Effects of replica running shoes upon external forces and muscle activity during running

Twelve participants ran (9 km . h(-1)) to test two types of running shoes: replica and original shoes. Ground reaction force, plantar pressure and electromyographic activity were recorded. The shoes were tested randomly and on different days. Comparisons between the two experimental conditions were made by analysis of variance (ANOVA) test (P <= 0.05). The time to first peak, loading rate of the first peak and impulse of the first 75 ms of stance were significantly different between the shoes (P <= 0.05), revealing an increase of impact forces for the replica shoes. The peak plantar pressure values were significantly higher (P <= 0.05) when wearing replica shoes. During running, the contact area was significantly smaller (P <= 0.05) for the replica shoe. The electromyographic activity of the analysed muscles did not show changes between the two shoes in running. These findings suggest that the use of replica running shoes can increase the external load applied to the human body, but may not change the muscle activity pattern during locomotion. This new mechanical situation may increase the risk of injuries in these movements.

Integration of processes induced air flotation and photo-Fenton for treatment of residual waters contaminated with xylene

Produced water in oil fields is one of the main sources of wastewater generated in the industry. It contains several organic compounds, such as benzene, toluene, ethyl benzene and xylene (BTEX), whose disposal is regulated by law. The aim of this study is to investigate a treatment of produced water integrating two processes, i.e., induced air flotation (IAF) and photo-Fenton. The experiments were conducted in a column flotation and annular lamp reactor for flotation and photodegradation steps, respectively. The first order kinetic constant of IAF for the wastewater studied was determined to be 0.1765 min(-1) for the surfactant EO 7. Degradation efficiencies of organic loading were assessed using factorial planning. Statistical data analysis shows that H2O2 concentration is a determining factor in process efficiency. Degradations above 90% were reached in all cases after 90 min of reaction, attaining 100% mineralization in the optimized concentrations of Fenton reagents. Process integration was adequate with 100% organic load removal in 20 min. The results of the integration of the IAF with the photo-Fenton allowed to meet the effluent limits established by Brazilian legislation for disposal. (C) 2011 Elsevier B.V. All rights reserved.

Processo biotecnologico a cellule immobilizzate per la produzione di acidi grassi volatili da acque di vegetazione

