of the various chromosomal protein subfractions, known to preferentially dissociate at the different ionic strengths in question, in the sealing of mononucleosomes and in the overall stability of polynucleosomes.

Cell Cycle↗
PubMed1992

Fluorescence cytometry of microtubules and nuclear DNA during cell-cycle and reverse-transformation.

Synchronized CHO-K1 cells and their dibutyryl c-AMP treated counterparts have been characterized by means of static and flow fluorescence cytometry at the level of nuclear DNA and cytoplasmic microtubules. In order to confirm earlier findings on synchronized population, Carnoy fixed and hydrolyzed, several new findings are here reported at the level of single intact cell. The fluorescence intensity of DAPI-stained glutaraldehyde fixed 2C cells correlates well with the average absorbance of the corresponding Feulgen-stained cells, thereby appearing also to be a measure of chromatin condensation during the G1 phase. In the early part of G1, the drastic alteration in anti-beta tubulin immunostaining is shown to parallel microtubule depolymerization induced by calcium or colcemide. The known 1-2 h lengthening of the G1 period after reverse-transformation appears to correlate with a similar delay in the abrupt chromatin decondensation. The above results are discussed in terms of the role of microtubules and nuclear morphometry (and their coupling) in the control of cell cycle progression of transformed vs. fibroblast-like cells.

Animals↗
PubMed1991

Changes in DNA superhelical density monitored by polarized light scattering.

Linear and circular lambda-DNA at different ethidium bromide concentrations have been studied by means of polarized light scattering, namely the S14, S34, S33 and S13 elements of Mueller matrix. While S33 at low angle appears well correlated with the total light scattering evaluated by optical density measurements at 632.8 nm for linear and circular DNA of the same mass, the magnitude and slope of the S14, S34 and S13 signals display significant changes for the circular lambda-DNA depending on the degree of negative superhelical density as induced by the different ethidium bromide concentrations. At the same time, for linear lambda-DNA the signal remains invariant, making explicit for the differential scattering of polarized light the possibility to obtain additional information by its angular dependence. Strikingly also the effect of 0.2% glutaraldehyde versus ethanol fixation on the native lambda-DNA structural properties appears to confirm earlier findings by other well-established probes. Results are discussed in terms of first physical principles and of their potential bearings towards our understanding of the mechanism controlling gene expression.

Bacteriophage lambda↗
PubMed1991

A qualitative approach to cell growth modeling and simulation for cancer chemotherapy.

A qualitative model of cell growth, based on qualitative process theory, is presented. The model can be used to analyze the effects of the interaction of antiproliferative drugs on cells when the effects of each specific drug are known, mainly when designing multi-drug protocols for optimal cancer treatment.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1991

Polarized light scattering of nucleosomes and polynucleosomes--in situ and in vitro studies.

Nucleosomes, chromatin and nuclei, extracted from rat hepatocytes, are studied by a new "in house" experimental configuration which measures circular intensity differential scattering (CIDS) and other elements of the polarized light scattering matrix. The Mueller matrix elements, S14 and S34, that are related to the geometric parameters of the superhelical arrangement of polynucleosomes point to the existence of a quaternary structure at low ionic strength for chromatin prepared by the cold-water method, which is lost by shearing, and is not found in the soluble chromatin prepared through the nuclease method. Only salt addition to a final concentration of 5 mM MgCl2, 150 mM NaCl and 10 mM Tris HCl (pH 7) yields a sizeable (S14 + S34) signal in the latter chromatin, which is however still different from the corresponding signal of native nuclei and of "cold-water" chromatin. Comfortingly, the (S14 + S34) signal from isolated nucleosomes is consistently very low (nearly zero) as predicted by multiple dipole simulation within the framework of classical electrodynamics. Results are discussed in terms of the topological constraints present in the native long chromatin fiber, which are lost after limited nuclease digestion and after shearing.

Animals↗
PubMed1990

The histone H1 globular region. A possible supersecondary structure from spectroscopic and statistical studies.

The central region of the basic nuclear protein, histone H1, has a highly conserved amino acid sequence and a globular structure which is still not known at atomic resolution. A possible secondary and supersecondary structure was predicted by combining experimental measurements of circular dichroism and NMR spectroscopy with a statistical method based on the amino acid sequence. Our results showed the protein fragment as being highly structured and having a total alpha-helix content of about 40%.

Amino Acid Sequence↗