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Biomedical subjects

G Bertoloni

Publications and source records attributed to G Bertoloni.

At least 19 recordsLinked to original sources
PubMed2006

S.A.M., the Italian Martian simulation chamber.

The Martian Environment Simulator (SAM "Simulatore di Ambiente Marziano") is a interdisciplinary project of Astrobiology done at University of Padua. The research is aimed to the study of the survival of the microorganisms exposed to the "extreme" planetary environment. The facility has been designed in order to simulate Mars' environmental conditions in terms of atmospheric pressure, temperature cycles and UV radiation dose. The bacterial cells, contained into dedicated capsules, will be exposed to thermal cycles simulating diurnal and seasonal Martian cycles. The metabolism of the different biological samples will be analysed at different phases of the experiment, to study their survival and eventual activity of protein synthesis (mortality, mutations and capability of DNA repairing). We describe the experimental facility and provide the perspectives of the biological experiments we will perform in order to provide hints on the possibility of life on Mars either autochthonous or imported from Earth.

Bacteria↗
PubMed2001

Detection of Chlamydophila pneumoniae DNA in peripheral blood mononuclear cells of blood donors in the north-east of Italy.

Recent studies have implicated Chlamydia pneumoniae (now Chlamydophila pneumoniae) in the pathogenesis of atherosclerosis and demonstrated its presence within human peripheral blood mononuclear cells (PBMCs). In this study the presence of C. pneumoniae DNA was assessed, using nested PCR, in PBMCs from 169 active blood donors as a function of age, of specific antibodies and C-reactive protein. The results obtained demonstrated a high degree of global positivity (46.15%), which was higher in females (52%) than in males (43.7%). Seroepidemiological studies showed a high percentage of positivity both in subjects positive by PCR (65.91%) and negative by PCR (71.74%). The clinical implication of such finding are under study.

Adolescent↗
PubMed2001

Demonstration of Chlamydia pneumoniae in atherosclerotic arteries from various vascular regions.

Chlamydia pneumoniae (CP) has been reported to be a pathogenic agent in the mechanism leading to atherosclerosis. The majority of available data is focused mainly on coronary artery disease whereas the distribution of CP in different areas, associated with atherosclerotic disorders, has not been completely clarified. In this study we investigated the presence of CP in atheromasic plaques from five different vascular areas (basilary artery, coronary artery, thoracic aorta, abdominal aorta, renal arteries) using nested polymerase chain reaction (PCR) and immunohistochemical staining (IHC), in order to establish the putative association of CP with atherosclerotic disease. The same atheromasic plaques were also tested for the presence of Helicobacter pylori (HP) and cytomegalovirus (CMV), other putative agents of atherosclerosis, using a nested PCR technique. Our data indicate that the presence of CP can be demonstrated in 100% of patients tested, considering globally the five areas of analysis. On the other hand the presence of HP has been demonstrated in four out of 18 patients (22.2%), and CMV only in three out of 18 (16.6%). Our results strongly suggest an association between CP and atherosclerosis and highlight the need for the detection of CP in multiple vascular areas of the same patient.

Aged↗
PubMed2000

Photosensitizing activity of hematoporphyrin on Staphylococcus aureus cells.

The photosensitizing action of hematoporphyrin (Hp) on two Staphylococcus aureus strains was investigated to determine if the photoprocess induces in vivo damage in DNA in addition to that occurring at the level of the cytoplasmic membrane. The results obtained demonstrate that the photokilling is dependent on the Hp dose even though the two strains, having a similar Hp-binding capacity, show different levels of photosensitivity. The electrophoretic analysis of cytoplasmic membrane proteins and DNA (chromosomal and plasmidial) suggests that the membrane represents the primary target of the photoprocess, while the DNA, that is damaged both in vivo and in vitro only at relatively long irradiation time, might be a secondary target. Moreover, the photoprocess results in mutagenesis for Salmonella typhimurium tester strains. This information is particularly important in view of the potential use of photodynamic therapy for the treatment of microbial infections.

Anti-Bacterial Agents↗
PubMed2000

Polylysine-porphycene conjugates as efficient photosensitizers for the inactivation of microbial pathogens.

Porphycenes are electronic isomers of porphyrins which, when neutral, display no appreciable photosensitizing action towards Gram-negative bacteria. The covalent binding of oligomeric polylysine moieties, which are cationic at physiological pH values, endows porphycenes with a significant phototoxic activity against Gram-negative bacteria while retaining their photoefficiency against a variety of microbial pathogens, including Gram-positive bacteria, fungi and mycoplasmas. The effect of the polylysine moiety is dependent on both the polylysine concentration and the degree of oligomerization. A suitable interplay among the various parameters opens the possibility to obtain either a broad spectrum of antimicrobial activity or a selective action toward a specific pathogen while minimizing the damage to human fibroblasts.

Anti-Bacterial Agents↗
PubMed1999

Role of ground and excited singlet state oxygen in the red light-induced stimulation of Escherichia coli cell growth.

