Scaling-up the production of thermostable lipolytic enzymes from Thermus aquaticus YT1

The lipolytic enzymes synthesized by Thermusaquaticus YT1 present extremely interesting properties of thermostability (more than 70% of activity after 12 days at 80°C and a half-life time of 1 h at 95°C), which point out the interest of proposing efficient strategies to successfully tackle the scale-up of the production process. In this study,viable scaling-up of the production process was implemented,and relevant aspects affecting the enzyme synthesis, such as the mineral composition of the culture medium, the aeration and the agitation have been evaluated.A strategy combining the modification of the culture medium and the aeration degree was also approached by adding perfluorocarbons, compounds which improve the availability of oxygen in the culture medium. An opposite response of biomass and lipolytic activity to the aeration conditions was found between scales (about 600 U L(-1) at high aeration levels in flask vs. 150 U L(-1) at high aeration rates in reactor), which further demonstrates the important role of the hydrodynamic conditions on the suitable development of the biological process. In all cases, the cultures were kinetically characterized and the Luedeking and Piret model turned out to be a valuable tool to conclude that the produced lipolytic enzyme is a growth-associated metabolite, no matter the medium and the scale.     

Seasonal carbon dynamics and water fluxes in an Amazon rainforest

Satellite‐based observations indicate that seasonal patterns in canopy greenness and productivity in the Amazon are negatively correlated with precipitation, with increased greenness occurring during the dry months. Flux tower measurements indicate that the canopy greening that occurs during the dry season is associated with increases in net ecosystem productivity (NEP) and evapotranspiration (ET). Land surface and terrestrial biosphere model simulations for the region have predicted the opposite of these observed patterns, with significant declines in greenness, NEP, and ET during the dry season. In this study, we address this issue mainly by developing an empirically constrained, light‐controlled phenology submodel within the Ecosystem Demography model version 2 (ED2). The constrained ED2 model with a suite of field observations shows markedly improved predictions of seasonal ecosystem dynamics, more accurately capturing the observed patterns of seasonality in water, carbon, and litter fluxes seen at the Tapajos National Forest, Brazil (2.86°S, 54.96°W). Long‐term simulations indicate that this light‐controlled phenology increases the resilience of Amazon forest NEP to interannual variability in climate forcing.     

Heart Rate Monitoring in Soccer: Interest and Limits During Competitive Match Play and Training, Practical Application

ABSTRACT: Alexandre, D, Da Silva, C, Hill-Haas, S, Wong, DP, Natali, AJ, De Lima, JRP, Filho, MGB, Marins, JCB, Garcia, ES, and Chamari, K. Heart rate monitoring in soccer: Interest and limits during competitive match play and training-Practical application. J Strength Cond Res 26(10): 2890-2906, 2012-The identification of physiological loads imposed by soccer training or match play reveals essential information, which may help improve training and recovery strategies. Until today, the use of heart rate (HR) monitoring is not standardized in soccer. Thus, the aim of this review was to analyze, determine and compare the exercise intensity (EI) monitored by HR in professional, youth, and recreational soccer players during matches and training sessions using a meta-analysis. Heart rate is one of the most common physiological variables used to determine exercise internal training load. The mean EI recorded during competitive matches was described as 70-80% of V[Combining Dot Above]O2max or 80-90% of maximal heart rate (HRmax), independent of the playing level. With respect to HR training zones, approximately 65% of the total match duration is spent at intensity of 70-90% HRmax and rarely below 65% HRmax. However, although HRmax is mostly employed in the literature, monitoring EI should be expressed in relation to reserve heart rate, as it was described as a more reliable indicator of HR, allowing interindividual comparisons. The HR response according to the playing position indicates that midfielders are characterized by the highest EI, followed by forwards and fullbacks. Moreover, in the second half of the match, the EI is lower than that observed during the first half; this reduction could be correlated with the level of the player's physical conditioning. Consequently, coaches may favor the use of interval training or small-sided training games because these are shown to improve both aerobic capacity and the ability to repeat high-intensity actions. Small-sided games allow reaching similar HR responses to those found during interval training and match play but with greater heterogeneity values. Future investigations should include a larger sample of players with special reference to playing position and the expression of EI in percentage of the reserve heart rate, analyzing the possible intergender differences in HR response.     

