Era uma vez...As sete casas da infortuna: relação actor - marioneta : Relatório de estágio

O presente relatório tem como objectivo analisar a realidade de uma Companhia de Marionetas em Portugal, Teatro e Marionetas de Mandrágora, incidindo sobre a relação entre o actor, marioneta e meio envolvente (site specific) no contexto de uma prática teatral concreta, a produção do espectáculo Era uma vez ...As Sete Casas da Infortuna, no Castelo de Santa Maria da Feira. Os resultados apurados acompanham os moldes em que se procurou passar de um actor convencional para a especialização de um micromundo teatral onde a aprendizagem foi, fulcral desde o processo de construção até, à manipulação em cena num universo de trabalho particularmente árduo e específico. A minha trajectória no seio da companhia começou pela familiarização e aprendizagem informal da prática das marionetas através da observação directa dos espectáculos e de diferentes projectos da companhia. A formação feita no local de estágio (atelier da Companhia) foi outra das modalidades de formação que acompanhou o meu trajecto ao longo do estágio através da minha colaboração em projectos como Teatro nas Instituições, sempre sob a égide da divisão de tarefas e de alguma autoaprendizagem com a devida supervisão e orientação de artistas especialistas, como o Director Criativo, enVide neFelibata. O laboratório de aprendizagem prosseguiu pela mão da marionetista Clara Ribeiro que orientou a minha formação no sentido de absorver princípios teórico-práticos como o Foco, o Movimento e o Olhar para uma melhor consciencialização do universo do teatro de marionetas e formas animadas. Ainda nessa aprendizagem, a figura de alguns artistas especialistas foi fulcral, nomeadamente a da cenógrafa Marta Fernandes da Silva, que revelou ser de extrema importância ao longo do estágio pois permitiu uma verdadeira intersecção entre as componentes teórica e prática. Dessa forma pude acompanhar todo o processo de criação do Era uma vez ...As Sete Casas da Infortuna, e colaborar na construção das marionetas que iria manipular como actor - marionetista. Relativamente à questão da componente da interpretação no estágio, esta foi assumida pela encenadora - marionetista, Filipa Alexandre que, com a sua orientação, permitiu um enfoque do grupo de trabalho num processo de criação colectiva. Assim, a nível pessoal, fui movido por uma necessidade premente de descodificar o papel do actor na sua relação com a marioneta e o meio envolvente (site specific). ABSTRACT: This study aims to examine the reality of a Puppet Company in Portugal, the Puppet Theater of Mandrágora, focusing on the relationship between actor, puppet and environment (site specific) in the context of an actual theatrical practice, Once upon a time ...Seven houses of infortune, in the Castle of Santa Maria da Feira. The results obtained follow the way in which one seeks to move from one "conventional" actor to the specialization of a micro theater where learning was central from the building process to manipulation on the scene in a universe of work particularly hard and specific. My initial course within the company began with the familiarization and informal learning of the practice of puppetry through direct observation of performances and various projects of the company. The training done at the company (workshop of the Company) was one of the training arrangements that accompanied my way along the training through assistance on projects such as Teatro nas Instituições, always under the auspices of the division of tasks and due self-teaching with appropriate supervision and guidance of expert artists, such as the Creative Director, enVide neFelibata. The learning laboratory continued by the hand of the puppeteer Clara Ribeiro who supervised my training in order to absorb theoretical and practical principles as the Focus, the Movement and the Look for better awareness of the world of puppetry and animated forms. Also at this level, the figure of some specialist artists were central, such as the scenographer Marta Fernandes da Silva, who proved to be extremely important during this training, allowing a true intersection between the theoretical and the practical components. I could experience the surroundings of the creation notebook of Era uma vez ...As Sete Casas da Infortuna, and collaborate in the preparation of puppets that would handle as an actor - puppeteer. As to the question of the interpretation component on training, this was in charge of stage director - puppeteer, and the Artistic Director, Filipa Alexandre, whose instructions allowed a focus of the working group in a process of collective creation. Accordingly, to a personal level, I was urged to decode the role of the actor in relation to the puppet and the environment (site specific).

