Full scale treatment of ASR wastes in a modified rotary kiln.

A plant, designed for the thermo-valorisation of tyres, was specifically modified in order to treat Automobile Shredder Residue (ASR). Results from two full-scale combustion experiments, carried out on large ASR feeding lots (thousands of tons) indicate the proposed technology as a potential route to help the fulfilling of impending 95% reuse and recovery target set by the End of life Vehicle (ELV) Directive (January 2015). The paper describes the main operational troubleshot occurred during the first experiment (emissions at the stack out of regulatory limits and problems of clogging on the conveyer belt) and the consequent upgrading solutions (pre-treatment, introduction of waste double low-flow screw feeder and a cyclone prior to the main fan, modification of rotatory kiln inlet) adopted to allow, during the second long-term experiment, a continuous basis operation of the plant in full compliance with the discharge limit to the atmosphere. Characterization of both ASR and combustion residues allowed to quantify a 18% of combustion residues as not dangerous waste while only the 2% as hazardous one. A pre-treatment for the reduction of fines in the ASR was recommended in order to achieve the required energy recovery efficiency.

On the ASR and ASR thermal residues characterization of full scale treatment plant.

In order to obtain 85% recycling, several procedures on Automotive Shredder Residue (ASR) could be implemented, such as advanced metal and polymer recovery, mechanical recycling, pyrolysis, the direct use of ASR in the cement industry, and/or the direct use of ASR as a secondary raw material. However, many of these recovery options appear to be limited, due to the possible low acceptability of ASR based products on the market. The recovery of bottom ash and slag after an ASR thermal treatment is an option that is not usually considered in most countries (e.g. Italy) due to the excessive amount of contaminants, especially metals. The purpose of this paper is to provide information on the characteristics of ASR and its full-scale incineration residues. Experiments have been carried out, in two different experimental campaigns, in a full-scale tyre incineration plant specifically modified to treat ASR waste. Detailed analysis of ASR samples and combustion residues were carried out and compared with literature data. On the basis of the analytical results, the slag and bottom ash from the combustion process have been classified as non-hazardous wastes, according to the EU waste acceptance criteria (WAC), and therefore after further tests could be used in future in the construction industry. It has also been concluded that ASR bottom ash (EWC - European Waste Catalogue - code 19 01 12) could be landfilled in SNRHW (stabilized non-reactive hazardous waste) cells or used as raw material for road construction, with or without further treatment for the removal of heavy metals. In the case of fly ash from boiler or Air Pollution Control (APC) residues, it has been found that the Cd, Pb and Zn concentrations exceeded regulatory leaching test limits therefore their removal, or a stabilization process, would be essential prior to landfilling the use of these residues as construction material.

Review of Italian experience on automotive shredder residue characterization and management.

Automotive Shredder Residue (ASR) is a special waste that can be classified as either hazardous or non hazardous depending on the amount of hazardous substances and on the features of leachate gathered from EN12457/2 test. However both the strict regulation concerning landfills and the EU targets related to End-of-Life Vehicles (ELVs) recovery and recycling rate to achieve by 2015 (Directive 2000/53/EC), will limit current landfilling practice and will impose an increased efficiency of ELVs valorization. The present paper considers ELVs context in Italy, taking into account ASRs physical-chemical features and current processing practice, focusing on the enhancement of secondary materials recovery. The application in waste-to-energy plants, cement kilns or metallurgical processes is also analyzed, with a particular attention to the possible connected environmental impacts. Pyrolysis and gasification are considered as emerging technologies although the only use of ASR is debatable; its mixing with other waste streams is gradually being applied in commercial processes. The environmental impacts of the processes are acceptable, but more supporting data are needed and the advantage over (co-)incineration remains to be proven.

Development and calibration of a model for biohydrogen production from organic waste.

