RESUMO
Tight relationships exist in the local Universe between the central stellar properties of galaxies and the mass of their supermassive black hole (SMBH)1-3. These suggest that galaxies and black holes co-evolve, with the main regulation mechanism being energetic feedback from accretion onto the black hole during its quasar phase4-6. A crucial question is how the relationship between black holes and galaxies evolves with time; a key epoch to examine this relationship is at the peaks of star formation and black hole growth 8-12 billion years ago (redshifts 1-3)7. Here we report a dynamical measurement of the mass of the black hole in a luminous quasar at a redshift of 2, with a look back in time of 11 billion years, by spatially resolving the broad-line region (BLR). We detect a 40-µas (0.31-pc) spatial offset between the red and blue photocentres of the Hα line that traces the velocity gradient of a rotating BLR. The flux and differential phase spectra are well reproduced by a thick, moderately inclined disk of gas clouds within the sphere of influence of a central black hole with a mass of 3.2 × 108 solar masses. Molecular gas data reveal a dynamical mass for the host galaxy of 6 × 1011 solar masses, which indicates an undermassive black hole accreting at a super-Eddington rate. This suggests a host galaxy that grew faster than the SMBH, indicating a delay between galaxy and black hole formation for some systems.
RESUMO
Cells use compartmentalization of enzymes as a strategy to regulate metabolic pathways and increase their efficiency1. The α- and ß-carboxysomes of cyanobacteria contain ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco)-a complex of eight large (RbcL) and eight small (RbcS) subunits-and carbonic anhydrase2-4. As HCO3- can diffuse through the proteinaceous carboxysome shell but CO2 cannot5, carbonic anhydrase generates high concentrations of CO2 for carbon fixation by Rubisco6. The shell also prevents access to reducing agents, generating an oxidizing environment7-9. The formation of ß-carboxysomes involves the aggregation of Rubisco by the protein CcmM10, which exists in two forms: full-length CcmM (M58 in Synechococcus elongatus PCC7942), which contains a carbonic anhydrase-like domain8 followed by three Rubisco small subunit-like (SSUL) modules connected by flexible linkers; and M35, which lacks the carbonic anhydrase-like domain11. It has long been speculated that the SSUL modules interact with Rubisco by replacing RbcS2-4. Here we have reconstituted the Rubisco-CcmM complex and solved its structure. Contrary to expectation, the SSUL modules do not replace RbcS, but bind close to the equatorial region of Rubisco between RbcL dimers, linking Rubisco molecules and inducing phase separation into a liquid-like matrix. Disulfide bond formation in SSUL increases the network flexibility and is required for carboxysome function in vivo. Notably, the formation of the liquid-like condensate of Rubisco is mediated by dynamic interactions with the SSUL domains, rather than by low-complexity sequences, which typically mediate liquid-liquid phase separation in eukaryotes12,13. Indeed, within the pyrenoids of eukaryotic algae, the functional homologues of carboxysomes, Rubisco adopts a liquid-like state by interacting with the intrinsically disordered protein EPYC114. Understanding carboxysome biogenesis will be important for efforts to engineer CO2-concentrating mechanisms in plants15-19.
