Search

An anticipatory stress response develops before an internal or external stimulus, which initiates a homeostasis process through a chain of responses that enable human organisms to face different threats, thus allowing them to adapt to a continuous and eliciting environment. In the current research, we analyzed the psychophysiological anticipatory anxiety response of professional soldiers prior to a real mission in an actual theater of operation. Autonomic modulation through the heart rate variability values, muscular strength manifestation, and psychological stress of 53 military personnel of Army Airmobile Forces (age: M = 35.4 years, SD = 5.88 years; height: M = 1.75 m, SD = 6.87 cm; body mass: M = 77.33 kg, SD = 11.95 kg; military duty = 14.44 years, SD = 6.43; military operation experience = 4 months, SD = 4.25 months) and a control group of 33 civil participants were analyzed. The military personnel presented significant differences in some HRV values related to the activation of sympathetic systems. We found that the military personnel presented an anticipatory anxiety response only at an autonomic level, showing an increased sympathetic modulation, but not at a psychological level, since their anxiety levels were not significantly different than those of the control civilians. In addition, this anticipatory anxiety response did not affect muscular strength manifestation, as it presented no significant differences between the military personnel and the control group.

In recent decades, enzyme-based biocatalytic systems have garnered increasing interest in industrial and applied research for catalysis and organic chemistry. Many enzymatic reactions have been applied to sustainable and environmentally friendly production processes, particularly in the pharmaceutical, fine chemicals, and flavor/fragrance industries. However, only a fraction of the enzymes available has been stepped up towards industrial-scale manufacturing due to low enzyme stability and challenging separation, recovery, and reusability. In this context, immobilization and co-immobilization in robust support materials have emerged as valuable strategies to overcome these inadequacies by facilitating repeated or continuous batch operations and downstream processes. To further reduce separations, it can be advantageous to use multiple enzymes at once in one pot. Enzyme co-immobilization enables biocatalytic synergism and reusability, boosting process efficiency and cost-effectiveness. Several studies on multi-enzyme immobilization and co-localization propose kinetic advantages of the enhanced turnover number for multiple enzymes. This review spotlights recent progress in developing versatile biocatalytic cascade systems by multi-enzyme co-immobilization on environmentally friendly biopolymers and nanostructured materials and their application scope in the chemical and biotechnological industries. After a succinct overview of carrier-based and carrier-free immobilization/co-immobilizations, co-immobilization of enzymes on a range of biopolymer and nanomaterials-based supports is thoroughly compiled with contemporary and state-of-the-art examples. This study provides a new horizon in developing effective and innovative multi-enzymatic systems with new possibilities to fully harness the adventure of biocatalytic systems.

Droplet microfluidics enables the ultrahigh-throughput screening of the natural or man-made genetic diversity for industrial enzymes, with reduced reagent consumption and lower costs than conventional robotic alternatives. Here we describe an example of metagenomic screening for nucleoside 2'-deoxyribosyl transferases using FACS as a more widespread and accessible alternative than microfluidic on-chip sorters. This protocol can be easily adapted to directed evolution libraries by replacing the library construction steps and to other enzyme activities, e.g., oxidases, by replacing the proposed coupled assay.

Droplet microfluidics enables the ultrahigh-throughput screening of the natural or man-made genetic diversity for industrial enzymes, with reduced reagent consumption and lower costs than conventional robotic alternatives. Here we describe an example of metagenomic screening for nucleoside 2'-deoxyribosyl transferases using FACS as a more widespread and accessible alternative than microfluidic on-chip sorters. This protocol can be easily adapted to directed evolution libraries by replacing the library construction steps and to other enzyme activities, e.g., oxidases, by replacing the proposed coupled assay.

