Transcriptome Analysis of Aspergillus flavus Reveals veA-Dependent Regulation of Secondary Metabolite Gene Clusters, Including the Novel Aflavarin Cluster.

The global regulatory veA gene governs development and secondary metabolism in numerous fungal species, including Aspergillus flavus. This is especially relevant since A. flavus infects crops of agricultural importance worldwide, contaminating them with potent mycotoxins. The most well-known are aflatoxins, which are cytotoxic and carcinogenic polyketide compounds. The production of aflatoxins and the expression of genes implicated in the production of these mycotoxins are veA dependent. The genes responsible for the synthesis of aflatoxins are clustered, a signature common for genes involved in fungal secondary metabolism. Studies of the A. flavus genome revealed many gene clusters possibly connected to the synthesis of secondary metabolites. Many of these metabolites are still unknown, or the association between a known metabolite and a particular gene cluster has not yet been established. In the present transcriptome study, we show that veA is necessary for the expression of a large number of genes. Twenty-eight out of the predicted 56 secondary metabolite gene clusters include at least one gene that is differentially expressed depending on presence or absence of veA. One of the clusters under the influence of veA is cluster 39. The absence of veA results in a downregulation of the five genes found within this cluster. Interestingly, our results indicate that the cluster is expressed mainly in sclerotia. Chemical analysis of sclerotial extracts revealed that cluster 39 is responsible for the production of aflavarin.

The exaggerated inflammatory response in Behçet's syndrome: identification of dysfunctional post-transcriptional regulation of the IFN-γ/CXCL10 IP-10 pathway.

The mechanisms underlying the exaggerated inflammatory response in Behçet's syndrome (BS) remain poorly understood. We investigated the response of CD14(+) blood monocytes to interferon (IFN)-γ, focusing on the chemokine CXCL10. Chemokine synthesis and release were analysed at a protein and mRNA level following stimulation with IFN-γ. Findings in BS patients were compared with 25 healthy controls (HC), 15 rheumatoid arthritis (RA) and 15 systemic lupus erythematosus (SLE) disease control patients. BS monocytes produced significantly more CXCL10 protein than HC monocytes from 2 h following IFN-γ stimulation, despite equivalent quantities of mRNA, suggesting more efficient translation. This was significantly more pronounced in BS with high disease activity and in those with ocular and neurological clinical manifestations. The imbalance between CXCL10 protein and mRNA expression was not observed in either RA or SLE patients, and was not seen with other chemokines studied (CXCL9, CXCL11 and CCL2). Furthermore, BS monocytes treated with an alternative stimulant (LPS) did not show abnormal tumour necrosis factor (TNF)-α release. Sucrose density gradients to segregate monocyte CXCL10 mRNA into free RNA or polysome-associated RNA showed equal proportions in BS and HC samples, suggesting that the difference between BS and HC may be due to reduced negative control of CXCL10 translation in BS at a post-initiation level. We conclude that BS monocytes have dysfunctional post-transcriptional regulation of CXCL10 mRNA, resulting in over-expression of CXCL10 protein upon IFN-γ stimulation. As CXCL10 is a chemokine that recruits mononuclear cells, this abnormality may contribute to the exaggerated inflammatory responses that characterizes BS.

One step bioconversion of waste precious metals into Serratia biofilm-immobilized catalyst for Cr(VI) reduction.