Assuntos
Proteínas de Bactérias/metabolismo , Organelas/metabolismo , Multimerização Proteica , Ribulose-Bifosfato Carboxilase/química , Ribulose-Bifosfato Carboxilase/metabolismo , Synechococcus/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/ultraestrutura , Ciclo do Carbono , Dióxido de Carbono/metabolismo , Anidrases Carbônicas/química , Anidrases Carbônicas/metabolismo , Anidrases Carbônicas/ultraestrutura , Microscopia Crioeletrônica , Dissulfetos/metabolismo , Modelos Moleculares , Oxirredução , Subunidades Proteicas/química , Subunidades Proteicas/metabolismo , Ribulose-Bifosfato Carboxilase/ultraestruturaRESUMO
Wnt/ß-Catenin signaling plays crucial roles in regenerative processes in eumetazoans. It also acts in regeneration and axial patterning in the simple freshwater polyp Hydra, whose morphallactic regenerative capacity is unparalleled in the animal kingdom. Previous studies have identified ß-catenin as an early response gene activated within the first 30min in Hydra head regeneration. Here, we have studied the role of ß-Catenin in more detail. First, we show that nuclear ß-Catenin signaling is required for head and foot regeneration. Loss of nuclear ß-Catenin function blocks head and foot regeneration. Transgenic Hydra tissue, in which ß-Catenin is over-expressed, regenerates more heads and feet. In addition, we have identified a set of putative ß-Catenin target genes by transcriptional profiling, and these genes exhibit distinct expression patterns in the hypostome, in the tentacles, or in an apical gradient in the body column. All of them are transcriptionally up-regulated in the tips of early head and foot regenerates. In foot regenerates, this is a transient response, and expression starts to disappear after 12-36h. ChIP experiments using an anti-HydraTcf antibody show Tcf binding at promoters of these targets. We propose that gene regulatory ß-Catenin activity in the pre-patterning phase is generally required as an early regeneration response. When regenerates are blocked with iCRT14, initial local transcriptional activation of ß-catenin and the target genes occurs, and all these genes remain upregulated at the site of both head and foot regeneration for the following 2-3 days. This indicates that the initial regulatory network is followed by position-specific programs that inactivate fractions of this network in order to proceed to differentiation of head or foot structures. brachyury1 (hybra1) has previously been described as early response gene in head and foot regeneration. The HyBra1 protein, however, appears in head regenerating tips not earlier than about twelve hours after decapitation, and HyBra1 translation does not occur in iCRT14-treated regenerates. Foot regenerates never show detectable levels of HyBra1 protein at all. These results suggest that translational control mechanisms may play a decisive role in the head- and foot-specific differentiation phase, and HyBra1 is an excellent candidate for such a key regulator of head specification.
Assuntos
Hydra/fisiologia , Regeneração/fisiologia , Via de Sinalização Wnt , beta Catenina/fisiologia , Animais , Padronização Corporal , Proteínas Fetais/fisiologia , Regulação da Expressão Gênica , Hibridização In Situ , Especificidade de Órgãos , Biossíntese de Proteínas , Regeneração/efeitos dos fármacos , Proteínas com Domínio T/fisiologia , beta Catenina/antagonistas & inibidores , beta Catenina/genéticaRESUMO
PURPOSE OF THE STUDY By 7th December 2016, 4,755 Bicon-Plus cups in total were implanted in the Czech Republic. Some of them have been continuously re-operated, while the most frequent reason of failure is polyethylene wear and aseptic loosening. The present study is focused on surface analysis of retrieved polyethylene Bicon-Plus cups and the determination of the roughness of their bearing surfaces. MATERIAL AND METHODS In this study, we had 13 high molecular weight polyethylene cups with the average time in situ of 8.11 years (3.6-13.7, SD 3.2) before the retrieval. The study population was composed of 3 men, 10 women, with the mean age of 53.31 years. An optical scanning method, based on the principle of active triangulation, was used to determine wear rate. The rate of wear was identified by means of an obtained scan subsequently processed with the use of the GOM Inspect software. The roughness of surfaces was analysed with the application of Contour GT-X8 profiler using the principle of phase shifting interferometry. Measurements of surface topography of the retrieved cups were performed on the entire bearing surfaces. For the individual surface changes, a typical range of surface roughness, describing the particular wear character, was determined. By means of morphology analysis of the tested implants, three areas were identified: unworn area; area representing the worn part of the cup; and the area roughened by parallel grooving. The total surface roughness was evaluated as an arithmetic mean of the measured values. Subsequently, the values were sorted based on frequency and were classified into categories defining the particular wear mechanisms. RESULTS Wear rate of the retrieved acetabular cups was evaluated based on the wear direction vector and the size of linear wear. The average linear wear was equal to 0.13 mm/year (ranging from 0.26 to 2.29 mm/year), and the mean value of total volumetric material loss was 44.37 mm3/year (the range being from 51.80 to 1,119.7 mm3/year). Using the optical profilometer, a map of roughness distribution of the individual cups was obtained. For each implant, 76 values of roughness were evaluated. With the respect to average roughness, the samples were sorted to various categories describing: surface polishing; abrasiveadhesive wear; surfaces with preserved grooving; substantial plastic deformation. DISCUSSION The results clearly showed an increase of wear depending on implant survival; however, the tendency is not linear. This fact can be attributed to a larger amount of abrasive particles, causing an increase of wear or occurrence of surface wear in terms of micro cracks and oxidation degradation of polyethylene. This study indicates that geometry, positioning, and cup alignment during the implantation have a fundamental impact on the cup durability. Further correlation, which was observed in the case of the cup with prevailing roughness in the range from 0.1 to 0.3 µm, is a relatively wide wear vector angle determined with the use of the optical method. Considering the implants with the longest survival time with no loosening of the acetabular cup, the mean angle of direction vector was 56.8° (SD 2.1°). CONCLUSIONS The present study provides the results of morphology analysis of the retrieved Bicon-Plus cups. In general, relatively high wear rate, mainly of abrasive-adhesive character was identified. The dependence between wear and implant in situ longevity was not clearly linear, which suggests the influence of other parameters on the polyethylene wear rate. An important role of implant positioning on survival was also revealed. Moreover, it seems that it can be a more important parameter than the characteristics of the patient. Key words: total hip arthroplasty, Bicon-Plus cup, retrieval analysis, surface analysis, wear measurement, roughness, deformation, survivorship.
