Microstructure and mechanical properties degradation at high temperature of tungsten alloys processed by forging

One of the challenges of science and engineering nowadays is to develop new ways to supply energy in a sustainable and ecological mode. The fussion energy could be the final answer but a myriad of problems must be solved previously.

Mechanical Degradation of Tungsten Alloys at Extreme Temperatures in Vacuum and Oxidation Atmospheres

Mechanical degradation of tungsten alloys at extreme temperatures in vacuum and oxidation atmospheres.

A visual framework to accelerate knowledge discovery based on dimensionality reduction minimizing degradation of quality

Tradicionalmente, el uso de técnicas de análisis de datos ha sido una de las principales vías para el descubrimiento de conocimiento oculto en grandes cantidades de datos, recopilados por expertos en diferentes dominios. Por otra parte, las técnicas de visualización también se han usado para mejorar y facilitar este proceso. Sin embargo, existen limitaciones serias en la obtención de conocimiento, ya que suele ser un proceso lento, tedioso y en muchas ocasiones infructífero, debido a la dificultad de las personas para comprender conjuntos de datos de grandes dimensiones. Otro gran inconveniente, pocas veces tenido en cuenta por los expertos que analizan grandes conjuntos de datos, es la degradación involuntaria a la que someten a los datos durante las tareas de análisis, previas a la obtención final de conclusiones. Por degradación quiere decirse que los datos pueden perder sus propiedades originales, y suele producirse por una reducción inapropiada de los datos, alterando así su naturaleza original y llevando en muchos casos a interpretaciones y conclusiones erróneas que podrían tener serias implicaciones. Además, este hecho adquiere una importancia trascendental cuando los datos pertenecen al dominio médico o biológico, y la vida de diferentes personas depende de esta toma final de decisiones, en algunas ocasiones llevada a cabo de forma inapropiada. Ésta es la motivación de la presente tesis, la cual propone un nuevo framework visual, llamado MedVir, que combina la potencia de técnicas avanzadas de visualización y minería de datos para tratar de dar solución a estos grandes inconvenientes existentes en el proceso de descubrimiento de información válida. El objetivo principal es hacer más fácil, comprensible, intuitivo y rápido el proceso de adquisición de conocimiento al que se enfrentan los expertos cuando trabajan con grandes conjuntos de datos en diferentes dominios. Para ello, en primer lugar, se lleva a cabo una fuerte disminución en el tamaño de los datos con el objetivo de facilitar al experto su manejo, y a la vez preservando intactas, en la medida de lo posible, sus propiedades originales. Después, se hace uso de efectivas técnicas de visualización para representar los datos obtenidos, permitiendo al experto interactuar de forma sencilla e intuitiva con los datos, llevar a cabo diferentes tareas de análisis de datos y así estimular visualmente su capacidad de comprensión. De este modo, el objetivo subyacente se basa en abstraer al experto, en la medida de lo posible, de la complejidad de sus datos originales para presentarle una versión más comprensible, que facilite y acelere la tarea final de descubrimiento de conocimiento. MedVir se ha aplicado satisfactoriamente, entre otros, al campo de la magnetoencefalografía (MEG), que consiste en la predicción en la rehabilitación de lesiones cerebrales traumáticas (Traumatic Brain Injury (TBI) rehabilitation prediction). Los resultados obtenidos demuestran la efectividad del framework a la hora de acelerar y facilitar el proceso de descubrimiento de conocimiento sobre conjuntos de datos reales. ABSTRACT Traditionally, the use of data analysis techniques has been one of the main ways of discovering knowledge hidden in large amounts of data, collected by experts in different domains. Moreover, visualization techniques have also been used to enhance and facilitate this process. However, there are serious limitations in the process of knowledge acquisition, as it is often a slow, tedious and many times fruitless process, due to the difficulty for human beings to understand large datasets. Another major drawback, rarely considered by experts that analyze large datasets, is the involuntary degradation to which they subject the data during analysis tasks, prior to obtaining the final conclusions. Degradation means that data can lose part of their original properties, and it is usually caused by improper data reduction, thereby altering their original nature and often leading to erroneous interpretations and conclusions that could have serious implications. Furthermore, this fact gains a trascendental importance when the data belong to medical or biological domain, and the lives of people depends on the final decision-making, which is sometimes conducted improperly. This is the motivation of this thesis, which proposes a new visual framework, called MedVir, which combines the power of advanced visualization techniques and data mining to try to solve these major problems existing in the process of discovery of valid information. Thus, the main objective is to facilitate and to make more understandable, intuitive and fast the process of knowledge acquisition that experts face when working with large datasets in different domains. To achieve this, first, a strong reduction in the size of the data is carried out in order to make the management of the data easier to the expert, while preserving intact, as far as possible, the original properties of the data. Then, effective visualization techniques are used to represent the obtained data, allowing the expert to interact easily and intuitively with the data, to carry out different data analysis tasks, and so visually stimulating their comprehension capacity. Therefore, the underlying objective is based on abstracting the expert, as far as possible, from the complexity of the original data to present him a more understandable version, thus facilitating and accelerating the task of knowledge discovery. MedVir has been succesfully applied to, among others, the field of magnetoencephalography (MEG), which consists in predicting the rehabilitation of Traumatic Brain Injury (TBI). The results obtained successfully demonstrate the effectiveness of the framework to accelerate and facilitate the process of knowledge discovery on real world datasets.

