Artificial micro RNA system in Dictyostelium discoideum

Dictyostelium discoideum is a social amoeba that serves as a model system for RNA interference and related mechanisms. Its position between plants and animals enables evolutionary snapshot of mechanisms and protein machinery involved in investigated subjects. MiRNAs are small regulatory RNAs that are evolutionary conserved and present in animals, plants, viruses and some prokaryotes. They have roles in development, cell growth and differentiation, apoptosis and their miss-regulation is associated with many diseases such as cancer, neurodegenerative disorders and diabetes. Recently, through sequencing of DNA libraries miRNAs have been discovered in D. discoideum. In this work, it has been shown that heterologues miRNA let-7 can be expressed and processed in D. discoideum. Expression of let-7 miRNA in social amoeba resulted in a strong developmental phenotype suggesting an overload of the processing/silencing system or/and endogenous targets. The various effects on prel-7 strain have been observed and characterized, serving as a background for postulation of miRNA roles. An artificial miRNA system has been established and imposed to D. discoideum, showing that miRNAs in Dictyostelium could mediate gene expression on the level of mRNA stability and on the posttranscriptional level. Furthermore, presence of translational inhibition as a type of gene control was shown for the first time in this organism. Due to it new structures representing co-localities of miRNA and target mRNA have been detected. Taken together, this work shows functional artificial miRNA system and postulates roles of endogenous small RNA in social amoeba.

Variability in the Stability and Productivity of Transfected Genes in Chinese Hamster Ovary (CHO) cells

In the field of biologics production, productivity and stability of the transfected gene of interest are two very important attributes that dictate if a production process is viable. To further understand and improve these two traits, we would need to further our understanding of the factors affecting them. These would include integration site of the gene, gene copy number, cell phenotypic variation and cell environment. As these factors play different parts in the development process, they lead to variable productivity and stability of the transfected gene between clones, the well-known phenomenon of “clonal variation”. A study of this phenomenon and how the various factors contribute to it will thus shed light on strategies to improve productivity and stability in the production cell line. Of the four factors, the site of gene integration appears to be one of the most important. Hence, it is proposed that work is done on studying how different integration sites affect the productivity and stability of transfected genes in the development process. For the study to be more industrially relevant, it is proposed that the Chinese Hamster Ovary dhfr-deficient cell line, CHO-DG44, is used as the model system.

Necessary and sufficient conditions for robust stability using modal intervals

In this paper, robustness of parametric systems is analyzed using a new approach to interval mathematics called Modal Interval Analysis. Modal Intervals are an interval extension that, instead of classic intervals, recovers some of the properties required by a numerical system. Modal Interval Analysis not only simplifies the computation of interval functions but allows semantic interpretation of their results. Necessary, sufficient and, in some cases, necessary and sufficient conditions for robust performance are presented

New delay-dependent stability criteria for uncertain neutral systems with mixed time-varying delays and nonlinear perturbations

The problem of stability analysis for a class of neutral systems with mixed time-varying neutral, discrete and distributed delays and nonlinear parameter perturbations is addressed. By introducing a novel Lyapunov-Krasovskii functional and combining the descriptor model transformation, the Leibniz-Newton formula, some free-weighting matrices, and a suitable change of variables, new sufficient conditions are established for the stability of the considered system, which are neutral-delay-dependent, discrete-delay-range dependent, and distributeddelay-dependent. The conditions are presented in terms of linear matrix inequalities (LMIs) and can be efficiently solved using convex programming techniques. Two numerical examples are given to illustrate the efficiency of the proposed method

Advances in animal blood processing: development of a biopreservation system and insights on the functional properties of plasma

