Fungal laccases as tools for biodegradation of industrial dyes

Laccases are blue copper oxidases, found in some plants and secreted by a wide range of ligninolytic fungi. These enzymes are well known for their ability in oxidizing several organic compounds, mainly phenolics and aromatic amines, at the expenses of molecular oxygen. Therefore, they could find application in the field of enzymatic bioremediation of many industrial wastewaters, and in particular to bleach and/or detoxify dye-containing effluents. Not all industrial dyes behave as laccase substrates, but this limitation is often overcome by the judicious use of redox mediators. These could substantially widen the application range of laccases as bioremediation tools. The present study encompasses the main properties of the most used industrial dyes as related to their chemical classification, fungal laccases and their molecular and catalytic features, the use of redox mediators, limitations and perspectives of the use of fungal laccases for industrial dye bleaching

Tyrosinase inhibition: general and applied aspects.

The active site of tyrosinase is described with a view to depicting its interactions with substrates and inhibitors. Occurrence and mechanism(s) of tyrosinase-mediated browning of agrofood products are reviewed, with regard to both enzymic and chemical reactions, and their control, modulation, and inhibition. Technical and applicational implications are discussed

Infectious diseases prevalence, vaccination coverage, and diagnostic challenges in a population of internationally adopted children referred to a Tertiary Care Children's Hospital from 2009 to 2015

Infectious diseases are common in internationally adopted children (IAC). With the objective to evaluate infectious diseases prevalence in a large cohort of IAC and to explore possible risk factors for tuberculosis (TB) and parasitic infections, clinical and laboratory data at first screening visit of all IAC (<18 years) consecutively referred to our Center in 2009 to 2015 were collected and analyzed. In total, 1612 children (median age: 5.40 years; interquartile range: 3.00–7.90) were enrolled, 123/1612 (7.60%) having medical conditions included in the special needs definition. The most frequent cutaneous infections were Molluscum contagiosum (42/1612; 2.60%) and Tinea capitis (37/1612; 2.30%). Viral hepatitis prevalence was <1% (hepatitis B virus [HBV]: 13 children, 0.80%; hepatitis C virus: 1 child, 0.10%; hepatitis A virus: 6 children, 0.40%). A parasitic infection was diagnosed in 372/1612 (23.10%) children. No risk factors for parasitosis were evidenced. Active TB was diagnosed in 4/1355 (0.3%) children, latent TB in 222/1355 (16.40%). Only 3.7% (51/1355) children had concordant positive tuberculin skin test (TST) and QuantiFERON-TB-Gold In-Tube (QFT-G-IT) results. Risk factors for TST+/QFT-G-IT− results were previous Bacille de Calmette-Guérin vaccination (adjusted odds ratio [aOR]: 2.18; 96% confidence interval [CI]: 1.26–3.79; P = 0.006), and age ≥5 years (aOR: 1.49; 95% CI: 1.06–2.11; P = 0.02). The proportion of children with nonprotective titers for vaccine-preventable diseases (VPD) ranged from 15.70% (208/1323) for tetanus to 35.10% (469/1337) for HBV. Infectious diseases were commonly observed in our cohort. The high rate of discordant TST/QFT-G results brings up questions regarding the optimal management of these children, and suggests that, at least in children older than 5 years, only QFT-G-IT results may be reliable. The low proportion of children protected for VPD, confirms importance of a timely screening

Biological activities and nutraceutical potentials of water extracts from different parts of Cynomorium coccineum L. (Maltese Mushroom)