Assuntos
Artroplastia de Quadril/instrumentação , Prótese de Quadril , Falha de Prótese , República Tcheca , Análise de Falha de Equipamento , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Polietileno , Desenho de Prótese , Propriedades de SuperfícieRESUMO
BACKGROUND: Oral challenges are the gold standard in food allergy diagnostic, but time-consuming. Aim of the study was to investigate the role of peanut- and hazelnut-component-specific IgE in the diagnostics of peanut and hazelnut allergy and to identify cutoff levels to make some challenges superfluous. METHODS: In a prospective and multicenter study, children with suspected peanut or hazelnut allergy underwent oral challenges. Specific IgE to peanut, hazelnut, and their components (Ara h 1, Ara h 2, Ara h 3, and Ara h 8, Cor a 1, Cor a 8, Cor a 9, and Cor a 14) were determined by ImmunoCAP-FEIA. RESULTS: A total of 210 children were challenged orally with peanut and 143 with hazelnut. 43% of the patients had a positive peanut and 31% a positive hazelnut challenge. With an area under the curve of 0.92 and 0.89, respectively, Ara h 2 and Cor a 14-specific IgE discriminated between allergic and tolerant children better than peanut- or hazelnut-specific IgE. For the first time, probability curves for peanut and hazelnut components have been calculated. A 90% probability for a positive peanut or hazelnut challenge was estimated for Ara h 2-specific IgE at 14.4 kU/l and for Cor a 14-specific IgE at 47.8 kU/l. A 95% probability could only be estimated for Ara h 2 at 42.2 kU/l. CONCLUSIONS: Ara h 2- and Cor a 14-specific IgE are useful to estimate the probability for a positive challenge outcome in the diagnostic work-up of peanut or hazelnut allergy making some food challenges superfluous.
Assuntos
Especificidade de Anticorpos , Arachis/efeitos adversos , Corylus/efeitos adversos , Hipersensibilidade Alimentar/diagnóstico , Hipersensibilidade Alimentar/imunologia , Imunoglobulina E/imunologia , Alérgenos , Antígenos de Plantas , Criança , Pré-Escolar , Comorbidade , Feminino , Humanos , Masculino , Hipersensibilidade a Amendoim/diagnóstico , Hipersensibilidade a Amendoim/imunologia , Estudos Prospectivos , Curva ROCRESUMO
We describe a new in operando approach for the investigation of heterogeneous processes at solid/liquid interfaces with elemental and chemical specificity which combines the preparation of thin liquid films using the meniscus method with standing wave ambient pressure X-ray photoelectron spectroscopy [Nemsák et al., Nat. Commun., 5, 5441 (2014)]. This technique provides information about the chemical composition across liquid/solid interfaces with sub-nanometer depth resolution and under realistic conditions of solution composition and concentration, pH, as well as electrical bias. In this article, we discuss the basics of the technique and present the first results of measurements on KOH/Ni interfaces.