The present work describes the functional and structural characterization of adenine phosphoribosyltransferase 2 from thermus thermophilus hb8 (ttaprt2). The combination of structural and substrate specificity data provided valuable information for immobilization studies. Dimeric ttaprt2 was immobilized onto glutaraldehydeactivated magresyn®amine magnetic iron oxide porous microparticles by two different strategies: a) an enzyme immobilization at ph 8.5 to encourage the immobilization process by n-termini (mttaprt2a, mttaprt2b, mttaprt2c) or b) an enzyme immobilization at ph 10.0 to encourage the immobilization process through surface exposed lysine residues (mttaprt2d, mttaprt2e, mttaprt2f). According to catalyst load experiments, mttaprt2b (activity: 480 iu g−1 biocatalyst, activity recovery: 52%) and mttaprt2f (activity: 507 iu g−1 biocatalyst, activity recovery: 44%) were chosen as optimal derivatives. The biochemical characterization studies demonstrated that immobilization process improved the thermostability of ttaprt2. Moreover, the potential reusability of mttaprt2b and mttaprt2f was also tested. Finally, mttaprt2f was employed in the synthesis of nucleoside-5′-monophosphate analogues. El presente trabajo describe la caracterización funcional y estructural de la adenina fosforribosiltransferasa 2 de thermus thermophilus hb8 (ttaprt2). La combinación de datos de especificidad estructural y de sustrato proporcionó información valiosa para los estudios de inmovilización. El ttaprt2 dimérico se inmovilizó en micropartículas porosas de óxido de hierro magnético magresyn®amine activado con glutaraldehido mediante dos estrategias diferentes: a) una inmovilización de enzima a ph 8.5 para alentar el proceso de inmovilización por n-termini (mttaprt2a, mttaprt2c) o bttaprt2c) o b) ph 10.0 para fomentar el proceso de inmovilización a través de residuos de lisina expuestos en la superficie (mttaprt2d, mttaprt2e, mttaprt2f). De acuerdo con los experimentos de carga de catalizador, mttaprt2b (actividad: 480 iu g − 1 biocatalizador, recuperación de actividad: 52%) y mttaprt2f (actividad: 507 iu g − 1 biocatalizador, recuperación de actividad: 44%) fueron elegidos como derivados óptimos. Los estudios de caracterización bioquímica demostraron que el proceso de inmovilización mejoró la termoestabilidad de ttaprt2. Además, también se probó la reutilización potencial de mttaprt2b y mttaprt2f. Finalmente, mttaprt2f se empleó en la síntesis de análogos de nucleósido-5'-monofosfato.

Insight into the catalytic mechanism of Lactobacillus leichmannii nucleoside 2′-deoxyribosyltransferase (LlNDT) has been gained by calculating a quantum mechanics-molecular mechanics (QM/MM) free-energy landscape of the reaction within the enzyme active site. Our results support an oxocarbenium species as the reaction intermediate and thus an S1 reaction mechanism in this family of bacterial enzymes. Our mechanistic proposal is validated by comparing experimental kinetic data on the impact of the single amino acid replacements Tyr7, Glu98 and Met125 with Ala, Asp and Ala/norLeu, respectively, and accounts for the specificity shown by this enzyme on a non-natural substrate. This work broadens our understanding of enzymatic C-N bond cleavage and C-N bond formation., Financial support from the FWF (Lise Meitner Program M 2260) to PAS-M, the Spanish Ministerio de Economía y Competitividad (SAF2015-64629-C2-2-R) to F. G. and Grants XSAN001906 from the Santander Foundation to J. F. L. is gratefully acknowledged. We thank the University of Vienna for computational resources at the Institute of Theoretical Chemistry.

Present research studied the psychophysiological response and shooting performance taking into consideration the experience of the soldier after a physical stress protocol, which leaded to acute fatigue conditions. Sixty soldiers were divided into experienced and non-experienced groups to conduct a high-intensity interval training (HIIT) based on endurance type exercises (10 intervals of 40 m run at maximum intensity with 2 changes of direction and 45 s of recovery. Psychophysiological and shooting performance variables were measured before, during and after the HIIT protocol. Significant increases in heart rate, blood lactate, lower limbs strength, as well as a significant decrease in blood oxygen saturation and body temperature were found in both groups. Shooting performance was affected by experience and fatigue, being more accentuated in non-experienced. Present HIIT protocol elicited a similar psychophysiological response than in previously reported combat manoeuvres, thus proving to be an optimal stimuli approach/intervention for soldier's specific physical preparation.

Fenneropenaeus merguiensis (commonly named banana shrimp) is one of the most important farmed crustacean worldwide species for the fisheries and aquaculture industry. Besides its nutritional value, it is a good source of chitinase, an enzyme with excellent biological and catalytic properties for many industrial applications. In the present study, a putative chitinase-encoding cDNA was synthesized from mRNA from F. merguiensis hepatopancreas tissue. Subsequently, the corresponding cDNA was cloned, sequenced and functionally expressed in Escherichia coli, and the recombinant F. merguiensis chitinase (rFmCHI) was purified by His-tag affinity chromatography. The bioinformatics analysis of aminoacid sequence of rFmCHI displayed a cannonical multidomain architecture in chitinases which belongs to glycoside hydrolase family 18 (GH18 chitinase). Biochemical characterization revealed rFmCHI as a monomeric enzyme of molecular weight 52 kDa with maximum activity at 40 °C and pH 6.0 Moreover, the recombinant enzyme is also stable up to 60 °C, and in the pH range 5.0-8.0. Steady-state kinetic studies for colloidal chitin revealed KM, Vmax and kcat values of 78.18 μM, 0.07261 μM. min−1 and 43.37 s−1, respectively. Overall, our results aim to demonstrate the potential of rFmCHI as suitable catalyst for bioconversion of chitin waste.

