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Biomedical subjects

E Sorrentino

Publications and source records attributed to E Sorrentino.

16 recordsLinked to original sources

DNA fragments induction in human fibroblasts by radiations of different qualities.

Experimental data on DNA double strand break (DSB) induction in human fibroblasts (AG1522), following irradiation with several radiation qualities, namely gamma rays, 0.84 MeV protons, 58.9 MeV u(-1) carbon ions, iron ions of 115 MeV u(-1), 414 MeV u(-1), 1 GeV u(-1), and 5 GeV u(-1), are presented. DSB yields were measured by calibrated Pulsed Field Gel Electrophoresis in the DNA fragment size range 0.023-5.7 Mbp. The DSB yields show little LET dependence, in spite of the large variation of the latter among the beams, and are slightly higher than that obtained using gamma rays. The highest yield was found for the 5 GeV u(-1) iron beam, that gave a value 30% higher than the 1 GeV u(-1) iron beam. A phenomenological method is used to parametrise deviation from randomness in fragment size spectra.

Cells, Cultured↗

A 244Cm irradiator for protracted exposure of cultured Mammalian cells with alpha particles.

A 244Cm alpha-particle irradiator was designed and constructed for radiobiological studies where protracted exposure at a low dose rate of cultured mammalian cells is required. It allows irradiation of a cell monolayer attached to the Mylar bottom of a specially designed Petri dish of 56 mm diameter (approximately 25 cm(2) area). The irradiator is based on a 20-mm-diameter stainless steel chamber containing a 148 kBq 244Cm source. The chamber, flushed with helium gas at a pressure kept slightly above the external pressure, is inserted into a cell incubator where temperature and CO2 concentration are controlled. Spectrometric and dosimetric characterization of the irradiator was carried out by means of an ion-implanted-silicon charged-particle detector, CR39 detectors, and Monte Carlo simulations with the TRIM code. Average LET of particles incident on the cells at the center of the Petri dish was evaluated to be 120 keV microm(-1) at 59 mm from the source, and the average dose rate was 5.69 x 10 Gy s(-1), with +12% and -8% variations at the center and the edge, respectively. The irradiator has been successfully tested and used for several experiments involving 16-d exposure of human fibroblasts monolayers.

Alpha Particles↗

Bile salt and acid tolerance of Lactobacillus rhamnosus strains isolated from Parmigiano Reggiano cheese.

This study aimed to compare phenotypic and genetic characteristics of Lactobacillus rhamnosus strains isolated at the end of the ripening of Parmigiano Reggiano cheese and to investigate an important prerequisite of probiotic interest, such as the capability to survive at low pH and in presence of bile salts. The use of API 50 CH, RAPD-PCR analysis and species-specific PCR allowed to ascertain the identity of 63 L. rhamnosus strains. Three L. rhamnosus strains isolated from Parmigiano Reggiano cheese, L. rhamnosus ATCC 7469T and the commercial strain L. GG were assayed to estimate the resistance to various stress factors reproducing in vitro some conditions of the gastro-intestinal environment such as low pH and different amounts of bile salts and acids. The behaviour of almost all the tested strains isolated from Parmigiano Reggiano cheese resulted analogous to that showed by L. GG.

Base Sequence↗

DNA DSB induced by iron ions in human fibroblasts: LET dependence and shielding efficiency.

This paper reports on DNA DSB induction in human fibroblasts by iron ions of different energies, namely 5, 1 GeV/u, 414 and 115 MeV/u, in absence or presence of different shields (PMMA, Al and Pb). Measure of DNA DSB was performed by calibrated Pulsed Field Gel Electrophoresis using the fragment counting method. The RBE-LET relationships for unshielded and shielded beams were obtained both in terms of dose average LET and of track average LET. Weak dependence on these parameters was observed for DSB induction. The shielding efficiency, evaluated by the ratio between the cross sections for unshielded and shielded beams, depends not only on the shield type and thickness, but also on the beam energy. Protection is only observed at high iron ions energy, especially at 5 GeV/u, where PMMA shield gives higher protection compared to Al or Pb shields of the same thickness expressed in g/cm2.

Aluminum↗

Shelf life of fresh sausages stored under modified atmospheres.

The aim of this study was to investigate differences in modified atmosphere packaging (MAP) for the improvement of the shelf life of fresh meat products. Three different conditions for preserving fresh sausages were tested: MAP1 (20% CO2, 70% O2, and 10% N2), MAP2 (40% CO2 and 60% O2), and MAP3 (40% CO2, 30% O2, and 30% N2). Samples from the MAP2 group had fewer spoilage bacteria, stable red color (no change of a* value), and good physical attributes (high water-holding capacity, little loss from cooking, and low shear force needed for cutting) compared with samples from other treatment groups. Thus, high concentrations of CO2 (40%) and O2 (60%) resulted in a longer shelf life for fresh sausages.

Aerobiosis↗

Presence of yeasts in southern Italian sourdoughs from Triticum aestivum flour.

