Characterization and application of two RANK-specific antibodies with different biological activities.

Antibodies play an important role in therapy and investigative biomedical research. The TNF-family member Receptor Activator of NF-κB (RANK) is known for its role in bone homeostasis and is increasingly recognized as a central player in immune regulation and epithelial cell activation. However, the study of RANK biology has been hampered by missing or insufficient characterization of high affinity tools that recognize RANK. Here, we present a careful description and comparison of two antibodies, RANK-02 obtained by phage display (Newa, 2014 [1]) and R12-31 generated by immunization (Kamijo, 2006 [2]). We found that both antibodies recognized mouse RANK with high affinity, while RANK-02 and R12-31 recognized human RANK with high and lower affinities, respectively. Using a cell apoptosis assay based on stimulation of a RANK:Fas fusion protein, and a cellular NF-κB signaling assay, we showed that R12-31 was agonist for both species. R12-31 interfered little or not at all with the binding of RANKL to RANK, in contrast to RANK-02 that efficiently prevented this interaction. Depending on the assay and species, RANK-02 was either a weak agonist or a partial antagonist of RANK. Both antibodies recognized human Langerhans cells, previously shown to express RANK, while dermal dendritic cells were poorly labeled. In vivo R12-31 agonist activity was demonstrated by its ability to induce the formation of intestinal villous microfold cells in mice. This characterization of two monoclonal antibodies should now allow better evaluation of their application as therapeutic reagents and investigative tools.

Enhancing actions of peptides derived from the γ-chain of fetal human hemoglobin on the immunostimulant activities of monophosphoryl lipid A.

Hemoglobin and its structures have been described since the 1990s to enhance a variety of biological activities of endotoxins (LPS) in a dose-dependent manner. To investigate the interaction processes in more detail, the system was extended by studying the interactions of newly designed peptides from the γ-chain of human hemoglobin with the adjuvant monophosphoryl lipid A (MPLA), a partial structure of lipid A lacking its 1-phosphate. It was found that some selected Hbg peptides, in particular two synthetic substructures designated Hbg32 and Hbg35, considerably increased the bioactivity of MPLA, which alone was only a weak activator of immune cells. These findings hold true for human mononuclar cells, monocytes and T lymphocytes. To understand the mechanisms of action in more detail, biophysical techniques were applied. These showed a peptide-induced change of the MPLA aggregate structure from multilamellar into a non-lamellar, probably inverted, cubic structure. Concomitantly, the peptides incorporated into the tightly packed MPLA aggregates into smaller units down to monomers. The fragmentation of the aggregates was an endothermic process, differing from a complex formation but rather typical for a catalytic reaction.

Effect of diet composition and ration size on key enzyme activities of glycolysis-gluconeogenesis, pentose phosphate pathway and amino acid metabolism in liver of Sparus aurata

The effects of diet composition and ration size on the activities of key enzymes involved in intermediary metabolism were studied in the liver of gilthead sea bream (Sparus aurata). Highcarbohydrate, low-protein diets stimulated 6-phosphofructo 1-kinase (EC 2.7.1.11), pyruvate kinase (EC 2.7.1.40), glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) enzyme activities, while they decreased alanine aminotransferase (EC 2.6.1.2) activity. A high degree of correlation was found between food ration size and the activity of the enzymes 6-phosphofructo 1-kinase, pyruvate kinase, glucose-6-phosphate dehydrogenase (positive correlations) and fructose-1,6-bisphosphatase (EC 3.1.3.11) (negative correlation). These correlations matched well with the high correlation also found between ration size and growth rate in starved fish refed for 22 d. Limited feeding (5 g/kg body weight) for 22 d decreased the activities of the key enzymes for glycolysis and lipogenesis, and alanine aminotransferase activity. The findings presented here indicate a high level of metabolic adaptation to both diet type and ration size. In particular, adaptation of enzyme activities to the consumption of a diet with a high carbohydrate level suggests that a carnivorous fish like Sparus aurata can tolerate partial replacement of protein by carbohydrate in the commercial diets supplied in culture. The relationship between enzyme activities, ration size and fish growth indicates that the enzymes quickly respond to dietary manipulations of cultured fish.