Sin financiación 11.889 Q1 JCR 2021 2.354 Q1 SJR 2021 No data IDR 2021 UEM

The “manufacture and modification” of molecules in nature are catalyzed by enzymes with exquisite selectivity, unparalleled rate acceleration and under “mild” reaction conditions. Nowadays, enzyme-catalyzed synthesis is an attractive and eco-friendly alternative to the traditional multi-step and contaminating chemical processes.

The use of magnetic biocatalysts is highly beneficial in bioprocesses technology, as it allows their easy recovering and enhances biocatalyst lifetime. Thus, it simplifies operational processing and increases efficiency, leading to more cost-effective processes. The use of small-size matrices as carriers for enzyme immobilization enables to maximize surface area and catalysts loading, also reducing diffusion limitations. As highly expensive nanoparticles (nm size) usually aggregate, their application at large scale is not recommended. In contrast, the use of magnetic micro-macro (µm-mm size) matrices leads to more homogeneous biocatalysts with null or very low aggregation, which facilitates an easy handling and recovery. The present review aims to highlight recent trends in the application of medium-to-high size magnetic biocatalysts in different areas (biodiesel production, food and pharma industries, protein purification or removal of environmental contaminants). The advantages and disadvantages of these above-mentioned magnetic biocatalysts in bioprocess technology will be also discussed. Santander Foundation (XSAN001906) 11.889 Q1 JCR 2021 2.354 Q1 SJR 2021 No data IDR 2021 UEM

There is a lack of information on the psychophysiological response of pilots under hypoxic conditions. The study of the physiological, psychological, cardiorespiratory, neurological, behavioural, sensory, and cognitive symptoms that may appear during training in hypobaric chambers is essential to optimize the training processes of aircrew members. Thus, the present study is aimed at analyzing the psychophysiological responses of aircrew members in an incremental hypoxia training protocol. Psychophysiological responses of 44 aircrew members (34 males and 10 females) in an incremental hypoxia training protocol (3 minutes at 0 meters, 8 minutes at 5,000 meters, and maximum time at 7500 meters) were measured. Results suggested that the incremental hypoxia training protocol did not affect cortical arousal and handgrip strength; however, it increased the sympathetic tone, perceived stress, perceived effort, and heart rate and decreased forced expiratory volume and blood oxygen saturation. Thus, we concluded that acute hypoxic hypobaric exposure leads to decreased parasympathetic tone, blood oxygen saturation, and maximal spirometry values, without negatively affecting handgrip strength and cortical arousal. This information will lead to find specific training systems that meet the real needs of aircrew. Project 2020/UEM43 3.246 Q3 JCR 2021 0.647 Q2 SJR 2021 No data IDR 2021 UEM

13 págs., 6 figs., 3 tabs., Nucleoside 2′-deoxyribosyltransferases (NDTs) catalyze the cleavage of glycosidic bonds of 2′-deoxynucleosides and the following transfer of the 2′-deoxyribose moiety to acceptor nucleobases. Here, we report the crystal structures and biochemical properties of the first tetrameric NDTs: the type I NDT from the mesophilic bacterium Enterococcus faecalis V583 (EfPDT) and the type II NDT from the bacterium Desulfotalea psychrophila (DpNDT), the first psychrophilic NDT. This novel structural and biochemical data permitted an exhaustive comparative analysis aimed to shed light into the basis of the high global stability of the psychrophilic DpNDT, which has a higher melting temperature than EfPDT (58.5 °C versus 54.4 °C) or other mesophilic NDTs. DpNDT possesses a combination of unusual structural motifs not present neither in EfPDT nor any other NDT that most probably contribute to its global stability, in particular, a large aliphatic isoleucine-leucine-valine (ILV) bundle accompanied by a vicinal disulfide bridge and also an intersubunit disulfide bridge, the first described for an NDT. The functional and structural features of DpNDT do not fit the standard features of psychrophilic enzymes, which lead us to consider the implication of (sub)cellular levels together with the protein level in the adaptation of enzymatic activity to low temperatures., This work was supported by grants CTQ2009-11543 (IDLM), PID2020-117025RB-I00 (JF-L) and AGL2017-84614-C2-2-R (JMM) from the Spanish Ministry of Science and Innovation, S2009/PPQ-1752 (IDLM) from Comunidad Autonoma de Madrid and XSAN192006 (IDLM) from the Santander Foundation. This research was funded in part by National Institutes of Health grant (GM115586 to AJ). The use of SBC beamlines at the Advanced Photon Source is supported by the U.S. Department of Energy (DOE) Office of Science and operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357 (AJ).