Maltese Mushroom (Cynomorium coccineum L.) is a non-photosynthetic plant that has been used in traditional medicine for many centuries. In this paper, water extracts from the whole plant, external layer and peeled plant were studied to determine the main components responsible for its biological activities, i.e., its antimicrobial, antioxidant, and anti-tyrosinase activities; its cytotoxicity against mouse melanoma B16F10 cells; and its pro-erectile activity in adult male rats. The results of electron transfer and hydrogen transfer assays showed that the antioxidant activity was mainly due to anthocyanins in the external layer, whereas the external layer and peeled plant extracts both inhibited the microbial growth of several Gram-positive strains. In contrast, the whole plant extract had the highest anti-tyrosinase activity and exhibited pro-erectile activity when administered subcutane-ously. Overall, this study elucidated which parts of Maltese Mushroom are responsible for its antimicrobial, antioxidant, and anti-tyrosinase activities and thus which extracts have potential for use in nutraceutical formulation

Intensified high yield production of GMMA based vaccines for high burden neglected disease

Vaccines are among the most important public health interventions currently available. The global demand for vaccines is growing due to global population growth, ongoing global immunization campaigns and the increase of antibiotic-resistant bacteria. Vaccines are of high importance in Low- and Middle-Income Countries (LMIC) where millions of deaths occur annually due to vaccines-preventable diseases. Global coverage would need at least 200 million doses annually of a two-dose vaccine to cover the at-risk birth cohort, and considerably more for catch up vaccination during a roll out phase. Especially for vaccines against diseases in LMIC, there is a major added constraint: the vaccine production cost per unit and manufacturing facilities investments must be minimized to ensure the vaccines will be both viable and sustainable. For diseases caused by Gram-negative bacteria, the vaccine platform based on the GMMA technology is promising. GMMA are highly immunogenic outer membrane exosomes produced from genetically engineered bacteria modified for high yield hyper-blebbing that can be purified with minimal downstream processing. Simple robust scalable processes make GMMA ideal for cost-conscious vaccines. Please click Download on the upper right corner to see the full abstract

Potential Role of M. tuberculosis Specific IFN-&#947; and IL-2 ELISPOT Assays in Discriminating Children with Active or Latent Tuberculosis

BACKGROUND: Although currently available IGRA have been reported to be promising markers for TB infection, they cannot distinguish active tuberculosis (TB) from latent infection (LTBI). OBJECTIVE: Children with LTBI, active TB disease or uninfected were prospectively evaluated by an in-house ELISPOT assay in order to investigate possible immunological markers for a differential diagnosis between LTBI and active TB. METHODS: Children at risk for TB infection prospectively enrolled in our infectious disease unit were evaluated by in-house IFN-γ and IL-2 based ELISPOT assays using a panel of Mycobacterium tuberculosis antigens. RESULTS: Twenty-nine children were classified as uninfected, 21 as LTBI and 25 as active TB cases (including 5 definite and 20 probable cases). Significantly higher IFN-γ ELISPOT responses were observed in infected vs. uninfected children for ESAT-6 (p<0.0001), CFP-10 (p<0.0001), TB 10.3 (p = 0.003), and AlaDH (p = 0.001), while differences were not significant considering Ag85B (p = 0.063), PstS1 (p = 0.512), and HspX (16 kDa) (p = 0.139). IL-2 ELISPOT assay responses were different for ESAT-6 (p<0.0001), CFP-10 (p<0.0001), TB 10.3 (p<0.0001), HspX (16 kDa) (p<0.0001), PstS1 (p<0.0001) and AlaDH (p = 0.001); but not for Ag85B (p = 0.063). Comparing results between children with LTBI and those with TB disease differences were significant for IFN-γ ELISPOT only for AlaDH antigen (p = 0.021) and for IL-2 ELISPOT assay for AlaDH (p<0.0001) and TB 10.3 antigen (p = 0.043). ROC analyses demonstrated sensitivity of 100% and specificity of 81% of AlaDH-IL-2 ELISPOT assay in discriminating between latent and active TB using a cut off of 12.5 SCF per million PBMCs. CONCLUSION: Our data suggest that IL-2 based ELISPOT with AlaDH antigen may be of help in discriminating children with active from those with latent TB

Analytical development to support manufacturing of a sustainable vaccine against Invasive Nontyphoidal Salmonellosis

