Fungal and bacterial species richness in biodeteriorated seventeenth century Venetian manuscripts.

Historical paper documents are susceptible to complex degradation processes, including biodeterioration, which can progressively compromise their aesthetic and structural integrity. This study analyses seventeenth century handwritten historical letters stored at the Correr Museum Library in Venice, Italy, exhibiting pronounced signs of biodeterioration. The techniques used encompassed traditional colony isolation on agar plates and proteomics analyses, employing nanoscale liquid chromatography coupled with high-resolution mass spectrometry (nano-LC-MS). Fluorescence microscopy was used for the first time in the historical paper biodeterioration context to supplement the conventional stereoscopic, optical, and scanning electron microscopic imaging techniques. This method enables the visualisation of microorganisms beyond and beneath the paper's surface through their natural intrinsic autofluorescence in a non-invasive and non-destructive way. The results demonstrate a diverse, complex, and abundant microbiota composed of coexisting fungal and bacterial species (Ascomycota, Mucoromycota, Basidiomycota, Proteobacteria, and Actinobacteria), along with mite carcasses, insects, parasites, and possibly protists. Furthermore, this study reveals certain species that were not previously documented in the biodeterioration of historical paper, including human pathogens, such as Histoplasma capsulatum, Brucella, Candida albicans, and species of Aspergillus (A. flavus, A. fumigatus, A. oryzae, A. terreus, A. niger) known to cause infections or produce mycotoxins, posing substantial risk to both artefacts and humans.

Biophysical Manipulation of the Extracellular Environment by Eurotium halophilicum.

Eurotium halophilicum is psychrotolerant, halophilic, and one of the most-extreme xerophiles in Earth's biosphere. We already know that this ascomycete grows close to 0 °C, at high NaCl, and-under some conditions-down to 0.651 water-activity. However, there is a paucity of information about how it achieves this extreme stress tolerance given the dynamic water regimes of the surface habitats on which it commonly occurs. Here, against the backdrop of global climate change, we investigated the biophysical interactions of E. halophilicum with its extracellular environment using samples taken from the surfaces of library books. The specific aims were to examine its morphology and extracellular environment (using scanning electron microscopy for visualisation and energy-dispersive X-ray spectrometry to identify chemical elements) and investigate interactions with water, ions, and minerals (including analyses of temperature and relative humidity conditions and determinations of salt deliquescence and water activity of extracellular brine). We observed crystals identified as eugsterite (Na4Ca(SO4)3·2H2O) and mirabilite (Na2SO4·10H2O) embedded within extracellular polymeric substances and provide evidence that E. halophilicum uses salt deliquescence to maintain conditions consistent with its water-activity window for growth. In addition, it utilizes a covering of hair-like microfilaments that likely absorb water and maintain a layer of humid air adjacent to the hyphae. We believe that, along with compatible solutes used for osmotic adjustment, these adaptations allow the fungus to maintain hydration in both space and time. We discuss these findings in relation to the conservation of books and other artifacts within the built environment, spoilage of foods and feeds, the ecology of E. halophilicum in natural habitats, and the current episode of climate change.

The role of bio-pollutants in the indoor air quality of old museum buildings: artworks biodeterioration as preview of human diseases.

Indoor air quality in buildings is strongly affected by chemical, physical, and biological agents. Long exposure to inadequate indoor air quality can be very dangerous for the building occupants and can lead to chronic diseases associated with the sick building syndrome (SBS). In this paper, the large presence of biological pollutants in the indoor rooms of an old building and its strict relationship with the outdoor/indoor air conditions were investigated studying Coronini Cronberg Palace Foundation, a historic house museum of the sixteenth century in Gorizia (Italy), where biological contamination affecting the artworks can soon become potentially harmful also for operators and visitors. Detailed aerobiological and microbiological analyses on organic natural materials, combined with a microclimate monitoring, were conducted to evaluate the influence of temperature and relative humidity levels within the Palace in the conspicuous growth and diffusion of microorganisms. Fungal and bacterial colonies damaging materials, mainly affected by the sudden fluctuations of hygrothermal values, were found to widely exceed Italian and international recommended levels for good air quality for both artworks and human beings. Understand their impact on human health would be strictly necessary to reduce biological risks for museum staff and cultural heritage users, but consequently to improve indoor air quality.

Milestones and recent discoveries on cell death mediated by mitochondria and their interactions with biologically active amines.