OBJECTIVES: For reduction of Cr(VI) the Pd-catalyst is excellent but costly. The objectives were to prove the robustness of a Serratia biofilm as a support for biogenic Pd-nanoparticles and to fabricate effective catalyst from precious metal waste. RESULTS: Nanoparticles (NPs) of palladium were immobilized on polyurethane reticulated foam and polypropylene supports via adhesive biofilm of a Serratia sp. The biofilm adhesion and cohesion strength were unaffected by palladization and catalytic biofilm integrity was also shown by magnetic resonance imaging. Biofilm-Pd and mixed precious metals on biofilm (biofilm-PM) reduced 5 mM Cr(VI) to Cr(III) when immobilized in a flow-through column reactor, at respective flow rates of 9 and 6 ml/h. The lower activity of the latter was attributed to fewer, larger, metal deposits on the bacteria. Activity was lost in each case at pH 7 but was restored by washing with 5 mM citrate solution or by exposure of columns to solution at pH 2, suggesting fouling by Cr(III) hydroxide product at neutral pH. CONCLUSION: A 'one pot' conversion of precious metal waste into new catalyst for waste decontamination was shown in a continuous flow system based on the use of Serratia biofilm to manufacture and support catalytic Pd-nanoparticles.

Interfacial tension measurements using MRI drop shape analysis.

Accurate interfacial tension data for fluid systems such as hydrocarbons and water is essential to many applications such as reservoir oil and gas recovery predictions. Conventional interfacial tension measurement techniques typically use optical images to analyze droplet shapes but require that the continuous-phase fluid be optically transparent and that the fluids are not refractive index matched. Magnetic resonance images obtain contrast between fluids using other mechanisms such as magnetic relaxation weighting, so systems that are impossible to measure with optical methods may be analyzed. In this article, we present high-field (9.4 T) MRI images of various droplets analyzed with axisymmetric drop shape analysis. The resultant interfacial tension data show good agreement with literature data. The method is subsequently demonstrated using both opaque continuous phases and refractive-index-matched fluids. We conclude with a brief consideration of the potential to extrapolate the methodology to lower magnetic fields (0.3 T), featuring more accessible hardware; although droplet imaging is possible, resolution and stability do not currently permit accurate interfacial tension measurements.

Attention Mobilization as a Modulator of Listening Effort: Evidence From Pupillometry.

Listening to speech in noise can require substantial mental effort, even among younger normal-hearing adults. The task-evoked pupil response (TEPR) has been shown to track the increased effort exerted to recognize words or sentences in increasing noise. However, few studies have examined the trajectory of listening effort across longer, more natural, stretches of speech, or the extent to which expectations about upcoming listening difficulty modulate the TEPR. Seventeen younger normal-hearing adults listened to 60-s-long audiobook passages, repeated three times in a row, at two different signal-to-noise ratios (SNRs) while pupil size was recorded. There was a significant interaction between SNR, repetition, and baseline pupil size on sustained listening effort. At lower baseline pupil sizes, potentially reflecting lower attention mobilization, TEPRs were more sustained in the harder SNR condition, particularly when attention mobilization remained low by the third presentation. At intermediate baseline pupil sizes, differences between conditions were largely absent, suggesting these listeners had optimally mobilized their attention for both SNRs. Lastly, at higher baseline pupil sizes, potentially reflecting overmobilization of attention, the effect of SNR was initially reversed for the second and third presentations: participants initially appeared to disengage in the harder SNR condition, resulting in reduced TEPRs that recovered in the second half of the story. Together, these findings suggest that the unfolding of listening effort over time depends critically on the extent to which individuals have successfully mobilized their attention in anticipation of difficult listening conditions.

A multicenter, randomized phase 2 study to establish combinations of CBP501, cisplatin and nivolumab for ≥3rd-line treatment of patients with advanced pancreatic adenocarcinoma.

