Alcohol production from cheese whey permeate using genetically modified flocculent yeast cells

Alcoholic fermentation of cheese whey permeate was investigated using a recombinant flocculating Saccharomyces cerevisiae, expressing the LAC4 (coding for β-galactosidase) and LAC12 (coding for lactose permease) genes of Kluyveromyces marxianus enabling for lactose metabolization. Data on yeast fermentation and growth on cheese whey permeate from a Portuguese dairy industry is presented. For cheese whey permeate having a lactose concentration of 50 gLˉ¹, total lactose consumption was observed with a conversion yield of ethanol close to the expected theoretical value. Using a continuously operating 5.5-L bioreactor, ethanol productivity near 10 g Lˉ¹ hˉ¹ (corresponding to 0.45 hˉ¹ dilution rate) was obtained, which raises new perspectives for the economic feasibility of whey alcoholic fermentation. The use of 2-times concentrated cheese whey permeate, corresponding to 100 gLˉ¹ of lactose concentration, was also considered allowing for obtaining a fermentation product with 5% (w/v) alcohol.Fundação para a Ciência e a Tecnologia (FCT) – PRAXIS XXI/BD/11306/97.Instituto de Biotecnologia e Química Fina (IBQF)

Industrial bees: the impact of apicultural intensification on local disease 1 prevalence

This is the author accepted manuscript. The final version is available from Wiley via the DOI in this record.1) It is generally thought that the intensification of farming will result in higher disease prevalences, although there is little specific modelling testing this idea. Focussing on honeybees, we build multi18 colony models to inform how ‘apicultural intensification’ is predicted to impact honeybee pathogen epidemiology at the apiary scale. 2) We used both agent-based and analytical models to show that three linked aspects of apicultural intensification (increased population sizes, changes in population network structure, and increased between-colony transmission) are unlikely to greatly increase disease prevalence in apiaries. Principally this is because even low-intensity apiculture exhibits high disease prevalence. 3) The greatest impacts of apicultural intensification are found for diseases with relatively low R0 (basic reproduction number), however, such diseases cause little overall disease prevalence and therefore the impacts of intensification are minor. Furthermore, the smallest impacts of intensification are for diseases with high R0 values, which we argue are typical of important honeybee diseases. 4) Policy Implications: Our findings contradict the idea that apicultural intensification by crowding honeybee colonies in large, dense apiaries leads to notably higher disease prevalences for established honeybee pathogens. More broadly, our work demonstrates the need for informative models of all agricultural systems and management practices in order to understand the implications of management changes on diseasesBiotechnology & Biological Sciences Research Council (BBSRC

Immune-mediated competition in rodent malaria is most likely caused by induced changes in innate immune clearance of merozoites

Malarial infections are often genetically diverse, leading to competitive interactions between parasites. A quantitative understanding of the competition between strains is essential to understand a wide range of issues, including the evolution of virulence and drug resistance. In this study, we use dynamical-model based Bayesian inference to investigate the cause of competitive suppression of an avirulent clone of Plasmodium chabaudi (AS) by a virulent clone (AJ) in immuno-deficient and competent mice. We test whether competitive suppression is caused by clone-specific differences in one or more of the following processes: adaptive immune clearance of merozoites and parasitised red blood cells (RBCs), background loss of merozoites and parasitised RBCs, RBC age preference, RBC infection rate, burst size, and within-RBC interference. These processes were parameterised in dynamical mathematical models and fitted to experimental data. We found that just one parameter μ, the ratio of background loss rate of merozoites to invasion rate of mature RBCs, needed to be clone-specific to predict the data. Interestingly, μ was found to be the same for both clones in single-clone infections, but different between the clones in mixed infections. The size of this difference was largest in immuno-competent mice and smallest in immuno-deficient mice. This explains why competitive suppression was alleviated in immuno-deficient mice. We found that competitive suppression acts early in infection, even before the day of peak parasitaemia. These results lead us to argue that the innate immune response clearing merozoites is the most likely, but not necessarily the only, mediator of competitive interactions between virulent and avirulent clones. Moreover, in mixed infections we predict there to be an interaction between the clones and the innate immune response which induces changes in the strength of its clearance of merozoites. What this interaction is unknown, but future refinement of the model, challenged with other datasets, may lead to its discovery

A single-column model intercomparison of a heavily drizzling stratocumulus-topped boundary layer