Le acque di vegetazione (AV) costituiscono un serio problema di carattere ambientale, sia a causa della loro elevata produzione sia per l’ elevato contenuto di COD che oscilla fra 50 e 150 g/l. Le AV sono considerate un refluo a tasso inquinante fra i più elevati nell’ambito dell’industria agroalimentare e la loro tossicità è determinata in massima parte dalla componente fenolica. Il presente lavoro si propone di studiare e ottimizzare un processo non solo di smaltimento di tale refluo ma anche di una sua valorizzazione, utlizzandolo come materia prima per la produzione di acidi grassi e quindi di PHA, polimeri biodegradabili utilizzabili in varie applicazioni. A tale scopo sono stati utilizzati due bioreattori anaerobici a biomassa adesa, di identica configurazione, con cui si sono condotti due esperimenti in continuo a diverse temperature e carichi organici al fine di studiare l’influenza di tali parametri sul processo. Il primo esperimento è stato condotto a 35°C e carico organico pari a 12,39 g/Ld, il secondo a 25°C e carico organico pari a 8,40 g/Ld. Si è scelto di allestire e mettere in opera un processo a cellule immobilizzate in quanto questa tecnologia si è rivelata vantaggiosa nel trattamento continuo di reflui ad alto contenuto di COD e carichi variabili. Inoltre si è scelto di lavorare in continuo poiché tale condizione, per debiti tempi di ritenzione idraulica, consente di minimizzare la metanogenesi, mediata da microrganismi con basse velocità specifiche di crescita. Per costituire il letto fisso dei due reattori si sono utilizzati due diversi tipi di supporto, in modo da poter studiare anche l’influenza di tale parametro, in particolare si è fatto uso di carbone attivo granulare (GAC) e filtri ceramici Vukopor S10 (VS). Confrontando i risultati si è visto che la massima quantità di VFA prodotta nell’ambito del presente studio si ha nel VS mantenuto a 25°C: in tale condizione si arriva infatti ad un valore di VFA prodotti pari a 524,668 mgCOD/L. Inoltre l’effluente in uscita risulta più concentrato in termini di VFA rispetto a quello in entrata: nell’alimentazione la percentuale di materiale organico presente sottoforma di acidi grassi volatili era del 54 % e tale percentuale, in uscita dai reattori, ha raggiunto il 59 %. Il VS25 rappresenta anche la condizione in cui il COD degradato si è trasformato in percentuale minore a metano (2,35 %) e questo a prova del fatto che l’acidogenesi ha prevalso sulla metanogenesi. Anche nella condizione più favorevole alla produzione di VFA però, si è riusciti ad ottenere una loro concentrazione in uscita (3,43 g/L) inferiore rispetto a quella di tentativo (8,5 g/L di VFA) per il processo di produzione di PHA, sviluppato da un gruppo di ricerca dell’università “La Sapienza” di Roma, relativa ad un medium sintetico. Si può constatare che la modesta produzione di VFA non è dovuta all’eccessiva degradazione del COD, essendo questa nel VS25 appena pari al 6,23%, ma piuttosto è dovuta a una scarsa concentrazione di VFA in uscita. Questo è di buon auspicio nell’ottica di ottimizzare il processo migliorandone le prestazioni, poiché è possibile aumentare tale concentrazione aumentando la conversione di COD in VFA che nel VS25 è pari a solo 5,87%. Per aumentare tale valore si può agire su vari parametri, quali la temperatura e il carico organico. Si è visto che il processo di acidogenesi è favorito, per il VS, per basse temperature e alti carichi organici. Per quanto riguarda il reattore impaccato con carbone attivo la produzione di VFA è molto ridotta per tutti i valori di temperatura e carichi organici utilizzati. Si può quindi pensare a un’applicazione diversa di tale tipo di reattore, ad esempio per la produzione di metano e quindi di energia.

Allestimenti di reattori multistadio per la digestione anaerobica dei reflui agro-industriali e confronto con processo convenzionale

This study fits into the context of activities aim at waste bioremediation and valorization through the production of energy according to principles of environmental sustainability. The experimental work was carried out at the laboratories of the Department of Civil Engineering, Environmental and Materials (DICAM) of the Faculty of Engineering. The main objective was to enhance the treatment of high organic loading waste, such as manure and cheese whey, through advanced anaerobic digestion systems in order to obtain biogas rich in methane. On the basis of the premise that the environmental conditions pertaining in most anaerobic wastewater digesters are not optimal for both fermentative and methanogenic microorganisms, the research was particularly focused on the implementation of two-phase anaerobic digesters. In fact a two-phase process permits selection and enrichment of different bacteria in each digester by independently controlling the digester operating conditions. Thus, the first phase (acidogenesis) can be operated to optimize acidogenic growth and the second phase (methanogenesis) to optimize methanogenic growth. (Ince O. , 1998). Before reactors’ set up, , some lab scale experiments were carried out to identify the best manure and whey ratio and the best conditions of temperature, pH, hydraulic retention time of acidogenesis an methanogenic phases.