Existing models for H2 production are capable of predicting digester failure caused by a specific disturbance. However, they are based on studies using simple sugars, while it is known that H2 production and fermentation kinetics vary with the composition and characteristics of the substrate used. Because the behaviour of biological processes may differ significantly when the digesting material is a complex matrix, such as organic waste, the aim of this study was to develop and calibrate a mathematical model for the prediction of hydrogen production on the basis of the results obtained from a laboratory scale experimental study using source-selected organic waste. The calibration was carried out for the most important kinetic parameters in mesophilic anaerobic digestion processes and also served as a sensitivity analysis for the influence of both the specific growth rate (µmax and the half velocity constant (k(s)), both of which are strongly dependant on the substrate used. High values of µmax led to a shorter lag-time and to an overestimate of the cumulative final H2 production relative to the experimentally measured production. Additionally, high values of ks associated with amino acid and sugar fermentation corresponded to a lower rate of substrate consumption and to a greater lag-time for growth of hydrogen-producing microorganisms. In this case, a lower final H2 production was predicted than that which was experimentally observed. Because the model development and calibration provided useful information concerning the role of the kinetic constants in the analysis of a fermentative H2 production process from organic wastes, they may also represent a good foundation for the analysis of fermentative H2 production from organic waste for pilot and full-scale applications.

Thermal process of fluff: preliminary tests on a full-scale treatment plant.

Until only recently fluff has been largely disposed of in controlled landfill sites. However, in Europe environmental regulations, including the EU Landfill Directive 1999/31/EC and ELV (End of Life Vehicle) Directive 2000/53/EC, have dramatically increased the pressure on all stakeholders to develop alternative solutions. As increasingly stringent legislation forces Shredder Residues (SR) to be diverted from landfilling, newly developed technologies will be in a position to compete for the market value of disposing of the waste. However, the fluff waste stream is so variable that it cannot be automatically assumed that processes developed for one type of fluff will prove to be suitable for other fluff streams. This situation has contributed towards convincing stakeholders to withhold investment funds or delay taking decisions as to how best to proceed; as a consequence, very few technologies have been fully developed on a commercial basis. It is of particular interest therefore that commercial alternatives to be used in dealing with this complex waste stream should be identified. The present paper illustrates the findings of a full-scale thermal treatment performed on SR samples obtained from various shredding plants. The outcome of the study provides an important contribution towards assessing the feasibility and reliability of the process, thus constituting a basic prerequisite for process performance evaluation. The full-scale plant, designed for the thermo-valorization of tyres, was purpose-modified to allow for fluff combustion. Three different fluff compositions (car fluff with different percentage of shredding, whites and 100% car fluff) were taken into consideration. Both the raw samples and solid products were thoroughly characterized. Combustion emissions were continuously analyzed during the test period, alternatively operating for tyre and fluff combustion. Classification of combustion residues for landfill disposal was carried out indicating only 2% (ashes) as hazardous waste. Preliminary results, obtained from a unsophisticated thermodynamic analysis of the process, indicated a value of 0.61 for energy efficiency parameter calculated in accordance with the Directive 2008/98/EC. To conclude, the thermal treatment investigated may be deemed an appropriate technique for use in managing fluff. Indeed, values obtained for all organic and inorganic contaminants released into the atmosphere were lower than legal limits prescribed, and a significant energy content was recovered from waste fractions.

[Antisepsis and disinfection in emerging and re-emerging viral diseases]. / Malattie virali emergenti e riemergenti: antisepsi e disinfezione mirate per prevenirne la diffusione.

Prevention and therapy of emerging and re-emerging viral diseases may be inefficient due the inavailability of specific vaccines, active chemotherapic drugs, acceptable pesticides, and vector control measures. To reduce contact spreading of viral infections, some biocides, as chlorine compounds or phenolic compounds, associated with detergents, may assume a leading position as safe substances with antiviral properties for the environmental decontamination. Detailed instructions on their use may be introduced in future guidelines related to emerging and re-emerging viral diseases.

[SARS: diagnosis, therapy, and especially prevention]. / SARS: diagnostica, terapia e soprattutto prevenzione.

