Lifetime prediction of encapsulated CdSexS1-x quantum platelets for color conversion in high luminance LED microdisplays.

The LED technology is seen today as the most promising approach to manufacture high luminance color microdisplays for augmented reality application. So far, it mostly involves blue micro-LED technology and quantum dots-based layers for green and red color generation by light down-conversion. Despite significant progress, the viability of this technology still raises many questions. Among them, the stability of the color conversion layer under nominal display operating conditions is still an issue which has not been thoroughly addressed yet. This paper provides experimental data on the aging behavior of CdSexS1-x quantum platelets (QP) for blue-to-red conversion, under a wide range of blue irradiation power. A modeling of the photoluminescence (PL) decrease versus aging time is proposed, that enables to reliably predict the lifetime of a color LED microdisplay in real operating conditions. At room temperature, the alumina encapsulated CdSexS1-x QPs exhibit a lifetime (t70) of 35,000 h under operating conditions representative of a microdisplay emitting 100,000 nits white light, in video mode. With an average daily use of 3 hours, it would represent for a microdisplay more than 30 years. In addition, the study highlights that display heating induces a lifetime decrease related to a thermally activated enhancement of the annihilation rate of PL emission centers. As a result, a display operated at 100,000 nits and 45°C would see its lifetime t70 reduced by a factor 4 (∼8 years), which remains acceptable for most micro-display applications.

Why antiplectic metachronal cilia waves are optimal to transport bronchial mucus.

The coordinated beating of epithelial cilia in human lungs is a fascinating problem from the hydrodynamics perspective. The phase lag between neighboring cilia is able to generate collective cilia motions, known as metachronal waves. Different kinds of waves can occur, antiplectic or symplectic, depending on the direction of the wave with respect to the flow direction. It is shown here, using a coupled lattice Boltzmann-immersed boundary solver, that the key mechanism responsible for their transport efficiency is a blowing-suction effect that displaces the interface between the periciliary liquid and the mucus phase. The contribution of this mechanism on the average flow generated by the cilia is compared to the contribution of the lubrication effect. The results reveal that the interface displacement is the main mechanism responsible for the better efficiency of antiplectic metachronal waves over symplectic ones to transport bronchial mucus. The conclusions drawn here can be extended to any two-layer fluid configuration having different viscosities, and put into motion by cilia-shaped or comb-plate structures, having a back-and-forth motion with phase lags.

Thermo-physical properties of synthetic mucus for the study of airway clearance.

In this article, dynamic viscosity, surface tension, density, heat capacity and thermal conductivity, of a bronchial mucus simulant proposed by Zahm et al., Eur Respir J 1991; 4: 311-315 were experiementally determined. This simulant is mainly composed of a galactomannan gum and a scleroglucan. It was shown that thermophysical properties of synthetic mucus are dependant of scleroglucan concentrations. More importantly and for some scleroglucan concentrations, the syntetic mucus, exhibits, somehow, comparable thermophysical properties to real bronchial mucus. An insight on the microstructure of this simulant is proposed and the different properties enounced previously have been measured for various scleroglucan concentrations and over a certain range of operating temperatures. This synthetic mucus is found to mimic well the rheological behavior and the surface tension of real mucus for different pathologies. Density and thermal properties have been measured for the first time. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3025-3033, 2017.

Expression of Tie-2 in human peripheral and autonomic nervous system.

Tie-2, a tyrosine kinase receptor, is essential for vascular integrity by regulating cellular adhesion between pericytes and endothelial cells. The aim of this study was to identify sites of expression of Tie-2 other than the vasculature. Tie-2 expression was first detected in human colon by Western blotting and reverse-transcription-polymerase chain reaction (RT-PCR) in tissue extracts. The presence of the Tie-2 mRNA and protein was detected by immunohistochemistry and in situ hybridization in cells of the colon myenteric and submucosal plexus, in both neuronal and Schwann cells. Tie-2 protein was also found in the nervous system of the female urogenital tract. In the human sciatic nerve and schwannoma, RT-PCR, Western blotting and immunohistochemistry analysis further confirmed the presence of Tie-2 mRNA and protein in non-autonomic peripheral nervous tissue. In conclusion, using several approaches and tissues we have demonstrated the presence of Tie-2 in human peripheral and autonomic nervous tissue, suggesting a role for Tie-2 in neural tissue. Thus, attempts to disrupt the tumour vessels by manipulation of the Tie-2 system in tumours may result in side-effects in peripheral nerves.