Irradiation of selected Escherichia coli defective strains with red-light induces a stimulation of the cell growth rate. Such effect is wavelength-dependent and is accompanied by a transient increase of the cell volume and some enzymic activities. The presence of oxygen appears to be essential for the occurrence of a significant photostimulatory effect. The results obtained upon irradiation in the presence of quenchers (tryptophan, histidine, azide) or enhancers (deuterium oxide) of singlet oxygen (1O2) strongly suggest that this activated oxygen derivative is generated by excitation of endocellular chromophores (possibly cytochromes). The reaction of 1O2 with nearby cellular targets could induce a sublethal cell damage which in turn promotes an accelerated cell metabolism.

Azides↗
PubMed1996

Studies on the mechanism of bacteria photosensitization by meso-substituted cationic porphyrins.

Cationic porphyrins have been shown to photoinduce the direct inactivation of Gram-positive (G+) and Gram-negative (G-) bacteria, thereby differing from anionic or neutral porphyrins which can photosensitize the G- bacteria only after permeabilization of their outer membrane. The present data show that the differences between these positively and negatively charged porphyrins are not related by a difference in the intrinsic photosensitizing efficiency, as determined by the photo-oxidation of model substrates or the yield of 1O2 generation; moreover, there are only minor differences in the quantum yield of porphyrin photobleaching. Rather, it appears that the positive charge promotes an electrostatic binding of the porphyrin to the outer cell surface inducing an initial limited damage which favours the penetration of the photosensitizer. Actually, the overall photoprocess is inhibited by the preincorporation of the porphyrin into liposomes, while it is enhanced by using amphiphilic dicationic porphyrins which bind to endocellular sites in larger amounts and in a more stable form.

Cell Survival↗
PubMed1996

Temporary axillo-femoral bypass graft for renal transplant protection during aortic aneurysm repair.

Aortic surgery in renal transplant recipients requires a method of maintaining intraoperative graft perfusion. Here we present a case in which temporary axillo-femoral bypass was used to perfuse a renal transplant during the aortic aneurysm repair; the rationale of inserting the temporary axillo-femoral bypass on transverse arteriotomies is pointed out. Other methods of renal graft perfusion are discussed.

Aortic Aneurysm, Abdominal↗
PubMed1994

Superoxide dismutase, catalase and cell dimorphism in Candida albicans cells exposed to methanol and different temperatures.

The combined effects of methanol and different temperatures on Candida albicans were studied. Growth curve, cell morphology, superoxide dismutase (SOD) and catalase activity levels have been determined. Cell growth in each medium was comparable to 28 degrees C and 37 degrees C. The growth rate was not affected by methanol, in the presence of glucose, while it was much lower in the absence of sugar. Cell dimorphism appeared after thermic stress and it was also dependent on the medium composition. In all media, both SOD and catalase levels were much higher at 37 degrees C. The presence of methanol per se did not affect the enzymatic levels, while the absence of glucose gave higher SOD levels.

Candida albicans↗
PubMed1993

Biochemical and morphological changes in Escherichia coli irradiated by coherent and non-coherent 632.8 nm light.

Irradiation of Escherichia coli cells with either coherent or non-coherent 632.8 nm light (4 J cm-2) causes a transient acceleration of cell proliferation, which is maximal about 60 min after the end of the phototreatment. The stimulatory effect is dose dependent and is especially evident in the case of defective E. coli strains which are in the logarithmic phase of growth, while it becomes less important when cells are exposed to non-coherent 600-700 nm light. Stimulated cells exhibit biochemical and morphological changes, such as an intensified synthesis of cytoplasmic membrane proteins, increased cell volume and ribosomal content, which are suggestive of an enhanced cell metabolism.

Cell Division↗
PubMed1993

Effect of extracellularly generated singlet oxygen on gram-positive and gram-negative bacteria.

In the separated surface-sensitizer system, a photosensitizer is physically separated from the substrate by a thin air layer under such conditions that only singlet oxygen can reach and oxidize the substrate, preventing the competition by type I photosensitized processes. This method has been used to study the reaction of singlet oxygen with Gram-positive (Streptococcus faecium) and Gram-negative (Escherichia coli) bacterial strains. Studies on cell samples exposed to singlet oxygen for different periods of time show a drastic decrease in survival for S. faecium, while E. coli becomes sensitive only when the integrity of the outer membrane is altered by treatment with CaCl2 or tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetic acid (Tris-EDTA). Biochemical and ultrastructural analyses suggest that the cytoplasmic membrane and the genetic material are the main sites damaged by singlet oxygen.

Enterococcus faecium↗
PubMed1992

Photosensitizing activity of water- and lipid-soluble phthalocyanines on prokaryotic and eukaryotic microbial cells.