The main purpose of this review is to analyze some aspects of the severe acute respiratory syndrome, SARS, in order to obtain useful data to suggest preventive actions to reduce the spreading of the disease. Many elements have been examined to reach some conclusions and to allow an updated discussion. Surgical masks protect more the patient than the caregiver. Simple or double surgical masks may be useful, as double gloving protects the hands of the surgical personnel against percutaneous transmission of HIV eventually present in contaminated blood. The frequent substitution of the external masks with a new one will improve the filtering activity against droplets produced by cough or sneezes of the patient. The use of respiratory masks may be suggested in hospitals or in restricted ventilated areas where, even if coronavirus variant is considered an environmental contaminant more than a respiratory risk, droplets nuclei may persist in the air and add consistent dangers to the heath-care givers. Considering that large and medium droplets may infect floors and surfaces, in addition to gloves, gowns, masks and eyes protection, the available list of viral and bacterial factors implicated in SARS ethiology suggests a better hand antisepsis using frequently the alcohol based gels (containing an high percentage of emollients substances), if available. A liquid soap with triclosan can also be used, if the health-care workers compliance to hand washing increases, as expected in this explosive situation. On the basis of the results of some experimental data, the environmental disinfection may be effected with ethyl alcohol 70% in water. Disinfection of floors or larger surfaces may be obtained with chlorine compounds solutions, after an accurate pre-cleaning. When corrosion, bleaching or gas production have to be avoided, chlorine compounds may be substituted by phenolic detergent disinfectants.

Experimental study on carbon removal in biological aerated filters.

The aim of the present work was to evaluate the performance of a pilot-scale BAF in terms of removal of organic matter and suspended solids to obtain a highly polished effluent. The first part of the research was the evaluation of the optimal filter media for a full scale BAF. Mechanical and biological tests were performed over four materials: glass, plastic, pozzolan and expanded clay (Arlita) and the results obtained showed that the plastic spheres and the Arlita particles were the optimal materials for both the mechanical and biological requirements. Hence, a down-flow pilot scale BAF was set up in the laboratory to treat a synthetic medium. As filter media first plastic spheres and then Arlita spheres were used. Carbon removal studies were carried out at several influent COD concentrations, specific removal efficiency and COD profiles along the height of the filter were determined and used to analyze the process. Validation and calibration of a mathematical model formulated for carbon removal, were also performed by using the experimental data obtained. The results showed that this system allows us to achieve the more strict limits on final effluent.

The plasma membrane calcium pump is the preferred calpain substrate within the erythrocyte.

The activation of calpain in normal human erythrocytes incubated in the presence of Ca2+ and the Ca2+ ionophore A23187 led to the decline of the Ca(2+)-dependent ATPase activity of the cells. Preloading of the erythrocyte with an anticalpain antibody prevented the decline. The pump was also inactivated by applied to isolated erythrocyte plasma membranes. The decline of the pump activity corresponded to the degradation of the pump protein and was inversely correlated to the amount of the natural inhibitor of calpain, calpastatin, present in the cells. In erythrocytes containing only 50% of the normal level the degradation started at a concentration of Ca2+ significantly lower than in normal cells. A comparison of the concentrations of Ca2+ required for the degradation of a number of erythrocyte membrane proteins showed that the Ca2+ pump and band 3 were the most sensitive. All other membrane proteins tested were attacked at higher levels of intracellular Ca2+. Thus, the degradation of the Ca2+ pump protein may be a simple and sensitive means to monitor calpain activation in vivo. Furthermore, the results have shown that the calpastatin level correlated directly with the amount of activable calpain and with the concentration of Ca2+ required to trigger the activation process.

Site-directed activation of calpain is promoted by a membrane-associated natural activator protein.