Observed degradation in photovoltaic plants affected by hot-spots

A number of findings have shown that the test procedures currently available to determine the reliability and durability of photovoltaic (PV) modules are insufficient to detect certain problems. To improve these procedures, ongoing research into the actual performance of the modules in the field is required. However, scientific literature contains but few references to field studies of defective modules. This article studies two different localized heating phenomena affecting the PV modules of two large-scale PV plants in Spain. The first problem relates to weak solder joints whilst the second is due to microcracks on the module cells. For both cases, the cause is identified, and consideration is given with regard to the effect on performance, the potential deterioration over time, and a way to detect the problems identified. The findings contained in this paper will prove to be of considerable interest to maintenance personnel at large-scale PV plants and also to those responsible for setting module quality standards and specifications, and even the PV module manufacturers themselves.

Sol-gel TiO2 nanoparticles prompt photocatalytic cement for pollution degradation

TiO2 nanoparticles (TiO2NPs) prepared by the sol–gel method have been incorporated to cement paste with the aim of creating a photocatalytic system capable of compensating, through degradation of hazardous molecules, the envi- ronmental impact associated to the production of the clinker. Doping was carried out at different mass ratios with TiO2NPs precursor solutions within a fresh ce- ment paste, which was then characterized using scanning electron microscopy (SEM). The photocatalytic performance was evaluated by the degradation of Methylene Blue (MB) using a 125W UV lamp as irradiating source. Main cement properties such as hydration degree and C-S-H content are affected by TiO2NPs doping level. Cement containing TiO2NPs exhibited an increasing photocatalytic activity for increasing doping, while the pure cement paste control could hardly degrade MB. The kinetics of the system where also studied and their second order behavior related to microstructural aspects of the system.

Unearthing the roots of degradation of Quercus pyrenaica coppices: an integrative perspective from clonal structure to carbon budgets