La sang és un subproducte amb un alt potencial de valorització que s'obté en quantitats importants en els escorxadors industrials. Actualment, la majoria de sistemes de recollida de la sang no segueixen unes mesures d'higiene estrictes, pel que esdevé un producte de baixa qualitat microbiològica. Conseqüentment, l'aprofitament de la sang és una sortida poc estimulant des del punt de vista econòmic, ja que acostuma a perdre les qualitats que permetrien l'obtenció de productes d'alt valor afegit. El capítol I del present treball s'inclou dins d'un projecte que proposa la inoculació de bacteris de l'àcid làctic (LAB) com un cultiu bioconservador de la sang, un sistema senzill i de baix cost que cerca l'estabilitat de la sang, tant microbiològica com fisicoquímica, durant el període del seu emmagatzematge. El capítol II s'emmarca dins d'un projecte que cerca la millora de l'aprofitament integral de la sang que, en el cas de la fracció plasmàtica, es centra en l'estudi de la funcionalitat dels seus principals constituents. Conèixer la contribució dels components majoritaris ha de permetre la millora de la funcionalitat dels ingredients alimentaris derivats. Els resultats presentats en aquesta tesi poden ajudar a la valorització de la sang porcina d'escorxadors industrials, mitjançant els coneixements adquirits pel que fa a la millora del seu sistema de recollida i del desenvolupament d'ingredients alimentaris amb interessants propietats funcionals.

An observing system simulation experiment for climate monitoring with GNSS radio occulation data: setup and testbed study

The long-term stability, high accuracy, all-weather capability, high vertical resolution, and global coverage of Global Navigation Satellite System (GNSS) radio occultation (RO) suggests it as a promising tool for global monitoring of atmospheric temperature change. With the aim to investigate and quantify how well a GNSS RO observing system is able to detect climate trends, we are currently performing an (climate) observing system simulation experiment over the 25-year period 2001 to 2025, which involves quasi-realistic modeling of the neutral atmosphere and the ionosphere. We carried out two climate simulations with the general circulation model MAECHAM5 (Middle Atmosphere European Centre/Hamburg Model Version 5) of the MPI-M Hamburg, covering the period 2001–2025: One control run with natural variability only and one run also including anthropogenic forcings due to greenhouse gases, sulfate aerosols, and tropospheric ozone. On the basis of this, we perform quasi-realistic simulations of RO observables for a small GNSS receiver constellation (six satellites), state-of-the-art data processing for atmospheric profiles retrieval, and a statistical analysis of temperature trends in both the “observed” climatology and the “true” climatology. Here we describe the setup of the experiment and results from a test bed study conducted to obtain a basic set of realistic estimates of observational errors (instrument- and retrieval processing-related errors) and sampling errors (due to spatial-temporal undersampling). The test bed results, obtained for a typical summer season and compared to the climatic 2001–2025 trends from the MAECHAM5 simulation including anthropogenic forcing, were found encouraging for performing the full 25-year experiment. They indicated that observational and sampling errors (both contributing about 0.2 K) are consistent with recent estimates of these errors from real RO data and that they should be sufficiently small for monitoring expected temperature trends in the global atmosphere over the next 10 to 20 years in most regions of the upper troposphere and lower stratosphere (UTLS). Inspection of the MAECHAM5 trends in different RO-accessible atmospheric parameters (microwave refractivity and pressure/geopotential height in addition to temperature) indicates complementary climate change sensitivity in different regions of the UTLS so that optimized climate monitoring shall combine information from all climatic key variables retrievable from GNSS RO data.

Fatty acids and the immune system: from basic science to clinical applications

Over the last 25 years, the effects of fatty acids on the immune system have been characterized using in vitro, animal and human studies. Advances in fatty acid biochemistry and molecular techniques have recently suggested new mechanisms by which fatty acids could potentially modify immune responses, including modification of the organization of cellular lipids and interaction with nuclear receptors. Possibilities for the clinical applications of n-3 PUFA are now developing. The present review focuses on the hypothesis that the anti-inflammatory properties of n-3 PUFA in the arterial wall may contribute to the protective effects of n-3 PUFA in CVD, as suggested by epidemiological and secondary prevention studies. Studies are just beginning to show that dietary n-3 PUFA can be incorporated into plaque lipid in human subjects, where they may influence the morphology and stability of the atherosclerotic lesion.

