Gene cloning, molecular modeling, and phylogenetics of serine protease P32 and serine carboxypeptidase SCP1 from nematophagous fungi Pochonia rubescens and Pochonia chlamydosporia

The fungi Pochonia chlamydosporia and Pochonia rubescens are parasites of nematode eggs and thus are biocontrol agents of nematodes. Proteolytic enzymes such as the S8 proteases VCP1 and P32, secreted during the pathogenesis of nematode eggs, are major virulence factors in these fungi. Recently, expression of these enzymes and of SCP1, a new putative S10 carboxypeptidase, was detected during endophytic colonization of barley roots by these fungi. In our study, we cloned the genomic and mRNA sequences encoding P32 from P. rubescens and SCP1 from P. chlamydosporia. P32 showed a high homology with the serine proteases Pr1A from the entomopathogenic fungus Metarhizium anisopliae and VCP1 from P. chlamydosporia (86% and 76% identity, respectively). However, the catalytic pocket of P32 showed differences in the amino acids of the substrate-recognition sites compared with the catalytic pockets of Pr1A and VCP1 proteases. Phylogenetic analysis of P32 suggests a common ancestor with protease Pr1A. SCP1 displays the characteristic features of a member of the S10 family of serine proteases. Phylogenetic comparisons show that SCP1 and other carboxypeptidases from filamentous fungi have an origin different from that of yeast vacuolar serine carboxypeptidases. Understanding protease genes from nematophagous fungi is crucial for enhancing the biocontrol potential of these organisms.

Synthesis of Chromen[4,3-b]pyrrolidines by Intramolecular 1,3-Dipolar Cycloadditions of Azomethine Ylides: An Experimental and Computational Assessment of the Origin of Stereocontrol

Azomethine ylides, generated from imine-derived O-cinnamyl or O-crotonyl salicylaldeyde and α-amino acids, undergo intramolecular 1,3-dipolar cycloaddition, leading to chromene[4,3-b]pyrrolidines. Two reaction conditions are used: (a) microwave-assisted heating (200 W, 185 °C) of a neat mixture of reagents, and (b) conventional heating (170 °C) in PEG-400 as solvent. In both cases, a mixture of two epimers at the α-position of the nitrogen atom in the pyrrolidine nucleus was formed through the less energetic endo-approach (B/C ring fusion). In many cases, the formation of the stereoisomer bearing a trans-arrangement into the B/C ring fusion was observed in high proportions. Comprehensive computational and kinetic simulation studies are detailed. An analysis of the stability of transient 1,3-dipoles, followed by an assessment of the intramolecular pathways and kinetics are also reported.

Using multilevel modeling and mixed methods to make theoretical progress in microfoundations for strategy research

The microfoundations research agenda presents an expanded theoretical perspective because it considers individuals, their characteristics, and their interactions as relevant variables to help us understand firm-level strategic issues. However, microfoundations empirical research faces unique challenges because processes take place at different levels of analysis and these multilevel processes must be considered simultaneously. We describe multilevel modeling and mixed methods as methodological approaches whose use will allow for theoretical advancements. We describe key issues regarding the use of these two types of methods and, more importantly, discuss pressing substantive questions and topics that can be addressed with each of these methodological approaches with the goal of making theoretical advancements regarding the microfoundations research agenda and strategic management studies in general.

Fixation pattern analysis with microperimetry in nystagmus patients

Objective: To assess the usefulness of microperimetry (MP) as an additional objective method for characterizing the fixation pattern in nystagmus. Design: Prospective study. Participants: Fifteen eyes of 8 subjects (age, 12–80 years) with nystagmus from the Lluís Alcanyís Foundation (University of Valencia, Spain) were included. Methods: All patients had a comprehensive ophthalmologic examination including a microperimetric examination (MAIA, CenterVue, Padova, Italy). The following microperimetric parameters were evaluated: average threshold (AT), macular integrity index (MI), fixating points within a circle of 1° (P1) and 2° of radius (P2), bivariate contour ellipse area (BCEA) considering 63% and 95% of fixating points, and horizontal and vertical axes of that ellipse. Results: In monocular conditions, 6 eyes showed a fixation classified as stable, 6 eyes showed a relatively unstable fixation, and 3 eyes showed an unstable fixation. Statistically significant differences were found between the horizontal and vertical components of movement (p = 0.001), as well as in their ranges (p < 0.001). Intereye comparison showed differences between eyes in some subjects, but only statistically significant differences were found in the fixation coordinates X and Y (p < 0.001). No significant intereye differences were found between microperimetric parameters. Between monocular and binocular conditions, statistically significant differences in the X and Y coordinates were found in all eyes (p < 0.02) except one. No significant differences were found between MP parameters for monocular or binocular conditions. Strong correlations of corrected distance visual acuity (CDVA) with AT (r = 0.812, p = 0.014), MI (r = –0.812, p = 0.014), P1 (r = 0.729, p = 0.002), horizontal diameter of BCEA (r = –0.700, p = 0.004), and X range (r = –0.722, p = 0.005) were found. Conclusions: MP seems to be a useful technology for the characterization of the fixation pattern in nystagmus, which seems to be related to the level of visual acuity achieved by the patient.

Analysis of physical and physiological requirements in soccer trainings in young soccer players (under -10 years)

The aim of this study is to analyse the physical and physiological factors in soccer training at different categories of training. The participants were 30 soccer players of 8-aside soccer in the under 10’s age group (9.93±0.25 years) who participated in the under 10 Provincial Tournament in Alicante. During training, the variables of covered distance, heart rate, speed (average and maximum values) as well as the methodology used and position were registered. After the statistical analysis and its related discussion, it was concluded that the players do not show differences in the covered total distance in relation to the category. Notwithstanding, there are differences with regards to speed and heart rate, which are caused by the greater physical development of the players in comparison to the under10’s age group category. Regarding the methodology employed, it is worth stressing that the coaches used, to a greater extend, the global method, followed by the mixed method.

Phrase-based statistical machine translation: explanation of its processes and statistical models and evaluation of the English to Spanish translations produced

Statistical machine translation (SMT) is an approach to Machine Translation (MT) that uses statistical models whose parameter estimation is based on the analysis of existing human translations (contained in bilingual corpora). From a translation student’s standpoint, this dissertation aims to explain how a phrase-based SMT system works, to determine the role of the statistical models it uses in the translation process and to assess the quality of the translations provided that system is trained with in-domain goodquality corpora. To that end, a phrase-based SMT system based on Moses has been trained and subsequently used for the English to Spanish translation of two texts related in topic to the training data. Finally, the quality of this output texts produced by the system has been assessed through a quantitative evaluation carried out with three different automatic evaluation measures and a qualitative evaluation based on the Multidimensional Quality Metrics (MQM).