Quercus pyrenaica es una especie rebrotadora de raíz intensa e históricamente aprovechada en monte bajo para la obtención de leñas, carbón y pastos. Debido al éxodo rural y a la aparición de nuevas fuentes energéticas, este aprovechamiento fue abandonado en la década de 1970. Desde entonces, las bajas producciones de madera y bellota y el puntisecado de los pies evidencian el generalizado estancamiento de estas masas. Uno de los mayores retos actuales de la selvicultura en el ámbito mediterráneo es encontrar usos alternativos para estos montes abandonados, siendo la conversión a monte alto una de las alternativas preferidas. Se han realizado resalveos de conversión, sin embrago, éstos se aplican sin un conocimiento integral de las causas de la degradación. En esta tesis doctoral, estudiamos un hipotético desequilibrio entre la parte radical y la parte aérea (R:S) de las cepas de rebollo como causa subyacente de su decaimiento. En una parcela experimental, aprovechada al menos desde el siglo XII, se realizaron análisis genéticos a priori para elucidar la estructura genética del rodal, y así estudiar la influencia del tamaño clonal en el funcionamiento de las cepas. Las cepas de mayor tamaño presentaron un menor crecimiento diametral de sus pies, así como mayores tasas de respiración radical, estimadas a partir de flujos internos de CO2 a través del xilema (FT) y de los flujos de CO2 del suelo. Estos resultados sugieren que el desequilibrio R:S aumenta con el tamaño clonal, dado que la eliminación periódica de órganos aéreos, al mismo tiempo que las raíces permanecen intactas, da lugar a un gran desarrollo del sistema radical que consume gran parte de los carbohidratos no estructurales (NSC) en respiración de mantenimiento, comprometiendo así el desarrollo de órganos aéreos. Se excavaron y pesaron dos cepas compuestas por cuatro y ocho pies, las cuales mostraron ratios R:S (0.5 y 1, respectivamente) superiores a los registrados en pies de origen sexual. Al igual que en otras especies rebrotadoras de raíz, se observaron altas concentraciones de NSC en las raíces (> 20% en primavera) y una gran proporción de albura en el sistema radical (52%) que alberga una notable reserva de NSC (87 kg en la cepa de mayor tamaño). En el sistema radical de dicha cepa, estimada mediante dataciones radiocarbónicas en 550 años de edad, se contaron 248 uniones radicales. La persistencia de sistemas radicales grandes, viejos, y altamente interconectados sugiere que la gran cantidad de recursos almacenados y consumidos en las raíces compensan un pobre desarrollo aéreo con una alta resiliencia vegetativa. Para un mejor entendimiento de los balances de carbono y del agotamiento de NSC en las cepas de rebollo, se midieron los flujos internos y externos de CO2 en troncos y los flujos de CO2 del suelo, y se estimó la respiración de órganos aéreos (RS) y subterráneos (RR). Estacionalmente, RS y RR reflejaron las dinámicas de flujo de savia y de crecimiento del tronco, y estuvieron determinadas principalmente por los flujos externos de CO2, dada la escasa contribución de FT a RS y RR (< 10% y < 2%, respectivamente). En una escala circadiana, la contribución de FT a RS aumentó hasta un 25% en momentos de alta transpiración. Las bajas concentraciones de CO2 en el xilema ([CO2] hasta un 0.11%) determinaron comparativamente unos bajos FT, probablemente causados por una limitada respiración del xilema y una baja resistencia a la difusión radial del CO2 impuestos por la sequía estival. Los pulsos de [CO2] observados tras las primeras lluvias de otoño apoyan esta idea. A lo largo del periodo vegetativo, el flujo medio de CO2 procedente del suelo (39 mol CO2 day-1) fue el mayor flujo respiratorio, tres y cuatro veces superior a RS (12 mol CO2 day-1) y RR (8-9 mol CO2 day-1), respectivamente. Ratios RR/RS menores que la unidad evidencian un importante peso de la respiración aérea como sumidero de carbono adicional. Finalmente, se ensayó el zanjado de raíces y el anillamiento de troncos como tratamientos selvícolas alternativos con el objetivo de aumentar las reservas de NSC en los troncos de las cepas. Los resultados preliminares desaconsejan el zanjado de raíces por el alto coste derivado posiblemente de la cicatrización de las heridas. El anillado de troncos imposibilitó el transporte de NSC a las raíces y aumentó la concentración de almidón por encima de la zona anillada, mientras que sistema radical se mantiene por los pies no anillados de la cepa. Son necesarias más mediciones y datos adicionales para comprobar el mantenimiento de esta respuesta positiva a largo plazo. Para concluir, destacamos la necesidad de estudios multidisciplinares que permitan una comprensión integral de la degradación de los rebollares ibéricos para poder aplicar a posteriori una gestión adecuada en estos montes bajos abandonados. ABSTRACT Quercus pyrenaica is a vigorous root-resprouting species intensively and historically coppiced for firewood, charcoal and woody pastures. Due to the rural exodus and the appearance of new energy sources, coppicing was abandoned towards 1970. Since then, tree overaging has resulted in stand stagnation displayed by slow stem growth, branch dieback, and scarce acorn production. The urgent need to find new alternative uses for abandoned coppices is recognized as one of the biggest challenges which currently faces Mediterranean silviculture; conversion into high forest by thinning is one of the preferred alternatives. For this aim, thinning has been broadly applied and seldom tested, although without a comprehensive understanding of the causes of stand stagnation. In this PhD study, we test the hypothesis of an imbalance between above- and below-ground organs, result of long term coppicing, as the underlying cause of Q. pyrenaica decay. In an experimental plot coppiced since at least the 12th century, genetic analyses were performed a priori to elucidate inconspicuous clonal structure of Q. pyrenaica to evaluate how clonal size affects the functioning of these multi-stemmed trees. Clonal size negatively affected diametric stem growth, whereas root respiration rates, measured by internal fluxes of CO2 through xylem (FT) and soil CO2 efflux, increased with clonal size. These results suggest root-to-shoot (R:S) imbalance intensifying with clonal size: periodic removal of aboveground organs whilst belowground organs remain undisturbed may have led to massive root systems which consume a great proportion of non-structural carbohydrates (NSC) for maintenance respiration, thus constraining aboveground performance. Furthermore, excavation of two multi-stemmed trees, composed by four and eight stems, revealed R:S ratios (0.5 and 1, respectively) greater than those reported for sexually regenerated trees. Moreover, as similarly observed in several root-resprouting species, NSC allocation to roots was favored ([NSC] > 20% in spring): a large proportion of sapwood maintained throughout the root system (52%) stored a remarkable NSC pool of 87 kg in the case of the largest clone. In this root system of the eight-stemmed tree, 248 root connections were counted and, by radiocarbon dating, its age was estimated to be 550-years-old. Persistence of massive, old and highly interconnected root systems suggests that enhanced belowground NSC storage and consumption reflects a trade-off between vegetative resilience and aboveground development. For a better understanding of tree carbon budget and the potential role of carbon starvation in Q. pyrenaica decay, internal and external stem CO2 fluxes and soil CO2 effluxes were monitored to evaluate respiratory costs above- and below-ground. On a seasonal scale, stem and root respiration (RS and RR) mirrored sap flow and stem growth dynamics. Respiration was determined to the greatest extent by external fluxes of CO2 to the atmosphere or soil, since FT accounted for a low proportion of RS and RR (< 10% and < 2%, respectively). On a diel scale, the contribution of FT to RS increased up to 25% at high transpiration rates. Comparatively low FT was determined by the low concentration of xylem CO2 registered ([CO2] as low as 0.11%), likely as a consequence of constrained xylem respiration and reduced resistance to CO2 radial diffusion imposed by summer drought. Xylem [CO2] pulses following first autumn rains support this idea. Averaged over the growing season, soil CO2 efflux was the greatest respiratory flux (39 mol CO2 day-1), three and four times greater than RS (12 mol CO2 day-1) and RR (8-9 mol CO2 day-1), respectively. Ratios of RR/RS below one evidence an additional and important weight of aboveground respiration as a tree carbon sink. Finally, root trenching and stem girdling were tested as complimentary treatments to thinning as a means to improve carbon reserves in stems of clonal trees. Preliminary results discouraged root trenching due to the high cost likely incurred for wound closure. Stem girdling successfully blocked NSC translocation downward, increasing starch concentrations above the girdled zone whilst the root system is fed by non-girdled stems within the clone. Further measurements and ancillary data are necessary to verify that this positive effect hold over time. To conclude, the need of multidisciplinary approaches for an integrative understanding on the functioning of abandoned Q pyrenaica coppices is highlighted for an appropriate management of these stands.