X-ray structure of HPr kinase: a bacterial protein kinase with a P-loop nucleotide-binding domain.

HPr kinase/phosphatase (HprK/P) is a key regulatory enzyme controlling carbon metabolism in Gram- positive bacteria. It catalyses the ATP-dependent phosphorylation of Ser46 in HPr, a protein of the phosphotransferase system, and also its dephosphorylation. HprK/P is unrelated to eukaryotic protein kinases, but contains the Walker motif A characteristic of nucleotide-binding proteins. We report here the X-ray structure of an active fragment of Lactobacillus casei HprK/P at 2.8 A resolution, solved by the multiwavelength anomalous dispersion method on a seleniated protein (PDB code 1jb1). The protein is a hexamer, with each subunit containing an ATP-binding domain similar to nucleoside/nucleotide kinases, and a putative HPr-binding domain unrelated to the substrate-binding domains of other kinases. The Walker motif A forms a typical P-loop which binds inorganic phosphate in the crystal. We modelled ATP binding by comparison with adenylate kinase, and designed a tentative model of the complex with HPr based on a docking simulation. The results confirm that HprK/P represents a new family of protein kinases, first identified in bacteria, but which may also have members in eukaryotes.

Mutations lowering the phosphatase activity of HPr kinase/phosphatase switch off carbon metabolism.

The oligomeric bifunctional HPr kinase/P-Ser-HPr phosphatase (HprK/P) regulates many metabolic functions in Gram-positive bacteria by phosphorylating the phosphocarrier protein HPr at Ser46. We isolated Lactobacillus casei hprK alleles encoding mutant HprK/Ps exhibiting strongly reduced phosphatase, but almost normal kinase activity. Two mutations affected the Walker motif A of HprK/P and four a conserved C-terminal region in contact with the ATP-binding site of an adjacent subunit in the hexamer. Kinase and phosphatase activity appeared to be closely associated and linked to the Walker motif A, but dephosphorylation of seryl-phosphorylated HPr (P-Ser-HPr) is not simply a reversal of the kinase reaction. When the hprKV267F allele was expressed in Bacillus subtilis, the strongly reduced phosphatase activity of the mutant enzyme led to increased amounts of P-Ser-HPr. The hprKV267F mutant was unable to grow on carbohydrates transported by the phosphoenolpyruvate:glycose phosphotransferase system (PTS) and on most non-PTS carbohydrates. Disrupting ccpA relieved the growth defect only on non-PTS sugars, whereas replacing Ser46 in HPr with alanine also restored growth on PTS substrates.

Efficient gene targeted random mutagenesis in genetically stable Escherichia coli strains.

We describe a method to generate in vivo collections of mutants orders of magnitude larger than previously possible. The method favors accumulation of mutations in the target gene, rather than in the host chromosome. This is achieved by propagating the target gene on a plasmid, in Escherichia coli cells, within the region preferentially replicated by DNA polymerase I (Pol I), which replicates only a minor fraction of the chromosome. Mutagenesis is enhanced by a conjunction of a Pol I variant that has a low replication fidelity and the absence of the mutHLS system that corrects replication errors. The method was tested with two reporter genes, encoding lactose repressor or lipase. The proportion of mutants in the collection was estimated to reach 1% after one cycle of growth and 10% upon prolonged cell cultivation, resulting in collections of 10(12)-10(13) mutants per liter of cell culture. The extended cultivation did not affect growth properties of the cells. We suggest that our method is well suited for generating protein variants too rare to be present in the collections established by methods used previously and for isolating the genes that encode such variants by submitting the cells of the collections to appropriate selection protocols.

Production of Cry11A and Cry11Ba toxins in Bacillus sphaericus confers toxicity towards Aedes aegypti and resistant Culex populations.