BACKGROUND: There is no standard of care for ≥ 3rd-line treatment of metastatic pancreatic adenocarcinoma (PDAC). CBP501 is a novel calmodulin-binding peptide that has been shown to enhance the influx of platinum agents into tumor cells and tumor immunogenicity. This study aimed to (1) confirm efficacy of CBP501/cisplatin/nivolumab for metastatic PDAC observed in a previous phase 1 study, (2) identify combinations that yield 35% 3-month progression-free survival rate (3MPFS) and (3) define the contribution of CBP501 to the effects of combination therapy. METHODS: CBP501 16 or 25 mg/m2 (CBP(16) or CBP(25)) was combined with 60 mg/m2 cisplatin (CDDP) and 240 mg nivolumab (nivo), administered at 3-week intervals. Patients were randomized 1:1:1:1 to (1) CBP(25)/CDDP/nivo, (2) CBP(16)/CDDP/nivo, (3) CBP(25)/CDDP and (4) CDDP/nivo, with randomization stratified by ECOG PS and liver metastases. A Fleming two-stage design was used, yielding a one-sided type I error rate of 2.5% and 80% power when the true 3MPFS is 35%. RESULTS: Among 36 patients, 3MPFS was 44.4% in arms 1 and 2, 11.1% in arm 3% and 33.3% in arm 4. Two patients achieved a partial response in arm 1 (ORR 22.2%; none in other arms). Median PFS and OS were 2.4, 2.1, 1.5 and 1.5 months and 6.3, 5.3, 3.7 and 4.9 months, respectively. Overall, all treatment combinations were well tolerated. Most treatment-related adverse events were grade 1-2. CONCLUSIONS: The combination CBP(25)/(16)/CDDP/nivo demonstrated promising signs of efficacy and a manageable safety profile for the treatment of advanced PDAC. CLINICAL TRIAL REGISTRATION: NCT04953962.

Microplastics fouling and interaction with polymeric membranes: A review.

The emergence and accumulation of microplastics (MPs) in various aquatic environments have recently raised significant concerns. Wastewater treatment plants (WWTPs) have been identified as one of the major sources of MPs discharge to the environment, implying a substantial need to improve advanced techniques for more efficient removal of MPs. Polymeric membranes have been proven effective in MPs removal. However, fouling is the main drawback of membrane processes and MPs can foul the membranes due to their small size and specific surface properties. Hence, it is important to investigate the impacts of MPs on membrane fouling to develop efficient membrane-based techniques for MPs removal. Although membrane technologies have a high potential for MPs removal, the interaction of MPs with membranes and their fouling effects have not been critically reviewed. The purpose of this paper is to provide a state-of-the-art review of MPs interaction with membranes and facilitate a better understanding of the relevant limitations and prospects of the membrane technologies. The first section of this paper is dedicated to a review of recent studies on MPs occurrence in WWTPs aiming to determine the most frequent MPs. This is followed by a summary of recent studies on MPs removal using membranes and discussions on the impact of MPs on membrane fouling and other probable issues (abrasion, concentration polarisation, biofouling, etc.). Finally, some recommendations for further research in this area are highlighted. This study serves as a valuable reference for future research on the development of anti-fouling membranes considering these new emerging contaminates.

Using non-invasive magnetic resonance imaging (MRI) to assess the reduction of Cr(VI) using a biofilm-palladium catalyst.

Industrial waste streams may contain contaminants that are valuable like Pd(II) and/or toxic and mutagenic like Cr(VI). Using Serratia sp. biofilm the former was biomineralized to produce a supported nanocrystalline Pd(0) catalyst, and this biofilm-Pd heterogeneous catalyst was then used to reduce Cr(VI) to less dangerous Cr(III) at room temperature, with formate as the electron donor. Cr(VI)((aq)) is non-paramagnetic while Cr(III)((aq)) is paramagnetic, which enabled spatial mapping of Cr species concentrations within the reactor cell using non-invasive magnetic resonance (MR) imaging experiments. Spatial reactivity heterogeneities were thus examined. In batch reactions, these could be attributed primarily to heterogeneity of Pd(0) distribution and to the development of gas bubbles within the reactor. In continuous flow reactions, spatial reactivity heterogeneities resulted primarily from heterogeneity of Cr(VI) delivery.

Towards optimum permeability reduction in porous media using biofilm growth simulations.