Eigenschaften von thermoresponsiven Bürstenpolymeren

In dieser Doktorarbeit werden die Eigenschaften von thermoresponsiven Bürstenpolymeren untersucht. Hierbei konnten erstmalig thermische Konformationsübergänge von zylindrischen Bürstenmolekülen auf Oberflächen beobachtet werden. Der Einfluss der Oberfläche auf die Umkehrbarkeit und die Kinetik der Übergänge wurde untersucht. Die dabei erhaltenen Erkenntnisse konnten verwendet werden, um das Verhalten der Polymere auf anderen Oberflächen vorherzusagen. Im zweiten Projekt wurde gezeigt, dass Einzelmolekül-Kraftspektroskopie eine gute Methode für die Untersuchung der mechanischen Eigenschaften von Bürstenpolymeren in guten Lösungsmitteln ist. Mit kleinen Substanzmengen kann die Persistenzlänge der Polymere bestimmt werden. Es ist möglich zu zeigen, dass die Persistenzlänge der Polymerbürsten von ihrer Seitenkettenlänge abhängt. Überraschenderweise funktioniert das Experiment nicht, wenn man die Bürsten aus einem Polymerfilm zieht anstatt ein Einzelmolekülexperiment durchzuführen. In diesem Fall zeigen die Kraft-Abstands-Kurven zu lange Kontur- und Persistenzlängen. Diese Beobachtung ist für lineare Polymere nicht gültig. Im dritten Teil der Doktorarbeit werden Kraft-Abstands-Experimente an einzelnen kollabierten Polymeren untersucht. In schlechten Lösungsmitteln zeigen die Bürsten ein moleküllängenabhängiges Kraft-Plateau, welches theoretisch vorausgesagt wurde und einen Phasenübergang von einem kollabierten zu einem entspannten Zustand der Polymerkette anzeigt. In Fly-Fishing-Experimenten kann man eine Hysterese zwischen den beiden Messkurven beobachten, welche bei mehrfachem Ziehen kleiner wird. Alle Experimente in schlechten Lösungsmitteln wurden mit linearen Polymeren reproduziert, um den Einfluss der Molekülarchitektur von den generellen Eigenschaften von Polymeren in schlechten Lösungsmitteln unterscheiden zu können. Zum Abschluss wird die Abhängigkeit der Polymerentfaltung von der Laderate des Experiments gemessen.

High Solids Co-Digestion of Food and Landscape Waste and the Potential for Ammonia Toxicity

A pilot-scale study was completed to determine the feasibility of high-solids anaerobic digestion (HSAD) of a mixture of food and landscape wastes at a university in central Pennsylvania (USA). HSAD was stable at low loadings (2g COD/L-day), but developed inhibitory ammonia concentrations at high loadings (15g COD/L-day). At low loadings, methane yields were 232L CH4/kg COD fed and 229L CH4/kg VS fed, and at high loadings yields were 211L CH4/kg COD fed and 272L CH4/kg VS fed. Based on characterization and biodegradability studies, food waste appears to be a good candidate for HSAD at low organic loading rates; however, the development of ammonia inhibition at high loading rates suggests that the C:N ratio is too low for use as a single substrate. The relatively low biodegradability of landscape waste as reported herein made it an unsuitable substrate to increase the C:N ratio. Codigestion of food waste with a substrate high in bioavailable carbon is recommended to increase the C:N ratio sufficiently to allow HSAD at loading rates of 15g COD/L-day. Copyright 2014 Elsevier Ltd. All rights reserved.

Shear band evolution in Zr/Hf-based bulk metallic glasses under static and dynamic indentations