Cry11A from Bacillus thuringiensis subsp. israelensis and Cry11Ba from Bacillus thuringiensis subsp. jegathesan were introduced, separately and in combination, into the chromosome of Bacillus sphaericus 2297 by in vivo recombination. Two loci on the B. sphaericus chromosome were chosen as target sites for recombination: the binary toxin locus and the gene encoding the 36-kDa protease that may be responsible for the cleavage of the Mtx protein. Disruption of the protease gene did not increase the larvicidal activity of the recombinant strain against Aedes aegypti and Culex pipiens. Synthesis of the Cry11A and Cry11Ba toxins made the recombinant strains toxic to A. aegypti larvae to which the parental strain was not toxic. The strain containing Cry11Ba was more toxic than strains containing the added Cry11A or both Cry11A and Cry11Ba. The production of the two toxins together with the binary toxin did not significantly increase the toxicity of the recombinant strain to susceptible C. pipiens larvae. However, the production of Cry11A and/or Cry11Ba partially overcame the resistance of C. pipiens SPHAE and Culex quinquefasciatus GeoR to B. sphaericus strain 2297.

Improvement of Bacillus sphaericus toxicity against dipteran larvae by integration, via homologous recombination, of the Cry11A toxin gene from Bacillus thuringiensis subsp. israelensis.

Integrative plasmids were constructed to enable integration of foreign DNA into the chromosome of Bacillus sphaericus 2297 by in vivo recombination. Integration of the aphA3 kanamycin resistance gene by a two-step procedure demonstrated that this strategy was applicable with antibiotic resistance selection. Hybridization experiments evidenced two copies of the operon encoding the binary toxin from B. sphaericus in the recipient strain. The Bacillus thuringiensis subsp. israelensis cry11Aal gene (referred to as cry11A), encoding a delta-endotoxin with toxicity against Culex, Aedes, and Anopheles larvae, was integrated either by a single crossover event [strain 2297 (::pHT5601), harboring the entire recombinant plasmid] or by two successive crossover events [strain 2297 (::cry11A)]. The level of the Cry11A production in B. sphaericus was high; two crystalline inclusions were produced in strain 2297 (::pHT5601). Synthesis of the Cry11A toxin conferred toxicity to the recombinant strains against Aedes aegypti larvae, for which the parental strain was not toxic. Interestingly, the level of larvicidal activity of strain 2297 (::pHT5601) against Anopheles stephensi was as high as that of B. thuringiensis subsp. israelensis and suggested synergy between the B. thuringiensis and B. sphaericus toxins. The toxicities of parental and recombinant B. sphaericus strains against Culex quinquefasciatus were similar, but the recombinant strains killed the larvae more rapidly. The production of the Cry11A toxin in B. sphaericus also partially restored toxicity for C. quinquefasciatus larvae from a population resistant to B. sphaericus 1593. In vivo recombination therefore appears to be a promising approach to the creation of new B. sphaericus strains for vector control.

Transcriptional regulation of the cryIVD gene operon from Bacillus thuringiensis subsp. israelensis.

The CryIVD protein is involved in the overall toxicity of the Bacillus thuringiensis subsp. israelensis parasporal inclusions and is one of the four major components of the crystals. Determination of the DNA sequence indicated that the cryIVD gene is the second gene of an operon which includes three genes. The first one encodes a 19-kDa polypeptide and has sequence homology with the orf1 gene of the Bacillus thuringiensis cryIIA and cryIIC operons. The second and third genes have already been identified and encode the CryIVD crystal protein and the P20 polypeptide, respectively. The promoter region was located by deletion analysis, and the 5' end of the mRNA was determined by primer extension mapping. Transcription of the cryIVD gene in B. thuringiensis subsp. israelensis strains is induced 9 h after the beginning of sporulation. Sequence analysis indicated two potential promoters, a strong one and a weak one, recognized respectively by the RNA polymerase associated with the sigma 35 or the sigma 28 factor of B. thuringiensis (sigma E and sigma K of Bacillus subtilis, respectively). Transcriptional lacZ fusion integrated in single copy into the chromosome of various B. subtilis sporulation mutants confirmed the sigma E dependence of cryIVD gene transcription.

