Biodegradation of phenol by free and encapsulated cells of a new Aspergillus sp. isolated from a contaminated site in southern Brazil

The aim of this study was to compare the biodegradation performance of phenol by using free and encapsulated cells of a new Aspergillus sp. strain isolated from a crude oil contaminated soil in southern Brazil. In batch cultures, maximum degradation rates were not significantly different between free and encapsulated cells, but a decrease in adaptation time for encapsulated ones was observed. This fact indicates the presence of a microenvironment that is more favorable to biodegradation inside encapsulated cells, because of the protector effect of gel matrix, which reduces abiotic stress. Encapsulated filamentous fungus Aspergillus sp. LEBM2 showed a promising application in bioaugmentation processes, reaching maximum phenol degradation rate of 7.71 ± 0.21 mg/l.h for an initial phenol concentration of 500 mg/l.Key words: Bioremediation, bioaugmentation, immobilization, phenol, filamentous fungi

Screening of yeasts capable of producing cellulase-free xylanase

Xylanases have largely been obtained from filamentous fungi and bacteria; few studies have investigated the production of this enzyme by yeasts. The aim of this study was to isolate yeasts from different sources, such as vegetables, cereal grains, fruits, and agro-industrial waste and to obtain yeasts capable of producing celulase-free xylanase. Samples were enriched using yeast malt broth, and yeasts were isolated on Wallerstein nutrient agar. In all, 119 yeast strains were isolated and evaluated in terms of their ability to degrade xylan, which was found in the medium by using agar degradation halos, the basis of this polysaccharide, and Congo red dye. Selected microorganisms were grown in complex medium and the enzymatic activities of endo-xylanase, β-xylosidase, carboxymetilcellulase, and filter paper cellulose were determined over 96 h of cultivation; the pH and biomass concentration were also evaluated. The yeast strain 18Y, which was isolated from chicory and later identified as Cryptococcus laurentii, showed the highest endo-xylanase activity (2.7 U.mL-1). This strain had the ability to produce xylanase with low levels of cellulase production (both CMCase [0.11 U.mL-1] and FPase [0.10 U.mL-1]). This result gives this strain great biotechnological potential since this enzyme can be used for industrial pulp and paper bleaching.Key words: Cryptococcus laurentii, endo-xylanase, xylan

Production of a rhamnolipid-type biosurfactant by Pseudomonas aeruginosa LBM10 grown on glycerol

The work herewith investigated the effect of the culture medium composition on rhamnolipid production by Pseudomonas aeruginosa LBM10, previously isolated from an estuarine environment in Southern Brazil. Experimental design and surface response methodology were used in order to improve biosurfactant production using glycerol, a renewable carbon source. The assays were carried out in a rotary shaker at 30°C and 180 rpm for 120 h and the parameters studied were glycerol concentration, C/N (carbon/nitrogen) and C/P (carbon/phosphorus) ratios. Low glycerol concentration and a phosphorus-limiting condition were favorable for rhamnolipid production. Contour plots constructed by predictive polynomial equations led to a glycerol concentration of 13.2 g/l, a C/N ratio of 12.8 and a C/P ratio of 40 in order to maximize rhamnolipid concentration (4.15 g/l) associated with a high emulsification index (61%).Keywords: Biosurfactant, surface-active compounds, experimental design, phosphorus limitatio

Different cell disruption methods for astaxanthin recovery by Phaffia rhodozyma

Astaxanthin (3,3'-dihydroxy-b,b'-carotene-4,4'-dione) is carotenoid of high market value whose demand has increased in such fields as aquaculture, pharmaceutical supplements and natural coloring. Cell disruption is the first step for isolating intracellular materials and it depends on the cell wall permeability. In order to maximize the  recovery of astaxanthin from Phaffia rhodozyma NRRL-Y17268, drying and freeze pretreatments were tested by different cell disruption methods: abrasion with celite, glass pearls in vortex agitator, ultrasonic waves, sodium  carbonate (Na2CO3) and dimethyl sulfoxide (DMSO). The method with Na2CO3 was not effective; meanwhile, the agitator with glass pearls, the abrasion with celite and the ultrasonic waves were found as promising for future  studies. As a result, the DMSO in freeze-dried biomass with 4 process cycles and biomass/DMSO relation of 0.025 g/ml was found to be the most efficient for analytical determination, increasing about up to 25 times the astaxanthin recovery.Key words: Carotenoids, yeast, chemical disruption, dimethyl sulfoxide