Bulk metallic glasses (BMGs) exhibit superior mechanical properties as compared with other conventional materials and have been proposed for numerous engineering and technological applications. Zr/Hf-based BMGs or tungsten reinforced BMG composites are considered as a potential replacement for depleted uranium armor-piercing projectiles because of their ability to form localized shear bands during impact, which has been known to be the dominant plastic deformation mechanism in BMGs. However, in conventional tensile, compressive and bending tests, limited ductility has been observed because of fracture initiation immediately following the shear band formation. To fully investigate shear band characteristics, indentation tests that can confine the deformation in a limited region have been pursued. In this thesis, a detailed investigation of thermal stability and mechanical deformation behavior of Zr/Hf-based BMGs is conducted. First, systematic studies had been implemented to understand the influence of relative compositions of Zr and Hf on thermal stability and mechanical property evolution. Second, shear band evolution under indentations were investigated experimentally and theoretically. Three kinds of indentation studies were conducted on BMGs in the current study. (a) Nano-indentation to determine the mechanical properties as a function of Hf/Zr content. (b) Static Vickers indentation on bonded split specimens to investigate the shear band evolution characteristics beneath the indention. (c) Dynamic Vickers indentation on bonded split specimens to investigate the influence of strain rate. It was found in the present work that gradually replacing Zr by Hf remarkably increases the density and improves the mechanical properties. However, a slight decrease in glass forming ability with increasing Hf content has also been identified through thermodynamic analysis although all the materials in the current study were still found to be amorphous. Many indentation studies have revealed only a few shear bands surrounding the indent on the top surface of the specimen. This small number of shear bands cannot account for the large plastic deformation beneath the indentations. Therefore, a bonded interface technique has been used to observe the slip-steps due to shear band evolution. Vickers indentations were performed along the interface of the bonded split specimen at increasing loads. At small indentation loads, the plastic deformation was primarily accommodated by semi-circular primary shear bands surrounding the indentation. At higher loads, secondary and tertiary shear bands were formed inside this plastic zone. A modified expanding cavity model was then used to predict the plastic zone size characterized by the shear bands and to identify the stress components responsible for the evolution of the various types of shear bands. The applicability of various hardness—yield-strength ( H −σγ ) relationships currently available in the literature for bulk metallic glasses (BMGs) is also investigated. Experimental data generated on ZrHf-based BMGs in the current study and those available elsewhere on other BMG compositions were used to validate the models. A modified expanding-cavity model, employed in earlier work, was extended to propose a new H −σγ relationship. Unlike previous models, the proposed model takes into account not only the indenter geometry and the material properties, but also the pressure sensitivity index of the BMGs. The influence of various model parameters is systematically analyzed. It is shown that there is a good correlation between the model predictions and the experimental data for a wide range of BMG compositions. Under dynamic Vickers indentation, a decrease in indentation hardness at high loading rate was observed compared to static indentation hardness. It was observed that at equivalent loads, dynamic indentations produced more severe deformation features on the loading surface than static indentations. Different from static indentation, two sets of widely spaced semi-circular shear bands with two different curvatures were observed. The observed shear band pattern and the strain rate softening in indentation hardness were rationalized based on the variations in the normal stress on the slip plane, the strain rate of shear and the temperature rise associated with the indentation deformation. Finally, a coupled thermo-mechanical model is proposed that utilizes a momentum diffusion mechanism for the growth and evolution of the final spacing of shear bands. The influence of strain rate, confinement pressure and critical shear displacement on the shear band spacing, temperature rise within the shear band, and the associated variation in flow stress have been captured and analyzed. Consistent with the known pressure sensitive behavior of BMGs, the current model clearly captures the influence of the normal stress in the formation of shear bands. The normal stress not only reduces the time to reach critical shear displacement but also causes a significant temperature rise during the shear band formation. Based on this observation, the variation of shear band spacing in a typical dynamic indentation test has been rationalized. The temperature rise within a shear band can be in excess of 2000K at high strain rate and high confinement pressure conditions. The associated drop in viscosity and flow stress may explain the observed decrease in fracture strength and indentation hardness. The above investigations provide valuable insight into the deformation behavior of BMGs under static and dynamic loading conditions. The shear band patterns observed in the above indentation studies can be helpful to understand and model the deformation features under complex loading scenarios such as the interaction of a penetrator with armor. Future work encompasses (1) extending and modifying the coupled thermo-mechanical model to account for the temperature rise in quasistatic deformation; and (2) expanding this model to account for the microstructural variation-crystallization and free volume migration associated with the deformation.