Effect of diet composition and ration size on key enzyme activities of glycolysis-gluconeogenesis, pentose phosphate pathway and amino acid metabolism in liver of Sparus aurata

The effects of diet composition and ration size on the activities of key enzymes involved in intermediary metabolism were studied in the liver of gilthead sea bream (Sparus aurata). Highcarbohydrate, low-protein diets stimulated 6-phosphofructo 1-kinase (EC 2.7.1.11), pyruvate kinase (EC 2.7.1.40), glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (EC 1.1.1.44) enzyme activities, while they decreased alanine aminotransferase (EC 2.6.1.2) activity. A high degree of correlation was found between food ration size and the activity of the enzymes 6-phosphofructo 1-kinase, pyruvate kinase, glucose-6-phosphate dehydrogenase (positive correlations) and fructose-1,6-bisphosphatase (EC 3.1.3.11) (negative correlation). These correlations matched well with the high correlation also found between ration size and growth rate in starved fish refed for 22 d. Limited feeding (5 g/kg body weight) for 22 d decreased the activities of the key enzymes for glycolysis and lipogenesis, and alanine aminotransferase activity. The findings presented here indicate a high level of metabolic adaptation to both diet type and ration size. In particular, adaptation of enzyme activities to the consumption of a diet with a high carbohydrate level suggests that a carnivorous fish like Sparus aurata can tolerate partial replacement of protein by carbohydrate in the commercial diets supplied in culture. The relationship between enzyme activities, ration size and fish growth indicates that the enzymes quickly respond to dietary manipulations of cultured fish.

Agglomeration vs. dispersion of economic activities in the districts of Paris

The spatial distribution of economic activity has often been analysed for wide geographical areas such as regions or metropolitan areas, but it has rarely been subject to microanalysis, especially outside the U.S. In this paper we focus on what happens within a large European city (Par is), and analyse how the industrial composition of its districts differs and how these districts evolve. We also analyse suburbanization process for both residents and the workforce and provide empirical evidence about the changing roles of the core and intramuros periphery. Keywords: agglomeration, suburbanization, Paris, micropolitan analysis

Learning in later life: participation in formal, non-formal and informal activities in a nationally representative Spanish sample

This article examines the participation of Spanish older people in formal, non-formal and informal learning activities and presents a profile of participants in each kind of learning activity. We used data from a nationally representative sample of Spanish people between 60 and 75 years old (n = 4,703). The data were extracted from the 2007 Encuesta sobre la Participación de la Población Adulta en Actividades de Aprendizaje (EADA, Survey on Adult Population Involvement in Learning Activities). Overall, only 22.8 % of the sample participated in a learning activity. However, there was wide variation in the participation rates for the different types of activity. Informal activities were far more common than formal ones. Multivariate logistic regression indicated that education level and involvement in social and cultural activities were associated with likelihood of participating, regardless of the type of learning activity. When these variables were taken into account, age did not predict decreasing participation, at least in non-formal and informal activities. Implications for further research, future trends and policies to promote older adult education are discussed.

Interactions of bacteria and fungi on decomposing litter: differential extracellular enzyme activities

Fungi and bacteria are key agents in plant litter decomposition in freshwater ecosystems. However, the specific roles of these two groups and their interactions during the decomposition process are unclear. We compared the growth and patterns of degradativeenzymes expressed by communities of bacteria and fungi grown separately and in coexistence on Phragmites leaves. The two groups displayed both synergistic and antagonistic interactions. Bacteria grew better together with fungi than alone. In addition, there was a negative effect of bacteria on fungi, which appeared to be caused by suppression of fungal growth and biomass accrual rather than specifically affecting enzyme activity. Fungi growing alone had a high capacity for the decomposition of plant polymers such as lignin, cellulose, and hemicellulose. In contrast, enzyme activities were in general low when bacteria grew alone, and the activity of key enzymes in the degradation of lignin and cellulose (phenol oxidase and cellobiohydrolase) was undetectable in the bacteria-only treatment. Still, biomass-specific activities of most enzymes were higher in bacteria than in fungi. The low total activity and growth of bacteria in the absence of fungi in spite of apparent high enzymatic efficiency during the degradation of many substrates suggest that fungi provide the bacteria with resources that the bacteria were not able to acquire on their own, most probably intermediate decomposition products released by fungi that could be used by bacteria