Pre-screening of filamentous fungi isolated from a contaminated site in Southern Brazil for bioaugmentation purposes

Four Aspergillus sp. strains were isolated from contaminated soil in Rio Grande, Southern Brazil. The biodegradation potential of these strains was evaluated using a simple method involving the determination of colony growth rates on plates containing a specific hydrocarbon or petroleumderivative as the only carbon source. The LEBM1 strain presented a high tolerance level to BTX. It was the only strain capable of growth on all the media, with growth rates varying from 1.3 to 2.2 mm/day. The LEBM2 strain presented the potential for phenol degradation, while the LEBM3 strain could be used for gasoline, diesel oil, hexane and chlorobenzene

A Mechanism for the Polarity Formation of Chemoreceptors at the Growth Cone Membrane for Gradient Amplification during Directional Sensing

Accurate response to external directional signals is essential for many physiological functions such as chemotaxis or axonal guidance. It relies on the detection and amplification of gradients of chemical cues, which, in eukaryotic cells, involves the asymmetric relocalization of signaling molecules. How molecular events coordinate to induce a polarity at the cell level remains however poorly understood, particularly for nerve chemotaxis. Here, we propose a model, inspired by single-molecule experiments, for the membrane dynamics of GABA chemoreceptors in nerve growth cones (GCs) during directional sensing. In our model, transient interactions between the receptors and the microtubules, coupled to GABA-induced signaling, provide a positive-feedback loop that leads to redistribution of the receptors towards the gradient source. Using numerical simulations with parameters derived from experiments, we find that the kinetics of polarization and the steady-state polarized distribution of GABA receptors are in remarkable agreement with experimental observations. Furthermore, we make predictions on the properties of the GC seen as a sensing, amplification and filtering module. In particular, the growth cone acts as a low-pass filter with a time constant ∼10 minutes determined by the Brownian diffusion of chemoreceptors in the membrane. This filtering makes the gradient amplification resistent to rapid fluctuations of the external signals, a beneficial feature to enhance the accuracy of neuronal wiring. Since the model is based on minimal assumptions on the receptor/cytoskeleton interactions, its validity extends to polarity formation beyond the case of GABA gradient sensing. Altogether, it constitutes an original positive-feedback mechanism by which cells can dynamically adapt their internal organization to external signals

A DNA Vaccine Encoding Multiple HIV CD4 Epitopes Elicits Vigorous Polyfunctional, Long-Lived CD4+ and CD8+ T Cell Responses

T-cell based vaccines against HIV have the goal of limiting both transmission and disease progression by inducing broad and functionally relevant T cell responses. Moreover, polyfunctional and long-lived specific memory T cells have been associated to vaccine-induced protection. CD4+ T cells are important for the generation and maintenance of functional CD8+ cytotoxic T cells. We have recently developed a DNA vaccine encoding 18 conserved multiple HLA-DR-binding HIV-1 CD4 epitopes (HIVBr18), capable of eliciting broad CD4+ T cell responses in multiple HLA class II transgenic mice. Here, we evaluated the breadth and functional profile of HIVBr18-induced immune responses in BALB/c mice. Immunized mice displayed high-magnitude, broad CD4+/CD8+ T cell responses, and 8/18 vaccine-encoded peptides were recognized. In addition, HIVBr18 immunization was able to induce polyfunctional CD4+ and CD8+ T cells that proliferate and produce any two cytokines (IFNγ/TNFα, IFNγ/IL-2 or TNFα/IL-2) simultaneously in response to HIV-1 peptides. For CD4+ T cells exclusively, we also detected cells that proliferate and produce all three tested cytokines simultaneously (IFNγ/TNFα/IL-2). The vaccine also generated long-lived central and effector memory CD4+ T cells, a desirable feature for T-cell based vaccines. By virtue of inducing broad, polyfunctional and long-lived T cell responses against conserved CD4+ T cell epitopes, combined administration of this vaccine concept may provide sustained help for CD8+ T cells and antibody responses- elicited by other HIV immunogens