This study presents an intercomparison of single-column model simulations of a nocturnal heavily drizzling marine stratocumulus-topped boundary layer. Initial conditions and forcings are based on nocturnal flight observations off the coast of California during the DYCOMS-II field experiment. Differences in turbulent and microphysical parameterizations between models were isolated by slightly idealizing and standardizing the specification of surface and radiative fluxes. For most participating models, the case was run at both typical operational vertical resolution of about 100 m and also at high vertical resolution of about 10 m. As in prior stratocumulus intercomparisons, the simulations quickly develop considerable scatter in liquid water path (LWP) between models. However, the simulated dependence of cloud base drizzle fluxes on LWP in most models is broadly consistent with recent observations. Sensitivity tests with drizzle turned off show that drizzle substantially decreases LWP for many models. The sensitivity of entrainment rate to drizzle is more muted. Simulated LWP and entrainment are also sensitive to the inclusion of cloud droplet sedimentation. Many models underestimate the fraction of drizzle that evaporates below cloud base, which may distort the simulated feedbacks of drizzle on turbulence, entrainment, and LWP

Supercritical CO2 Extraction as a Tool to Isolate Anti-Inflammatory Sesquiterpene Lactones from Cichorium intybus L. Roots

Funding: This work was conducted under the “CHIC” project (H2020-NMBP-BIO-2017) with financial support received from the EU Horizon 2020 research and innovation programme under grant agreement N. 760891.Cichorium intybus L. or chicory plants are a natural source of health-promoting compounds in the form of supplements such as inulin, as well as other bioactive compounds such as sesquiterpene lactones (SLs). After inulin extraction, chicory roots are considered waste, with most SLs not being harnessed. We developed and optimized a new strategy for SL extraction that can contribute to the conversion of chicory root waste into valuable products to be used in human health-promoting applications. In our work, rich fractions of SLs were recovered from chicory roots using supercritical CO2. A response surface methodology was used to optimize the process parameters (pressure, temperature, flow rate, and co-solvent percentage) for the extraction performance. The best operating conditions were achieved at 350 bar, 40 °C, and 10% EtOH as a co-solvent in a 15 g/min flow rate for 120 min. The extraction with supercritical CO2 revealed to be more selective for the SLs than the conventional solid-liquid extraction with ethyl acetate. In our work, 1.68% mass and a 0.09% sesquiterpenes yield extraction were obtained, including the recovery of two sesquiterpene lactones (8-deoxylactucin and 11β,13-dihydro-8-deoxylactucin), which, to the best of our knowledge, are not commercially available. A mixture of the abovementioned compounds were tested at different concentrations for their toxic profile and anti-inflammatory potential towards a human calcineurin/NFAT orthologue pathway in a yeast model, the calcineurin/Crz1 pathway. The SFE extract obtained, rich in SLs, yielded results of inhibition of 61.74 ± 6.87% with 50 µg/mL, and the purified fraction containing 8-deoxylactucin and 11β,13-dihydro-8-deoxylactucin inhibited the activation of the reporter gene up to 53.38 ± 3.9% at 10 µg/mL. The potential activity of the purified fraction was also validated by the ability to inhibit Crz1 nuclear translocation and accumulation. These results reveal a possible exploitable green technology to recover potential anti-inflammatory compounds from chicory roots waste after inulin extraction.publishersversionpublishe

Formulation of the Dutch Atmospheric Large-Eddy Simulation (DALES) and Overview of Its Applications

The current version of the Dutch Atmospheric Large-Eddy Simulation (DALES) is presented. DALES is a large-eddy simulation code designed for studies of the physics of the atmospheric boundary layer, including convective and stable boundary layers as well as cloudy boundary layers. In addition, DALES can be used for studies of more specific cases, such as flow over sloping or heterogeneous terrain, and dispersion of inert and chemically active species. This paper contains an extensive description of the physical and numerical formulation of the code, and gives an overview of its applications and accomplishments in recent years

Identification of compounds responsible for the anthelmintic effects of chicory (Cichorium intybus) by molecular networking and bio-guided fractionation