RESUMO
PURPOSE OF THE STUDY: The wear of articular surfaces is considered one of the most important factors limiting the life of total hip arthroplasty (THA). It is assumed that the particles released from the surface of a softer material induce a complex inflammatory response, which will eventually result in osteolysis and aseptic loosening. Implant wear is related to a friction coefficient which depends on combination of the materials used, roughness of the articulating surfaces, internal clearance, and dimensions of the prosthesis. MATERIAL AND METHODS: The selected parameters of the bearing couples tested were studied using an experimental device based on the principle of a pendulum. Bovine serum was used as a lubricant and the load corresponded to a human body mass of 75 kg. The friction coefficient was derived from a curve of slowdown of pendulum oscillations. Roughness was measured with a device working on the principle of interferometry. Clearance was assessed by measuring diameters of the acetabular and femoral heads with a 3D optical scanner. The specimens tested included unused metal-on-highly cross-linked polyethylene, ceramic-on-highly cross-linked polyethylene and ceramic-on-ceramic bearing couples with the diameters of 28 mm and 36 mm. For each measured parameter, an arithmetic mean was calculated from 10 measurements. RESULTS: 1) The roughness of polyethylene surfaces was higher by about one order of magnitude than the roughness of metal and ceramic components. The Protasul metal head had the least rough surface (0.003 µm). 2) The ceramic-on-ceramic couples had the lowest clearance. Bearing couples with polyethylene acetabular liners had markedly higher clearances ranging from 150 µm to 545 µm. A clearance increased with large femoral heads (up to 4-fold in one of the couple tested). 3) The friction coefficient was related to the combination of materials; it was lowest in ceramic-on-ceramic surfaces (0.11 to 0.12) and then in ceramic-on-polyethylene implants (0.13 to 0.14). The friction coefficient is supposed to increase with a decreasing femoral head diameter. However, in the bearing couples with polyethylene liners manufactured by one company, paradoxically, the friction coefficient slightly increased with an increase in femoral head size from 28 mm to 36 mm. 4) The lowest friction moment (< 3.5 Nm) was found for ceramic-on-ceramic implants 28 mm in diameter; the highest values were recorded in metal-on-polyethylene bearing couples 36 mm in diameter (> 7 Nm). DISCUSSION: Although our study confirmed that the bearing couples produced by different manufacturers varied to some extent in the parameters studied, in our opinion, this variability was not significant because it was not within an order of magnitude in any of the tests. CONCLUSIONS: The study showed that both the friction coefficient and the friction moment are affected more by the combination of materials than by the diameter of a femoral head. The best results were achieved in ceramic-on-ceramic implants.
Assuntos
Artroplastia de Quadril/instrumentação , Prótese de Quadril , Cerâmica , Fricção , Humanos , Teste de Materiais/métodos , Desenho de Prótese , Falha de Prótese , Propriedades de SuperfícieRESUMO
The Oncologic and Palliative Network Landshut is a problem-solving approach to structurally weak rural areas to improve an a dequate care of critically ill patients, especially by a close involvement of outpatient and inpatient care providers. These networks not only improve the medical and nursing care of patients, but can also be cost-effective.
Assuntos
Assistência Ambulatorial/organização & administração , Acessibilidade aos Serviços de Saúde/organização & administração , Necessidades e Demandas de Serviços de Saúde/organização & administração , Comunicação Interdisciplinar , Colaboração Intersetorial , Oncologia/organização & administração , Neoplasias/terapia , Cuidados Paliativos/organização & administração , Admissão do Paciente , Resolução de Problemas , Serviços de Saúde Rural/organização & administração , Alemanha , Humanos , Unidades Móveis de Saúde/organização & administraçãoRESUMO
The Oncologic and Palliative Network Landshut is a problem-solving approach to structurally weak rural areas to improve an adequate care of critically ill patients, especially by a close involvement of outpatient and inpatient care providers. These networks not only improve the medical and nursing care of patients, but can also be cost-effective.
Assuntos
Assistência Ambulatorial/organização & administração , Comportamento Cooperativo , Comunicação Interdisciplinar , Oncologia/organização & administração , Neoplasias/terapia , Cuidados Paliativos/organização & administração , Admissão do Paciente , Equipe de Assistência ao Paciente/organização & administração , Resolução de Problemas , Serviços de Saúde Rural/organização & administração , Alemanha , Acessibilidade aos Serviços de Saúde/organização & administração , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Necessidades e Demandas de Serviços de Saúde/organização & administração , Necessidades e Demandas de Serviços de Saúde/estatística & dados numéricos , Humanos , Unidades Móveis de Saúde/organização & administração , Unidades Móveis de Saúde/provisão & distribuição , Neoplasias/epidemiologia , Serviços de Saúde Rural/provisão & distribuição , Recursos HumanosRESUMO
The objective of this study is to develop a reliable tribological model to enable a more thorough investigation of the frictional behavior of fascia tissues connected to non-specific lower back pain. Several models were designed and evaluated based on their coefficient of friction, using a low-frequency, low-load reciprocating motion. The study found that two technical elastomers, layered on PDMS to simulate the fascia and underlying muscle, are suitable substitutes for biological tissue in the model. The influence of tribopair geometry was also examined, and the results showed that greater conformity of contact leads to a lower COF, regardless of the material combination used. Finally, the friction properties of HA of various molecular weights and concentrations were tested.