Sviluppo di processi di comunicazione aptici tra uomo e macchina

La ricerca si focalizza sul rapporto tra tecnologie abilitanti e corpo umano. La miniaturizzazione delle tecnologie, unita alla loro maggiore diffusione negli ambienti, porta ad interrogarsi sull’efficacia dell’integrazione di esse con corpo e attività ad esso connesse. Il contesto problematico della ricerca riguarda i dispositivi indossabili e il progetto di soluzioni destinate a risolvere inediti bisogni o potenziare i sensi umani. La letteratura scientifica e i casi studio circoscrivono il piede come efficace piattaforma per la sperimentazione di interfacce aptiche di comunicazione uomo/macchina, atte a connettere il corpo con informazioni referenziate all’ambiente. Il piede, elemento motorio duplice e simmetrico, ha un’elevata qualità percettiva ed è morfologicamente adeguato all’applicazione di tecnologie emergenti. La posizione di soglia, tra spazio e corpo, consente la raccolta di stimoli da entrambe le aree. La bibliografia evidenzia quanto la pressione, rispetto alla vibrazione, sia preferibile nella comunicazione aptica in quanto componente naturale dei linguaggi relazionali del corpo. Dall’analisi multidisciplinare emerge infine l’opportunità di sviluppo del ritmo come componente strutturale dei messaggi. I legami relazionali tra ritmo, corpo e comportamenti umani sono evidenti in molteplici meccanismi: trascinamento ritmico, mimesi ritmica, sincronia. La messa in relazione di piede, pressione e ritmo diventa affordance dello spazio, capace di suggerire, enfatizzare o attivare determinati comportamenti. L’unione di questi elementi è qui definita ritmica podotattile ed esplicitata nella tesi della descrizione delle sue caratteristiche, dalla circoscrizione di campi e azioni applicative e dalla raccolta dati sui test effettuati con i prototipi costruiti. Le analisi quantitative e qualitative dei dati di lettura del movimento e delle emozioni dimostrano quanto l’utilizzo di un linguaggio ritmico aptico nel piede esprima elevate potenzialità di integrazione con il corpo nel rispetto del comfort e dell’equilibrio attentivo nei flussi di azione preesistenti. I risultati aprono riflessioni su nuove applicazioni progettuali nel campo museale, lavorativo e urbano.

Metocean Climate Impact on the Fatigue Assessment of a Jacket-Type Offshore Platform

When the offshore oil and gas supplies exhaust, most offshore platforms are decommissioned and removed. The purpose of this paper is to evaluate the fatigue damage that will occur during the service life of a jacket-type offshore platform using different fatigue approaches in particular locations. The locations considered for this metocean climate impact study were Norway (North Sea), Portugal (Atlantic Ocean - Leixões) and Italy (Adriatic Sea). A finite element model was created by the means of Sesam and two different fatigue analysis, deterministic and spectral, were applied. For the fatigue assessment, an appropriate description of the site-specific wave environment, during the jacket platform service life, must be accomplished. This description is usually provided by a wave scatter diagram. Wave scatter diagrams usually represent the long-term wave environment during a (typical) year and are based on several years of site-specific data to ensure that they adequately represent the wave environment at the location of the structure. In this thesis, the comparison between these fatigue approaches will serve as a pilot study for planned reliability analysis in decommissioned offshore platforms in order to maximize the reuse of these platforms for future wind generation systems.

Adattamento morfologico e variazione dell'espressione genica dopo pneumonectomia: osservazioni su modello sperimentale animale

