An In-Silico Comparative Study of Lipases from the Antarctic Psychrophilic Ciliate Euplotes focardii and the Mesophilic Congeneric Species Euplotes crassus: Insight into Molecular Cold-Adaptation

Cold-adapted enzymes produced by psychrophilic organisms have elevated catalytic activities at low temperatures compared to their mesophilic counterparts. This is largely due to amino acids changes in the protein sequence that often confer increased molecular flexibility in the cold. Comparison of structural changes between psychrophilic and mesophilic enzymes often reveal molecular cold adaptation. In the present study, we performed an in-silico comparative analysis of 104 hydrolytic enzymes belonging to the family of lipases from two evolutionary close marine ciliate species: The Antarctic psychrophilic Euplotes focardii and the mesophilic Euplotes crassus. By applying bioinformatics approaches, we compared amino acid composition and predicted secondary and tertiary structures of these lipases to extract relevant information relative to cold adaptation. Our results not only confirm the importance of several previous recognized amino acid substitutions for cold adaptation, as the preference for small amino acid, but also identify some new factors correlated with the secondary structure possibly responsible for enhanced enzyme activity at low temperatures. This study emphasizes the subtle sequence and structural modifications that may help to transform mesophilic into psychrophilic enzymes for industrial applications by protein engineering.     

Homogeneous self-cleaning coatings on cellulose materials derived from TIP/TiO2 P25

The aim of this research was to study TiO₂ nanocoatings formation and to investigate their self-cleaning effects when applied on cellulose materials. Two different approaches for achieving nanocoatings were used. First, coatings were generated in situ through an acid and alkaline catalyzed sol-gel process with or without added water. Another type of coatings was prepared starting from commercial TiO₂ P25 powder. In order to acquire homogeneous coatings from TiO₂ P25 nanoparticles with uniform nanoparticles size distribution, pH of aqueous TiO₂ P25 dispersions was varied. The dispersion preparation conditions were studied by dynamic light scattering (DLS) and zeta potential (ζ-potential) analysis. The resulting TiO₂ nanocoatings were analyzed in terms of their surface morphology using scanning electron microscopy (SEM). Nanocoatings obtained from pure aqueous dispersions of TiO₂ P25 nanoparticles were inhomogeneous with huge agglomerates; however by changing the pH of dispersion and consequently changing the surface charge of TiO₂ P25 nanoparticles as well, more homogeneous nanocoatings with uniform TiO₂ nanoparticles distribution were prepared. Significant differences between solgel derived coatings were observed. Sol-gel process without added water yielded more homogeneous coatings than sol-gel process with addition of water. Completely different surface morphologies were obtained using alkaline or acid catalyst. Acid catalyzed sol-gel process yielded nanocoatings with long, extended, thin structures; contrary, under alkaline conditions particles grow in size with decrease in number. Fourier transform infrared (FTIR) spectroscopy was used to study the coatings’ microstructure. Furthermore the formation of mono-disperse nanoparticles on the fiber surface resulted in enhanced photocatalytic activity. Degradation of colored stain applied on TiO₂-treated samples was investigated by colorimetric measurements. Photocatalytic activity of nanocoatings prepared via acid catalyzed sol-gel process without water addition was comparable to that of nanocoatings derived from aqueous dispersions of commercial TiO₂ P25 nanoparticles.     

Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism

The DJ-1 gene encodes a ubiquitous, highly conserved protein. Here, we show that DJ-1 mutations are associated with PARK7, a monogenic form of human parkinsonism. The function of the DJ-1 protein remains unknown, but evidence suggests its involvement in the oxidative stress response. Our findings indicate that loss of DJ-1 function leads to neurodegeneration. Elucidating the physiological role of DJ-1 protein may promote understanding of the mechanisms of brain neuronal maintenance and pathogenesis of Parkinson's disease.     

Standardization of canine meningioma grading: Validation of new guidelines for reproducible histopathologic criteria

Canine meningiomas are currently graded using the human grading system. Recently published guidelines have adapted the human grading system for use in dogs. The goal of this study was to validate the new guidelines for canine meningiomas. To evaluate the inter-observer agreement, 5 veterinary surgical pathologists graded 158 canine meningiomas following the human grading system alone or with the new guidelines. The inter-observer agreement for histologic grade and each of the grading criteria (mitotic grade, invasion, spontaneous necrosis, macronucleoli, small cells, hypercellularity, pattern loss and anaplasia) was evaluated using the Fleiss kappa index. The diagnostic accuracy (sensitivity and specificity) was assessed by comparing the diagnoses obtained with the 2 grading systems with a consensus grade (considered the reference classification). The consensus histologic grade was obtained by agreement between 4 experienced veterinary neuropathologists following the guidelines. Compared with the human grading alone, the canine-specific guidelines increased the inter-observer agreement for: histologic grade (κ = 0.52); invasion (κ = 0.67); necrosis (κ = 0.62); small cells (κ = 0.36); pattern loss (κ = 0.49) and anaplasia (κ = 0.55). Mitotic grade agreement remained substantial (κ = 0.63). The guidelines improved the sensitivity in identifying grade 1 (95.6%) and the specificity in identifying grade 2 (96.2%) meningiomas. In conclusion, the new grading guidelines for canine meningiomas are associated with an overall improvement in the inter-observer agreement and higher diagnostic accuracy in diagnosing grade 1 and grade 2 meningiomas.