956 resultados para Stabilization of looking


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This essay raises problems around the possible relationships of Semiotics, understood as science that investigates the life of signs, with Physical Education. It emphasizes the Semiotics or the General Logic of Signs elaborated by Charles S. Peirce (1839-1914), and presents its fundamentals, applications and implications for research in the field of Physical Education. It concludes that Semiotics allows launching new epistemic looks at the analysis and intervention in Physical Education, in order to: comprehend all its manifestations as expressive, meaningful and communicative; review the existing dichotomies in Physical Education; claim for the enhancement of looking at the subjects, their conducts and living motion situations as producers of language, and, therefore, of communication with the world.

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Pós-graduação em Agronomia (Produção Vegetal) - FCAV

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The research aimed to evaluate the relationship between the thermal environment of the rest pens in a pig slaughterhouse and the surface temperature (ST) of the animals as well as the effect of intermittent water spraying on thermal comfort. There were two visits to slaughter industrial being in each selected three pens of rest, one for control treatment (no spray throughout the period) and two for the treatment of intermittent spraying water (30 minutes intervals with constant spray followed by 30 minutes then turned off). Assessments began at the time of discharging animals in which are taken the thermographic images from lots using an infrared thermographic camera. After housing in the pens, were registered the thermographic images and the respiratory frequency (RF) every five minutes for four hours. Were evaluated the temperature, relative humidity and temperature-humidity index (THI). Considering the values of THI was observed that the animals remained in a condition of comfort only at the time of his arrival at the slaughterhouse. The ST and RF of the animals kept in the pens without sprinkler system (control) showed a linear increase in function of time of day, with increasing temperature. The treatment with intermittent sprinkling of water was observed oscillatory behavior of the ST and RF, however it was always lower than the control treatment, even in times when the water sprinklers were turned off. The time required of spray to reduction and stabilization of the ST of the animals was limited to the combination of temperature and relative humidity, being estimated at 25 minutes at times of higher THI. The maximum environmental temperature during evaluation was 28.4 oC.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Identifying new uses for residues of industries that process large quantities of biomass, as in bioethanol production, is essential for a sustainable development with reduced impact on the environment, which is the reason why many efforts have been devoted to find noble uses for lignins. in this study, a lignin obtained from sugarcane bagasse in a bioethanol producing plant was carboxymethylated to yield the water-soluble carboxymethyl lignin (CML), which was then used as stabilizing agent in aqueous alumina (Al2O3) suspensions. CML had a degree of substitution 0.46 +/- 0.01, in relation to the C9 unit of lignin, and behaved as a polyelectrolyte in a large pH range owing to the dissociation of carboxylic groups. The action of CML as stabilizing agent of alumina aqueous suspensions was investigated using viscometry, zeta potential, and photon correlation spectroscopy (PCS) measurements, mainly as a function of pH and time. Overall, the results showed that CML had a good performance as a deflocculating agent, because it led to dispersions with low viscosity and small change in particle size as a function of time. The positive effect from the addition of CML was confirmed in the morphological features of the material obtained from the alumina suspensions after elimination of water, as indicated by scanning electron microscopy. The stabilization of alumina suspensions afforded by CML opens the way for similar applications of modified lignins, whose electrical and structural properties may be tuned for specific uses in various industries, including the ceramic industry. (C) 2011 Elsevier B.V. All rights reserved.

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PURPOSE: To investigate the effect of lovastatin on renal ischemia followed by reperfusion. METHODS: Thirty one Wistar rats submitted to left renal ischemia for 60 minutes followed by contralateral nephrectomy were divided into two groups: A (n = 17, control, no treatment), and B (n = 14, lovastatin 15 mg/kg/day p.o. ten days before ischemia). The animals were sacrificed at the end of ischemia, after 24 hours and at seven days after reperfusion. Survival, serum urea and creatinine levels and renal mitochondrial function were evaluated. RESULTS: Mortality was 29.4% in group A and 0.7% in group B. Urea and creatinine levels were increased in both groups, but the values were significantly lower in group B. Mitochondrial function showed decoupling in 83.4% of group A, as opposed to 38.4/% of group B. CONCLUSIONS: The result shows a protective action of renal function by lovastatin administered before ischemia/reperfusion. Since most of the mitochondrial fraction presented membranes with the ability to maintain ATP production in group B, stabilization of the mitochondrial membrane should be considered as part of the protective action of lovastatin on renal function in ischemia/reperfusion.