Introduzione La pneumonectomia su modello animale potrebbe essere un’utile piattaforma di studio per approfondire i meccanismi della risposta compensatoria al danno polmonare. Scopo dello studio è determinare la presenza di variazioni morfologiche e di espressione del trascrittoma dopo pneumonectomia. Materiali e metodi Undici suini sono stati sottoposti a pneumonectomia sinistra. Sono stati eseguiti prelievi sito-specifici intraoperatori su polmone sinistro e successivamente confrontati con prelievi sito-specifici su polmone destro dopo eutanasia a 60 giorni. I prelievi degli animali con decorso regolare sono stati sottoposti a RNA-sequencing e successiva analisi computazionale per valutare il peso funzionale del singolo gene o di clusters di geni. Risultati Un animale è stato escluso per insorgenza di ernia diaframmatica. In 7/10 è stata riscontrata apertura della pleura mediastinica con parziale erniazione del polmone controlaterale e shift mediastinico. L’istologia ha mostrato dilatazione degli spazi aerei, rottura dei setti interalveolari, lieve infiammazione, assenza di fibrosi, stiramento radiale dei bronchi e riduzione del letto capillare. L’analisi di bulk RNA-sequencing ha identificato 553 geni espressi in modo differenziale (DEG)(P<0,001) tra pre e post-pneumonectomia. I primi 10 DEG up-regolati: Edn1, Areg, Havcr2, Gadd45g, Depp1, Cldn4, Atf3, Myc, Gadd45b, Socs3; i primi 10 geni down-regolati: Obscn, Cdkn2b, ENSSSCG00000015738, Prrt2, Amer1, Flrt3, Efnb2, Tox3, Znf793, Znf365. Tra i DEG è stata riscontrata una predominanza di geni specifici dei macrofagi. L’analisi di gene ontology basata su DAVID ha mostrato un significativo arricchimento della "via di segnalazione apoptotica estrinseca"(FDR q=7,60x10 -3), della via di “Risposta all’insulina”(FDR q=7,60x10 -3) ed un arricchimento di geni “Regolatori negativi del segnale DDX58/IFIH1”(FDR q=7.50x10 -4). Conclusioni Il presente studio conferma la presenza di variazioni macroscopiche e microscopiche fenotipiche dopo pneumonectomia. L’RNA sequencing e lo studio di genomica traslazionale hanno mostrato l’esistenza di geni singoli e di network di geni disregolati dopo pneumonectomia, prevalentemente in determinate popolazioni cellulari.

Percorsi di valorizzazione artistica di ambienti naturali per una nuova consapevolezza ecologica

L’idea di questa tesi nasce da due riflessioni maturate in parallelo nel corso del mio percorso di tirocinio e nate, più in generale, durante quello di studi. La prima è relativa ai tentativi di convergenza tra arte, scienza e cultura in linea con le strategie di pianificazione comunitarie del futuro, e la seconda sul tema del Service Design e in particolare dello Human Centered Design, considerato, nell’analisi di questo lavoro, insufficiente a inquadrare e cogliere interamente il sistema relazionale entro cui è contestualizzato l’utente-cittadino, o più in generale il soggetto per cui si progetta e si pianifica. Un primo passaggio concettuale e progettuale derivato da queste riflessioni è stato quindi quello dello spostamento del focus del servizio dall’esclusività dell’utente all’inclusività dell’utente inserito in un ambiente, in considerazione del bisogno latente e ultimo di ogni essere umano: una convivenza e una relazione sana e cosciente con ciò che lo circonda, sottolineando l’importanza della molteplicità di relazioni che legano ogni individuo al contesto in cui è inserito. Il progetto di tesi si pone infatti l’obiettivo di ribaltare la narrazione negativa e il graduale allontanamento degli abitanti di Ravenna dalle grandi aree naturali che circondano la città, attraverso un servizio di promozione che integri attività di produzione artistica e culturale con percorsi di ingaggio, sensibilizzazione e confronto dei cittadini e degli artisti stessi. Il servizio progettato è stato concretizzato in un sistema di valorizzazione artistica tramite residenze per ambienti naturali mirato alla diffusione di una nuova consapevolezza ecologica e territoriale nella popolazione. È stato sviluppato partendo da un evento di lancio site-specific per giungere alla definizione di un format idealmente replicabile ed espandibile per qualsiasi territorio in cui ve ne fosse bisogno.

Relationship between B-Cell-specific moloney murine leukemia virus integration site 1 (BMI-1) and homologous recombination regulatory genes in invasive ductal breast carcinomas