Assuntos
Fáscia , Fricção , Teste de Materiais , Fáscia/fisiologia , Dimetilpolisiloxanos/química , Fenômenos Biomecânicos , Modelos Biológicos , Elastômeros/químicaRESUMO
The objective of this paper is to briefly review the engineering biotribology of total hip arthroplasty (THA). At present the issue of lubrication, wear and friction within a THA is being studied in detail and the results have been published in a number of papers. The reason is obvious since the rate at which the articulating surfaces wear off affects the longevity of a THA to a great extent. Seeking an optimal biotribological configuration remains an important task for both the researchers and orthopaedic surgeons. In order to enable the orthopaedic community to think over THA biotribology and, at the same time, communicate with the representatives of technical professions, knowledge of the basic principles of biotribology and their understanding is necessary.
Assuntos
Artroplastia de Quadril , Engenharia Biomédica/métodos , Prótese de Quadril , Propriedades de Superfície , Artroplastia de Quadril/instrumentação , Artroplastia de Quadril/métodos , Prótese de Quadril/classificação , Prótese de Quadril/normas , Humanos , Comunicação Interdisciplinar , Falha de PróteseRESUMO
There is an urgent need to understand how organisms respond to multiple, potentially interacting drivers in today's world. The effects of the pollutants anthropogenic sound (pile driving sound playbacks) and waterborne cadmium were investigated across multiple levels of biology in larval and juvenile Norway lobster, Nephrops norvegicus under controlled laboratory conditions. The combination of pile driving playbacks (170 dBpk-pk re 1 µPa) and cadmium combined synergistically at concentrations >9.62 µg[Cd] L-1 resulting in increased larval mortality, with sound playbacks otherwise being antagonistic to cadmium toxicity. Exposure to 63.52 µg[Cd] L-1 caused significant delays in larval development, dropping to 6.48 µg[Cd] L-1 in the presence of piling playbacks. Pre-exposure to the combination of piling playbacks and 6.48 µg[Cd] L-1 led to significant differences in the swimming behaviour of the first juvenile stage. Biomarker analysis suggested oxidative stress as the mechanism resultant deleterious effects, with cellular metallothionein (MT) being the predominant protective mechanism.
Assuntos
Cádmio , Nephropidae , Animais , Cádmio/toxicidade , Metalotioneína , Ruído , Som , Espectrografia do SomRESUMO
In Saccharomyces cerevisiae, impairment of protein phosphatase PP2ARts1 leads to temperature and hyperosmotic stress sensitivity, yet the underlying mechanism and the scope of action of the phosphatase in the stress response remain elusive. Using a quantitative mass spectrometry-based approach we have identified a set of putative substrate proteins that show both hyperosmotic stress- and PP2ARts1-dependent changes in their phosphorylation pattern. A comparative analysis with published MS-shotgun data revealed that the phosphorylation status of many of these sites is regulated by the MAPKAP kinase Rck2, suggesting that the phosphatase antagonizes Rck2 signaling. Detailed gel mobility shift assays and protein-protein interaction analysis strongly indicate that Rck2 activity is directly regulated by PP2ARts1 via a SLiM B56-family interaction motif, revealing how PP2ARts1 influences the response to hyperosmotic stress in Yeast.