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Two parametrically-induced phenomena are addressed in the context of a double pendulum subject to a vertical base excitation. First, the parametric resonances that cause the stable downward vertical equilibrium to bifurcate into large-amplitude periodic solutions are investigated extensively. Then the stabilization of the unstable upward equilibrium states through the parametric action of the high-frequency base motion is documented in the experiments and in the simulations. It is shown that there is a region in the plane of the excitation frequency and amplitude where all four unstable equilibrium states can be stabilized simultaneously in the double pendulum. The parametric resonances of the two modes of the base-excited double pendulum are studied both theoretically and experimentally. The transition curves (i.e., boundaries of the dynamic instability regions) are constructed asymptotically via the method of multiple scales including higher-order effects. The bifurcations characterizing the transitions from the trivial equilibrium to the periodic solutions are computed by either continuation methods and or by time integration and compared with the theoretical and experimental results.

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Objective: To evaluate the effect of different chewing gum brands on the salivary pH of children with primary dentition. Method: Forty children were selected and assigned to four groups: control (no chewing gum); sugarless chewing gum; chewing gum with casein phosphopeptide-amorphous calcium phosphate; and chewing gum with xylitol. The first saliva collection was made after supervised tooth brushing for stabilization of the oral pH. Next, all children were instructed to drink slowly 100 mL of a cola-based soft drink (Coca-Cola®) and a new saliva collection was made 10 min later. Then, each group chewed on the chewing gum for 5 min and discarded it after this time. Saliva was collected again at 5, 10 and 15 min intervals after start using the chewing gum. Measurement of salivary pH was made with colorimetric test papers and a digital pH-meter. Data were analyzed statistically by analysis of variance and Tukey’s test at a 5% significance level. Results: The use of chewing gums accelerated the increase of salivary pH to considerably alkaline levels after consumption of an acidic beverage, especially within the first minutes. The highest levels were obtained in the groups of children that used chewing gums containing xylitol and casein phosphopeptide-amorphous calcium phosphate. Conclusion: Children that used the chewing gums after ingestion of an acidic soft drink presented an increase in salivary pH, with the best results in the groups that used chewing gums containing casein phosphopeptide-amorphous calcium phosphate and xylitol.

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We have previously shown the differential expression of versican in the mouse uterus under ovarian hormone influence. We also demonstrated there is not a direct correlation between mRNA levels and protein expression, suggesting posttranscriptional events, such as alteration in mRNA stability. This posttranscriptional effect may result in the elongation and stabilization of transcripts poly(A) tail. Thus, the aim of this study was to analyze whether estradiol (E2) regulates versican mRNA stability and expression in a dose-related and time-dependent manner. For this purpose female mice were ovariectomized and treated with a single injection of 0.1 or 10 μg E2. To block transcription a group of females received a single injection of alpha-amanitin before hormone administration. Uterine tissues were collected 30 min, 1, 3, 6, 12 and 24 h after treatments and processed for quantitative real time PCR (qPCR), RACE-PAT Assay and immunohistochemistry. qPCR showed that versican mRNA levels are higher than control from 3 to 24 h after E2 administration, whereas after transcription inhibition versican mRNA unexpectedly increases within 3 h, which can be explained when transcriptional blockers alter the degradation rate of the transcript, resulting in the superinduction of this mRNA. Accordingly, analysis of versican transcript poly(A) tail evidenced a longer product 3 h after treatment, but not after 12 h. Versican immunoreaction becomes conspicuous in the superficial stroma only 3 h after E2 injection, whereas the whole stroma is immunoreactive from 6 h onward. These results demonstrate that E2 modulates versican at the transcriptional and posttranscriptional levels in a time-dependent manner.