Bench-scale evaluation of the activated sludge process for treatment of a high -strength chemical plant wastewater

A bench-scale treatability study was conducted on a high-strength wastewater from a chemical plant to develop an alternative for the existing waste stabilization pond treatment system. The objective of this study was to determine the treatability of the wastewater by the activated sludge process and, if treatable, to determine appropriate operating conditions, and to evaluate the degradability of bis(2-chloroethyl)ether (Chlorex) and benzene in the activated sludge system. Four 4-L Plexi-glass, complete mixing, continuous flow activated sludge reactors were operated in parallel under different operating conditions over a 6-month period. The operating conditions examined were hydraulic retention time (HRT), sludge retention time (SRT), nutrient supplement, and Chlorex/benzene spikes. Generally the activated sludge system treating high-strength wastewater was stable under large variations of organic loading and operating conditions. At an HRT of 2 days, more than 90% removal efficiency with good sludge settleability was achieved when the organic loading was less than 0.4 g BOD$\sb5$/g MLVSS/d or 0.8 g COD/g MLVSS/d. At least 20 days of SRT was required to maintain steady operation. Phosphorus addition enhanced the performance of the system especially during stressed operation. On the average, removals of benzene and Chlorex were 73-86% and 37-65%, respectively. In addition, the low-strength wastewater was treatable by activated sludge process, showing more than 90% BOD removal at a HRT of 0.5 days. In general, the sludge had poor settling characteristics. The aerated lagoon process treating high-strength wastewater also provided significant organic reduction, but did not produce an acceptable effluent concentration. ^

Tratamiento de vinazas provenientes de etanol en un reactor de lecho fluidizado inverso