Introducing online teaching in Humanities: A case study about the acceptance of online activities by the academic staff of classical languages

The purpose of this paper is to show the perceptions of the academic staff of classical languages (ancient Greek and Latin) concerning use of online activities during their courses. The study was carried out in three countries: Greece (three major Universities), Spain (University of Barcelona) and the United States (University of California, Berkeley) with the participation of thirty-three academic instructors. Depending on the level of use and acceptance of the ICT and following G. Moore¿s classification, we separated the participating academics in three groups: the conservatives, the mainstream and the early adopters. The fact that the smallest group is the third clearly shows the necessity for teachers¿ preparation and training before introducing innovative projects in the classroom. Since the starting point for the application of innovation in the classroom is the teacher, policy makers should focus on helping them become conscious of changes in teaching methods and include their opinion during the design of innovative projects.

Introducing online teaching in Humanities: A case study about the acceptance of online activities by the academic staff of classical languages

The purpose of this paper is to show the perceptions of the academic staff of classical languages (ancient Greek and Latin) concerning use of online activities during their courses. The study was carried out in three countries: Greece (three major Universities), Spain (University of Barcelona) and the United States (University of California, Berkeley) with the participation of thirty-three academic instructors. Depending on the level of use and acceptance of the ICT and following G. Moore¿s classification, we separated the participating academics in three groups: the conservatives, the mainstream and the early adopters. The fact that the smallest group is the third clearly shows the necessity for teachers¿ preparation and training before introducing innovative projects in the classroom. Since the starting point for the application of innovation in the classroom is the teacher, policy makers should focus on helping them become conscious of changes in teaching methods and include their opinion during the design of innovative projects.

Enseñar a aprender a pensar en los centros educativos, incluso en las actividades de evaluación.= Teaching to learn how to think at schools, even within assessment activities

El auténtico protagonismo de los centros educativostiene que dirigirse a ayudar a pensar a sus alumnos y aenseñarlos a aprender, es decir, el docente tiene queenseñar estrategias de aprendizaje y debe promover elesfuerzo del estudiante para facilitar la construcción deesquemas y el aprendizaje permanente.El profesor debe utilizar cualquier situación deaprendizaje para enseñar dichas estrategias deaprendizaje, incluso en las situaciones de evaluación;por lo tanto, en este trabajo se sugiere que en lasevaluaciones de los alumnos y alumnas se tenga encuenta la metacognición como factor fundamental en elaprendizaje y la enseñanza

Natural occurrence, biological activities and synthesis of eight-, nine-, and eleven-membered ring lactones

The natural occurrence, biological activities and synthetic approaches to natural eight-, nine-, and eleven-membered lactones is reviewed. These medium ring lactones are grouped according to ring size, and their syntheses are discussed. The structures of some natural products early identified as medium-ring lactones were revised after total synthesis.