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Because of its electronic properties, sulfur plays a major role in a variety of metabolic processes and, more in general, in the chemistry of life. In particular, S-S bridges between cysteines are present in the amino acid backbone of proteins. Protein disulfur radical anions may decay following different paths through competing intra and intermolecular routes, including bond cleavage, disproportionation, protein-protein cross linking, and electron transfer. Indeed, mass spectrometry ECD (electron capture dissociation massspectroscopy) studies have shown that capture of low-energy (<0.2 eV) electrons by multiply protonated proteins is followed by dissociation of S-S bonds holding two peptide chains together. In view of the importance of organic sulfur chemistry, we report on electron interactions with disulphide bridges. To study these interactions we used as prototypes the molecules dimethyl sulfide [(CH3)2S] and dimethyl disulfide [(H3C)S2(CH3)]. We seek to better understand the electron-induced cleavage of the disulfide bond. To explore dissociative processes we performed electron scattering calculations with the Schwinger Multichannel Method with pseudopotentials (SMCPP), recently parallelized with OpenMP directives and optimized with subroutines for linear algebra (BLAS) and LAPACK routines. Elastic cross sections obtained for different S-S bond lengths indicate stabilization of the anion formed by electron attachment to a σ*SS antibonding orbital, such that dissociation would be expected.

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The objectives of this thesis are to establish a chronological framework for environmental changes during the last 15,000 years in northwest Romania, to reconstruct the vegetation development, and to evaluate the underlying processes for forest dynamics. Furthermore, an overview of earlier and ongoing pollenstratigraphic work in Romania is provided. Sediments from two former crater lakes, Preluca Tiganului and Steregoiu, situated in the Gutaiului Mountains, on the western extremity of the Eastern Carpathians at 730 m and 790 m a.s.l., respectively were obtained and analysed for high-resolution pollen, macrofossils, charcoal, mineral magnetic parameters and organic matter. The chronostratigraphic framework was provided by dense AMS 14C measurements. Cold and dry climatic conditions are indicated by the occurrence of open vegetation with shrubs and herbs, and cold lake water prior to 14,700 cal. yr BP. The climatic improvement at the beginning of the Lateglacial interstadial (around 14,700 cal. yr BP) is seen by the development of open forests. These were dominated by Pinus and Betula, but contained also new arriving tree taxa, such as Populus, Alnus and Prunus. The gradual establishment of forests may have led to a stabilization of the soils in the catchment. Between ca. 14,100 and 13,800 cal. yr BP the forest density became reduced to stands of Pinus, Betula, Alnus, Larix and Populus trees and grassland expanded, suggesting colder climatic conditions. Picea arrived as a new taxon at around 13,800 cal. yr BP, and between 13,800 and 12,900 cal. yr BP, the surroundings of the sites were predominantly covered by Picea forest. This forest included Betula, Pinus, Alnus, Larix and Populus and, from 13,200 cal. yr BP onwards also Ulmus. At ca. 12,900 cal. yr BP, the forest became significantly reduced and at 12,600 cal. yr BP, a recurrence of open vegetation with stands of Larix, Pinus, Betula, Salix and Alnus is documented, lasting until 11,500 cal. yr BP. This distinct change in vegetation may by taken as a strong decline in temperature and moisture availability. At the transition to the Holocene, at ca. 11,500 cal. yr BP, Pinus, Betula and Larix quickly expanded (from small local stands) and formed open forests, probably as a response to warmer and more humid climatic conditions. At 11,250 cal. yr BP Ulmus and Picea expanded and the landscape became completely forested. The rapid increase of Ulmus and Picea after 11,500 cal. yr BP may suggest the existence of small residual populations close to the study sites during the preceding cold interval. Ulmus was the first and most prominent deciduous taxa in the early Holocene in the Gutaiului Mountains. From ca. 10,750 cal. yr BP onwards Quercus, Tilia, Fraxinus and Acer expanded and Corylus arrived. A highly diverse, predominantly deciduous forest with Ulmus, Quercus, Tilia, Fraxinus, Acer, Corylus and Picea developed between 10,700 and 8200 cal. yr BP, which possibly signifies more continental climatic conditions. The development of a Picea-Corylus dominated forest between 8200 and 5700 cal. yr BP is likely connected to a more humid and cooler climate. The establishment of Carpinus and Fagus was dated to 5750 cal. yr BP and 5200 cal. yr BP, respectively. The dominance of Fagus during the late Holocene, from 4000 cal. yr BP onwards, may have been related to cooler and more humid climatic conditions. First signs of human activities are recorded around 2300 cal. yr BP, but only during the last 300 years did local human impact become significant. The vegetation development recorded in the Gutaiului Mountains during the Lateglacial is very similar to reconstructions based on lowland sites, whereas higher elevation sites seem not to have always experienced visible vegetation changes. The time of tree arrival and expansion during the past 11,500 cal. yr BP seems to have occurred almost synchronously across Romania. The composition of the forests during the Holocene in the Gutaiului Mountains is consistent with that reconstructed at mid-elevation sites, but differs from the forest composition at higher elevations. Important differences between the Gutaiului Mountains and other studied sites in Romania are a low representation of Carpinus and a late and weak human impact. The available data sets for Romania give evidence for the presence of coniferous and cold-tolerant deciduous trees before 14,700 cal. yr BP. Glacial refugia for Ulmus may have occurred in different parts of Romania, whereas the existence of Quercus, Tilia, Corylus and Fraxinus has not been corroborated.

