Monitoring of Carboxypeptidase Digestion by Matrix-Assisted Laser Desorption and Ionization Mass Spectrometry

The potential of matrix-assisted laser desorption and ionization mass spectrometry (LDI-MS) is demonstrated by monitoring and analyzing the digestion of (human) pTH (1–34), a synthetic peptide with carboxypeptidases Y and B. All occurring ion signals in the mass spectra could be identified as degraded peptides. By calculating the mass differences between successive degraded peptides, it was possible to identify the released amino acids and to determine 8 amino acids of the C-terminus of the original peptide. For a single MS measurement, only 2 pmol of substrate was needed. Time-course analysis of the cleavage of the first amino acid residue gave insight into the kinetics involved. These measurements strongly support the hope that quantitative information about concentrations can be extracted from LDI-MS

Variations in SME Characteristics and the Use of Service Intermediaries for R&D

[EN] Large companies increasingly look externally for opportunities to enhance innovation, which has resulted in closer study of innovation systems. We examined the role of service intermediaries (universities, technology centers, and consultants) within these systems using a sample of predominately small- and medium-sized enterprises located in Valencia, Spain. As with studies of large companies, absorptive capacity (e.g., research and development expenditures) was positively related to the likelihood that a firm would engage in service collaborations. However, the rate of collaboration was higher relative to studies dominated by larger companies. Also, there were differences in the pattern of firm characteristics associated with the use of specific services. For example, the level of SME export activity related positively to the use of technology centers, but negatively to university collaborationsThe authors thank the Alto Consejo Consultivo en I + D of the Generalitat Valenciana and IMPIVA for facilitating the access to the survey on which this paper is based and, especially, Manuel López Estornell and Emilio Cubel for preparing the questionnaire and the sample. This work has been partially supported by the Universitat Politecnica de Valencia under grants PAID-06-10/2424 and PAID 06-10/2392, and Generalitat Valenciana under grants GV/2011/002 and GV/2011/049Castrogiovanni, GJ.; Doménech I De Soria, J.; Mas Verdú, F. (2012). Variations in SME Characteristics and the Use of Service Intermediaries for R&D. Canadian Journal of Administrative Sciences / Revue Canadienne des Sciences de l'Administration. 1-11. doi:10.1002/CJAS.231S11

Genome-Wide Identification and Mapping of NBS-Encoding Resistance Genes in Solanum tuberosum Group Phureja

The majority of disease resistance (R) genes identified to date in plants encode a nucleotide-binding site (NBS) and leucine-rich repeat (LRR) domain containing protein. Additional domains such as coiled-coil (CC) and TOLL/interleukin-1 receptor (TIR) domains can also be present. In the recently sequenced Solanum tuberosum group phureja genome we used HMM models and manual curation to annotate 435 NBS-encoding R gene homologs and 142 NBS-derived genes that lack the NBS domain. Highly similar homologs for most previously documented Solanaceae R genes were identified. A surprising ∼41% (179) of the 435 NBS-encoding genes are pseudogenes primarily caused by premature stop codons or frameshift mutations. Alignment of 81.80% of the 577 homologs to S. tuberosum group phureja pseudomolecules revealed non-random distribution of the R-genes; 362 of 470 genes were found in high density clusters on 11 chromosomes

Changing perspectives on the internationalization of R&D and innovation by multinational enterprises: a review of the literature

Internationalization of R&D and innovation by Multinational Enterprises (MNEs) has undergone a gradual and comprehensive change in perspective over the past 50 years. From sporadic works in the late 1950s and in the 1960s, it became a systematically analysed topic in the 1970s, starting with pioneering reports and “foundation texts”. Our review unfolds the theoretical and empirical evolution of the literature from dyadic interpretations of centralization versus decentralization of R&D by MNEs to more comprehensive frameworks, wherein established MNEs from Advanced Economies still play a pivotal role, but new players and places also emerge in the global generation and diffusion of knowledge. Hence views of R&D internationalization increasingly rely on concepts, ideas and methods from IB and other related disciplines such as industrial organization, international economics and economic geography. Two main findings are highlighted. First, scholarly research pays an increasing attention to the network-like characteristics of international R&D activities. Second, different streams of literature have emphasized the role of location- specific factors in R&D internationalization. The increasing emphasis on these aspects has created new research opportunities in some key areas, including inter alia: cross-border knowledge sourcing strategies, changes in the geography of R&D and innovation, and the international fragmentation of production and R&D activities

Preparative LC/MS Technology: A Key Component of the Existing High Speed Synthesis Platform at Syngenta

Sizable investments in laboratory automation and state-of-the-art purification equipment were made at Syngenta Crop Protection with the objective to increase synthesis capacity and effectively support typical lead optimization activities. The successful implementation of a high-speed-synthesis platform consisted of the modular assembly of various commercially available liquid-handling workstations in combination with in-house developed parallel synthesis reaction manifolds. Preparative LC/MS technology was identified as a key success factor for optimizing overall throughput by addressing existing bottlenecks in parallel synthesis such as the fast and unattended isolation of compounds. Automated data analysis is crucial for high-throughput quality control and to allow all compounds of interprets to be sorted rapidly, i.e. those having defined structures and passing predefined purity criteria. The integrated application of the tools as described is intended to support chemists in their task to derive reliable structure–activity relationships more quickly and hopefully to shorten the time needed to identify innovative drug candidates

Stability measurements of antisense oligonucleotides by capillary gel electrophoresis.

The approach of using antisense oligonucleotides as potential drugs is based on hybridization of a short chemically-modified oligonucleotide with complementary cellular DNA or RNA sequences. A critical question is the stability of chemically modified antisense oligonucleotides in cellular environments. In a model system, resistance against various nucleases was evaluated by capillary gel electrophoresis (CGE). For some of the samples, matrix assisted laser desorption and ionization mass spectrometry (MALDI-MS) was used as an additional analytical tool to perform stability measurements. Using CGE, the enzymatic degradation of single nucleotides from the oligomer can be followed after different incubation times. 10% T polyacrylamide gels give baseline resolution for oligonucleotides ranging between 5 and 30 bases in length. The kinetic influence of a specific nuclease concentration and the antisense oligonucleotide structure on the cleavage reaction are discussed. Also, a simple desalting method to improve the injection efficiency and sensitivity of the method are described. Examples of measurements of chemically modified antisense 19-mers are presented