Sunflower meal and spring pea ruminal degradation protection using malic acid or orthophosphoric acid-heat treatments

The effects of solutions of malic or orthophosphoric acids (0.752 Eqg/kg of feed) and heat to protect proteins of sunflower meal (SFM) and spring pea (SP) against ruminal degradation were studied using particle transit, 15N infusion, in situ and electrophoretic techniques. Three wethers fitted with rumen and duodenum cannulae were successively fed three isoproteic diets including SFM and SP, untreated or treated with malic or orthophosphoric acids. Incubations of tested meals were only performed while feeding the respective diet. Estimates of the ruminally undegraded fraction (RU) and its intestinal digestibility of dry matter, organic matter (only for RU), crude protein and starch (only in SP) were obtained considering ruminal microbial contamination and particle comminution and outflow rates. When corrected for microbial contamination, estimates of RU and intestinal digestibility decreased in all tested fractions for both feeds. All RU estimates increased with the protective treatments, whereas intestinal digestibility-dry matter also increased in SFM. Low intestinal digestibility-crude protein values suggested the presence of antitrypsin factors in SP. Protective treatments of both feeds led to consistent increases in the intestinal digested fraction of dry matter and crude protein, being only numerically different for SP-starch (60.5% as average). However, treatments also reduced the organic matter fermentation, which may decrease ruminal microbial protein synthesis. Electrophoretic studies showed albumin disappearance in both SFM and SP, whereas changes in other RU proteins were more pronounced in SP than SFM.