The photosensitizing activity of lipophilic zinc-phthalocyanine (Zn-Pc) and its water-soluble sulphonated derivative (Zn-PcS) towards Streptococcus faecium and Candida albicans was studied and correlated with the amount of cell-bound photosensitizer. With both micro-organisms Zn-PcS was more tightly bound in larger amounts than Zn-Pc in the protoplasts of the cytoplasmic membrane. As a consequence, the photoinduced damage in S. faecium initially involved membrane proteins, while DNA was modified only upon prolonged irradiation. For C. albicans only Zn-PcS showed a preferential affinity for the spheroplasts and the decrease in cell survival was not accompanied by detectable modifications of the electrophoretic pattern of membrane proteins. The photoinduced ultrastructural alteration of both micro-organisms suggests damage at membrane level. This would indicate the involvement of different targets in bacteria and yeast for phthalocyanine photosensitization.

Candida albicans↗
PubMed1990

Photosensitizing activity of water- and lipid-soluble phthalocyanines on Escherichia coli.

Escherichia coli, as most Gram-negative bacteria, is insensitive to the photosensitizing action of both lipid-soluble Zinc-phthalocyanine (Zn-Pc) and water-soluble Zinc-mono/disulfonated phthalocyanine (Zn-PcS). Photosensitivity can be induced by alteration of the outer membrane, as obtained by either induction of competence or treatment with Tris-EDTA. Both phthalocyanines largely bind at the level of the cytoplasmic membrane; however, Zn-PcS shows a superior photosensitizing activity as compared with Zn-Pc. Biochemical analyses performed on irradiated cells suggest that the cytoplasmic membrane is an important target of the photoprocess, while DNA is not involved.

Bacterial Outer Membrane Proteins↗
PubMed1989

Factors influencing the haematoporphyrin-sensitized photoinactivation of Candida albicans.

Photosensitizing activity of haematoporphyrin (Hp) on Candida albicans cells is mainly promoted by unbound dye molecules in the bulk aqueous medium. Moreover, the death of photosensitized cells is dependent on the dye concentration, irradiation time, irradiation temperature, and the composition of the growth media. Morphological and biochemical studies indicate that the photoprocess involves an initial limited alteration of the cytoplasmic membrane, which allows the penetration of the dye into the cell with consequent photodamage of intracellular targets. In this respect, the Hp-sensitized photoinactivation of eukaryotic microbial cells differs from that in prokaryotic cells.

Candida albicans↗
PubMed1988

A further insight into the mechanism of action of anthracycline antibiotics.

The uptake of Adriamycin (ADM) by eucaryotic and procaryotic cells and the interaction of the antibiotic with structurally organized DNA were investigated. The aim of this work was to understand why ADM is endowed with very low antibiotic activity in spite of being highly cytotoxic for mammalian cells, and to get a further insight into the mechanism of action of this compound. Spectrophotometric, fluorometric, chiroptical, and electron microscopic techniques were used. The drug was shown to concentrate in mammalian cells but failed to do so in bacteria and penetrated rather poorly in Candida albicans. The chromatin binding affinity of ADM appeared to be substantially reduced with respect to the affinity for free DNA. Complex stereochemistry was also influenced by the presence of nucleosomal arrangements in the polynucleotide. In addition, evidence was obtained that the antibiotic tended to accumulate at specific (linker) chromatin regions and to cause extensive compaction of organized DNA. The observed phenomena may play a relevant role in the mode of drug action in eucaryotic cells and help to explain the lack of antibacterial activity by ADM in spite of the apparent attainment of great enough intracellular levels to affect DNA template function.

Bacteria↗
PubMed1986

Phagocytosis of Mycoplasma pneumoniae and Acholeplasma laidlawii measured as inhibition of [3H]uridine uptake by macrophages.

Many studies of the interaction between phagocytes and mycoplasmas have given controversial results. This is probably due both to the small size of the microorganisms and their ability to attach to the cell membrane, making it difficult to distinguish between adsorption and ingestion. To overcome these difficulties we took advantage of a phenomenon we noted occurring concomitantly with phase-contrast microscope-monitored phagocytosis of heat-killed C. albicans, i.e., a reduction of [3H]uridine uptake by macrophages from culture medium. This approach allowed us to measure the ability of mouse peritoneal macrophages and the macrophage-like P 388 D 1 continuous cell line to phagocytose Mycoplasma pneumoniae and Acholeplasma laidlawii. Live, UV-killed and specific antiserum-opsonized mycoplasmas were tested. A. laidlawii was ingested under all the conditions mentioned above, while live M. pneumoniae was not phagocytosed unless UV-killed. Phagocytosis of UV-killed M. pneumoniae was directly verified by transmission electron microscopy studies. Data obtained with opsonized M. pneumoniae indicated no ingestion by mouse peritoneal macrophages and incomplete phagocytosis with P388 D 1 macrophages, suggesting that different responses by different types of phagocytes can be observed. In spite of a lack of information concerning the biological meaning of the inhibition of macrophage RNA metabolism during phagocytosis, our data suggest that this phenomenon may be used to study the phagocytosis of microorganisms which are difficult to visualize.

Acholeplasma laidlawii↗