While biological clogging of porous systems can be problematic in numerous processes (e.g., microbial enhanced oil recovery-MEOR), it is targeted during bio-barrier formation to control sub-surface pollution plumes in ground water. In this simulation study, constant pressure drop (CPD) and constant volumetric flow rate (CVF) operational modes for nutrient provision for biofilm growth in a porous system are considered with respect to optimum (minimum energy requirement for nutrient provision) permeability reduction for bio-barrier applications. Biofilm growth is simulated using a Lattice-Boltzmann (LB) simulation platform complemented with an individual-based biofilm model (IbM). A biomass detachment technique has been included using a fast marching level set (FMLS) method that models the propagation of the biofilm-liquid interface with a speed proportional to the adjacent velocity shear field. The porous medium permeability reduction is simulated for both operational modes using a range of biofilm strengths. For stronger biofilms, less biomass deposition and energy input are required to reduce the system permeability during CPD operation, whereas CVF is more efficient at reducing the permeability of systems containing weaker biofilms.

Biofouling of spiral-wound nanofiltration and reverse osmosis membranes: a feed spacer problem.

Biofouling was studied in full-scale and pilot-scale installations, test-rigs and membrane fouling monitors by conventional methods as well as Magnetic Resonance Imaging (MRI). Independent of permeate production, the feed spacer channel pressure drop and biomass concentration increased similarly in a nanofiltration pilot installation. In the presence of a feed spacer the absolute feed channel pressure drop increase caused by biomass accumulation was much higher than when a feed spacer was absent: in both spiral-wound nanofiltration and reverse osmosis systems biofouling is dominantly a feed spacer problem. This conclusion is based on (i) in-situ visual observations of the fouling accumulation, (ii) in-situ non-destructive observations of the fouling accumulation and velocity distribution profiles using MRI, and (iii) differences in pressure drop and biomass development in monitors with and without feed spacer. MRI studies showed that even a restricted biofilm accumulation on the feed channel spacer influenced the velocity distribution profile strongly. Biofouling control should be focused on the development of low fouling feed spacers and hydrodynamic conditions to restrict the impact of biomass accumulation on the feed channel pressure drop increase.

Does the association between substance use and sexual risk behaviors among high school students vary by sexual identity?

OBJECTIVE: Limited information exists on whether associations between substance use behaviors (SUBs) and sexual risk behaviors (SRBs) vary by sexual identity. METHODS: Data from the 2015 national Youth Risk Behavior Survey (n = 15,624), were analyzed to assess associations between SUBs (cigarette smoking, alcohol use, binge drinking, marijuana use, prescription drug misuse, injection drug use, illicit drug use) and SRBs (sexual activity, number of partners, condom use). Logistic regression models calculated adjusted prevalence ratios (aPR), stratified by sexual identity, and interaction effects for sexual identity were introduced to models to determine if associations varied by sexual identity. RESULTS: All SUBs had significant associations with current sexual activity and 4+ sexual partners for both heterosexual and LGB students. No condom use during last sexual intercourse was significantly associated with all SUBs except alcohol use among heterosexual students, while no condom use was only significantly associated with injection drug use among LGB students. Associations between current sexual activity and SUBs were significantly stronger among heterosexual compared to LGB students for smoking (aPR = 2.39;95% CI:2.15,2.65 vs aPR = 1.49;95% CI:1.14,1.95), marijuana use (2.41;2.15,2.71 vs 1.86;1.58,2.19) and prescription drug misuse (2.10;1.93,2.28 vs 1.60;1.28,2.00). Associations between no condom use and SUBs were significantly stronger for heterosexual compared to LGB students only for smoking (1.32;1.16,1.50 vs 0.96;0.73,1.25) and marijuana use (1.22;1.07,1.38 vs 0.90;0.72,1.12). CONCLUSIONS: The relationship between most SUBs and SRBs did not vary significantly by sexual identity. These findings underscore the importance coordinating school-based programs to prevent substance use and promote sexual health.