Challenges and Means for the Front End Activities of Software Development

The front end of innovation is regarded as one of the most important steps in building new software products or services, and the most significant benefits in software development can be achieved through improvements in the front end activities. Problems in the front end phase have an impact on customer dissatisfaction with delivered software, and on the effectiveness of the entire software development process. When these processes are improved, the likelihood of delivering high quality software and business success increases. This thesis highlights the challenges and problems related to the early phases of software development, and provides new methods and tools for improving performance in the front end activities of software development. The theoretical framework of this study comprises two fields of research. The first section belongs to the field of innovation management, and especially to the management of the early phases of the innovation process, i.e. the front end of innovation. The second section of the framework is closely linked to the processes of software engineering, especially to the early phases of the software development process, i.e. the practice of requirements engineering. Thus, this study extends the theoretical knowledge and discloses the differences and similarities in these two fields of research. In addition, this study opens up a new strand for academic discussion by connecting these research directions. Several qualitative business research methodologies have been utilized in the individual publications to solve the research questions. The theoretical and managerial contribution of the study can be divided into three areas: 1) processes and concepts, 2) challenges and development needs, and 3) means and methods for the front end activities of software development. First, the study discloses the difference and similarities between the concepts of the front end of innovation and requirements engineering, and proposes a new framework for managing the front end of the software innovation process, bringing business and innovation perspectives into software development. Furthermore, the study discloses managerial perceptions of the similarities and differences in the concept of the front end of innovation between the software industry and the traditional industrial sector. Second, the study highlights the challenges and development needs in the front end phase of software development, especially challenges in communication, such as linguistic problems, ineffective communication channels, a communication gap between users/customers and software developers, and participation of multiple persons in software development. Third, the study proposes new group methods for improving the front end activities of software development, especially customer need assessment, and the elicitation of software requirements.

Antioxidant and cytotoxic activities of 'açaí' (Euterpe precatoria Mart.)

Decoction of roots of the Amazonian palm açaí (Euterpe precatoria Mart.) is widely used by Brazilian and Peruvian people as an anti-inflammatory, to heal kidney and liver diseases and against snake bites. In this study, the antioxidant activity of extracts and flavonoids (quercetin, catechin, epicatechin, rutin and astilbin) isolated from roots and leaf stalks of E. precatoria was investigated using β-carotene in TLC plates and DPPH radical scavenging in a spectrophotometric bioassay. All extracts and flavonoids showed activity. Also, the cytotoxic activity of these extracts was evaluated by the brine shrimp (Artemia salina) larvicide bioassay and was lower than that of lapachol, used as control. The presence of flavonoids and sitosterol-3-O-β-D-glucopyranoside in the extracts can justify the use of the plant in traditional medicine.

In vitro antimicrobial and antioxidant activities of a crude extract and fractions from Buddleja thyrsoides Lam. Leaves

Crude extract and fractions of Buddleja thyrsoides were investigated regarding antioxidant activities by DPPH, total phenolic contents by Folin-Ciocalteau and antimicrobial activity by the broth microdilution method. Total phenolics varied from 214.07 ± 3.6 to 438.4 ± 0.3 mg g-1. Crude extract, ethyl acetate, dichloromethane and butanolic fractions exhibited a weak scavenging activity (SC50=186.04 ± 10.8, 137.70 ± 8.5, 146.89 ± 9.0 and 165.71 ± 3.2 µg mL-1, respectively). A correlation between the antioxidant activities and total phenolic contents could be shown (r=0.857, p<0.01). The lowest value of MIC was observed with butanolic fraction against Saccharomyces cerevisiae (MIC and MFC at 62.5 µg mL-1). Dichloromethane and ethyl acetate fractions were effective against Staphylococcus aureus with MIC value at 250 and 500 µg mL-1 respectively.

Chemical composition and antibacterial activities from the essential oils of myrtaceae species planted in Brazil

The essential oils of seven Myrtaceae species were investigated for its chemical composition and antibacterial activity. The volatile oils were characterized by a high content of monoterpenoids of which 1,8-cineole (88.0, 65.0 and 77.0% for Melaleuca hypericifolia, Callistemon viminalis and Callistemon citrinus respectively), terpinen-4-ol (47.0 and 49.8% for Melaleuca thymifolia and Callistemon polandii respectively) and α-pinene (54.5% for Kunzea ericoides) were the major components. The oil from M. linariifolia was characterized by a high concentration of methyleugenol (87.2%). The oil from Melaleuca thymifolia was the most active, exhibiting high antimicrobial activity against all tested bacteria.