Mitochondria represent cell "powerhouses," being involved in energy transduction from the electrochemical gradient to ATP synthesis. The morphology of their cell types may change, according to various metabolic processes or osmotic pressure. A new morphology of the inner membrane and mitochondrial cristae, significantly different from the previous one, has been proposed for the inner membrane and mitochondrial cristae, based on the technique of electron tomography. Mitochondrial Ca(2+) transport (the transporter has been isolated) generates reactive oxygen species and induces the mitochondrial permeability transition of both inner and outer mitochondrial membranes, leading to induction of necrosis and apoptosis. In the mitochondria of several cell types (liver, kidney, and heart), mitochondrial oxidative stress is an essential step in the induction of cell death, although not in brain, in which the phenomenon is caused by a different mechanism. Mitochondrial permeability transition drives both apoptosis and necrosis, whereas mitochondrial outer membrane permeability is characteristic of apoptosis. Adenine nucleotide translocase remains the most important component involved in membrane permeability, with the opening of the transition pore, although other proteins, such as ATP synthase or phosphate carriers, have been proposed. Intrinsic cell death is triggered by the release from mitochondria of proteic factors, such as cytochrome c, apoptosis inducing factor, and Smac/DIABLO, with the activation of caspases upon mitochondrial permeability transition or mitochondrial outer membrane permeability induction. Mitochondrial permeability transition induces the permeability of the inner membrane in sites in contact with the outer membrane; mitochondrial outer membrane permeability forms channels on the outer membrane by means of various stimuli involving Bcl-2 family proteins. The biologically active amines, spermine, and agmatine, have specific functions on mitochondria which distinguish them from other amines. Enzymatic oxidative deamination of spermine by amine oxidases in tumor cells may produce reactive oxygen species, leading to transition pore opening and apoptosis. This process could be exploited as a new therapeutic strategy to combat cancer.

Detection of volatile metabolites of moulds isolated from a contaminated library.

The principal fungal species isolated from a contaminated library environment were tested for their microbial volatile organic compound (MVOC) production ability. Aspergillus creber, A. penicillioides, Cladosporium cladosporioides, Eurotium chevalieri, E. halophilicum, Penicillium brevicompactum and P. chrysogenum were cultivated on suitable culture media inside sample bottles specifically designed and created for direct MVOC injection to a GC-MS instrument. The fungal emissions were monitored over several weeks to detect changes with the aging of the colonies, monitored also by respirometric tests. A total of 55 different MVOCs were detected and isopropyl alcohol, 3-methyl-1-butanol and 2-butanone were the principal compounds in common between the selected fungal species. Moreover, 2,4-dimethylheptane, 1,4-pentadiene, styrene, ethanol, 2-methyl-1-butanol, acetone, furan and 2-methylfuran were the most detected compounds. For the first time, the MVOC production for particular fungal species was detected. The species A. creber, which belongs to the recently revised group Aspergillus section Versicolores, was characterized by the production of ethanol, furan and 1,4-pentadiene. For the xerophilic fungus E. halophilicum, specific production of acetone, 2-butanone and 1,4-pentadiene was detected, supported also by respirometric data. The results demonstrated the potential use of this method for the detection of fungal contamination phenomena inside Cultural Heritage's preservation environments.

Spermine cycling in mitochondria is mediated by adenine nucleotide translocase activity: mechanism and pathophysiological implications.

Spermine, besides to be transported in mitochondria by an energy dependent electrophoretic mechanism, can be also released by two different mechanisms. The first one is induced in deenergizing conditions by FCCP or antimycin A and it is mediated by an electroneutral exchange spermine protons. The second one takes place in energizing conditions during the activity of the adenine nucleotide translocase and is mediated by an electroneutral symport mechanism involving the efflux in co-transport of spermine and phosphate and the exchange of exogenous ADP with endogenous ATP. The triggering of this mechanism permits an alternating cycling of spermine across the mitochondrial membrane, that is spermine is transported or released by energized mitochondria in the absence or presence of ATP synthesis, respectively. The physiological implications of this cycling of spermine are related to the induction or prevention of mitochondrial permeability transition and, consequently, on apoptosis or its prevention.

Bidirectional fluxes of spermine across the mitochondrial membrane.

