label free fluorescence detection of kinase activity using a gold nanoparticle based indicator displacement assay

A straightforward fluorescence indicator-displacement assay (IDA) has been developed for the quantitative analysis of ATP→ADP conversion

Sustained Exendin-4 Secretion through Gene Therapy Targeting Salivary Glands in Two Different Rodent Models of Obesity/Type 2 Diabetes.

Exendin-4 (Ex-4) is a Glucagon-like peptide 1 (GLP-1) receptor agonist approved for the treatment of Type 2 Diabetes (T2DM), which requires daily subcutaneous administration. In T2DM patients, GLP-1 administration is reported to reduce glycaemia and HbA1c in association with a modest, but significant weight loss. The aim of present study was to characterize the site-specific profile and metabolic effects of Ex-4 levels expressed from salivary glands (SG) in vivo, following adeno-associated virus-mediated (AAV) gene therapy in two different animal models of obesity prone to impaired glucose tolerance and T2DM, specifically, Zucker fa/fa rats and high fed diet (HFD) mice. Following percutaneous injection of AAV5 into the salivary glands, biologically active Ex-4 was detected in the blood of both animal models and expression persisted in salivary gland ductal cell until the end of the study. In treated mice, Ex-4 levels averaged 138.9±42.3 pmol/L on week 6 and in treated rats, mean circulating Ex-4 levels were 238.2±72 pmol/L on week 4 and continued to increase through week 8. Expression of Ex-4 resulted in a significant decreased weight gain in both mice and rats, significant improvement in glycemic control and/or insulin sensitivity as well as visceral adipose tissue adipokine profile. In conclusion, these results suggest that sustained site-specific expression of Ex-4 following AAV5-mediated gene therapy is feasible and may be useful in the treatment of obesity as well as trigger improved metabolic profile

Prevalence of the <i>Rhizobium etli</i>-like allele in genes coding for 16S rRNA among the indigenous rhizobial populations found associated with wild beans from the Southern Andes in Argentina

A collection of rhizobial isolates from nodules of wild beans,Phaseolus vulgaris var. aborigineus, found growing in virgin lands in 17 geographically separate sites in northwest Argentina was characterized on the basis of host range, growth, hybridization to a nifH probe, analysis of genes coding for 16S rRNA (16S rDNA), DNA fingerprinting, and plasmid profiles. Nodules in field-collected wild bean plants were largely dominated by rhizobia carrying the 16S rDNA allele of Rhizobium etli. A similar prevalence of the R. etli allele was observed among rhizobia trapped from nearby soil. Intragroup diversity of wild bean isolates with either R. etli-like or Rhizobium leguminosarum bv. phaseoli-like alleles was generally found across northwest Argentina. The predominance of the R. etli allele suggests that in this center of origin of P. vulgaris the coevolution of Rhizobium spp. and primitive beans has resulted in this preferential symbiotic associationInstituto de Biotecnologia y Biologia Molecula

Genetic PTX3 deficiency and aspergillosis in stem-cell transplantation

BACKGROUND: The soluble pattern-recognition receptor known as long pentraxin 3 (PTX3) has a nonredundant role in antifungal immunity. The contribution of single-nucleotide polymorphisms (SNPs) in PTX3 to the development of invasive aspergillosis is unknown. METHODS: We screened an initial cohort of 268 patients undergoing hematopoietic stem-cell transplantation (HSCT) and their donors for PTX3 SNPs modifying the risk of invasive aspergillosis. The analysis was also performed in a multicenter study involving 107 patients with invasive aspergillosis and 223 matched controls. The functional consequences of PTX3 SNPs were investigated in vitro and in lung specimens from transplant recipients. RESULTS: Receipt of a transplant from a donor with a homozygous haplotype (h2/h2) in PTX3 was associated with an increased risk of infection, in both the discovery study (cumulative incidence, 37% vs. 15%; adjusted hazard ratio, 3.08; P=0.003) and the confirmation study (adjusted odds ratio, 2.78; P=0.03), as well as with defective expression of PTX3. Functionally, PTX3 deficiency in h2/h2 neutrophils, presumably due to messenger RNA instability, led to impaired phagocytosis and clearance of the fungus. CONCLUSIONS: Genetic deficiency of PTX3 affects the antifungal capacity of neutrophils and may contribute to the risk of invasive aspergillosis in patients treated (Funded by the European Society of Clinical Microbiology and Infectious Diseases and others) .with HSCT.Supported by grants from the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) (to Dr. Carvalho); the German Ministry for Education and Science (03Z2JN21, to Dr. Kurzai); the European Commission (FP7-HEALTH-2009-260338, to Dr. Romani; FP7-HEALTH-2011-280873, to Dr. Mantovani), the European Research Council (ERC-2008-AdG-233417, to Dr. Mantovani; ERC-2011-AdG-293714, to Dr. Romani), Associazione Italiana per la Ricerca sul Cancro (99629, to Dr. Mantovani); and Fundacao para a Ciencia e Tecnologia, Portugal (SFRH/BPD/46292/2008, to Dr. Carvalho; SFRH/BD/65962/2009, to Dr. Cunha; and SFRH/BPD/70783/2010, to Dr. Almeida)

An NBD derivative of the selective rat toxicant norbormide as a new probe for living cell imaging

