In search of lost time constants and of non-Michaelis–Menten parameters

SummaryUpon completing 100 years since it was published, the work Die Kinetik der Invertinwirkung by Michaelis and Menten (MM) was celebrated during the 6th Beilstein ESCEC Symposium 2013. As the 7th Beilstein ESCEC Symposium 2015 debates enzymology in the context of complex biological systems, a post-MM approach is required to address cell-like conditions that are well beyond the steady-state limitations. The present contribution specifically addresses two hitherto ambiguous constants whose interest was, however, intuited in the original MM paper: (i) the characteristic time constant τ∞, which can be determined using the late stages of any progress curve independently of the substrate concentration adopted; and (ii) the dissociation constant KS, which is indicative of the enzyme–substrate affinity and completes the kinetic portrayal of the Briggs–Haldane reaction scheme. The rationale behind τ∞ and KS prompted us to revise widespread concepts of enzyme's efficiency, defined by the specificity constant kcat/KM, and of the Michaelis constant KM seen as the substrate concentration yielding half-maximal rates. The alternative definitions here presented should help recovering the wealth of published kcat/KM and KM data from the criticism that they are subjected. Finally, a practical method is envisaged for objectively determining enzyme's activity, efficiency and affinity – (EA)2 – from single progress curves. The (EA)2 assay can be conveniently applied even when the concentrations of substrate and enzyme are not accurately known

New strategies for the enhancement and promotion of local identity in coastal areas. The case of the Sorrento peninsula

The paper proposes to consider the port areas as strategic nodes for the valorisation of the territory. In these areas the reuse of buildings represents the opportunity to integrate new economic and cultural activities for tourist promotion and enhancement of local resources. The localisation of new cultural, recreational and promotional activities can also increase territorial competitiveness and attractiveness and employment rate, related to functional mix and economic activities. In many contemporary cities, recent rehabilitation processes carried out this strategy, including the reuse of existing buildings in the ports by new activities especially connected with the maritime tradition of the city. The proposal aims to verify the opportunity to build a strategy based on valorisation and management of excellent coastal areas characterized by a network of small ports in small cities. The Sorrento Peninsula (Italy) represents an emblematic case study in order to enhance the role of port areas for the promotion of natural, cultural and historical resources in a systemic vision of the territory. The reuse of existing buildings for tourism promotion together with the rehabilitation of the pedestrian paths system can contribute to realize a network that – starting from port areas – improves accessibility and usability

Work satisfaction within community pharmacy professionals

Work is a profession, a craft, or an activity in which man produces something central to his life. The job satisfaction is reflected simultaneously on the worker and the functioning of organizations. Workers more satisfied with their work have higher productivity levels, higher subjective well-being and well-being with life in general. Community pharmacy professionals are categorized in three professional categories: pharmacists, pharmacy technicians and technical pharmacy assistants. Work satisfaction was previously associated with gender, age, job category, years of service, remuneration, security concerning the future of the profession, relationships with colleagues and satisfaction with the professional category. The objectives of this study are to evaluate the satisfaction level of community pharmacy professionals and to identify associated factors. These professionals have revealed, in general, a level of moderated satisfaction. There were no significant differences between job satisfaction and gender, age, years of service, professional category and remuneration

Testes de conforto térmico aplicados a uma malha multifuncional desenvolvida com novos materiais