B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi-1) is a Polycomb group protein that is able to induce telomerase activity, enabling the immortalization of epithelial cells. Immortalized cells are more susceptible to double-strand breaks (DSB), which are subsequently repaired by homologous recombination (HR). BRCA1 is among the HR regulatory genes involved in the response to DNA damage associated with the RAD51 protein, which accumulates in DNA damage foci after signaling H2AX, another important marker of DNA damage. Topoisomerase III beta (topoIII beta) removes HR intermediates before chromosomal segregation, preventing damage to cellular DNA structure. In breast carcinomas positive for BMI-1 the role of proteins involved in HR remains to be investigated. The aim of this study was to evaluate the association between BMI-1 and homologous recombination proteins. Using tissue microarrays containing 239 cases of primary breast tumors, the expression of Bmi-1, BRCA-1, H2AX, Rad51, p53, Ki-67, topoIII beta, estrogen receptors (ER), progesterone receptors (PR), and HER-2 was analyzed by immunohistochemistry. We observed high Bmi-1 expression in 66 cases (27.6%). Immunohistochemical overexpression of BMI-1 was related to ER (p=0.004), PR (p<0.001), Ki-67 (p<0.001), p53 (p=0.003), BRCA1 (p=0.003), H2AX (p=0.024) and topoIII beta (p<0,001). Our results show a relationship between the expression of BMI-1 and HR regulatory genes, suggesting that Bmi-1 overexpression might be an important event in HR regulation. However, further studies are necessary to understand the mechanisms in which Bmi-1 could regulate HR pathways in invasive ductal breast carcinomas.

The BCL2 major breakpoint region is a sequence- and cell-cycle-specific binding site of the Ku antigen.

The majority of translocations involving BCL2 are very narrowly targeted to three breakpoint clusters evenly spaced over a 100-bp region of the gene's terminal exon. We have recently shown that the immediate upstream boundary of this major breakpoint region (mbr) is a specific recognition site for single-strand DNA (ssDNA) binding proteins on the sense and antisense strands. The downstream flank of the mbr is a helicase binding site. In this report we demonstrate that the helicase and ssDNA binding proteins show reciprocal changes in binding activity over the cell cycle. The helicase is maximally active in G1 and early S phases; the ssDNA binding proteins are maximally active in late S and G2/M phases. An inhibitor of helicase binding appears in late S and G2/M. Finally, at least one component of the helicase binding complex is the Ku antigen. Thus, a protein with helicase activity implicated in repair of double-strand breaks, variable (diversity) joining recombination, and, potentially, cell-cycle regulation is targeted to the BCL2 mbr.

Ataxia telangiectasia mutated (ATM) kinase and ATM and Rad3 related kinase mediate phosphorylation of Brca1 at distinct and overlapping sites - In vivo assessment using phospho-specific antibodies

Recent studies have provided evidence that breast cancer susceptibility gene products (Brca1 and Brca2) suppress cancer, at least in part, by participating in DNA damage signaling and DNA repair. Brca1 is hyperphosphorylated in response to DNA damage and co-localizes with Rad51, a protein involved in homologous-recombination, and Nbs1·Mre11·Rad50, a complex required for both homologous-recombination and nonhomologous end joining repair of damaged DNA. Here, we report that there is a qualitative difference in the phosphorylation states of Brca1 between ionizing radiation (IR) and UV radiation. Brca1 is phosphorylated at Ser-1423 and Ser-1524 after IR and UV; however, Ser-1387 is specifically phosphorylated after IR, and Ser-1457 is predominantly phosphorylated after UV. These results suggest that different types of DNA-damaging agents might signal to Brca1 in different ways. We also provide evidence that the rapid phosphorylation of Brca1 at Ser-1423 and Ser-1524 after IR (but not after UV) is largely ataxia telangiectasia mutated (ATM) kinase-dependent. The overexpression of catalytically inactive ATM and Rad3 related (ATR) kinase inhibited the UV-induced phosphorylation of Brca1 at these sites, indicating that ATR controls Brca1 phosphorylation in vivo after the exposure of cells to UV light. Moreover, ATR associates with Brca1; ATR and Brca1 foci co-localize both in cells synchronized in S phase and after exposure of cells to DNA-damaging agents. ATR can itself phosphorylate the region of Brca1 phosphorylated by ATM (Ser-Gln cluster in the C terminus of Brca1, amino acids 1241-1530). However, there are additional uncharacterized ATR phosphorylation site(s) between residues 521 and 757 of Brca1. Taken together, our results support a model in which ATM and ATR act in parallel but somewhat overlapping pathways of DNA damage signaling but respond primarily to different types of DNA lesion.