Carbohydrate binding modules enhance cellulose enzymatic hydrolysis by increasing access of cellulases to the substrate.

Plant biomass is a low-cost and abundant source of carbohydrates for production of fuels, "green" chemicals and materials. Currently, biochemical conversion of the biomass into sugars via enzymatic hydrolysis is the most viable technology. Here, the role of carbohydrate binding modules (CBMs) in the disruption of insoluble polysaccharide structures and their capacity to enhance cellulase-promoted lignocellulosic biomass hydrolysis was investigated. We show that CBM addition promotes generation of additional reducing ends in the insoluble substrate by cellulases. On the contrary, bovine serum albumin (BSA), widely used in prevention of a non-specific protein binding, causes an increase in soluble reducing-end production, when applied jointly with cellulases. We demonstrate that binding of CBMs to cellulose is non-homogeneous, irreversible and leads to its amorphisation. Our results also reveal effects of CBM-promoted amorphogenesis on cellulose hydrolysis by cellulases.

Non-invasive mass transfer measurements in complex biofilm-coated structures.

We demonstrate a novel application of (13)C pulsed field gradient (PFG) NMR to monitor mass transfer, due to both flow and diffusion, in a 3D complex porous support structure modified by biofilm growth. This enables timescales an order of magnitude larger than previously possible to be accessed with respect to displacement probability distribution (propagator) measurements. The evolution in the propagator shape with observation time to the Gaussian asymptote (constant dispersion coefficient) is consequently well resolved. We also simulated the measured displacement propagators with good agreement between experiment and prediction. The methodology has significant potential for the selective characterization of the transport of nutrients, metabolic products, pollutants and biocides in such complex biofilm-containing structures.

Spatially resolved quantification of metal ion concentration in a biofilm-mediated ion exchanger.

A bioremediation process to remove Co(2+) from aqueous solution is investigated in this study using a magnetic resonance imaging (MRI) protocol to rapidly obtain multiple 2D spatially resolved Co(2+) ion concentration maps. The MRI technique is described in detail and its ability to determine the evolution in both axial and radial concentration profiles demonstrated, from which total column capacity can be determined. The final ion exchange column design allows operation in the 'plug flow' regime, hence making use of its full capacity before breakthrough. Conventional techniques for such process optimization are either restricted to the analysis of the exchanger outlet, which provides no information on the spatial heterogeneity of the system, or are invasive and need a variety of sample points to obtain 1D concentration information. To the best of our knowledge, our results represent the first concentration maps describing the bioremediation of metal ion contaminated water.

A rapid measurement of flow propagators in porous rocks.

NMR flow propagators have been obtained for brine flowing through Bentheimer sandstone using the rapid DiffTrain pulse sequence. In this way, 8 flow propagators at different observation times Delta were acquired in 67 mins, compared to 7 h for the same measurements implemented with conventional pulsed field gradient (PFG) sequences. DiffTrain allows this time saving to be achieved through the acquisition of multiple displacement probability distributions over a range of Delta in a single measurement. If only the propagator moments are required, this experiment time can be further reduced to 9 mins through appropriate sparse sampling at low q values. The propagator moments obtained from DiffTrain measurements with dense and sparse q-space sampling are shown to be equivalent to those obtained from conventional PFG measurements.

Changes in the proteome of Escherichia coli during growth at 15 degrees C after incubation at 2, 6 or 8 degrees C for 4 days.