Challenge testing the lactoperoxidase system against a range of bacteria using different activation agents

Lactoperoxidase (LP) exerts antimicrobial effects in combination with H2O2 and either thiocyanate (SCN-) or a halide (e. g., I-). Garlic extract in the presence of ethanol has also been used to activate the LP system. This study aimed to determine the effects of 3 LP activation systems (LP+SCN-+H2O2; LP+I-+H2O2; LP + garlic extract + ethanol) on the growth and activity of 3 test organisms (Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus). Sterilized milk was used as the reaction medium, and the growth pattern of the organisms and a range of keeping quality (KQ) indicators (pH, titratable acidity, ethanol stability, clot on boiling) were monitored during storage at the respective optimum growth temperature for each organism. The LP+I-+H2O2 system reduced bacterial counts below the detection limit shortly after treatment for all 3 organisms, and no bacteria could be detected for the duration of the experiment (35 to 55 h). The KQ data confirmed that the milk remained unspoiled at the end of the experiments. The LP + garlic extract + ethanol system, on the other hand, had no effect on the growth or KQ with P. aeruginosa, but showed a small retardation of growth of the other 2 organisms, accompanied by small increases (5 to 10 h) in KQ. The effects of the LP+SCN-+H2O2 system were intermediate between those of the other 2 systems and differed between organisms. With P. aeruginosa, the system exerted total inhibition within 10 h of incubation, but the bacteria regained viability after a further 5 h, following a logarithmic growth curve. This was reflected in the KQ indicators, which implied an extension of 15 h. With the other 2 bacterial species, LP+SCN-+H2O2 exerted an obvious inhibitory effect, giving a lag phase in the growth curve of 5 to 10 h and KQ extension of 10 to 15 h. When used in combination, I- and SCN- displayed negative synergy.

Alternative strategies for activation of the natural lactoperoxidase system in cows' milk: trials in Tanzania

Thiocyanate content and lactoperoxidase activity of individual cow's milk of different breeds were determined, and the effects of different lactoperoxidase system (LP-s) activation strategies were compared. Lactoperoxidase activity varied significantly between Friesian and both Ayrshire and Tanzania Short Horn Zebu (TSHZ), but differences between Ayrshire and TSHZ were not significant. There was no significant variation in SCN- content between breeds. The LP-s was activated using three strategies based on SCN-: namely; equal concentrations of SCN- and H2O2 (7:7, 10:10, 15 :15 mg/l), excess SCN- concentrations (15:10, 20:10, 25:10 mg SCN-: H2O2/I), and excess H2O2 concentrations (10:15, 10:20, 10:25 mg SCN-: H2O2/I), plus a fourth strategy based on I- (15 : 15 mg I- : H2O2/I). The keeping quality (KQ) was assessed using pH, titratable acidity, clot on boiling and alcohol stability tests. All activation strategies enhanced the shelf life of milk (typically increasing KQ from around 10 to around 20 h), but it was clear that the effectiveness of the LP-s depends on the type and concentrations of the activators of the system. The LP-s activated using I- as an electron donor was more effective than the LP-s activated using SCN- as an electron donor, increasing the KQ by a further 6-8 h compared with SCN-.

Effect of pH on the antimicrobial activity and oxidative stability of oil-in-water emulsions containing caffeic acid

Antioxidant properties in food are dependent on various parameters. These include the pH value and interactions with food components, including proteins or metal ions. food components affect antioxidant stability and also influence the properties of microorganisms and their viability. This paper describes an investigation of the effect of pH on the antioxidant and antibacterial properties of caffeic acid in different media. The pH values studied, using an oil-in-water emulsion as model system, were 3, 5 (with and without phosphate buffer), and 9. Effects of mixtures of caffeic acid, bovine serum albumin (BSA), and Fe (III) on oxidative deterioration in the emulsion samples were studied. The results show that the antioxidant activity of caffeic acid was increased by the presence of BSA. This effect was pH dependent and was affected by the presence of iron Ions. Antibacterial properties were also pH dependent. The minimum concentration of caffeic acid required to inhibit some microorganisms in the pH range of 5 to 7 was determined. A concentration of 0.41% (w/w) caffeic acid was enough to inhibit the growth of some of the studied microorganisms in the pH range of 5 to 7. However, near-neutral pH concentrations higher than 0.4% were needed to inhibit some microorganisms, including Listeria monocytogenes, E. coli, and Staphylococcus aureus, in the medium.