The polyamine spermine is transported into the mitochondrial matrix by an electrophoretic mechanism having as driving force the negative electrical membrane potential (ΔΨ). The presence of phosphate increases spermine uptake by reducing ΔpH and enhancing ΔΨ. The transport system is a specific uniporter constituted by a protein channel exhibiting two asymmetric energy barriers with the spermine binding site located in the energy well between the two barriers. Although spermine transport is electrophoretic in origin, its accumulation does not follow the Nernst equation for the presence of an efflux pathway. Spermine efflux may be induced by different agents, such as FCCP, antimycin A and mersalyl, able to completely or partially reduce the ΔΨ value and, consequently, suppress or weaken the force necessary to maintain spermine in the matrix. However this efflux may also take place in normal conditions when the electrophoretic accumulation of the polycationic polyamine induces a sufficient drop in ΔΨ able to trigger the efflux pathway. The release of the polyamine is most probably electroneutral in origin and can take place in exchange with protons or in symport with phosphate anion. The activity of both the uptake and efflux pathways induces a continuous cycling of spermine across the mitochondrial membrane, the rate of which may be prominent in imposing the concentrations of spermine in the inner and outer compartment. Thus, this event has a significant role on mitochondrial permeability transition modulation and consequently on the triggering of intrinsic apoptosis.

A comparison between the responses of neutral red and acridine orange: acridine orange should be preferential and alternative to neutral red as a dye for the monitoring of contaminants by means of biological sensors.

The acridine orange (AO) and neutral red (NR) dyes, commonly used as probes to measure the internal pH in acidic vesicles, are compared in this article. The comparison between the two dyes (arising from calculations taking into account their analytical constants) illustrated that the use of AO is preferential to that of NR because the AO response is sensitive over the whole pH range between 4.0 and 7.4, whereas the NR response is effective only between pHs 4.0 and 6.0. In addition, it became evident from the mitochondrial respiration response that NR, unlike AO, is a protonophore. When taken into consideration, these two properties suggest that AO is more suitable than NR as an indicator of toxicity measurements in water samples because the environmental toxic compounds induce pH changes in the acidic vesicles of biological structures that are used as environmental biosensors.

TCMS inhibits ATP synthesis in mitochondria: a systematic analysis of the inhibitory mechanism.

The interactions of the antifouling compound TCMS (2,3,5,6-tetrachloro-4-methylsulphonyl pyridine) with rat liver mitochondria have been investigated. The results indicate that the compound inhibits ATP synthesis. Further investigations regarding the ATP synthesis mechanism suggest that TCMS inhibits succinic dehydrogenase of the mitochondrial respiratory chain. As the respiratory chain is similar in all living organisms, it can be concluded that the toxic effect of TCMS most likely depend on the different bioavailability of the compound and on the different importance of mitochondria in the ATP production in the animal species.

The interactions of cobalt(II) with mitochondria from rat liver.

The interactions of Co(2+) with mitochondria have been investigated. The results indicate that Co(2+) inhibits ATP synthesis. Further investigations into ATP synthesis mechanisms indicated that inhibition is due to the opening of a transmembrane pore. The opening of this pore causes the collapse of the high-energy intermediate where, under a pH and a potential gradient, the energy is stored and subsequently utilized to form ATP from ADP.

Trialkyllead compounds induce the opening of the MTP pore in rat liver mitochondria.

The interactions of the tributyl, triethyl and trimethyllead compounds with energized mitochondria have been investigated in this paper. It has been shown that the (alkyl)(3)Pb-Cl compounds induce swelling in mitochondria suspended in a sucrose medium. The phenomenon is more marked the higher the lipophilicity and occurs in the following order: (Bu)(3)Pb>(Et)(3)Pb>(Me)(3)Pb. As swelling is inhibited by cyclosporine, this suggests that the swelling is due to the opening of a trans-membrane pore (MTP pore) in the mitochondria. As this pore can be responsible for the inhibition of the ATP synthesis, and, consequently for cell death, the opening of the pore could be one of the reasons for the toxicity of the (alkyl)(3)Pb-X compounds.

Irgarol inhibits the synthesis of ATP in mitochondria from rat liver.

The interactions of Irgarol with rat liver mithocondrial have been investigated. The results indicate that Irgarol inhibits the ATP synthesis. The analysis of the various steps involved in the ATP synthesis suggests that the inhibition is due to the opening of small-size pores.

An in vitro study of the interaction of sea-nine with rat liver mitochondria.