Due to their versatility and ability to provide tunable chemo-, regio-, and enantio-selectivitites, enzyme-mediated transformations offer an effective and eco-friendly alternative to the traditional multistep chemical synthesis methods, which often require organic solvents, hazardous reagents, and tedious purification processes. While enzymes provide high catalytic and specific activity at mild conditions, several operational drawbacks hamper the application of enzymes as industrial biocatalysts. These include poor performance when confronting non-natural substrates, inhibition by certain components in the reaction media, short lifetime, lack of reusability, and recovery postreaction. Among these, achieving enzyme stabilization is most challenging from a practical standpoint.

Nowadays, pharmaceutical industry demands competitive and eco-friendly processes for active pharmaceutical ingredients (APIs) manufacturing. In this context, enzyme and whole-cell mediated processes offer an efficient, sustainable and cost-effective alternative to the traditional multi-step and environmentally-harmful chemical processes. Particularly, 2'-deoxyribosyltransferases (NDTs) have emerged as a novel synthetic alternative, not only to chemical but also to other enzyme-mediated synthetic processes. This review describes recent findings in the development and scaling up of NDTs as industrial biocatalysts, including the most relevant and recent examples of single enzymatic steps, multienzyme cascades, chemo-enzymatic approaches, and engineered biocatalysts. Finally, to reflect the inventive and innovative steps of NDT-mediated bioprocesses, a detailed analysis of recently granted patents, with specific focus on industrial synthesis of nucleoside-based APIs, is hereunder presented. Sin financiación 17.681 Q1 JCR 2021 2.482 Q1 SJR 2021 No data IDR 2021 UEM

Many Gram-negative bacteria produce N-acyl-homoserine lactones (AHLs), quorum sensing (QS) molecules that can be enzymatically inactivated by quorum quenching (QQ) processes, this approach is considered an emerging antimicrobial alternative. In this study, kinetic parameters of several AHLs hydrolyzed by penicillin acylase from Streptomyces lavendulae (SlPA) and aculeacin A acylase from Actinoplanes utahensis (AuAAC) have been determined. Both enzymes catalyze efficiently the amide bond hydrolysis in AHLs with different acyl chain moieties (with or without 3-oxo modification) and exhibit a clear preference for AHLs with long acyl chains (C12-HSL &gt, C14-HSL &gt, C10-HSL &gt, C8-HSL for SlPA, whereas C14-HSL &gt, C12-HSL &gt, C8-HSL for AuAAC). Involvement of SlPA and AuAAC in QQ processes was demonstrated by Chromobacterium violaceum CV026-based bioassays and inhibition of biofilm formation by Pseudomonas aeruginosa, a process controlled by QS molecules, suggesting the application of these multifunctional enzymes as quorum quenching agents, this being the first time that quorum quenching activity was shown by an aculeacin A acylase. In addition, a phylogenetic study suggests that SlPA and AuAAC could be part of a new family of actinomycete acylases, with a preference for substrates with long aliphatic acyl chains, and likely involved in QQ processes.

Nowadays enzymatic synthesis of nucleic acid derivatives is gaining momentum over traditional chemical synthetic processes. Biotransformations catalyzed by whole cells or enzymes offer an ecofriendly and efficient alternative to the traditional multistep chemical methods, avoiding the use of chemical reagents and organic solvents that are expensive and environmentally harmful. Herein we report for the first time the covalent immobilization a uracil phosphoribosyltransferase (UPRT). In this sense, UPRT from Thermus thermophilus HB8 was immobilized onto glutaraldehyde-activated MagReSyn®Amine magnetic iron oxide porous microparticles (MTtUPRT). According to the catalyst load experiments, MTtUPRT3 was selected as optimal biocatalyst for further studies. MTtUPRT3 was active and stable in a broad range of temperature (70–100 °C) and in the pH interval 6–8, displaying maximum activity at 100 °C and pH 7 (activity 968 IU/gsupport, retained activity 100%). In addition, MTtUPRT3 could be reused up to 8 times in the synthesis of uridine-5′-monophosphate (UMP). Finally, MTtUPRT3 was successfully applied in the sustainable synthesis of different 5-modified uridine-5′-monophosphates at short times. Taking into account these results, MTtUPRT3 would emerge as a valuable biocatalyst for the synthesis of nucleoside monophosphates through an efficient and environmentally friendly methodology. Sin financiación 3.036 JCR (2020) Q2, 35/72 Biophysics 0.878 SJR (2020) Q1, 29/121 Analytical Chemistry No data IDR 2019 UEM