En el estado de Veracruz, al sur de México, se ubican empresas dedicadas a la obtención de etanol a partir de melaza de azúcar de caña. Las más pequeñas, tienen una producción promedio de 20,000 L de alcohol/día. Los efluentes de la producción de etanol incluyen agua de enfriamiento de condensadores, agua del lavado de tanques de fermentación y vinazas, estas últimas son los efluentes más contaminantes en las destilerías, por su concentración de material orgánico biodegradable y no biodegradable. Las vinazas se generan en grandes volúmenes, produciéndose de 12 a 15 litros de vinazas por cada litro de alcohol destilado. Estos efluentes se caracterizan por tener altas temperaturas, pH ácido y una elevada concentración de DQO así como de sólidos totales. La determinación de la biodegradabilidad anaerobia de un agua residual, permite estimar la fracción de DQO que puede ser transformada potencialmente en metano y la DQO recalcitrante que queda en el efluente. Para el desarrollo de una prueba de biodegradabilidad, es importante considerar diversos factores relacionados con la composición del agua a tratar, composición de los lodos y las condiciones bajo las cuales se lleva a cabo la prueba. La digestión anaerobia de aguas residuales industriales es comúnmente usada en todo el mundo, ofrece significativas ventajas para el tratamiento de efluentes altamente cargados. Los sistemas anaerobios de tratamiento de aguas residuales industriales incluyen tecnologías con biopelículas, estos sistemas de tratamiento anaerobio con biopelícula son una tecnología bien establecida para el tratamiento de efluentes industriales. El Reactor de Lecho Fluidizado Inverso Anaerobio (LFI) ha sido diseñado para el tratamiento de aguas residuales de alta carga, teniendo como ventajas el empleo de un soporte que proporciona una gran superficie y un bajo requerimiento de energía para la fluidización del lecho. En el presente trabajo, se lleva a cabo el análisis de un proceso de producción de etanol, identificando a los efluentes que se generan en el mismo. Se encuentra que el efluente final está compuesto principalmente por las vinazas provenientes del proceso de destilación. En la caracterización de las vinazas provenientes del proceso de producción de etanol a partir de melaza de azúcar de caña, se encontraron valores promedio de DQO de 193.35 gDQO/L, para los sólidos totales 109.78 gST/L y pH de 4.64. Así mismo, en esta investigación se llevó a cabo una prueba de biodegradabilidad anaerobia, aplicada a la vinaza proveniente de la producción de etanol. En la caracterización de los lodos empleados en el ensayo se obtiene una Actividad Metanogénica Especifica de 0.14 g DQO/gSSV.d. El porcentaje de remoción de DQO de la vinaza fue de 62.7%, obteniéndose una k igual a 0.031 h-1 y una taza de consumo de sustrato de 1.26 gDQO/d. El rendimiento de metano fue de 0.19 LCH4/g DQOremovida y el porcentaje de biodegradabilidad de 54.1%. El presente trabajo también evalúa el desempeño de un LFI, empleando Extendospher® como soporte y tratando efluentes provenientes de la producción de etanol. El reactor se arrancó en batch y posteriormente se operó en continuo a diferentes Cargas Orgánicas Volumétricas de 0.5, 1.0, 3.3, 6.8 y 10.4 g DQO/L.d. Además, se evaluaron diferentes Tiempos de Residencia Hidráulica de 10, 5 y 1 días. El sistema alcanzó las siguientes eficiencias promedio de remoción de DQO: 81% para la operación en batch; 58, 67, 59 y 50 % para las cargas de 0.5, 1.0, 3.3, 6.8 g DQO/L.d respectivamente. Para la carga de 10.4 g DQO/L.d, la eficiencia promedio de remoción de DQO fue 38%, en esta condición el reactor presentó inestabilidad y disminución del rendimiento de metano. La generación de metano inició hasta los 110 días de operación del reactor a una carga de 1.0 g DQO/L.d. El sistema alcanzó un rendimiento de metano desde 0.15 hasta 0.34 LCH4/g DQO. Durante la operación del reactor a una carga constante de 6.4 g DQO/L.d, y un TRH de 1 día, se alcanzó una eficiencia promedio de remoción de DQO de 52%. In the state of Veracruz, to the south of Mexico, there are located companies dedicated to the production of ethanol from molasses of cane sugar. The smallest, have a average production of 20,000 L ethanol/day. The effluent of production of ethanol include water of condensers, water originated from the cleanliness of tanks of fermentation and vinasses, the above mentioned are more effluent pollutants in the distilleries, for the poor organic matter degradability. The vinasses are generated in high volumes, producing from 12 to 15 L of vinasses per every liter of distilled ethanol. These effluent are characterized by its high temperature, pH acid and a high concentration of DQO as well as high concentration of TS. The determination of the anaerobic degradability of a waste water, it allows to estimate the fraction of DQO that can be transformed potentially into methane and the recalcitrant DQO that stays in the effluent. For the development of degradability test, it is important to consider factors related to the composition of the water to be treated, composition of the sludge and the conditions under which the test is carried out. The anaerobic digestion of industrial wastes water is used commonly in the whole world, it offers significant advantages for the treatment of effluent highly loaded. The anaerobic treatment of industrial wastes water include technologies with biofilms, this anaerobic treatment whit biofilms systems, is a well-established technology for treatment of industrial effluents. The Anaerobic Inverse Fluidized Bed Reactor (IFBR) has been developed to provide biological treatment of high strength organic wastewater for their large specific surface and their low energy requirements for fluidization. In this work, there is carried out the analysis of a process of production of ethanol, identifying the effluent ones that are generated in the process. One determined that the effluent end is composed principally by the vinasses originated from the process of distillation. In the characterization of the vinasses originated from the process of production of ethanol from cane sugar molasses, there were average values of DQO of 193.35 gDQO/L, average values of solid of 109.78 gST/L and pH of 4.64. In this investigation there was carried out a anaerobic degradability test of the vinasses generated in the production of ethanol. In the characterization of the sludge used in the essay, the specific methanogenic activity (SMA) was 0.14 gDQO/gSSV.d. The average removal of DQO of the vinasses was 62.7 %, k equal to 0.031 h-1 was obtained one and a rate of removal substrate of 1.26 gDQO/d. The methane yield was 0.19 LCH4/gDQO removed and the anaerobic biodegradability was a 54.1 %. This study describes the performance of IFBR with Extendospher®, for the treatment of vinasses. The start-up was made in batch, increasing gradually the Organic Load Rate (OLR): 0.5, 1.0, 3.3, 6.8 and 10.4 g COD/L.d. Different Hydraulic Retention Times (HRT) were evaluated: 10, 5 and 1 days. During the operation in batch, the COD removal obtained was of 81 %, and for OLR of 0.5, 1.0, 3.3, 6.8 g COD/L.d the removal obtained was 58, 67, 59 and 50 % respectively. For a maximum OLR of 10.4 g COD/L.d, the COD removal was 38 %, and the system presented instability and decrease of the yield methane. The methane production initiated after 110 days of the start-up of the IFBR, to organic load rate of 1.0 g COD/L.d. The system reached values in the methane yield from 0.15 up to 0.34 LCH4/g CODremoved, for the different organic load rates. During the operation to a constant OLR of 6.4 g COD/L.d, and a HRT of 1 day, the Anaerobic Inverse Fluidized Bed Reactor reached a maximum efficiency of removal of 52 %.