Examination of the Cytotoxic and Embryotoxic Potential and Underlying Mechanisms of Next-Generation Synthetic Trioxolane and Tetraoxane Antimalarials

Semisynthetic artemisinin-based therapies are the first-line treatment for P. falciparum malaria, but next-generation synthetic drug candidates are urgently required to improve availability and respond to the emergence of artemisinin-resistant parasites. Artemisinins are embryotoxic in animal models and induce apoptosis in sensitive mammalian cells. Understanding the cytotoxic propensities of antimalarial drug candidates is crucial to their successful development and utilization. Here, we demonstrate that, similarly to the model artemisinin artesunate (ARS), a synthetic tetraoxane drug candidate (RKA182) and a trioxolane equivalent (FBEG100) induce embryotoxicity and depletion of primitive erythroblasts in a rodent model. We also show that RKA182, FBEG100 and ARS are cytotoxic toward a panel of established and primary human cell lines, with caspase-dependent apoptosis and caspase-independent necrosis underlying the induction of cell death. Although the toxic effects of RKA182 and FBEG100 proceed more rapidly and are relatively less cell-selective than that of ARS, all three compounds are shown to be dependent upon heme, iron and oxidative stress for their ability to induce cell death. However, in contrast to previously studied artemisinins, the toxicity of RKA182 and FBEG100 is shown to be independent of general chemical decomposition. Although tetraoxanes and trioxolanes have shown promise as next-generation antimalarials, the data described here indicate that adverse effects associated with artemisinins, including embryotoxicity, cannot be ruled out with these novel compounds, and a full understanding of their toxicological actions will be central to the continuing design and development of safe and effective drug candidates which could prove important in the fight against malaria.     

Visitor influence on the behavior of captive greater rheas (Rhea americana, Rheidae Aves)

Visitors can affect and reduce the welfare of nonhuman animals. The Belo Horizonte Zoo, Brazil, had a group of greater rheas intended for reintroduction to the wild. Because this group received public visitation, evaluating its effect on the birds' behavior and welfare was important. The study conducted 60 hr of behavioral observations: 30 with, and 30 without, visitors in front of the birds' enclosure. The study, conducted April-December 2009, collected data using scan sampling with instantaneous recording of behavior every minute. The study collected data on public behavior, visitor density, and time spent observing the birds. More than 4,000 persons visited the birds' enclosure: 9.86 s average time spent. Public behaviors most expressed were walking-watching-talking and stopped-watching-talking; visitors or not, greater rheas' most expressed behaviors (inversed between treatments) were foraging and walking alert; defecating/urinating and other behaviors differed statistically between treatments. Walking alert was most expressed in the presence of visitors; defecating/urinating and other behaviors were most expressed in their absence. Greater rheas seemed to habituate to visitors. Birds' behaviors differed little in visitors' presence or absence.     

Screening for Intellectual Disability Using High-Resolution CMA Technology in a Retrospective Cohort from Central Brazil

Intellectual disability is a complex, variable, and heterogeneous disorder, representing a disabling condition diagnosed worldwide, and the etiologies are multiple and highly heterogeneous. Microscopic chromosomal abnormalities and well-characterized genetic conditions are the most common causes of intellectual disability. Chromosomal Microarray Analysis analyses have made it possible to identify putatively pathogenic copy number variation that could explain the molecular etiology of intellectual disability. The aim of the current study was to identify possible submicroscopic genomic alterations using a high-density chromosomal microarray in a retrospective cohort of patients with otherwise undiagnosable intellectual disabilities referred by doctors from the public health system in Central Brazil. The CytoScan HD technology was used to detect changes in the genome copy number variation of patients who had intellectual disability and a normal karyotype. The analysis detected 18 CNVs in 60% of patients. Pathogenic CNVs represented about 22%, so it was possible to propose the etiology of intellectual disability for these patients. Likely pathogenic and unknown clinical significance CNVs represented 28% and 50%, respectively. Inherited and de novo CNVs were equally distributed. We report the nature of CNVs in patients from Central Brazil, representing a population not yet screened by microarray technologies.