Chitinolytic enzymes are capable to catalyze the chitin hydrolysis. Due to their biomedical and biotechnological applications, nowadays chitinolytic enzymes have attracted worldwide attention. Chitinolytic enzymes have provided numerous useful materials in many different industries, such as food, pharmaceutical, cosmetic, or biomedical industry. Marine enzymes are commonly employed in industry because they display better operational properties than animal, plant, or bacterial homologs. In this mini-review, we want to describe marine chitinolytic enzymes as versatile enzymes in different biotechnological fields. In this regard, interesting comments about their biological role, reaction mechanism, production, functional characterization, immobilization, and biotechnological application are shown in this work. Sin financiación 3.670 JCR (2018) Q2, 41/162 Biotechnology & Applied Microbiology 1.127 SJR (2018) Q1, 53/342 Biotechnology, 22/114 Applied Microbiology and Biotechnology, 451/2844 Medicine (miscellaneous) No data IDR 2018 UEM

Background: In recent years, enzymatic methods have shown to be an efficient and sustainable alternative for the synthesis of nucleosides and nucleoside-5'-monophosphates (NMPs) to the traditional multistep chemical methods, since chemical glycosylation reactions include several protection-deprotection steps and the use of chemical reagents and organic solvents that are expensive and environmentally harmful. Results: In this mini-review, we want to illustrate the application of phosphoribosyltransferases (PRTs) in enzymatic synthesis of NMPs. In this sense, many different examples about the use of PRTs as biocatalysts, as whole cells or enzymes, are described. In addition, it also includes detailed comments about structure and catalytic mechanism of described PRTs, as well as their possible biological role and therapeutic use, substrate specificity and advances in detection of new enzyme specificities towards different substrates. In addition, several examples about the use of PRTs in mono or multi-enzymatic synthesis of NMP analogues are shown. Finally, a brief discussion about advantages and drawbacks of the use of PRTs as industrial biocatalyst of NMPs has been commented. Conclusion: Despite the great potential of PRTs as biocatalysts for industrial synthesis of NMPs, several drawbacks must be overcome before reaching a suitable industrial application. In this sense, multi-enzyme systems provide an appropriate framework for this purpose. Moreover, future advances in different disciplines as protein engineering, bioinformatics and -omics will help to reach this goal. Sin financiación 2.757 JCR (2017) Q2, 114/261 Pharmacology & Pharmacy 0.612 SJR (2017) Q2, 62/164 Drug Discovery No data IDR 2017 UEM

Nowadays, enzymatic synthesis of nucleotides is an efficient and sustainable alternative to chemical methodologies. In this regard, after the biochemical characterization of wild-type and mutant uracil phosphoribosyltransferases from Toxoplasma gondii (TgUPRT, TgUPRT2, and TgUPRT3), TgUPRT2 was selected as the optimal candidate (69.5 IU mg-1, UMP synthesis) for structure-guided immobilization onto Ni2+ chelate (MNiUPRT2) and onto glutaraldehyde-activated microparticles (MGlUPRT2). Among resulting derivatives, MNiUPRT23 (6127 IU g-1biocat; 92% retained activity; 3-5 fold enhanced stability at 50-60 °C) and MGlUPRT2N (3711 IU g-1biocat; 27% retained activity; 8-20 fold enhanced stability at 50-60 °C) displayed the best operability. Moreover, the enzymatic synthesis of different pyrimidine NMPs was performed. Finally, the reusability of both derivatives in 5-FUMP synthesis (MNiUPRT23, 80% retained activity after 7 cycles, 5 min; MGlUPRT2N, 70% retained activity after 10 cycles, 20 min) was carried out at short times. Santander Foundation (SAN151610) Universidad Europea de Madrid (2020/UEM42) National Institutes of Health (5R21MH116415) 11.889 Q1 JCR 2021 2.354 Q1 SJR 2021 No data IDR 2021 UEM