MIF Participates in Toxoplasma gondii-Induced Pathology Following Oral Infection

BACKGROUND: Macrophage migration inhibitory factor (MIF) is essential for controlling parasite burden and survival in a model of systemic Toxoplasma gondii infection. Peroral T. gondii infection induces small intestine necrosis and death in susceptible hosts, and in many aspects resembles inflammatory bowel disease (IBD). Considering the critical role of MIF in the pathogenesis of IBD, we hypothesized that MIF participates in the inflammatory response induced by oral infection with T. gondii. METHODOLOGY/PRINCIPAL FINDINGS: Mif deficient (Mif(-/-)) and wild-type mice in the C57Bl/6 background were orally infected with T. gondii strain ME49. Mif(-/-) mice had reduced lethality, ileal inflammation and tissue damage despite of an increased intestinal parasite load compared to wt mice. Lack of MIF caused a reduction of TNF-α, IL-12, IFN-γ and IL-23 and an increased expression of IL-22 in ileal mucosa. Moreover, suppressed pro-inflammatory responses at the ileal mucosa observed in Mif(-/-) mice was not due to upregulation of IL-4, IL-10 or TGF-β. MIF also affected the expression of matrix metalloproteinase-9 (MMP-9) but not MMP-2 in the intestine of infected mice. Signs of systemic inflammation including the increased concentrations of inflammatory cytokines in the plasma and liver damage were less pronounced in Mif(-/-) mice compared to wild-type mice. CONCLUSION/SIGNIFICANCE: In conclusion, our data suggested that in susceptible hosts MIF controls T. gondii infection with the cost of increasing local and systemic inflammation, tissue damage and death

Enhancing Social-Emotional Outcomes in Early Years (E-SEE): Randomized Pilot Study of Incredible Years Infant and Toddler Programs

Abstract: Social emotional development in infancy is a predictor of outcomes in later life, yet there is little evidence of effectiveness for parenting interventions designed to enhance social emotional wellbeing in infancy. An 18-month two-arm randomized controlled pilot trial evaluated the feasibility of a definitive trial of Incredible Years (IY) Infant and Toddler parent programs delivered in a proportionate universal model, called Enhancing Social-Emotional Health and Wellbeing in the Early Years (E-SEE) Steps. Intervention families received an IY Babies book (universal dose), followed by the IY Infant and/or the Toddler group-based programs, based on parent depression (PHQ-9) and/or child social emotional development (ASQ:SE-2) scores. Control parents received services as usual. Parents from two English local authorities with a child eight-weeks-old or younger participated, and were block randomized using a web-based system. Primary endpoints for the study were feasibility parameters relating to recruitment, retention, intervention fidelity and appropriateness of measures. 205 participants were randomized (152:53, intervention:control). Our target was 288 parents. Trial retention rate was higher than expected, with a completion rate of 88% (n = 181, 137:44) at follow-up 3; equating to 94% of 192 expected participants. Intervention uptake was lower than expected. Fidelity of delivery was acceptable and measures were deemed appropriate. A definitive trial is feasible with design amendments to include: introduction of a child screener for intervention eligibility; enhanced intervention material; revised sample size and random allocation ratio. Our internal pilot became an external pilot due to these changes