Ataxia telangiectasia mutated (ATM) kinase and ATM and Rad3 related kinase mediate phosphorylation of Brca1 at distinct and overlapping sites - In vivo assessment using phospho-specific antibodies

Recent studies have provided evidence that breast cancer susceptibility gene products (Brca1 and Brca2) suppress cancer, at least in part, by participating in DNA damage signaling and DNA repair. Brca1 is hyperphosphorylated in response to DNA damage and co-localizes with Rad51, a protein involved in homologous-recombination, and Nbs1.Mre11.Rad50, a complex required for both homologous-recombination and nonhomologous end joining repair of damaged DNA. Here, we report that there is a qualitative difference in the phosphorylation states of Brca1 between ionizing radiation (IR) and UV radiation. Brca1 is phosphorylated at Ser-1423 and Ser-1524 after IR and W; however, Ser-1387 is specifically phosphorylated after IR, and Ser-1457 is predominantly phosphorylated after W. These results suggest that different types of DNA-damaging agents might signal to Brca1 in different ways. We also provide evidence that the rapid phosphorylation of Brca1 at Ser-1423 and Ser-1524 after IR (but not after W) is largely ataxia telangiectasia mutated (ATM) kinase-dependent. The overexpression of catalytically inactive ATM and Rad3 related (ATR) kinase inhibited the UV-induced phosphorylation of Brca1 at these sites, indicating that ATR controls Brca1 phosphorylation in vivo after the exposure of cells to UV light. Moreover, ATR associates with Brca1; ATR and Brca1 foci co-localize both in cells synchronized in S phase and after exposure of cells to DNA-damaging agents. ATR can itself phosphorylate the region of Brca1 phosphorylated by ATM (Ser-Gln cluster in the C terminus of Brca1, amino acids 1241-1530), However, there are additional uncharacterized ATR phosphorylation site(s) between residues 521 and 757 of Brca1, Taken together, our results support a model in which ATM and ATR act in parallel but somewhat overlapping pathways of DNA damage signaling but respond primarily to different types of DNA lesion.

Hypomorphic mutation in the site-1 protease Mbtps1 endows resistance to persistent viral infection in a cell-specific manner.

The prototypic arenavirus lymphocytic choriomeningitis virus (LCMV), which naturally persists in rodents, represents a model for HIV, HBV, and HCV. Cleavage of the viral glycoprotein precursor by membrane-bound transcription factor peptidase, site 1 (Mbtps1 or site-1 protease), is crucial for the life cycle of arenaviruses and therefore represents a potential target for therapy. Recently, we reported a viable hypomorphic allele of Mbtps1 (woodrat) encoding a protease with diminished enzymatic activity. Using the woodrat allele, we examine the role of Mbtps1 during persistent LCMV infection. Surprisingly, Mbtps1 inhibition limits persistent but not acute viral infection and is associated with an organ/cell type-specific decrease in viral titers. Analysis of bone marrow-derived dendritic cells from woodrat mice supports their specific role in resolving persistent viral infection. These results support in vivo targeting of Mbtps1 in the treatment of arenavirus infections and demonstrate a critical role for dendritic cells in persistent viral infections.

Accumulation of human immunodeficiency virus-specific cytotoxic T lymphocytes away from the predominant site of virus replication during primary infection.

Down-regulation of the initial burst of viremia during primary human immunodeficiency virus (HIV) infection is thought to be mediated predominantly by HIV-specific CD8+ cytotoxic T lymphocytes (CTL). This response is associated with major perturbations in the T cell receptor (TCR) repertoire. To investigate the failure of the cellular immune response to adequately control viral spread and replication and to prevent establishment of HIV infection, changes in the TCR repertoire and in the distribution of virus-specific CTL between blood and lymph node were analyzed in three patients with primary infection. By the combined use of clonotype-specific polymerase chain reaction and analysis of the frequency of in vivo activated HIV-specific CTL, it was shown that HIV-specific CTL clones preferentially accumulated in blood as opposed to lymph node. Accumulation of HIV-specific CTL in blood occurred prior to effective down-regulation of virus replication in both blood and lymph node. These findings should provide new insights into how HIV, and possibly other viruses, elude the immune response of the host during primary infection.

New generation vaccine induces effective melanoma-specific CD8+ T cells in the circulation but not in the tumor site.