Purine phosphoribosyltransferases, purine PRTs, are essential enzymes in the purine salvage pathway of living organisms. They are involved in the formation of C-N glycosidic bonds in purine nucleosides-50-monophosphate (NMPs) through the transfer of the 5-phosphoribosyl group from 5-phospho-a-D-ribosyl-1-pyrophosphate (PRPP) to purine nucleobases in the presence of Mg2þ. Herein, we report a simple and thermostable process for the one-pot, one-step synthesis of some purine NMPs using xanthine phosphoribosyltransferase, XPRT or adenine phosphoribosyltransferase, APRT2, from Thermus thermophilus HB8. In this sense, the cloning, expression and purification of TtXPRT and TtAPRT2 is described for the first time. Both genes, xprt and aprt2 were expressed as his-tagged enzymes in E. coli BL21(DE3) and purified by a heat-shock treatment, followed by Ni-affinity chromatography and a final, polishing gel-filtration chromatography. Biochemical characterization revealed TtXPRT as a tetramer and TtAPRT2 as a dimer. In addition, both enzymes displayed a strong temperature dependence (relative activity >75% in a temperature range from 70 to 90 C), but they also showed very different behaviour under the influence of pH. While TtXPRT is active in a pH range from 5 to 7, TtAPRT2 has a high dependence of alkaline conditions, showing highest activity values in a pH range from 8 to 10. Finally, substrate specificity studies were performed in order to explore their potential as industrial biocatalyst for NMPs synthesis. European University of Madrid and Santander Foundation 1.627 JCR (2018) Q3, 121/162 Biotechnology & Applied Microbiology; Q4, 252/299 Biochemistry & Molecular Biology 0.299 SJR (2018) Q3, 184/342 Biotechnology; Q4, 347/462 Biochemistry, 44/60 Catalysis No data IDR 2018 UEM

We aimed to analyse the effect of experience level in the psychophysiological response and specific fine motor skills of novel and expert parachute warfighters during a tactical combat parachute jump. We analysed blood oxygen saturation, heart rate, salivary cortisol, blood glucose, lactate and creatinkinase, leg strength, isometric hand-grip strength, cortical arousal, specific fine motor skills and cognitive anxiety, somatic anxiety and self-confident before and after a tactical combat parachute jump in 40 warfighters divided in two group, novel (n = 17) and expert group (n = 23). Novels presented a higher heart rate, lactate, cognitive anxiety, somatic anxiety and a lower self-confident than experts during the jump. We concluded that experience level has a direct effect on the psychophysiological response since novel paratroopers presented a higher psychophysiological response than compared to the expert ones, however this result neither affected the specific fine motor skills nor the muscle structure after a tactical combat parachute jump. Sin financiación 2.582 JCR (2016) Q2, 34/84 Physiology, 25/77 Psychology; Q3, 145/259 Neurosciences 1.403 SJR (2016) Q2, 18/69 Neuropsychology and Physiological Psychology, 49/158 Neuroscience (miscellaneous), 33/107 Physiology (medical) No data IDR 2016 UEM

The present work aims to develop a magnetic biocatalyst for customized production of nucleoside analogues using mutant His-tagged purine 2'-deoxyribosyltransferase from Trypanosoma brucei (TbPDT

The use of nucleoside 2′-deoxyribosyltransferases (NDTs) as biocatalysts for the industrial synthesis of nucleoside analogues is often hindered by their strict preference for 2′-deoxyribonucleosides. It is shown herein that a highly versatile purine NDT from Trypanosoma brucei (TbPDT) can also accept ribonucleosides as substrates; this is most likely because of the distinct role played by Asn53 at a position that is usually occupied by Asp in other NDTs. Moreover, this unusual activity was improved about threefold by introducing a single amino acid replacement at position 5, following a structure-guided approach. Biophysical and biochemical characterization revealed that the TbPDT variant is a homodimer that displays maximum activity at 50 °C and pH 6.5 and shows a remarkably high melting temperature of 69 °C. Substrate specificity studies demonstrate that 6-oxopurine ribonucleosides are the best donors (inosine>guanosine≫adenosine), whereas no significant preferences exist between 6-aminopurines and 6-oxopurines as base acceptors. In contrast, no transferase activity could be detected on xanthine and 7-deazapurines. TbPDT was successfully employed in the synthesis of a wide range of modified ribonucleosides containing different purine analogues., This work was supported by grant XSAN001906 from the Santander Foundation (to J.F.L.) and grant SAF2015-64629-C2-2-R from the Spanish MEC/MICINN (to F.G.)