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In this thesis we focussed on the characterization of the reaction center (RC) protein purified from the photosynthetic bacterium Rhodobacter sphaeroides. In particular, we discussed the effects of native and artificial environment on the light-induced electron transfer processes. The native environment consist of the inner antenna LH1 complex that copurifies with the RC forming the so called core complex, and the lipid phase tightly associated with it. In parallel, we analyzed the role of saccharidic glassy matrices on the interplay between electron transfer processes and internal protein dynamics. As a different artificial matrix, we incorporated the RC protein in a layer-by-layer structure with a twofold aim: to check the behaviour of the protein in such an unusual environment and to test the response of the system to herbicides. By examining the RC in its native environment, we found that the light-induced charge separated state P+QB - is markedly stabilized (by about 40 meV) in the core complex as compared to the RC-only system over a physiological pH range. We also verified that, as compared to the average composition of the membrane, the core complex copurifies with a tightly bound lipid complement of about 90 phospholipid molecules per RC, which is strongly enriched in cardiolipin. In parallel, a large ubiquinone pool was found in association with the core complex, giving rise to a quinone concentration about ten times larger than the average one in the membrane. Moreover, this quinone pool is fully functional, i.e. it is promptly available at the QB site during multiple turnover excitation of the RC. The latter two observations suggest important heterogeneities and anisotropies in the native membranes which can in principle account for the stabilization of the charge separated state in the core complex. The thermodynamic and kinetic parameters obtained in the RC-LH1 complex are very close to those measured in intact membranes, indicating that the electron transfer properties of the RC in vivo are essentially determined by its local environment. The studies performed by incorporating the RC into saccharidic matrices evidenced the relevance of solvent-protein interactions and dynamical coupling in determining the kinetics of electron transfer processes. The usual approach when studying the interplay between internal motions and protein function consists in freezing the degrees of freedom of the protein at cryogenic temperature. We proved that the “trehalose approach” offers distinct advantages with respect to this traditional methodology. We showed, in fact, that the RC conformational dynamics, coupled to specific electron transfer processes, can be modulated by varying the hydration level of the trehalose matrix at room temperature, thus allowing to disentangle solvent from temperature effects. The comparison between different saccharidic matrices has revealed that the structural and dynamical protein-matrix coupling depends strongly upon the sugar. The analyses performed in RCs embedded in polyelectrolyte multilayers (PEM) structures have shown that the electron transfer from QA - to QB, a conformationally gated process extremely sensitive to the RC environment, can be strongly modulated by the hydration level of the matrix, confirming analogous results obtained for this electron transfer reaction in sugar matrices. We found that PEM-RCs are a very stable system, particularly suitable to study the thermodynamics and kinetics of herbicide binding to the QB site. These features make PEM-RC structures quite promising in the development of herbicide biosensors. The studies discussed in the present thesis have shown that, although the effects on electron transfer induced by the native and artificial environments tested are markedly different, they can be described on the basis of a common kinetic model which takes into account the static conformational heterogeneity of the RC and the interconversion between conformational substates. Interestingly, the same distribution of rate constants (i.e. a Gamma distribution function) can describe charge recombination processes in solutions of purified RC, in RC-LH1 complexes, in wet and dry RC-PEM structures and in glassy saccharidic matrices over a wide range of hydration levels. In conclusion, the results obtained for RCs in different physico-chemical environments emphasize the relevance of the structure/dynamics solvent/protein coupling in determining the energetics and the kinetics of electron transfer processes in a membrane protein complex.