10 páginas, 7 figuras.Increasing resistance towards anthelmintic drugs has necessitated the search for alternative treatments for the control of gastrointestinal nematode parasites. Animals fed on chicory (Cichorium intybus L.), a temperate (pasture) crop, have reduced parasite burdens, hence making C. intybus a potentially useful source for novel anthelmintic compounds or a diet-based preventive/therapeutic option. Here, we utilized in vitro bioassays with the parasitic nematode Ascaris suum and molecular networking techniques with five chicory cultivars to identify putative active compounds. Network analysis predicted sesquiterpene lactones (SL) as the most likely group of anthelmintic compounds. Further bioassay-guided fractionation supported these predictions, and isolation of pure compounds demonstrated that the SL 8-deoxylactucin (8-DOL) is the compound most strongly associated with anti-parasitic activity. Furthermore, we showed that 8-DOL acts in a synergistic combination with other SL to exert the anti-parasitic effects. Finally, we established that chicory-derived extracts also showed activity against two ruminant nematodes (Teladorsagia circumcincta and Cooperia oncophora) in in vitro assays. Collectively, our results confirm the anti-parasitic activity of chicory against a range of nematodes, and pave the way for targeted extraction of active compounds or selective breeding of specific cultivars to optimize its future use in human and veterinary medicine.We are very grateful for the guidance and support by Dr. Thomas Ostenfeld Larsen and Christopher Phippen, Technical University of Denmark, Natural product discovery, and the laboratory assistance of Mette Schjelde, University of Copenhagen. This work was funded by the Danish Council for Independent Research (Grant DFF–6111-00394). Fractionation and purification of compounds were further supported by the Green Development and Demonstration Program (GUDP) (Project No. 34009-17-1220). MPE was supported by CONICYT Chile (FONDE-CYT Postdoctorado #317087

Evaluation of large-Eddy simulations via observations of nocturnal marine stratocumulus

Data from the first research flight (RF01) of the second Dynamics and Chemistry of Marine Stratocumulus (DYCOMS-II) field study are used to evaluate the fidelity with which large-eddy simulations (LESs) can represent the turbulent structure of stratocumulus-topped boundary layers. The initial data and forcings for this case placed it in an interesting part of parameter space, near the boundary where cloud-top mixing is thought to render the cloud layer unstable on the one hand, or tending toward a decoupled structure on the other hand. The basis of this evaluation consists of sixteen 4-h simulations from 10 modeling centers over grids whose vertical spacing was 5 m at the cloud-top interface and whose horizontal spacing was 35 m. Extensive sensitivity studies of both the configuration of the case and the numerical setup also enhanced the analysis. Overall it was found that (i) if efforts are made to reduce spurious mixing at cloud top, either by refining the vertical grid or limiting the effects of the subgrid model in this region, then the observed turbulent and thermodynamic structure of the layer can be reproduced with some fidelity; (ii) the base, or native configuration of most simulations greatly overestimated mixing at cloud top, tending toward a decoupled layer in which cloud liquid water path and turbulent intensities were grossly underestimated; (iii) the sensitivity of the simulations to the representation of mixing at cloud top is, to a certain extent, amplified by particulars of this case. Overall the results suggest that the use of LESs to map out the behavior of the stratocumulus-topped boundary layer in this interesting region of parameter space requires a more compelling representation of processes at cloud top. In the absence of significant leaps in the understanding of subgrid-scale (SGS) physics, such a representation can only be achieved by a significant refinement in resolution—a refinement that, while conceivable given existing resources, is probably still beyond the reach of most centers

Tailoring and evaluating treatment with the Patient-Specific Needs Evaluation:A Patient-Centered Approach

BACKGROUND: No patient-reported instrument assesses patient-specific information needs, treatment goals, and Personal Meaningful Gain (PMG, a novel construct evaluating individualized, clinically relevant improvement). This study reports the development of the Patient-Specific Needs Evaluation (PSN) and examines its discriminative validity (i.e., its ability to distinguish satisfied from dissatisfied patients) and test-retest reliability in patients with hand or wrist conditions.METHODS: A mixed-methods approach was used to develop and validate the PSN, following COSMIN guidelines, including pilot testing, a survey (pilot: n=223, final PSN: n=275), cognitive debriefing (n=16), expert input, and validation. Discriminative validity was assessed by comparing the satisfaction level of patients who did or did not achieve their PMG (n=1,985) and test-retest reliability using absolute agreement, Cohen's kappa, and ICCs (n=102). We used a sample of 2,860 patients to describe responses to the final PSN.RESULTS: The PSN has only five questions (completion time ±3 minutes) and is freely accessible online. The items and response options were considered understandable by 90-92% and complete by 84-89% of the end-users. The PSN had excellent discriminative validity (Cramer's V: 0.48, p&lt;0.001) and moderate to high test-retest reliability (Kappa: 0.46-0.68, ICCs: 0.53-0.73).CONCLUSIONS: The PSN is a freely available patient-centered decision-support tool that helps clinicians tailor their consultations to the patient's individual needs and goals. It contains the PMG, a novel construct evaluating individualized, clinically relevant treatment outcomes. The PSN may function as a conversation starter, facilitate expectation management, and aid shared decision-making. The PSN is implementation-ready and can be readily adapted to other patient populations.LEVEL OF EVIDENCE: I.</p