Systematic distortions of perceptual stability investigated using immersive virtual reality

Using an immersive virtual reality system, we measured the ability of observers to detect the rotation of an object when its movement was yoked to the observer's own translation. Most subjects had a large bias such that a static object appeared to rotate away from them as they moved. Thresholds for detecting target rotation were similar to those for an equivalent speed discrimination task carried out by static observers, suggesting that visual discrimination is the predominant limiting factor in detecting target rotation. Adding a stable visual reference frame almost eliminated the bias. Varying the viewing distance of the target had little effect, consistent with observers underestimating distance walked. However, accuracy of walking to a briefly presented visual target was high and not consistent with an underestimation of distance walked. We discuss implications for theories of a task-independent representation of visual space. © 2005 Elsevier Ltd. All rights reserved.

Information system evolvability, feedback and pattern languages

Information systems for business are frequently heavily reliant on software. Two important feedback-related effects of embedding software in a business process are identified. First, the system dynamics of the software maintenance process can become complex, particularly in the number and scope of the feedback loops. Secondly, responsiveness to feedback can have a big effect on the evolvability of the information system. Ways have been explored to provide an effective mechanism for improving the quality of feedback between stakeholders during software maintenance. Understanding can be improved by using representations of information systems that are both service-based and architectural in scope. The conflicting forces that encourage change or stability can be resolved using patterns and pattern languages. A morphology of information systems pattern languages has been described to facilitate the identification and reuse of patterns and pattern languages. The kind of planning process needed to achieve consensus on a system's evolution is also considered.

Subspace system identification framework for the analysis of multimoded propagation of THz-transient signals

We provide a system identification framework for the analysis of THz-transient data. The subspace identification algorithm for both deterministic and stochastic systems is used to model the time-domain responses of structures under broadband excitation. Structures with additional time delays can be modelled within the state-space framework using additional state variables. We compare the numerical stability of the commonly used least-squares ARX models to that of the subspace N4SID algorithm by using examples of fourth-order and eighth-order systems under pulse and chirp excitation conditions. These models correspond to structures having two and four modes simultaneously propagating respectively. We show that chirp excitation combined with the subspace identification algorithm can provide a better identification of the underlying mode dynamics than the ARX model does as the complexity of the system increases. The use of an identified state-space model for mode demixing, upon transformation to a decoupled realization form is illustrated. Applications of state-space models and the N4SID algorithm to THz transient spectroscopy as well as to optical systems are highlighted.

Robust pole assignment and optimal stability margins

A robust pole assignment by linear state feedback is achieved in state-space representation by selecting a feedback which minimises the conditioning of the assigned eigenvalues of the closed-loop system. It is shown here that when this conditioning is minimised, a lower bound on the stability margin in the frequency domain is maximised.

On the existence of two-dimensional Euler flows satisfying energy-Casimir stability criteria

The energy-Casimir stability method, also known as the Arnold stability method, has been widely used in fluid dynamical applications to derive sufficient conditions for nonlinear stability. The most commonly studied system is two-dimensional Euler flow. It is shown that the set of two-dimensional Euler flows satisfying the energy-Casimir stability criteria is empty for two important cases: (i) domains having the topology of the sphere, and (ii) simply-connected bounded domains with zero net vorticity. The results apply to both the first and the second of Arnold’s stability theorems. In the spirit of Andrews’ theorem, this puts a further limitation on the applicability of the method. © 2000 American Institute of Physics.