Norbormide (NRB) is a unique compound that acts directly on rat vascular myocytes to trigger a contractile process, through an as yet unknown mechanism, which results in the selective contraction of rat peripheral arteries. To gain insight into the mechanisms involved in NRB rat-selective activity, we investigated the subcellular distribution of NRB-AF12, a nitrobenzoxadiazole (NBD)-derivative of NRB, in living NRB-sensitive and NRB-insensitive cells. In both cell types, NRB-AF12 localized to the endoplasmic reticulum (ER), Golgi apparatus, mitochondria, lysosomes, and endosomes; however, in NRB-sensitive cells, the fluorescence also extended to the plasma membrane. NRB-AF12 was rapidly internalized into the cells, could easily be washed out and then reloaded back into the same cells, all with a high degree of reproducibility. Cells exposed for 24 h to NRB-AF12 did not show apparent signs of toxicity, even at concentrations of the dye (10 μM) much higher than those required for fluorescence labeling (500 ηM). The distribution pattern of NRB-AF12 fluorescence was near identical to that of ER-Tracker® (Er-Tr), a fluorescent derivative of glibenclamide, a known KATP channel blocker. Displacement tests did not demonstrate, but at the same time did not rule out the possibility of a common target for ER-Tr, NRB-AF12, NRB, and glibenclamide. On the basis of these results we hypothesize a common target site for NRB-AF12 and ER-Tr, and a similar target profile for NRB and glibenclamide, and propose NRB-AF12 as an alternative fluorescence probe to ER-Tracker. Furthermore, NRB-based fluorescence derivatives could be designed to selectively label single cellular structures

Empirical modelling of translation and interpreting

Empirical research is carried out in a cyclic way: approaching a research area bottom-up, data lead to interpretations and ideally to the abstraction of laws, on the basis of which a theory can be derived. Deductive research is based on a theory, on the basis of which hypotheses can be formulated and tested against the background of empirical data. Looking at the state-of-the-art in translation studies, either theories as well as models are designed or empirical data are collected and interpreted. However, the final step is still lacking: so far, empirical data has not lead to the formulation of theories or models, whereas existing theories and models have not yet been comprehensively tested with empirical methods. This publication addresses these issues from several perspectives: multi-method product- as well as process-based research may gain insights into translation as well as interpreting phenomena. These phenomena may include cognitive and organizational processes, procedures and strategies, competence and performance, translation properties and universals, etc. Empirical findings about the deeper structures of translation and interpreting will reduce the gap between translation and interpreting practice and model and theory building. Furthermore, the availability of more large-scale empirical testing triggers the development of models and theories concerning translation and interpreting phenomena and behavior based on quantifiable, replicable and transparent data

Empirical modelling of translation and interpreting

Empirical research is carried out in a cyclic way: approaching a research area bottom-up, data lead to interpretations and ideally to the abstraction of laws, on the basis of which a theory can be derived. Deductive research is based on a theory, on the basis of which hypotheses can be formulated and tested against the background of empirical data. Looking at the state-of-the-art in translation studies, either theories as well as models are designed or empirical data are collected and interpreted. However, the final step is still lacking: so far, empirical data has not lead to the formulation of theories or models, whereas existing theories and models have not yet been comprehensively tested with empirical methods. This publication addresses these issues from several perspectives: multi-method product- as well as process-based research may gain insights into translation as well as interpreting phenomena. These phenomena may include cognitive and organizational processes, procedures and strategies, competence and performance, translation properties and universals, etc. Empirical findings about the deeper structures of translation and interpreting will reduce the gap between translation and interpreting practice and model and theory building. Furthermore, the availability of more large-scale empirical testing triggers the development of models and theories concerning translation and interpreting phenomena and behavior based on quantifiable, replicable and transparent data

Empirical modelling of translation and interpreting

Empirical research is carried out in a cyclic way: approaching a research area bottom-up, data lead to interpretations and ideally to the abstraction of laws, on the basis of which a theory can be derived. Deductive research is based on a theory, on the basis of which hypotheses can be formulated and tested against the background of empirical data. Looking at the state-of-the-art in translation studies, either theories as well as models are designed or empirical data are collected and interpreted. However, the final step is still lacking: so far, empirical data has not lead to the formulation of theories or models, whereas existing theories and models have not yet been comprehensively tested with empirical methods. This publication addresses these issues from several perspectives: multi-method product- as well as process-based research may gain insights into translation as well as interpreting phenomena. These phenomena may include cognitive and organizational processes, procedures and strategies, competence and performance, translation properties and universals, etc. Empirical findings about the deeper structures of translation and interpreting will reduce the gap between translation and interpreting practice and model and theory building. Furthermore, the availability of more large-scale empirical testing triggers the development of models and theories concerning translation and interpreting phenomena and behavior based on quantifiable, replicable and transparent data

Empirical modelling of translation and interpreting

Empirical research is carried out in a cyclic way: approaching a research area bottom-up, data lead to interpretations and ideally to the abstraction of laws, on the basis of which a theory can be derived. Deductive research is based on a theory, on the basis of which hypotheses can be formulated and tested against the background of empirical data. Looking at the state-of-the-art in translation studies, either theories as well as models are designed or empirical data are collected and interpreted. However, the final step is still lacking: so far, empirical data has not lead to the formulation of theories or models, whereas existing theories and models have not yet been comprehensively tested with empirical methods. This publication addresses these issues from several perspectives: multi-method product- as well as process-based research may gain insights into translation as well as interpreting phenomena. These phenomena may include cognitive and organizational processes, procedures and strategies, competence and performance, translation properties and universals, etc. Empirical findings about the deeper structures of translation and interpreting will reduce the gap between translation and interpreting practice and model and theory building. Furthermore, the availability of more large-scale empirical testing triggers the development of models and theories concerning translation and interpreting phenomena and behavior based on quantifiable, replicable and transparent data