For better understanding of the complex behaviour of Escherichia coli at chiller temperatures, log phase E. coli grown at 15 degrees C were incubated at 8, 6, or 2 degrees C for 4 days, and were then incubated at 15 degrees C for 12 h. Cultures were sampled after incubation at the lower temperatures, and during subsequent incubation at 15 degrees C. Proteins extracted from the samples were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Spots of 45 previously identified proteins that were differentially expressed at 15 or < or =8 degrees C were quantified by image analysis. After incubation at 8 or 6 degrees C for 4 days cells were growing with or without formation of elongated cells (filaments), respectively, but growth did not occur at 2 degrees C. In cells incubated at 8 or 6 degrees C proteins associated with the stress response and energy generation were upregulated and proteins associated with protein synthesis were downregulated, while protein levels in cells incubated at 2 degrees C were little changed. When cells were then incubated at 15 degrees C, the levels of differentially expressed proteins in cells that had been incubated at 8 or 6 degrees C decreased or increased towards the levels found in cells growing at 15 degrees C, but some proteins were still under or over expressed after 12 h. In cells incubated at 15 degrees C after incubation at 2 degrees C, the levels of many of the proteins declined but the levels of proteins associated with protein synthesis increased. The findings indicate that the physiological states of log phase E. coli incubated at < or =2 degrees C or at higher chiller temperature are different, but that for both states incubation at an above chiller temperature for >3 generations is required before protein levels adjusted to those usual for the higher temperature. Cells in these different physiological states may respond differently to other stresses encountered during warming of chilled foods.

A resistive Q-switch for low-field NMR systems.

An NMR Q-switch was designed and constructed specifically for use with low-field NMR apparatus. This featured a comparatively simple resistive damping design. It served to reduce the r.f. probe ring-down time, and hence reduced the signal acquisition delay from 25 ms to 9 ms, on an Earth's magnetic field NMR system. The advantage of this earlier acquisition was demonstrated for both an aqueous suspension of iron oxide particles and using an NMR flow meter.

Intrinsic dispersivity of randomly packed monodisperse spheres.

We determine the intrinsic longitudinal dispersivity l(d) of randomly packed monodisperse spheres by separating the intrinsic stochastic dispersivity l(d) from dispersion by unavoidable sample dependent flow heterogeneities. The measured l(d), scaled by the hydrodynamic radius r(h), coincide with theoretical predictions [Saffman, J. Fluid Mech. 7, 194 (1960)] for dispersion in an isotropic random network of identical capillaries of length l and radius a, for l/a=3.82, and with rescaled simulation results [Maier et al., Phys. Fluids 12, 2065 (2000)].

Evolving micro-structures in drying detergent pastes quantified using NMR.

Magnetic resonance methods have been used to probe the evolution of the internal micro-structure of an industrially important detergent mixture, as a function of total water content. Measurements of the apparent diffusion coefficient of the water content were obtained as a function of diffusion observation time. These data were interpreted to render the surface-to-volume ratio and tortuosity of the pore space in which the water resided. Pore dimensions were found to decrease as moisture content was reduced, partially as a consequence of solute deposition. Deposition of solute material was confirmed through the application of NMR T(1) relaxation measurements. These were analysed using regularisation techniques to yield T(1) population distributions. Average pore sizes as a function of water content were extracted from this data and were in good agreement with the results of the diffusion analysis.

Modelling biofilm-modified hydrodynamics in 3D.

A simulation-based study to predict the impact of biofilm growth on displacement distributions for flow of water through a supporting packed bed is presented. The lattice Boltzmann method and a directed random walk algorithm are used, and are applied to the system with and without biofilm being present. The aim of this simulation study is to model the anomalous transport dynamics induced by biofilm, as reported in the literature, and thus to study the impact of observation time, delta, on the shape of the displacement distributions (propagators). We believe that this is the first demonstration of a propagator simulation for flow through a complex porous structure modulated by biofilm growth. The propagator distributions undergo a transition from a pre-asymptotic to a Gaussian-shaped distribution with increasing delta. The propagators were simulated for a wide range of delta going up to 500 seconds. This transition occurs with and without biofilm, but is very significantly delayed when biofilm is present due to the consequential development of essentially stagnant regions. The transition can be classified into three stages: a diffusion-dominated stage, a "twin-peak" stage and an advection-dominated stage.