Samples of sourdoughs obtained from 13 artisanal bakeries located in the Molise and Campania regions were analysed. The sourdoughs were produced with the exclusive use of Triticum aestivum wheat flour. pH values of sourdoughs from Molise were generally lower than those from Campania. The number of yeasts in the samples of sourdoughs from Molise was generally higher than in those from Campania, which in two cases evidenced counts about 2 log cfu x g(-1). By utilising and comparing traditional and biomolecular techniques of identification a complete picture of the isolates was obtained: 58 strains were identified as Saccharomyces cerevisiae, five as Candida colliculosa, four as C. lambica, three as C. krusei, three as C. valida and two as C. glabrata.

Bread↗

Influence of a low background radiation environment on biochemical and biological responses in V79 cells.

We present the results of an experiment aimed at comparing the effects of different background radiation environments on metabolism and responses to gamma-rays and cycloheximide of cultured mammalian cells. Chinese hamster V79 cells were maintained in exponential growth in parallel for up to 9 months at the Istituto Superiore di Sanità (ISS) and at the INFN-Gran Sasso underground Laboratory (LNGS) where exposure due to gamma-rays and to radon was reduced by factors of about 70 and 25, respectively. After 9 months the cells grown at the LNGS (cumulative gamma dose about 30 microGy, average radon concentration around 5 Bq/m(3)), compared to the cells grown at the ISS (cumulative gamma-ray dose about 2 mGy, average radon concentration around 120 Bq/m(3)), exhibited i). a significant increase of the cell density at confluence, ii). a significantly higher capacity to scavenge organic and inorganic hydroperoxides but a reduced scavenging capacity towards superoxide anions and iii). an increase in both the basal hprt mutation frequency and sensitivity to the mutagenic effect of gamma-rays. The cells grown at the LNGS also showed a greater apoptotic sensitivity starting at the third month of culture, that was no longer detected after 9 months. Overall, these data suggest a role of background ionizing radiation in determining an adaptive response, although they cannot be considered conclusive.

Air Pollution, Indoor↗

DNA fragmentation in mammalian cells exposed to various light ions.

Elucidation of how effects of densely ionizing radiation at cellular level are linked to DNA damage is fundamental for a better understanding of the mechanisms leading to genomic damage (especially chromosome aberrations) and developing biophysical models to predict space radiation effects. We have investigated the DNA fragmentation patterns induced in Chinese hamster V79 cells by 31 keV/micrometer protons, 123 keV/micrometer helium-4 ions and gamma rays in the size range 0.023-5.7 Mbp, using calibrated Pulsed Field Gel Electrophoresis (PFGE). The frequency distributions of fragments induced by the charged particles were shifted towards smaller sizes with respect to that induced by comparable doses of gamma rays. The DSB yields, evaluated from the fragments induced in the size range studied, were higher for protons and helium ions than for gamma rays by a factor of about 1.9 and 1.2, respectively. However, these ratios do not adequately reflect the RBE observed on the same cells for inactivation and mutation induced by these beams. This is a further indication for the lack of correlation between the effects exerted at cellular level and the initial yield of DSB. The dependence on radiation quality of the fragmentation pattern suggests that it may have a role in damage repairability. We have analyzed these patterns with a "random breakage" model generalized in order to consider the initial non-random distribution of the DNA molecules. Our results suggest that a random breakage mechanism can describe with a reasonable approximation the DNA fragmentation induced by gamma rays, while the approximation is not so good for light ions, likely due to the interplay between ion tracks and chromatin organization at the loop level.

Animals↗

Inactivation of human cells exposed to fractionated doses of low energy protons: relationship between cell sensitivity and recovery efficiency.

Within the framework of radiation biophysics research in the hadrontherapy field, split-dose studies have been performed on four human cell lines with different radiation sensitivity (SCC25, HF19, H184B5 F5-1 M10, and SQ20B). Low energy protons of about 8 and 20 keV/micron LET and gamma-rays were used to study the relationship between the recovery ratio and the radiation quality. Each cell line was irradiated with two dose values corresponding to survival levels of about 5% and 1%. The same total dose was also delivered in two equal fractions separated by 1.5, 3, and 4.5 hours. A higher maximum recovery ratio was observed for radiosensitive cell lines as compared to radioresistant cells. The recovery potential after split doses was small for slow protons, compared to low-LET radiation. These data show that radiosensitivity may not be related to a deficient recovery, and suggest a possible involvement of inducible repair mechanisms.

Cell Line↗

Rapid detection of meso-diaminopimelic acid in lactic acid bacteria by microwave cell wall hydrolysis.

Two innovative microwave hydrolysis procedures for rapidly detecting meso-diaminopimelic acid in lactic acid bacteria have been developed. Whole-cell hydrolysis was performed both in liquid phase and vapor phase with 6 N HCl in sealed vessels using a microwave oven equipped with pressure and temperature probes. The presence or absence of meso-diaminopimelic acid determined by TLC, after the application of liquid- and vapor-phase microwave hydrolysis procedures, gave the same qualitative results as those obtained by traditional hydrolysis. These standardized microwave hydrolysis procedures permit a drastic reduction in hydrolysis time, from 16-20 h to less than 10 min and, consequently, in the total time of meso-diaminopimelic acid analysis (less than 90 min). Microwave hydrolysis in vapor phase is particularly convenient because, besides reducing hydrolysis time, it also eliminates the successive troublesome step of HCl removal.