Human erythrocytes contain a calpain activator protein with a molecular mass of approx. 40 kDa. The activator is present in association with the plasma membrane and promotes expression of calpain activity at a concentration of Ca2+ close to physiological values. The initial step of the activating mechanism involves association of the activator with calpain, followed by autoproteolytic activation of the proteinase in the presence of 1 microM Ca2+, at a rate identical to that induced by 1 mM Ca2+. In a reconstituted system, the activator binds to erythrocyte membranes, but not to phospholipid vesicles, suggesting the participation of an intrinsic membrane protein(s). In its membrane-associated form the activator selectively binds calpain, thus favouring interaction of the proteinase with the inner surface of plasma membranes. These results further confirm the importance of a natural activator protein in promoting intracellular activation of calpain under physiological conditions through a site-directed mechanism, which explains the high specificity of the proteinase for membrane of cytoskeletal proteins.

Different susceptibility of red cell membrane proteins to calpain degradation.

The presence of low levels of calpastatin activity in erythrocytes of hypertensive rats affects regulation of calpain activity so it is highly susceptible to activation within physiological fluctuations in [Ca2+]. Under identical conditions, in red cells of normotensive rats, calpain activation is efficiently controlled by the high levels of calpastatin activity, and a progressive increase in proteinase activity can only be observed in parallel with a decrease in the level of calpastatin. In intact erythrocytes from hypertensive rats exposed to small variations in [Ca2+], degradation of anion transport protein (band 3) and Ca(2+)-ATPase appears as a primary event indicating that these two transmembrane proteins are probably early recognized as targets of intracellular calpain activity. Furthermore, band 3 protein seems to be structurally modified in erythrocytes from hypertensive rats, as indicated by its increased susceptibility to degradation in the presence of 10-50 microM Ca2+. In addition, when exposed to progressive and limited increases in [Ca2+], erythrocytes from hypertensive rats, but not those from normotensive rats, show a high degree of fragility that can be restored to normal values by inhibition of calpain. These results indicate that, within fluctuations in [Ca2+] close to physiological values, regulation of calpain activity is efficiently accomplished in normal erythrocytes but is completely lost in cells from hypertensive animals. Regulation is of critical importance in maintaining normal structural and functional properties of selective red cell membrane and cytoskeletal proteins, among which band 3 and Ca(2+)-ATPase appear to be the substrates with highest susceptibility to digestion by calpain.

Modulation of inhibitory efficiency of rat skeletal muscle calpastatin by phosphorylation.

Rat skeletal muscle calpastatin form is markedly modified in its inhibitory properties by means of a reverse reaction which involves both phosphorylation and dephosphorylation. Dephospho-calpastatin shows greater inhibitory efficiency versus mu-calpain, whereas phospho-calpastatin shows maximal inhibition versus m-calpain. Both forms are present in fresh rat muscle. Phosphorylation has been reproduced "in vitro" using a homologous Ca2+ independent protein kinase and found to result in the incorporation of approximately one mole of 32P per mole of protein. Dephosphorylation was induced by treatment with alkaline phosphatase and 32P release shown found to correlate with modifications of the inhibitory properties. This reversible covalent modification of calpastatin is considered an important advancement in the understanding of how different calpain isoforms can be more efficiently controlled by a single inhibitor isozyme form.

Mechanism of action of the calpain activator protein in rat skeletal muscle.

Rat skeletal muscle contains a calpain activator protein characterized by a high specificity for calpain II, the high Ca(2+)-requiring isoform of this class of proteinases. The activator protein increases the rate of intramolecular conversion of the native 80-kDa catalytic subunit of calpain into the autolysed 75-kDa forms with maximal rate at concentrations of calcium approximately 25 times lower than those required by the native proteinase. The activator protein interacts with native calpain II forming a 1:1 complex; interaction does not occur with the fully activated form, produced by autoproteolysis. Even after immobilization to membranes, the activator binds to calpain, which then undergoes sequential activation and release from its bound form. The activator is itself resistant to digestion by calpain II, whereas it increases the rate at which homologous calpastatin is degraded by the proteinase. Taken together, these results are indicative of the existence in rat skeletal muscle of an activating system specific for calpain II which is potentially involved in the regulation of the inhibitory efficiency of calpastatin, through modulation of its intracellular level.

Identification of two calpastatin forms in rat skeletal muscle and their susceptibility to digestion by homologous calpains.