Although increasing evidence suggests that CTL are important to fight the development of some cancers, the frequency of detectable tumor-specific T cells is low in cancer patients, and these cells have generally poor functional capacities, compared with virus-specific CD8(+) T cells. The generation with a vaccine of potent CTL responses against tumor Ags therefore remains a major challenge. In the present study, ex vivo analyses of Melan-A-specific CD8(+) T cells following vaccination with Melan-A peptide and CpG oligodeoxynucleotides revealed the successful induction in the circulation of effective melanoma-specific T cells, i.e., with phenotypic and functional characteristics similar to those of CTL specific for immunodominant viral Ags. Nonetheless, the eventual impact on tumor development in vaccinated melanoma donors remained limited. The comprehensive study of vaccinated patient metastasis shows that vaccine-driven tumor-infiltrating lymphocytes, although activated, still differed in functional capacities compared with blood counterparts. This coincided with a significant increase of FoxP3(+) regulatory T cell activity within the tumor. The consistent induction of effective tumor-specific CD8(+) T cells in the circulation with a vaccine represents a major achievement; however, clinical benefit may not be achieved unless the tumor environment can be altered to enable CD8(+) T cell efficacy.

Activation of human meiosis-specific recombinase Dmc1 by Ca2+.

Rad51 and its meiotic homolog Dmc1 are key proteins of homologous recombination in eukaryotes. These proteins form nucleoprotein complexes on single-stranded DNA that promote a search for homology and that perform DNA strand exchange, the two essential steps of genetic recombination. Previously, we demonstrated that Ca2+ greatly stimulates the DNA strand exchange activity of human (h) Rad51 protein (Bugreev, D. V., and Mazin, A. V. (2004) Proc. Natl. Acad. Sci. U. S. A. 101, 9988-9993). Here, we show that the DNA strand exchange activity of hDmc1 protein is also stimulated by Ca2+. However, the mechanism of stimulation of hDmc1 protein appears to be different from that of hRad51 protein. In the case of hRad51 protein, Ca2+ acts primarily by inhibiting its ATPase activity, thereby preventing self-conversion into an inactive ADP-bound complex. In contrast, we demonstrate that hDmc1 protein does not self-convert into a stable ADP-bound complex. The results indicate that activation of hDmc1 is mediated through conformational changes induced by free Ca2+ ion binding to a protein site that is distinct from the Mg2+.ATP-binding center. These conformational changes are manifested by formation of more stable filamentous hDmc1.single-stranded DNA complexes. Our results demonstrate a universal role of Ca2+ in stimulation of mammalian DNA strand exchange proteins and reveal diversity in the mechanisms of this stimulation.

Homing phenotypes of tumor-specific CD8 T cells are predetermined at the tumor site by crosspresenting APCs.

Expression of tissue-specific homing molecules directs antigen-experienced T cells to particular peripheral tissues. In studies using soluble antigens that focused on skin and gut, antigen-presenting cells (APCs) within regional lymphoid tissues were proposed to be responsible for imprinting homing phenotypes. Whether this occurs in other sites and after physiologic antigen processing and presentation is unknown. We define in vivo imprinting of distinct homing phenotypes on monospecific T cells responding to antigens expressed by tumors in intracerebral, subcutaneous, and intraperitoneal sites with efficient brain-tropism of CD8 T cells crossprimed in the cervical lymph nodes (LNs). Multiple imprinting programs could occur simultaneously in the same LN when tumors were present in more than one site. Thus, the identity of the LN is not paramount in determining the homing phenotype; this critical functional parameter is dictated upstream at the site of antigen capture by crosspresenting APCs.

Specific detection of Lettuce mosaic virus isolates belonging to the Most type

Lettuce mosaic virus (LMV)-Most isolates can infect and are seed-borne in cultivars containing the mol gene. A reverse transcription and polymerase chain reaction (RT-PCR)-based test was developed for the specific detection of LMV-Most isolates. Based on the complete genome sequences of three LMV isolates belonging respectively to the Most type, the Common type and neither of these two types, three different assays were compared: (i) presence of a diagnostic restriction site in the region of the genome encoding the variable N-terminus of the capsid protein, in the 3' end of the genome, (ii) RT-PCR using primers designed to amplify a cDNA corresponding to a portion of the P1 coding region, in the 5' end of the genome and (iii) RT-PCR using primers designed to amplify a central region of the genome. The assays were performed against a collection of 21 isolates from different geographical origins and representing the molecular variability of LMV. RT-PCR of the central region of the genome was preferred because its results are expected to be less affected by natural recombination between LMV isolates, and it allows sensitive detection of LMV-Most in situations of single as well as mixed contamination. (C) 2004 Elsevier B.V. All rights reserved.