Os produtores europeus, os engenheiros têxteis e os designers devem, no desenvolvimento de novas peças de vestuário, terem em consideração as novas exigências dos consumidores em relação ao conforto. Uma grande variedade de áreas de investigação foram, nos últimos tempos, criadas e desenvolvidas permitindo atingir as exigências relacionadas com o conforto, nomeadamente, considerando os aspectos funcionais e estéticos, entre outros. O conforto térmico humano é, de certa forma, um dos componentes mais importantes a considerar, podendo ser definido como a principal troca de calor e massa e calor do corpo humano com o meio ambiente, através da roupa. Um dos objectivos deste trabalho foi, através do trabalho de uma equipa multidisciplinar, desenvolver uma malha funcional de forma a permitir o conforto termo-fisiológico a quem a utilizar. O presente trabalho permite assim investigar as várias etapas relacionadas com o estudo do conforto no desenvolvimento de novas peças de vestuário: o desenvolvimento de novas malhas, a execução de testes num manequim térmico; a simulação numérica do corpo humano com e sem roupa e o desenvolvimento de testes subjectivos de conforto térmico. A nova malha produzida foi caracterizada e testada em laboratório, em termos de quantidade de vapor de água e da permeabilidade ao ar e outros parâmetros físicos. A malha produzida foi aplicada como forro de botas de caminhada e para o fabrico de t-shirts. Os protótipos das botas de caminhada foram avaliados de forma subjectiva através de questionários. Um manequim térmico foi utilizado para medir o isolamento térmico das t-shirts e a distribuição de temperatura na pele. Juntamente com este trabalho experimental, um modelo de termo regulação do corpo humano foi desenvolvido para validar os dados experimentais obtidos. Alguns dos resultados são apresentados e discutidos

Build-Up an Economical Tool for Machining Operations Cost Estimation

Currently, there is a lack of affordable and simple tools for the estimation of these costs, especially for machining operations. This is particularly true for manufacturing SMEs, in which the cost estimation of machined parts is usually performed based only on required material for part production, or involves a time-consuming, non-standardized technical analysis. Therefore, a cost estimation tool was developed, based on the calculated machining times and amount of required material, based on the final drawing of the requested workpiece. The tool was developed primarily for milling machines, considering milling, drilling, and boring/threading operations. Regarding the considered materials, these were primarily aluminum alloys. However, some polymer materials were also considered. The tool first estimates the required time for total part production and then calculates the total cost. The total production time is estimated based on the required machining operations, as well as drawing, programming, and machine setup time. A part complexity level was also introduced, based on the number of details and operations required for each workpiece, which will inflate the estimated times. The estimation tool was tested in a company setting, comparing the estimated operation time values with the real ones, for a wide variety of parts of differing complexity. An average error of 14% for machining operation times was registered, which is quite satisfactory, as this time is the most impactful in terms of machining cost. However, there are still some problems regarding the accuracy in estimating finishing operation timesinfo:eu-repo/semantics/publishedVersio

Using an Artificial Neural Network Approach to Predict Machining Time

One of the most critical factors in producing plastic injection molds is the cost estimation of machining services, which significantly affects the final mold price. These services’ costs are determined according to the machining time, which is usually a long and expensive operation. If it is considered that the injection mold parts are all different, it can be understood that the correct and quick estimation of machining times is of great importance for a company’s success. This article presents a proposal to apply artificial neural networks in machining time estimation for standard injection mold parts. For this purpose, a large set of parts was considered to shape the artificial intelligence model, and machining times were calculated to collect enough data for training the neural networks. The influences of the network architecture, input data, and the variables used in the network’s training were studied to find the neural network with greatest prediction accuracy. The application of neural networks in this work proved to be a quick and efficient way to predict cutting times with a percent error of 2.52% in the best case. The present work can strongly contribute to the research in this and similar sectors, as recent research does not usually focus on the direct prediction of machining times relating to overall production cost. This tool can be used in a quick and efficient manner to obtain information on the total machining cost of mold parts, with the possibility of being applied to other industry sectorsinfo:eu-repo/semantics/publishedVersio

Advanced and clean technologies for chromium tanned leather waste recycling and green energy production

Chromium tanned leather wastes are difficult to valorise by chemical and biological processes dueto the strong bonds established between collagen and chromium. Thus a one-step fast clean pressureassisted alkaline hydrolysis method has been studied to disrupt recalcitrant bonds. The effects of calciumoxide, temperature, time, liquid to solid ratio and leather scrap size on organic matrix destruction, chromiumdissolution and anaerobic biodegradability of hydrolyzates obtained were evaluated. The results show thatpressure-assisted hydrolysis with CaO may be a good alternative to reduce leather waste volume, obtainbiodegradable solutions with low Cr concentration and final residues usable as a chromium bearingresource. In the optimised conditions, about 50% to 55% of the leather is dissolved. The slurries obtainedcontain above 90% of the chromium. The hydrolyzates show good anaerobic biodegradability mostly in therange of 50% to 70 %, indicating them as a source of biogas