Two forms of calpastatin, differing in their specificity for the homologous calpain isozymes I and II, have been separated from rat skeletal muscle extracts and purified to homogeneity. Calpastatin I, the first form to elute in chromatography on DE32, is more effective against calpain I, while calpastatin II is more effective as an inhibitor of calpain II. Based on their molecular mass (approximately 105 kDa) both calpastatin forms belong to the high molecular mass class found in muscles of other animal species (Murachi, T., 1989, Biochem. Int. 18, 263-294). For calpain I, which is active with low (mu-M) concentrations of Ca2+, maximum inhibition with either calpastatin form was observed over a wide range of Ca2+ concentrations. With calpain II, which requires high (mM) concentrations of Ca2+ for activity, maximum inhibition required Ca2+ concentrations above 1 mM. Both calpastatin forms were found to be highly sensitive to degradation by calpain II, but almost completely resistant to degradation by calpain I. Degradation of calpastatin by calpain II is competitively inhibited by the addition of a calpain substrate. Isovaleryl carnitine (IVC), an intermediate product of L-leucine catabolism, previously demonstrated to be a potent and specific activator of rat skeletal muscle calpain II (Pontremoli, S., Melloni, E., Viotti, P. L., Michetti, M., Di Lisa, F., and Siliprandi, N., 1990. Biochem. Biophys. Res. Commun. 167, 373-380) greatly enhances the rate of degradation of calpastatins by calpain II. IVC, which decreases the Ca2+ requirement for maximal calpain II activity, also decreases the concentration of Ca2+ required for digestion of the inhibitor. For calpain II, regulation by either calpastatins may occur only in the presence of high [Ca2+].

Inactivation of hepatocyte protein kinase C by carbon tetrachloride: involvement of drug's metabolic activation and prooxidant effect.

The involvement of CCl4 biotransformation mechanism in decreasing the Protein Kinase C activity has been analyzed in hepatocytes isolated from phenobarbital-pretreated rats. A significant inhibition (55%) and an almost total disappearance (87%) of the enzyme activity were observed at 15 min and at 30 min incubation with CCl4, respectively. Cell preincubation with Trolox or desferrioxamine allowed a marked whilst not complete protection of both cytosolic and particulate Protein Kinase C activity. These results show that the CCl4 reactive metabolites play a primary role in hepatocyte Protein Kinase C impairment and suggest that besides lipid peroxidation other mechanisms -possibly a derangement of Ca2+ homeostasis- may be involved in this process.

Identification of an endogenous activator of calpain in rat skeletal muscle.

An additional component of the regulatory system of rat skeletal muscle calpain has been identified. It exerts a potent activating effect on calpain activity and is a heat stable small molecular weight protein. Of the two calpain isozymes present in muscle, the activator is specific for calpain II, being uneffective with calpain I. It promotes activation of the proteinase by reducing 50 fold, from 1 mM to of 20 microM, the requirement of Ca2+ for maximum catalytic activity of the proteinase. However in the presence of the activator calpain II expresses a consistent fraction of the maximum activity even at significantly lower concentrations of Ca2+ (below 5 microM Ca2+). The activator effect follows kinetics that are consistent with the presence of specific binding sites on the calpain molecules. The activator not only removes in a dose dependent fashion the inhibition of calpain by calpastatin, but also prevents inhibition of the proteinase upon the addition of calpastatin. Competition experiments revealed that the proteinase contains distinct sites for the activator and the inhibitor, and that both ligands can bind to calpain with the formation of an almost fully active ternary complex.

Murine erythroleukemia cell differentiation: possible involvement of a calcium dependent neutral proteinase.

Murine erythroleukemia cells contain a single type of calpain classified, on the basis of its calcium requirement, as a type I proteinase. The enzyme is practically inactive at concentrations of calcium below 10 microM and expresses maximal activity in the presence of 0.12-0.15 mM Ca2+. The affinity for Ca2+ cannot be reduced by exposure of the enzyme to conditions known to promote autoproteolysis of calpain. Expression of catalytic activity at lower concentrations of Ca2+, is promoted by the interaction with phospholipid vesicles or plasma membranes. Conditions that promote activation of calpain, induce also the self-inactivation of the enzyme. During terminal differentiation of murine erythroleukemia cells induced by HMBA, the intracellular level of calpain activity undergoes significative reduction. Similar decrease in calpain activity progressively occurs during the loss of sensitivity to HMBA as a result of density growth arrest.