The psychophysiological response due to stress of soldiers in actual combat operations have been determined by recent researchers, but there is a lack of knowledge about the most effective training methodologies to prepare these population for these new psychophysiological and tactical requirements. The aim of this study was to analyze the effect of an operative high intensity interval training (HIIT) in the psychophysiological response and shooting performance of professional soldiers. We analyzed 20 soldiers of the Spanish Army which performed an operative HIIT composed by 3 series of 7 repetitions of exercises based on operative military procedures with 30 s of workload and 30 s of resting between repetitions and 5 min resting between series. Blood lactate, rate of perceived exertion and stress, upper and lower limbs, and respiratory muscle strength, skin temperature, blood oxygen saturation, heart rate, cortical arousal, short-term memory and anxiety response were measured before and after the training. After the training rated of perceived exertion and stress, leg strength, heart rate and lactate presented a significant increase and blood oxygen saturation and cortical arousal significantly decreased. An operative HIIT achieves similar psychophysical response than the evaluated in combat simulations in professional soldiers, producing a decrease in cortical arousal and lactate values over the anaerobic threshold. Investigadores recientes han determinado la respuesta psicofisiológica debida al estrés de los soldados en las operaciones de combate reales, pero existe una falta de conocimiento sobre las metodologías de entrenamiento más efectivas para preparar a esta población para estos nuevos requisitos psicofisiológicos y tácticos. El objetivo de este estudio fue analizar el efecto de un entrenamiento operativo de intervalo de alta intensidad (HIIT) en la respuesta psicofisiológica y el rendimiento de tiro de los soldados profesionales. Analizamos 20 soldados del Ejército español que realizaron un HIIT operativo compuesto por 3 series de 7 repeticiones de ejercicios basados ​​en procedimientos militares operativos con 30 s de carga de trabajo y 30 s de descanso entre repeticiones y 5 min de descanso entre series. El lactato sanguíneo, la tasa de esfuerzo percibido y el estrés, las extremidades superiores e inferiores y la fuerza muscular respiratoria, la temperatura de la piel, la saturación de oxígeno en la sangre, la frecuencia cardíaca, la activación cortical, la memoria a corto plazo y la respuesta de ansiedad se midieron antes y después del entrenamiento. Después del entrenamiento, se evaluó el esfuerzo y esfuerzo percibidos, la fuerza de las piernas, la frecuencia cardíaca y el lactato presentaron un aumento significativo y la saturación de oxígeno en la sangre y la excitación cortical disminuyeron significativamente.

The study of facial emotional expressions not only gives information about communicative and adaptive processes, but also provides valuable knowledge about some aspects of human behaviour, such as emotions and intentions. Taking into account the proved evidence that deficits in processing emotional information shows difficulties in executive functions and social behavior, the present work aims to compare the perceptual and dimensional similarities between the emotions transmitted by the faces (facial emotions) and those typically associated with two particular emotional contexts (offenders and non-offenders). To evaluate the contextual influence on emotional face categorization, the emotional recognition in 69 male young (35 offenders (17.22 ± 1.5 years) and 34 non-offenders (16.90 ± 1.56 years) from Barranquilla (Colombia) was analyzed. Experimental results displayed significant differences among the two groups, mainly focused in the recognition of anger and disgust faces. In this sense, the analysis of abovementioned data may lead to the development of more specific and cost-effective therapeutic treatments for offender population. Sin financiación 2.826 JCR (2019) Q2, 16/53 Behavioral Sciences 0.993 SJR (2019) Q2, 26/82 Behavioral Neuroscience No data IDR 2019 UEM

Nowadays, enzyme-mediated processes offer an eco-friendly and efficient alternative to the traditional multistep and environmentally harmful chemical processes. Herein we report the enzymatic synthesis of cladribine by a novel 20 -deoxyribosyltransferase (NDT)-based combined biocatalyst. To this end, Lactobacillus delbrueckii NDT (LdNDT) was successfully immobilized through a two-step immobilization methodology, including a covalent immobilization onto glutaraldehydeactivated biomimetic silica nanoparticles followed by biocatalyst entrapment in calcium alginate. The resulting immobilized derivative, SiGPEI 25000-LdNDT-Alg, displayed 98% retained activity and was shown to be active and stable in a broad range of pH (5–9) and temperature (30–60 ◦C), but also displayed an extremely high reusability (up to 2100 reuses without negligible loss of activity) in the enzymatic production of cladribine. Finally, as a proof of concept, SiGPEI 25000-LdNDT-Alg was successfully employed in the green production of cladribine at mg scale. Agencia Nacional de Pro-mociónCientífica y Tecnológica (PICT 2014–3438) Consejo Nacional de InvestigacionesCientíficas y Técnicas (PIP 2014- 11220130100805CO) Universidad Nacional de Quilmes (PUNQ 1305/19, PPROF 2017- EXPTE.895/18) Santander Foundation (XSAN192006) European University of Madrid (2020/UEM42) 6.064 Q2 JCR 2021 1.019 Q1 SJR 2021 No data IDR 2021 UEM