Ubiquitin-dependent degradation of IκBα is mediated by a ubiquitin ligase Skp1/Cul 1/F-box protein FWD1

Activation of the transcription factor nuclear factor kappa B (NF-κB) is controlled by proteolysis of its inhibitory subunit (IκB) via the ubiquitin-proteasome pathway. Signal-induced phosphorylation of IκBα by a large multisubunit complex containing IκB kinases is a prerequisite for ubiquitination. Here, we show that FWD1 (a mouse homologue of Slimb/βTrCP), a member of the F-box/WD40-repeat proteins, is associated specifically with IκBα only when IκBα is phosphorylated. The introduction of FWD1 into cells significantly promotes ubiquitination and degradation of IκBα in concert with IκB kinases, resulting in nuclear translocation of NF-κB. In addition, FWD1 strikingly evoked the ubiquitination of IκBα in the in vitro system. In contrast, a dominant-negative form of FWD1 inhibits the ubiquitination, leading to stabilization of IκBα. These results suggest that the substrate-specific degradation of IκBα is mediated by a Skp1/Cull 1/F-box protein (SCF) FWD1 ubiquitin-ligase complex and that FWD1 serves as an intracellular receptor for phosphorylated IκBα. Skp1/Cullin/F-box protein FWD1 might play a critical role in transcriptional regulation of NF-κB through control of IκB protein stability.

Mass spectrometric and capillary electrophoretic investigation of the enzymatic degradation of heparin-like glycosaminoglycans

Difficulties in determining composition and sequence of glycosaminoglycans, such as those related to heparin, have limited the investigation of these biologically important molecules. Here, we report methodology, based on matrix-assisted laser desorption ionization MS and capillary electrophoresis, to follow the time course of the enzymatic degradation of heparin-like glycosaminoglycans through the intermediate stages to the end products. MS allows the determination of the molecular weights of the sulfated carbohydrate intermediates and their approximate relative abundances at different time points of the experiment. Capillary electrophoresis subsequently is used to follow more accurately the abundance of the components and also to measure sulfated disaccharides for which MS is not well applicable. For those substrates that produce identical or isomeric intermediates, the reducing end of the carbohydrate chain was converted to the semicarbazone. This conversion increases the molecular weight of all products retaining the reducing terminus by the “mass tag” (in this case 56 Da) and thus distinguishes them from other products. A few picomoles of heparin-derived, sulfated hexa- to decasaccharides of known structure were subjected to heparinase I digestion and analyzed. The results indicate that the enzyme acts primarily exolytically and in a processive mode. The methodology described should be equally useful for other enzymes, including those modified by site-directed mutagenesis, and may lead to the development of an approach to the sequencing of complex glycosaminoglycans.

Role of calpain in skeletal-muscle protein degradation

Although protein degradation is enhanced in muscle-wasting conditions and limits the rate of muscle growth in domestic animals, the proteolytic system responsible for degrading myofibrillar proteins in skeletal muscle is not well defined. The goals of this study were to evaluate the roles of the calpains (calcium-activated cysteine proteases) in mediating muscle protein degradation and the extent to which these proteases participate in protein turnover in muscle. Two strategies to regulate intracellular calpain activities were developed: overexpression of dominant-negative m-calpain and overexpression of calpastatin inhibitory domain. To express these constructs, L8 myoblast cell lines were transfected with LacSwitch plasmids, which allowed for isopropyl β-d-thiogalactoside-dependent expression of the gene of interest. Inhibition of calpain stabilized fodrin, a well characterized calpain substrate. Under conditions of accelerated degradation (serum withdrawal), inhibition of m-calpain reduced protein degradation by 30%, whereas calpastatin inhibitory domain expression reduced degradation by 63%. Inhibition of calpain also stabilized nebulin. These observations indicate that calpains play key roles in the disassembly of sarcomeric proteins. Inhibition of calpain activity may have therapeutic value in treatment of muscle-wasting conditions and may enhance muscle growth in domestic animals.