The interactions of the antifouling compound Sea-Nine with rat liver mitochondria have been studied. The results indicate that low doses of this compound inhibit adenosine 5'-triphosphate (ATP) synthesis. Further investigations indicate that ATP synthesis inhibition should be due to an interaction of Sea-Nine with the succinic dehydrogenase in the mitochondrial respiratory chain.

A possible transport mechanism for aluminum in biological membranes.

The transport mechanism of aluminum in lysosomes extracted from rat liver has been investigated in this paper. The experimental evidence supports the hypothesis that aluminum is transported inside lysosomes in the form of an Al(OH)(3) electroneutral compound, the driving force being the internal acidic pH. This mechanism could help to explain the presence of aluminum in cells in many illnesses.

Frozen mitochondria as rapid water quality bioassay.

A rapid and relatively low cost bioassay, usable in routine screening water test has been developed modifying the beef heart mitochondria test. In our experiments, mitochondria (FM22) were frozen at -22 degrees C, instead of -80 degrees C (FM80), and their applicability and sensitivity was verified. The oxygen consumption was measured by a Clark electrode that was interfaced to a PC to collect test analysis data. Blank tests were carried out to verify the oxygen consumption linear fitting. Toxicity tests were performed using pure organic and inorganic compounds, such to verify the FM22 sensitivity. A piecewise regression, through an Excel Macro, identified the break-point in the oxygen consumption and calculated the toxicity. The IC50s of the tested compounds were calculated and ranged from 0.123 to 0.173 mg/l for heavy metals (Cd, Cr, Cu, Ni, Pb and Zn) and from 0.572 to 10.545 mg/l for organics (benzene, DMSO, DDE, endrin, dichloromethane, chlorobenzene, 1,2-dichlorobenzene and 1,3-dichlorobenzene). Water effluent samples were then tested. The FM22 gave different toxic reactions to them. Water samples were characterised for heavy metals. The FM22 bioassay had a higher sensitivity than the FM80 and a high reproducibility in the toxicity test with pure compounds. The FM22 test was a good predictor of toxicity for water samples; the bioassay is easy, low cost and rapid, then usable for routine tests.

The mitochondria from beef heart as biosensors for the selective monitoring of phenols.

We propose a simple and rapid procedure which allows for the selective monitoring in solution of toxic compounds which behave as uncouplers of the oxidative phosphorylation. Since all phenols, are uncouplers of the oxidative phosphorylation, the procedure allows for the selective monitoring in solution of phenols even in presence of other toxic compounds. The biological sensor are the mitochondria from beef heart. This biosensor is easily available without a stabular and therefore the biosensor and the whole procedure is very simple and not expensive. By linear regression analysis, it results that the procedure well predicts the response of the standard fish method to phenols. Therefore the procedure can be utilized as prescreening analysis for the monitoring the phenols in aqueous samples.

Methylmercury induces the opening of the permeability transition pore in rat liver mitochondria.

Interactions of methylmercury (CH(3)HgCl) with non-energized mitochondria from rat liver (non-respiring mitochondria) have been investigated in this paper. It has been shown that CH(3)HgCl induces swelling in mitochondria suspended in a sucrose medium. Swelling has also been induced by detergent compounds and by phenylarsine, a chemical compound which induces opening of the permeant transition pore (MTP). Opening of the MTP is inhibited by means of cyclosporine A. Results indicate that the swelling induced by CH(3)HgCl, as in the case of phenylarsine, is inhibited by cyclosporine A and Mg(2+), while swelling induced by detergent compounds is not cyclosporine sensitive. This comparison suggests that CH(3)HgCl induces opening of a permeability transition pore (MTP). Since the opening of an MTP induces cell death, this interaction with MTP could be one of the causes of toxicity of CH(3)HgCl.

The interactions of cetyltrimethylammonium with mitochondria: an uncoupler or a detergent?

The interactions of cetyltrimethylammonium (CTA) with mitochondria have been investigated. We confirm, as already observed in a previous paper, that this compound behaves as proton carrier (or uncoupler) of the oxidative phosphorylation, but evidences suggest that this compound enhances the membrane permeability to many other compounds such as sucrose. We conclude therefore that CTA as a detergent enhances membrane permeability to all ions including protons. Some evidences are also given that the inhibitory effect of CTA on the mitochondrial respiratory chain is a consequence of the swelling induced.