GVGH is developing a candidate trivalent Salmonella vaccine to fight invasive nontyphoidal Salmonellosis (iNTS) and typhoid fever, especially aimed for sub-Saharan Africa to impact disease burden and to reduce anti-microbial resistance spread. This trivalent vaccine may be the only viable option for a sustainable iNTS vaccine in sub-Saharan Africa over the separate administration of Typhoid Conjugate Vaccines (TCV) and a vaccine against iNTS. GVGH generated the iNTS-TCV formulation by combining the GMMA technology for the iNTS components, S. Typhimurium (STm) and S. Enteritidis (SEn) GMMA adsorbed on Alhydrogel, and the Vi-CRM197 glycoconjugate, originally developed by GVGH and recently WHO prequalified as TCV TYPHIBEV by Biological E Ltd (Hyderabad, India). A set of analytical methods to support the vaccine lot release and characterization have been developed by GVGH. In particular, to quantify the key active ingredients of iNTS components a competitive ELISA-based method (FAcE, Formulated Alhydrogel competitive ELISA assay) has been setup and characterized in terms of specificity, accuracy and precision. Vi component is instead characterized by means of HPAEC-PAD method, able to specifically identify and quantify the total polysaccharide in the final drug product. With regard to safety assessment, a Monocyte Activation Test (MAT) has been developed as to monitor the intrinsic pyrogenicity of GMMA-based vaccines and applied as surveillance test for the Phase 1 clinical lot, with the plan to set release criteria based on clinical experience. In vivo potency assay has been set to characterize the immunogenicity of vaccine lots in comparison to freshly formulated material at the time of release and during real-time stability. A significant antibody response to each of the active ingredients of the trivalent vaccine is raised in mice and assessed by Parallel Line Assay. Overall, the applied analytical panel and the results support the development of an iNTS-TCV vaccine as a viable option for a sustainable iNTS vaccine in sub-Saharan Africa

Bioinspired versus enzymatic oxidation of some homologous thionine dyes in the presence of immobilized metalloporphyrin catalysts and ligninolytic enzymes

Thionines are recalcitrant and polluting textile dyes presenting various degrees of N-methylation. In this paper, a complete series of homologous thionines was used as the substrates for oxidation in the presence of a bioinspired commercial iron-porphyrin immobilized on to imidazoleand pyridine-functionalized fumed silica, to emulate the active site of ligninolytic peroxidases. The obtained catalytic adducts showed a remarkable ability to catalyze thionine dye oxidation in the presence of different oxidants such as potassium monopersulfate and hydrogen peroxide. Different oxidation patterns were obtained and mechanistically discussed, in comparison with those observed in the presence of some ligninolytic oxidizing enzymes

Techno-Economic Analysis of a Small-Scale Biomass-to-Energy BFB Gasification-Based System

In order to limit global warming to around 1.5&#8315;2.0 &#176;C by the end of the 21st century, there is the need to drastically limit the emissions of CO2. This goal can be pursued by promoting the diffusion of advanced technologies for power generation from renewable energy sources. In this field, biomass can play a very important role since, differently from solar and wind, it can be considered a programmable source. This paper reports a techno-economic analysis on the possible commercial application of gasification technologies for small-scale (2 MWe) power generation from biomass. The analysis is based on the preliminary experimental performance of a 500 kWth pilot-scale air-blown bubbling fluidized-bed (BFB) gasification plant, recently installed at the Sotacarbo Research Centre (Italy) and commissioned in December 2017. The analysis confirms that air-blown BFB biomass gasification can be profitable for the applications with low-cost biomass, such as agricultural waste, with a net present value up to about 6 M&#8364; as long as the biomass is provided for free; on the contrary, the technology is not competitive for high-quality biomass (wood chips, as those used for the preliminary experimental tests). In parallel, an analysis of the financial risk was carried out, in order to estimate the probability of a profitable investment if a variation of the key financial parameters occurs. In particular, the analysis shows a probability of 90% of a NPV at 15 years between 1.4 and 5.1 M&#8364; and an IRR between 11.6% and 23.7%