Unbinding process of adsorbed proteins under external stress studied by atomic force microscopy spectroscopy

We report the study of the dynamics of the unbinding process under a force load f of adsorbed proteins (fibrinogen) on a solid surface (hydrophilic silica) by means of atomic force microscopy spectroscopy. By varying the loading rate rf, defined by f = rf t, t being the time, we find that, as for specific interactions, the mean rupture force increases with rf. This unbinding process is analyzed in the framework of the widely used Bell model. The typical dissociation rate at zero force entering in the model lies between 0.02 and 0.6 s−1. Each measured rupture is characterized by a force f0, which appears to be quantized in integer multiples of 180–200 pN.

Unbinding forces of single antibody-antigen complexes correlate with their thermal dissociation rates

Point mutants of three unrelated antifluorescein antibodies were constructed to obtain nine different single-chain Fv fragments, whose on-rates, off-rates, and equilibrium binding affinities were determined in solution. Additionally, activation energies for unbinding were estimated from the temperature dependence of the off-rate in solution. Loading rate-dependent unbinding forces were determined for single molecules by atomic force microscopy, which extrapolated at zero force to a value close to the off-rate measured in solution, without any indication for multiple transition states. The measured unbinding forces of all nine mutants correlated well with the off-rate in solution, but not with the temperature dependence of the reaction, indicating that the same transition state must be crossed in spontaneous and forced unbinding and that the unbinding path under load cannot be too different from the one at zero force. The distance of the transition state from the ground state along the unbinding pathway is directly proportional to the barrier height, regardless of the details of the binding site, which most likely reflects the elasticity of the protein in the unbinding process. Atomic force microscopy thus can be a valuable tool for the characterization of solution properties of protein-ligand systems at the single molecule level, predicting relative off-rates, potentially of great value for combinatorial chemistry and biology.

Chemically distinct transition states govern rapid dissociation of single L-selectin bonds under force

Carbohydrate–protein bonds interrupt the rapid flow of leukocytes in the circulation by initiation of rolling and tethering at vessel walls. The cell surface carbohydrate ligands are glycosylated proteins like the mucin P-selectin glycoprotein ligand-1 (PSGL-1), which bind ubiquitously to the family of E-, P-, and L-selectin proteins in membranes of leukocytes and endothelium. The current view is that carbohydrate–selectin bonds dissociate a few times per second, and the unbinding rate increases weakly with force. However, such studies have provided little insight into how numerous hydrogen bonds, a Ca2+ metal ion bond, and other interactions contribute to the mechanical strength of these attachments. Decorating a force probe with very dilute ligands and controlling touch to achieve rare single-bond events, we have varied the unbinding rates of carbohydrate–selectin bonds by detachment with ramps of force/time from 10 to 100,000 pN/sec. Testing PSGL-1, its outer 19 aa (19FT), and sialyl LewisX (sLeX) against L-selectin in vitro on glass microspheres and in situ on neutrophils, we found that the unbinding rates followed the same dependence on force and increased by nearly 1,000-fold as rupture forces rose from a few to ≈200 pN. Plotted on a logarithmic scale of loading rate, the rupture forces reveal two prominent energy barriers along the unbinding pathway. Strengths above 75 pN arise from rapid detachment (<0.01 sec) impeded by an inner barrier that requires a Ca2+ bond between a single sLeX and the lectin domain. Strengths below 75 pN occur under slow detachment (>0.01 sec) impeded by the outer barrier, which appears to involve an array of weak (putatively hydrogen) bonds.