The actual COVID-19 pandemic scenario has generated a context of uncertainty, helplessness, and inequality. Yet, the perception of COVID-19 risk has influenced nutritional, psychological, and physical activity patterns depending on gender. We conducted the present research with the aim of studying gender differences of university students in the perceived risk of the COVID-19 pandemic, and in psychological, nutritional, oral health, and physical activity habits. To reach the study’s aim, 300 volunteer university students completed an online questionnaire which analyzed variables of perceived risk of the COVID-19 pandemic, psychological profiles, and nutritional, oral health, and physical activity habits. Results showed that females presented a higher perception of danger to the COVID-19 virus than males but showed no differences in how the pandemic has affected personal lives. Females showed higher values of anxiety, conscientiousness, neuroticism, and openness to experience, while males presented higher values of extraversion. Nutritionally, males presented greater consumption of soft drinks, meat, and pasta or rice, and lower buccal hygiene. Yet, no differences were found regarding physical activity patterns. Results from the present study could be used by various educational institutions to implement multidisciplinary interventions to reduce the stress and risk perception.

With the aim to explore cultural differences in stress-related psychological, nutrition, physical activity, and oral health factors between Spanish and Latin American professors, we analysed stress-related factors in 598 professors (39.9% male, 60.1% female, 41.3 &plusmn, 9.8 years) by a collection of questionnaires, which involved psychological, nutritional, physical activity and oral health items. Results showed how Spanish professors presented significantly (p &le, 0.05) higher scores than Latin American professors in perceived stress (Spanish: 21.40 &plusmn, 4.32 vs. Latin American: 20.36 &plusmn, 4.31), teaching stress (Spanish: 6.59 &plusmn, 2.28 vs. Latin American: 6.00 &plusmn, 2.99) and neuroticism (Spanish: 5.40 &plusmn, 2.10 vs. Latin American: 4.58 &plusmn, 1.72). Spanish professors also showed healthier nutritional and physical activity habits than their Latin American counterparts, presenting higher consumption of milk products and a higher numbers of meals per day, greater weekly meat and fish consumption and higher weekly resistance training, as well as less eating between hours and snacking consumption. Nevertheless, Spanish professors brushed their teeth less and showed a higher smoking habit than Latin American professors. We concluded that there were cultural differences between Spanish and Latin American professors. In the present research, Spanish professors showed significantly higher burnout levels, teaching stress, perceived stress, and neuroticism than Latin American professors, and several differences were also found around health behaviours. These differences in perceived stress, teaching stress and burnout syndrome may be due to the habituation process of Latin American professors, and probably are associated with a higher stressful and demanding socio-cultural context.

In this work, a mono- and a bi-enzymatic analytical immobilized enzyme reactors (IMERs) were developed as prototypes for biosynthetic purposes and their performances in the in-flow synthesis of nucleoside analogues of pharmaceutical interest were evaluated. Two biocatalytic routes based on nucleoside 2′-deoxyribosyltransferase from Lactobacillus reuteri (LrNDT) and uridine phosphorylase from Clostridium perfrigens (CpUP)/purine nucleoside phosphorylase from Aeromonas hydrophila (AhPNP) were investigated in the synthesis of 2′-deoxy, 2′,3′-dideoxy and arabinonucleoside derivatives. LrNDT-IMER catalyzed the synthesis of 5-fluoro-2′-deoxyuridine and 5-iodo-2′-deoxyuridine in 65–59% conversion yield, while CpUP/AhPNP-IMER provided the best results for the preparation of arabinosyladenine (60% conversion yield). Both IMERs proved to be promising alternatives to chemical routes for the synthesis of nucleoside analogues. The developed in-flow system represents a powerful tool for the fast production on analytical scale of nucleosides for preliminary biological tests. Sin financiación 9.642 JCR (2020) Q1, 1/14 Agricultural Engineering 2.489 SJR (2020) Q1, 13/168 Bioengineering No data IDR 2020 UEM