Isovalerylcarnitine is a specific activator of the high calcium requiring calpain forms.

Isovalerylcarnitine, a product of the catabolism of L-leucine, is a potent activator of rat calpains isolated from erythrocytes, kidney, liver, skeletal and heart muscle. Only calpains II, but not calpains I, are activated by IVC, with the only exception of rat erythrocyte calpain I, the only species present in these cells which has a Ca2+ requirement higher than that of most calpain I isoenzymes. Activation by IVC involves a dual effect: 1) a ten fold increase in the affinity of calpain for Ca2+, and 2) an increase in the Vmax 1.3-1.6 fold above the values observed with the native enzymes at saturating [Ca2+] as well as with the autolyzed fully active calpain form at 5 microM Ca2+. The increased affinity for calcium results in an increased rate of autoproteolysis of calpain II. Activation by IVC is additive to that promoted by interaction (or association) to phospholipids vesicles. Together these results suggest that IVC may operate as a selective activator of calpain both in the cytosol and at the membrane level; in the latter case in synergism with the activation induced by association of the proteinase to the cell membrane.

Differential expression of protein kinase C isozymes and erythroleukemia cell differentiation.

Hexamethylene bisacetamide (HMBA) and other polar/apolar chemical agents are potent inducers of erythroid differentiation in murine erythroleukemia cells (MELC), as well as other transformed cell lines. Although the mechanism of action of HMBA is not yet known, evidence has been obtained that protein kinase C (PKC) plays a role in this process. In this study we provide further evidence that establishes this relationship. MELC contain two principal PKC activities, PKC beta and PKC alpha. MELC variants, selected for resistance to vincristine (VC), which display acceleration of their rates of induced differentiation, are enriched in PKC beta activity. When MELC are exposed to HMBA there is a fall in PKC activity, largely accounted for by a decline in PKC beta. This decline in PKC activity is faster in the VC-resistant, rapidly differentiating MELC. We previously demonstrated that VC-resistant MELC are resistant to the inhibition of differentiation by the phorbol ester, phorbol 12-myristate 13-acetate (PMA). In both VC-sensitive and -resistant MELC, PMA causes rapid membrane translocation and then a decline in PKC activity, accompanied by a generation of a Ca2+- and phospholipid-independent protein kinase activity. In VC/PMA-resistant variants, this Ca2+/phospholipid-independent protein kinase activity persists considerably longer than in the VC-sensitive variants. This correlates with the resistance to PMA and provides additional evidence for a role for the Ca2+/phospholipid-independent protein kinase activity during induced differentiation.

Vincristine-resistant erythroleukemia cell line has marked increased sensitivity to hexamethylenebisacetamide-induced differentiation.

Hexamethylenebisacetamide (HMBA)-induced murine erythroleukemia (MEL) differentiation is a multistep process. Commitment is the capacity to express terminal cell division and characteristics of the differentiated phenotype even after the cells are removed from culture with inducer. Culture of MEL cell line 745A.DS19 (DS19) with HMBA causes commitment to terminal differentiation after a latent period of about 10-12 hr. Previous studies have shown that during this latent period, HMBA causes a number of metabolic changes, including modulation in expression of certain protooncogenes. We now report the development of a MEL cell line (designated V3.17) derived from DS19 that is resistant to vincristine and is (i) markedly more sensitive to HMBA, (ii) induced to commitment without a detectable latent period, and (iii) resistant to the effects of phorbol ester and dexamethasone, which are potent inhibitors of HMBA-mediated DS19 differentiation. We suggest that this V3.17 MEL cell line may express a factor that circumvents HMBA-mediated early events, which prepare the cells for commitment to terminal differentiation.