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Although nickel is a toxic metal for living organisms in its soluble form, its importance in many biological processes recently emerged. In this view, the investigation of the nickel-dependent enzymes urease and [NiFe]-hydrogenase, especially the mechanism of nickel insertion into their active sites, represent two intriguing case studies to understand other analogous systems and therefore to lead to a comprehension of the nickel trafficking inside the cell. Moreover, these two enzymes have been demonstrated to ensure survival and colonization of the human pathogen H. pylori, the only known microorganism able to proliferate in the gastric niche. The right nickel delivering into the urease active site requires the presence of at least four accessory proteins, UreD, UreE, UreF and UreG. Similarly, analogous process is principally mediated by HypA and HypB proteins in the [NiFe]-hydrogenase system. Indeed, HpHypA and HpHypB also have been proposed to act in the activation of the urease enzyme from H. pylori, probably mobilizing nickel ions from HpHypA to the HpUreE-HpUreG complex. A complete comprehension of the interaction mechanism between the accessory proteins and the crosstalk between urease and hydrogenase accessory systems requires the determination of the role of each protein chaperone that strictly depends on their structural and biochemical properties. The availability of HpUreE, HpUreG and HpHypA proteins in a pure form is a pre-requisite to perform all the subsequent protein characterizations, thus their purification was the first aim of this work. Subsequently, the structural and biochemical properties of HpUreE were investigated using multi-angle and quasi-elastic light scattering, as well as NMR and circular dichroism spectroscopy. The thermodynamic parameters of Ni2+ and Zn2+ binding to HpUreE were principally established using isothermal titration calorimetry and the importance of key histidine residues in the process of binding metal ions was studied using site-directed mutagenesis. The molecular details of the HpUreE-HpUreG and HpUreE-HpHypA protein-protein assemblies were also elucidated. The interaction between HpUreE and HpUreG was investigated using ITC and NMR spectroscopy, and the influence of Ni2+ and Zn2+ metal ions on the stabilization of this association was established using native gel electrophoresis, light scattering and thermal denaturation scanning followed by CD spectroscopy. Preliminary HpUreE-HpHypA interaction studies were conducted using ITC. Finally, the possible structural architectures of the two protein-protein assemblies were rationalized using homology modeling and docking computational approaches. All the obtained data were interpreted in order to achieve a more exhaustive picture of the urease activation process, and the correlation with the accessory system of the hydrogenase enzyme, considering the specific role and activity of the involved protein players. A possible function for Zn2+ in the chaperone network involved in Ni2+ trafficking and urease activation is also envisaged.

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This work shows for the first time that native CSTB polymerizes on addition of Cu2+ and DnaK (Hsp70). Cysteines are involved in the polymerization process and in particular at least one cysteine is necessary. We propose that Cu2+ interacts with the thiol group of cysteine and oxidize it. The oxidized cysteine modifies the CSTB structure allowing interaction with DnaK/Hsp70 to occur. Thus, Cu2+ binding to CSTB exposes a site for DnaK and such interaction allows the polymerization of CSTB. The polymers generated from native CSTB monomers, are DTT sensitive and they may represent physiological polymers. Denatured CSTB does not require Cu2+ and polymerizes simply on addition of DnaK. The polymers generated from denatured CSTB do not respond to DTT. They have characteristics similar to those of the CSTB toxic aggregates described in vivo in eukaryotic cells following CSTB over-expression. Interaction between CSTB and Hsp70 is shown by IP experiments. The interaction occurs with WT CSTB and not with the cys mutant. This suggests that disulphur bonds are involved. Methal-cathalyzed oxidation of proteins involves reduction of the metal ion(s) bound to the protein itself and oxidation of neighboring ammino acid residues resulting in structural modification and de-stabilization of the molecule. In this work we propose that the cysteine thyol residue of CSTB in the presence of Cu2+ is oxidized, and cathalyzes the formation of disulphide bonds with Hsp70, that, once bound to CSTB, mediates its polymerization. In vivo this molecular mechanism of CSTB polymerization could be regulated by redox environment through the cysteine residue. This may imply that CSTB physiological polymers have a specific cellular function, different from that of the protease inhibitor known for the CSTB monomer. This hypothesis is interesting in relation to Progressive Myoclonus Epilepsy of type 1 (EPM1). This pathology is usually caused by mutations in the CSTB gene. CSTB is a ubiquitous protein, but EPM1 patients have problems only in the central nervous system. Maybe physiological CSTB polymers have a specific function altered in people affected by EPM1.