Cell Wall↗

Inactivation of human normal and tumour cells irradiated with low energy protons.

PURPOSE: To analyse the cell inactivation frequencies induced by low energy protons in human cells with different sensitivity to photon radiation. MATERIALS AND METHODS: Four human cell lines with various sensitivities to photon irradiation were used: the SCC25 and SQ20B derived from human epithelium tumours of the tongue and larynx, respectively, and the normal lines M/10, derived from human mammary epithelium, and HF19 derived from a lung fibroblast. The cells were irradiated with y-rays and proton beams with linear energy transfer (LET) from 7 to 33 keV/microm. Clonogenic survival was assessed. RESULTS: Survival curves are reported for each cell line following irradiation with gamma-rays and with various proton LETs. The surviving fraction after 2 Gy of gamma-rays was 0.72 for SQ20B cells, and 0.28-0.35 for the other cell lines. The maximum LET proton effectiveness was generally greater than that of gamma-rays. In particular there was a marked increase in beam effectiveness with increasing LET for the most resistant cells (SQ20B) whose 2 Gy-survival varied from 0.72 with gamma-radiation down to 0.37 with 30 keV/microm protons. The relative biological effectiveness (RBE(2 Gy gamma)) with the 30 keV/microm beam, evaluated as the ratio of 2 Gy to the proton dose producing the same inactivation level as that given by 2 Gy of gamma-rays, was 3.2, 1.8, 1.3 and 0.8 for SQ20B, M/10, SCC25, and HF19, respectively. CONCLUSIONS: RBE for inactivation with high-LET protons increased with the cellular radioresistance to gamma-rays. The cell line with the greatest resistance to gamma-rays was the most responsive to the highest LET proton beam. A similar trend has also been found in studies reported in the literature with He, C, N ions with LET in the range 20-125 keV/microm on human tumour cell lines.

Cell Line↗

Acetyl-L-carnitine in Alzheimer disease: a short-term study on CSF neurotransmitters and neuropeptides.

Acetyl-L-carnitine (ALCAR) is a drug currently under investigation for Alzheimer disease (AD) therapy. ALCAR seems to exert a number of central nervous system (CNS)-related effects, even though a clear pharmacological action that could explain clinical results in AD has not been identified yet. The aim of this study was to determine cerebrospinal fluid (CSF) and plasma biological correlates of ALCAR effects in AD after a short-term, high-dose, intravenous, open treatment. Results show that ALCAR CSF levels achieved under treatment were significantly higher than the ones at baseline, reflecting a good penetration through the blood-brain barrier and thus a direct CNS challenge. ALCAR treatment produced no apparent change on CSF classic neurotransmitters and their metabolite levels (homovanillic acid, 5-hydroxyindoleacetic acid, MHPG, dopamine, choline). Among CSF peptides, while corticotropin-releasing hormone and adrenocorticotropic hormone remained unchanged, beta-endorphins significantly decreased after treatment; plasma cortisol levels matched this reduction. Since both CSF beta-endorphins and plasma cortisol decreased, one possible explanation is that ALCAR reduced the AD-dependent hypothalamic-pituitary-adrenocortical (HPA) axis hyperactivity. At present, no clear explanation can be proposed for the specific mechanism of this action.

Aged↗

Enterocin 226NWC, a bacteriocin produced by Enterococcus faecalis 226, active against Listeria monocytogenes.

Enterococcus faecalis 226, isolated from natural whey cultures utilized as starters in the manufacture of mozzarella cheese from water-buffalo milk, produces a bacteriocin designated enterocin 226NWC. The bacteriocin was isolated from culture supernatant fluids of the producer strain and was active against strains of the same species and Listeria monocytogenes, but not against useful lactic acid bacteria. Enterocin 226NWC is a protein with an apparent molecular weight of about 5800; it is relatively heat-stable and has a bactericidal mode of action. Listeria monocytogenes, growing in the presence of the enterocin 226NWC producer strain in broth and in reconstituted skim milk, was inhibited.

Animals↗

Conjugal transfer of plasmid-borne bacteriocin production in Enterococcus faecalis 226 NWC.

Enterococcus faecalis 226 NWC, isolated from natural whey cultures utilized as starter in water-buffalo Mozzarella cheese manufacture, produces a bacteriocin, designated Enterocin 226 NWC, which is inhibitory to Listeria monocytogenes. Plasmid analysis of E. faecalis 226 NWC showed a single 5.2-kb plasmid, pEF226. In conjugation experiments, pEF226 was transferred into a plasmid-free strain of E. faecalis JH2-2. The transfer required direct cell-to-cell contact and was not inhibited by DNase. The identity of conjugation was confirmed by digestion with SmaI restriction endonuclease and subsequent pulsed-field gel electrophoresis (PFGE) of the genomic DNA of E. faecalis 226, E. faecalis JH2-2 and of the isolates after the mating. The data indicate that the ability of E. faecalis 226 NWC to produce the bacteriocin is linked to the 5.2-kb conjugative plasmid pEF226.

Bacteriocins↗