Force and kinetic barriers to unzipping of the DNA double helix

A theory of the unzipping of double-stranded DNA is presented and is compared to recent micromanipulation experiments. It is shown that the interactions that stabilize the double helix and the elastic rigidity of single strands simply determine the sequence-dependent ≈12-pN force threshold for DNA strand separation. Using a semimicroscopic model of the binding between nucleotide strands, we show that the greater rigidity of the strands when formed into double-stranded DNA, relative to that of isolated strands, gives rise to a potential barrier to unzipping. The effects of this barrier are derived analytically. The force to keep the extremities of the molecule at a fixed distance, the kinetic rates for strand unpairing at fixed applied force, and the rupture force as a function of loading rate are calculated. The dependence of the kinetics and of the rupture force on molecule length is also analyzed.

Reator anaeróbio em batelada seqüencial contendo biomassa imobilizada submetido a aumento de carga orgânica tratando água residuária sintética

Nesse estudo, procurou-se investigar a influência do aumento de carga orgânica em um ASBR operado a 30ºC, agitado mecanicamente e contendo microrganismos imobilizados em suporte inerte. Para tal, foi aplicado um carregamento orgânico volumétrico variando de 1,5 a 6,0 g DQO/L.dia, alimentando-se um reator construído em acrílico (5,4 L) com 2 L de água residuária sintética com concentrações de 500 a 2000 mg DQO/L com fontes de carboidrato/proteína/lipídio, em bateladas de 8 a 12 h. O sistema apresentou eficiências de remoção de material orgânico entre 73% e 88% para as condições estudadas. Entretanto, quando o reator foi alimentado com uma concentração de 2000 mg DQO/L em bateladas de 8 h, verificou-se o acúmulo de ácidos voláteis totais, refletindo na redução da eficiência de remoção de material orgânico para 55%, para amostras filtradas. Os perfis dinâmicos ao longo da batelada permitiram concluir que, para a condição com o mesmo carregamento orgânico, porém com concentrações afluentes e tempos de ciclo diferentes, a produção inicial de ácidos voláteis totais foi mais acentuada para o caso de alimentação com maior concentração. Para condições com concentrações afluentes iguais, o tempo a mais para o ciclo foi fundamental para obter-se efluente de melhor qualidade em termos de remoção de matéria orgânica.

Strain and damage sensing properties on multifunctional cement composites with CNF admixture

Both strain and damage sensing properties on carbon nanofiber cement composites (CNFCC) are reported in the present paper. Strain sensing tests were first made on the material’s elastic range. The applied loading levels have been previously calculated from mechanical strength tests. The effect of several variables on the strain-sensing function was studied, e.g. cement pastes curing age, current density, loading rate or maximum stress applied. All these parameters were discussed using the gage factor as reference. After this first set of elastic experiments, the same specimens were gradually loaded until material’s failure. At the same time both strain and resistivity were measured. The former was controlled using strain gages, and the latter using a multimeter on a four probe setup. The aim of these tests was to prove the sensitivity of these CNF composites to sense their own damage, i.e. check the possibility of fabricating structural damage sensors with CNFCC’s. All samples with different CNF dosages showed good strain-sensing capacities for curing periods of 28 days. Furthermore, a 2%CNF reinforced cement paste has been sensitive to its own structural damage.