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Zusammenfassung:Im Infektionszyklus des Hepatitis-B-Virus spielt das große L-Hüllprotein mit seiner einzigartigen PräS1-Domäne eine zentrale Rolle. Es vermittelt die Bindung und Aufnahme in die Leberzelle, die Verpackung der Nukleokapside in die Virushülle, die Regulation der cccDNA-Amplifikation und eine transkriptionelle Aktivierung in der Wirtszelle. Zur Erfüllung seiner vielfältigen Aufgaben benötigt das L-Protein Unterstützung durch Wirtzellfaktoren, von denen einige im Rahmen dieser Untersuchung durch Verwendung von PräS1-Konstrukten als Fängerproteine im Hefe-Zwei-Hybrid-System identifiziert wurden. Mehrere Klone, die im Hefe-Zwei-Hybrid-Test mit dem C-terminalen PräS1-Fängerprotein (Aminosäure 44-108) isoliert worden waren, enthielten Teile der cDNA von gamma2-Adaptin, einem mutmaßlichen Mitglied der Clathrin-Adaptor-Proteine. Diese sind für intrazelluläre Membrantransportprozesse mittels clathrinumhüllter Vesikel verantwortlich. Unter den interagierenden Klonen, die mit dem N-terminalen Konstrukt des L-Proteins (Aminosäure 1-70) isoliert worden waren, befand sich überproportional häufig eine cDNA, die der schweren Kette H4 der Inter-Alpha-Trypsin-Inhibitor-Familie homolog war. H4 besitzt vermutlich bei der 'Akute-Phase-Reaktion', die Entzündungen folgt, und bei der Stabilisierung der extrazellulären Matrix physiologische Bedeutung. Weitere Klone kodierten für die Serinprotease C1r. Diese ist Bestandteil des C1-Komplex, der ersten Komponente des klassischen Komplementsystems. Die Spezifität der Bindung zwischen den positiven Klonen und der PräS1-Domäne wurde in weiteren biochemischen Interaktionstests bestätigt, sodaß H4, C1r und gamma2-Adaptin als Wirtszellfaktoren in der Physiologie des Hepatitis-B-Virus wahrscheinlich eine Rolle spielen.Abstract:Little is known about host cell factors necessary for hepatitis B virus assembly and infectivity. Central to virogenesis is the large L envelope protein that mediates hepatocyte receptor binding, envelopment of viral capsids, regulation of supercoiled DNA amplification and transcriptional transactivation. To assess its multiple functions and host-protein assistance involved, we here initiated a yeast two-hybrid screen using the L-specific preS1 domain as bait to screen a human liver cDNA library for L-interacting proteins. One of the most prominent cDNAs interacting with aminoacid sequence 44-108 of L-protein encodes for gamma2-adaptin, a novel clathrin adaptor-related protein responsible for protein sorting and trafficking. Among the clones interacting with the N-terminal construct of L-protein (aminoacid sequence 1-70), a frequently isolated cDNA corresponds to the gene for inter-alpha-trypsin family heavy chain H4, likely to be involved in acute inflammatory phase response and stabilization of extracellular matrices. Some other interacting clones were found to carry the cDNA for the serine protease C1r, a subunit of the C1 complex which initiates the classical complement cascade. The specificity of the interaction between the positive clones and the preS1 domain was further confirmed in independent biochemical experiments. Taken together, the results suggest a role for H4, C1r and gamma2-adaptin as host-cell factors in L-mediated process of viral biogenesis and/or pathogenesis.