Assuntos
Pressão Osmótica , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Monoéster Fosfórico Hidrolases/metabolismo , Fosforilação , Proteína Fosfatase 2 , Proteínas Serina-Treonina Quinases , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Transdução de Sinais/fisiologiaRESUMO
The structure and spectral signatures of the protonated homodimer of pyridine in its complex with a poorly coordinating anion have been studied in solution in CDF(3)/CDClF(2) down to 120 K and in a single crystal. In both phases, the hydrogen bond is asymmetric. In the solution, the proton is involved in a fast reversible transfer that determines the multiplicity of NMR signals and the sign of the primary H/D isotope effect of --0.95 ppm. The proton resonates at 21.73 ppm that is above any value reported in the past and is indicative of a very short hydrogen bond. By combining X-ray diffraction analysis with model computations, the position of the proton in the crystal has been defined as d(N-H) = 1.123 Å and d(H···N) = 1.532 Å. The same distances have been estimated using a (15)N NMR correlation. The frequency of the protonic out-of-plane bending mode is 822 cm(-1) in agreement with Novak's correlation.
Assuntos
Dimerização , Piridinas/química , Análise Espectral/métodos , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Difração de Nêutrons , Prótons , Difração de Raios XRESUMO
Total knee arthroplasty has become a routine procedure for patients suffering from joint diseases. Although the number of operations continuously increases, a limited service-life of implants represents a persisting challenge for scientists. Understanding of lubrication may help to suitably explain tribological processes on the way to replacements that become durable well into the third decade of service. The aim of the present study is to assess the formation of protein lubricating film in the knee implant. A developed knee simulator was used to observe the contact of real femoral and transparent polymer tibial component using fluorescent microscopy. The contact was lubricated by various protein solutions with attention to the behaviour of albumin and γ-globulin. In order to suitably mimic a human synovial fluid, hyaluronic acid and phospholipids were subsequently added to the solutions. Further, the change in shape and the migration of the contact zone were studied. The results showed considerable appearance differences of the contact over the swing phase of the simplified gait cycle. Regarding film formation, a strong interaction of the various molecules of synovial fluid was observed. It was found that the thickness of the lubricating layer stabilizes within around 50 s. Throughout the contact zone, protein agglomerations were present and could be clearly visualised using the applied optical technique.
Assuntos
Artroplastia do Joelho , Líquido Sinovial , Albuminas , Humanos , Articulação do Joelho , Lubrificação , gama-GlobulinasRESUMO
The chaperonins GroEL and GroES of Escherichia coli facilitate protein folding in an adenosine triphosphate (ATP)-dependent reaction cycle. The kinetic parameters for the formation and dissociation of GroEL-GroES complexes were analyzed by surface plasmon resonance. Association of GroES and subsequent ATP hydrolysis in the interacting GroEL toroid resulted in the formation of a stable GroEL:ADP:GroES complex. The complex dissociated as a result of ATP hydrolysis in the opposite GroEL toroid, without formation of a symmetrical GroEL:(GroES)2 intermediate. Dissociation was accelerated by the addition of unfolded polypeptide. Thus, the functional chaperonin unit is an asymmetrical GroEL:GroES complex, and substrate protein plays an active role in modulating the chaperonin reaction cycle.
Assuntos
Adenosina Trifosfatases/metabolismo , Chaperonina 10/metabolismo , Chaperonina 60/metabolismo , Dobramento de Proteína , Difosfato de Adenosina/metabolismo , Trifosfato de Adenosina/metabolismo , Chaperonina 10/química , Chaperonina 60/química , Endopeptidase K , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Magnésio/farmacologia , Serina Endopeptidases/metabolismoRESUMO
The crystal structure of the adenine nucleotide exchange factor GrpE in complex with the adenosine triphosphatase (ATPase) domain of Escherichia coli DnaK [heat shock protein 70 (Hsp70)] was determined at 2.8 angstrom resolution. A dimer of GrpE binds asymmetrically to a single molecule of DnaK. The structure of the nucleotide-free ATPase domain in complex with GrpE resembles closely that of the nucleotide-bound mammalian Hsp70 homolog, except for an outward rotation of one of the subdomains of the protein. This conformational change is not consistent with tight nucleotide binding. Two long alpha helices extend away from the GrpE dimer and suggest a role for GrpE in peptide release from DnaK.