Role of the CryIVD polypeptide in the overall toxicity of Bacillus thuringiensis subsp. israelensis.

The gene encoding the CryIVD protein of B. thuringiensis subsp. israelensis crystals was disrupted by in vivo recombination. The toxicity of the CryIVD protein-free inclusions was similar to that of the wild-type crystals on Anopheles stephensi larvae but was half the wild-type toxicity on Culex pipiens and Aedes aegypti larvae.

Expression of cryIVA and cryIVB Genes, Independently or in Combination, in a Crystal-Negative Strain of Bacillus thuringiensis subsp. israelensis.

The cryIVA and cryIVB genes, encoding the 125- and 135-kDa proteins, respectively, of Bacillus thuringiensis subsp. israelensis, were cloned either alone or together into a shuttle vector and expressed in a nontoxic strain of B. thuringiensis subsp. israelensis. The CryIVB protein was produced at a high level during sporulation and accumulated as inclusions; in contrast, the CryIVA polypeptide did not form such structures unless it was cloned on a higher-copy-number plasmid. Transcriptional fusions between the cryIVA or cryIVB gene promoter and the lacZ gene were constructed. The poor synthesis of CryIVA was not due to a poor efficiency of transcription from the cryIVA gene promoter. Mosquitocidal assays performed with purified inclusions showed that CryIVA was toxic for larvae of the species Aedes aegypti, Anopheles stephensi, and Culex pipiens, whereas CryIVB displayed activity only toward Aedes aegypti and Anopheles stephensi. The activity of inclusions containing both polypeptides was higher than that of single-peptide inclusions but was not as high as that of the native crystals, which contain at least four polypeptides.

[Alcoholic fermentation of inulin by various strains of yeasts]. / Fermentation alcoolique de l'inuline par quelques souches de levures.

Strains of fourteen species of yeasts able to ferment inulin without previous chemical or physical hydrolysis were studied on semi-synthetic medium by evaluation of CO2 production under anaerobic conditions. Among them, Kluyveromyces cicerisporus, Candida macedoniensis and Candida utilis showed the best kinetic characteristics of fermentation. Experiments were carried out to specify the action of different parameters such as temperature, pH and exogenous ethanol concentration. The results obtained on semi-synthetic medium were confirmed on Jerusalem artichoke juice. The optimal temperature for fermentation was 35 degrees C for the three strains, but K. cicerisporus alone conserved its fermentative capacity at 40 degrees C, at low pH (3.5) and with 6% (v/v) exogenous ethanol concentration. This strain appears to be a good yeast for industrial production of ethanol from inulin substrate.

[DNA-DNA hybridization in several species of Hansenula]. / Hybridation ADN-ADN chez quelques espèces du genre Hansenula.

The genus Hansenula was considered a long time ago as a good pattern for phylogenetic research. In 1969, Wickerham proposed an evolutive scheme based upon morphological, physiological and ecological criteria. Recently, relatedness among yeasts were analysed by DNA-DNA hybridization in liquid medium. H. anomala var. anomala (G + C content: 37.1%) was compared with H. anomala var. schneggii (37.6%), H. subpelliculosa (33.8%) line 3, H. sydowiorum (40.1%) and H. muscicola (37.1%). These results showed little relatedness between H. anomala var. anomala/H. ciferrii and H. anomala var. anomala/H. subpelliculosa. On the other hand, H. anomala var. schneggii shared 89.5% of its nucleotide sequences with H. anomala var. anomala. These 2 strains were considered to represent the same species. H. holstii showed 67.1% complementarity with H. anomala var. anomala: this strain is considered to represent valid species, different from H. anomala var. anomala, but H. muscicola with 72.5% relatedness to H. anomala var. anomala could be considered as a 'limit species'. An unexpected finding was that H. beckii was closely related to H. anomala var. anomala (84.8%). These data suggested the inadequacy of current criteria used to establish the phylogenetic lines in genus Hansenula.