Development of fast dissolving polymer-based microneedles for delivery of an antigenic melanoma cell membrane

Delivery of cancer cell membranes (CM) is a new approach for the activation of the immune system and the induction of immunotherapy of cancer. Local delivery of melanoma CM into skin can induce efficient immune stimulation of antigen presenting cells (APCs), such as dendritic cells. In the current study, fast dissolving microneedles (MNs) were developed for the delivery of melanoma B16F10 CM. Two polymers were tested for the fabrication of MNs: poly(methyl vinyl ether-co-maleic acid) (PMVE-MA) and hyaluronic acid (HA). The incorporation of CM in MNs was achieved through coating of the MNs using a multi-step layering procedure or the micromolding technique. The CM loading and its stabilization were improved by adding sugars (sucrose and trehalose) and a surfactant (Poloxamer 188), respectively. In an ex vivo experiment, both PMVE-MA and HA showed fast dissolutions (<30 s) after insertion into porcine skin. However, HA-MA showed better mechanical properties, namely improved resistance to fracture when submitted to a compression force. Overall, a B16F10 melanoma CM-dissolving MN system was efficiently developed as a promising device suggesting further studies in immunotherapy and melanoma applications

Development of fast dissolving polymer-based microneedles for delivery of an antigenic melanoma cell membrane

Delivery of cancer cell membranes (CM) is a new approach for the activation of the immune system and the induction of immunotherapy of cancer. Local delivery of melanoma CM into skin can induce efficient immune stimulation of antigen presenting cells (APCs), such as dendritic cells. In the current study, fast dissolving microneedles (MNs) were developed for the delivery of melanoma B16F10 CM. Two polymers were tested for the fabrication of MNs: poly(methyl vinyl ether-co-maleic acid) (PMVE-MA) and hyaluronic acid (HA). The incorporation of CM in MNs was achieved through coating of the MNs using a multi-step layering procedure or the micromolding technique. The CM loading and its stabilization were improved by adding sugars (sucrose and trehalose) and a surfactant (Poloxamer 188), respectively. In an ex vivo experiment, both PMVE-MA and HA showed fast dissolutions (<30 s) after insertion into porcine skin. However, HA-MA showed better mechanical properties, namely improved resistance to fracture when submitted to a compression force. Overall, a B16F10 melanoma CM-dissolving MN system was efficiently developed as a promising device suggesting further studies in immunotherapy and melanoma applications

Development of fast dissolving polymer-based microneedles for delivery of an antigenic melanoma cell membrane

Delivery of cancer cell membranes (CM) is a new approach for the activation of the immune system and the induction of immunotherapy of cancer. Local delivery of melanoma CM into skin can induce efficient immune stimulation of antigen presenting cells (APCs), such as dendritic cells. In the current study, fast dissolving microneedles (MNs) were developed for the delivery of melanoma B16F10 CM. Two polymers were tested for the fabrication of MNs: poly(methyl vinyl ether-co-maleic acid) (PMVE-MA) and hyaluronic acid (HA). The incorporation of CM in MNs was achieved through coating of the MNs using a multi-step layering procedure or the micromolding technique. The CM loading and its stabilization were improved by adding sugars (sucrose and trehalose) and a surfactant (Poloxamer 188), respectively. In an ex vivo experiment, both PMVE-MA and HA showed fast dissolutions (<30 s) after insertion into porcine skin. However, HA-MA showed better mechanical properties, namely improved resistance to fracture when submitted to a compression force. Overall, a B16F10 melanoma CM-dissolving MN system was efficiently developed as a promising device suggesting further studies in immunotherapy and melanoma applications