Assuntos
Adenosina Trifosfatases/química , Proteínas de Bactérias/química , Proteínas de Escherichia coli , Proteínas de Choque Térmico HSP70/química , Proteínas de Choque Térmico/química , Chaperonas Moleculares/química , Conformação Proteica , Difosfato de Adenosina/metabolismo , Adenosina Trifosfatases/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/metabolismo , Sítios de Ligação , Cristalografia por Raios X , Dimerização , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico/metabolismo , Ligação de Hidrogênio , Modelos Moleculares , Chaperonas Moleculares/metabolismo , Dados de Sequência Molecular , Estrutura Secundária de ProteínaRESUMO
The Escherichia coli chaperonin GroEL and its regulator GroES are thought to mediate adenosine triphosphate-dependent protein folding as an asymmetrical complex, with substrate protein bound within the GroEL cylinder. In contrast, a symmetrical complex formed between one GroEL and two GroES oligomers, with substrate protein binding to the outer surface of GroEL, was recently proposed to be the functional chaperonin unit. Electron microscopic and biochemical analyses have now shown that unphysiologically high magnesium concentrations and increased pH are required to assemble symmetrical complexes, the formation of which precludes the association of unfolded polypeptide. Thus, the functional significance of GroEL:(GroES)2 particles remains to be demonstrated.
Assuntos
Chaperonina 10/metabolismo , Chaperonina 60/metabolismo , Trifosfato de Adenosina/farmacologia , Adenilil Imidodifosfato/farmacologia , Chaperonina 10/química , Chaperonina 10/ultraestrutura , Chaperonina 60/química , Chaperonina 60/ultraestrutura , Concentração de Íons de Hidrogênio , Magnésio/farmacologia , Microscopia Eletrônica de Transmissão e Varredura , Dobramento de ProteínaRESUMO
Hatchery-reared juvenile turbot (Scophthalmus maximus L.) were exposed for 3 weeks, under laboratory conditions, to inter-tidal sediments collected from polluted sites in Cork Harbour (Whitegate and Agahda) and a reference site at Ballymacoda Co., Cork, Ireland. The potential of the sediment exposure to induce cytochrome P450 activities and CYP1A1 in the fish was assessed. Chemical analysis revealed that the sediments originating from the reference and harbour sites were contaminated principally with PAHs-the harbour sites having double the levels of those at the reference site. Following 3 weeks exposure to the sediments western blotting demonstrated a strong immunogenic response for CYP1A1 in the liver, but not for gill or intestine. P450 activities were generally significantly higher than those exposed to reference site sediment. Liver was the most responsive tissue with significantly greater P450 activities compared with gill and intestinal tissues.
Assuntos
Citocromo P-450 CYP1A1/efeitos dos fármacos , Monitoramento Ambiental , Linguados/metabolismo , Sedimentos Geológicos/química , Poluentes Químicos da Água/toxicidade , Animais , Western Blotting , Citocromo P-450 CYP1A1/biossíntese , Indução Enzimática/efeitos dos fármacos , Brânquias/efeitos dos fármacos , Brânquias/enzimologia , Intestinos/efeitos dos fármacos , Intestinos/enzimologia , Irlanda , Fígado/efeitos dos fármacos , Fígado/enzimologia , Hidrocarbonetos Policíclicos Aromáticos/análise , Água do Mar/química , Poluentes Químicos da Água/análiseRESUMO
In the context of natural-based wastewater treatment technologies (such as constructed wetlands - CW) the use of a low-cost, continuous-like biosensor tool for the assessment of operational conditions is of key importance for plant management optimization. The objective of the present study was to assess the potential use of constructed wetland microbial fuel cells (CW-MFC) as a domestic wastewater COD assessment tool. For the purpose of this work four lab-scale CW-MFCs were set up and fed with pre-settled domestic wastewater at different COD concentrations. Under laboratory conditions two different anodic materials were tested (graphite rods and gravel). Furthermore, a pilot-plant based experiment was also conducted to confirm the findings previously recorded for lab-scale experiments. Results showed that in spite of the low coulombic efficiencies recorded, either gravel or graphite-based anodes were suitable for the purposes of domestic wastewater COD assessment. Significant linear relationships could be stablished between inlet COD concentrations and CW-MFC Ecell whenever contact time was above 10â¯h. Results also showed that the accuracy of the CW-MFC was greatly compromised after several weeks of operation. Pilot experiments showed that CW-MFC presents a good bio-indication response between week 3 and 7 of operation (equivalent to an accumulated organic loading between 100 and 200â¯g COD/m2, respectively). Main conclusion of this work is that of CW-MFC could be used as an "alarm-tool" for qualitative continuous influent water quality assessment rather than a precise COD assessment tool due to a loss of precision after several weeks of operation.