Listeria monocytogenes infection of P388D1 macrophages results in a biphasic NF-κB (RelA/p50) activation induced by lipoteichoic acid and bacterial phospholipases and mediated by IκBα and IκBβ degradation

As previously reported, Listeria monocytogenes infection of P388D1 macrophages results in a rapid induction of NF-κB DNA-binding activity. Here we show that this induction of NF-κB activity occurs in a biphasic mode: first, a transient, IκBα degradation-dependent phase of activity, also induced by the nonvirulent species Listeria innocua, which is mediated by binding of the bacteria to the macrophage, or by adding Listeria-derived lipoteichoic acid to the macrophage; the second persistent phase of activation is only markedly induced when the bacteria enter the cytoplasm of the host cell and express the virulence genes plcA and plcB, encoding two phospholipases. We suggest that products of the enzymatic activity of phospholipases directly interfere with host cell signal transduction pathways, thus leading to persistent NF-κB activation via persistent IκBβ degradation.

A Toolbox for Quantitative Gene Expression in Varroa destructor : RNA Degradation in Field Samples and Systematic Analysis of Reference Gene Stability

Funding: This project has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no. 613960 (SMARTBEES) (http://www.smartbees-fp7.eu/) and Veterinary Medicines Directorate, Department for Environment Food & Rural Affairs (Project # VM0517) (https://www.gov.uk/government/organisations/veterinary-medicines-directorate). CHM was supported by a Biosciences Knowledge Transfer Network Biotechnology and Biological Sciences Research Council (KTN-BBSRC CASE) Studentship (BB/L502467/1) (http://www.bbsrc.ac.uk/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Acknowledgments We gratefully acknowledge Mr Sebastian Bacz’s expert help and advice with beekeeping.

Def1 Promotes the Degradation of Pol3 for Polymerase Exchange to Occur During DNA-Damage–Induced Mutagenesis in Saccharomyces cerevisiae

Funding: Wellcome Trust, 070247/Z/03/A. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Heme is an effector molecule for iron-dependent degradation of the bacterial iron response regulator (Irr) protein

The bacterial iron response regulator (Irr) protein mediates iron-dependent regulation of heme biosynthesis. Pulse–chase and immunoprecipitation experiments showed that Irr degraded in response to 6 μM iron with a half-life of ≈30 min and that this regulated stability was the principal determinant of control by iron. Irr contains a heme regulatory motif (HRM) near its amino terminus. A role for heme in regulation was implicated by the retention of Irr in heme synthesis mutants in the presence of iron. Addition of heme to low iron (0.3 μM) cultures was sufficient for the disappearance of Irr in cells of the wild-type and heme mutant strains. Spectral and binding analyses of purified recombinant Irr showed that the protein bound heme with high affinity and caused a blue shift in the absorption spectrum of heme to a shorter wavelength. A Cys29 → Ala substitution within the HRM of Irr (IrrC29A) abrogated both high affinity binding to heme and the spectral blue shift. In vivo turnover experiments showed that, unlike wild-type Irr, IrrC29A was stable in the presence of iron. We conclude that iron-dependent degradation of Irr involves direct binding of heme to the protein at the HRM. The findings implicate a regulatory role for heme in protein degradation and provide direct evidence for a functional HRM in a prokaryote.

Development of an in vitro mRNA decay system for Escherichia coli: Poly(A) polymerase I is necessary to trigger degradation

Using a novel Escherichia coli in vitro decay system in which polysomes are the source of both enzymes and mRNA, we demonstrate a requirement for poly(A) polymerase I (PAP I) in mRNA turnover. The in vitro decay of two different mRNAs (trxA and lpp) is triggered by the addition of ATP only when polysomes are prepared from a strain carrying the wild-type gene for PAP I (pcnB+). The relative decay rates of these two messages are similar in vitro and in vivo. Poly(A) tails are formed on both mRNAs, but no poly(A) tails are detected on the 3′ end of mature 23S rRNA. The size distribution of poly(A) tails generated in vitro, averaging 50 nt in length, is comparable to that previously reported in vivo. PAP I activity is associated exclusively with the polysomes. Exogenously added PAP I does not restore mRNA decay to PAP I− polysomes, suggesting that, in vivo, PAP I may be part of a multiprotein complex. The potential of this in vitro system for analyzing mRNA decay in E. coli is discussed.