Thermoelectric performance of a metastable thin-film Heusler alloy.

Thermoelectric materials transform a thermal gradient into electricity. The efficiency of this process relies on three material-dependent parameters: the Seebeck coefficient, the electrical resistivity and the thermal conductivity, summarized in the thermoelectric figure of merit. A large figure of merit is beneficial for potential applications such as thermoelectric generators. Here we report the thermal and electronic properties of thin-film Heusler alloys based on Fe2V0.8W0.2Al prepared by magnetron sputtering. Density functional theory calculations suggest that the thin films are metastable states, and measurements of the power factor-the ratio of the Seebeck coefficient squared divided by the electrical resistivity-suggest a high intrinsic figure of merit for these thin films. This may arise from a large differential density of states at the Fermi level and a Weyl-like electron dispersion close to the Fermi level, which indicates a high mobility of charge carriers owing to linear crossing in the electronic bands.

Two-Dimensional Conical Dispersion in ZrTe_{5} Evidenced by Optical Spectroscopy.

Zirconium pentatelluride was recently reported to be a 3D Dirac semimetal, with a single conical band, located at the center of the Brillouin zone. The cone's lack of protection by the lattice symmetry immediately sparked vast discussions about the size and topological or trivial nature of a possible gap opening. Here, we report on a combined optical and transport study of ZrTe_{5}, which reveals an alternative view of electronic bands in this material. We conclude that the dispersion is approximately linear only in the a-c plane, while remaining relatively flat and parabolic in the third direction (along the b axis). Therefore, the electronic states in ZrTe_{5} cannot be described using the model of 3D Dirac massless electrons, even when staying at energies well above the band gap 2Δ=6 meV found in our experiments at low temperatures.

Kondo Insulator to Semimetal Transformation Tuned by Spin-Orbit Coupling.

Recent theoretical studies of topologically nontrivial electronic states in Kondo insulators have pointed to the importance of spin-orbit coupling (SOC) for stabilizing these states. However, systematic experimental studies that tune the SOC parameter λ_{SOC} in Kondo insulators remain elusive. The main reason is that variations of (chemical) pressure or doping strongly influence the Kondo coupling J_{K} and the chemical potential µ-both essential parameters determining the ground state of the material-and thus possible λ_{SOC} tuning effects have remained unnoticed. Here, we present the successful growth of the substitution series Ce_{3}Bi_{4}(Pt_{1-x}Pd_{x})_{3} (0≤x≤1) of the archetypal (noncentrosymmetric) Kondo insulator Ce_{3}Bi_{4}Pt_{3}. The Pt-Pd substitution is isostructural, isoelectronic, and isosize, and it therefore is likely to leave J_{K} and µ essentially unchanged. By contrast, the large mass difference between the 5d element Pt and the 4d element Pd leads to a large difference in λ_{SOC}, which thus is the dominating tuning parameter in the series. Surprisingly, with increasing x (decreasing λ_{SOC}), we observe a Kondo insulator to semimetal transition, demonstrating an unprecedented drastic influence of the SOC. The fully substituted end compound Ce_{3}Bi_{4}Pd_{3} shows thermodynamic signatures of a recently predicted Weyl-Kondo semimetal.

Transdifferentiation.

Recent progress in studies of development and differentiation has greatly stimulated analysis of transdifferentiation, and more cell types capable of transdifferentiation have been documented. Growth factors must be essential, key factors in the regulation of the transdifferentiation process, in cooperation with components of the extracellular matrix, which helps to stabilize the differentiated state of tissues. Trials to induce transdifferentiation artificially by transfection of genes have also begun.

Increase in actin contents and elongation of apical projections in retinal pigmented epithelial cells during development of the chicken eye.

The structural and biochemical changes of cytoskeletal components of retinal pigmented epithelial cells were studied during the development of chicken eyes. When the cytoskeletal components of the pigmented epithelial cells from various stages of development were examined by SDS PAGE, actin contents in the cells markedly increased between the 15-d-old and hatching stages. Immunofluorescence microscopy showed that chicken pigmented epithelial cells have two types of actin bundles. One is the circumferential bundle associated with the zonula adherens region as previously reported (Owaribe, K., and H. Masuda, 1982, J. Cell Biol., 95:310-315). The other is the paracrystalline bundle forming the core of the apical projections. The increase in actin contents after the 15-d-old stage is accompanied by the formation and elongation of core filaments of apical projections in the cells. During this period the apical projections extend into extracellular space among outer and inner segments of photoreceptor cells. Accompanying this change is an elongation of the paracrystalline bundles of actin filaments in the core of the projection. By electron microscopy, the bundles decorated with muscle heavy meromyosin showed unidirectional polarity, and had transverse striations with approximately 12-nm intervals, as determined by optical diffraction of electron micrographs. Since the shape of these bundles was not altered in the presence or absence of Ca2+, they seemed not to have villin-like proteins. Unlike the circumferential bundles, the paracrystalline bundles did not contract when exposed to Mg-ATP. These observations indicate that the paracrystalline bundles are structurally and functionally different from the circumferential actin bundles.

Demonstration of contractility of circumferential actin bundles and its morphogenetic significance in pigmented epithelium in vitro and in vivo.

Each pigmented epithelial cell bears circumferential actin bundles at its apical level when the pigmented epithelium is established in eyes in situ or in culture in vitro. Well-differentiated pigmented epithelia in culture were treated with a 50% glycerol solution containing 0.1 M KCl, 5 mM EDTA, and 10 mM sodium phosphate buffer, pH 7.2, for 24 h or more at 4 degrees C. When the glycerinated epithelium was transferred to the ATP solution, each cell constituting the epithelium began to contract. The epithelium was cleaved into many cell groups as a result of contraction of each cell. The periphery of each cell group was lifted to form a cup or vesicle and eventually detached from the substratum. However, those cells that had not adhered tightly and not formed a monolayer epithelium with typical polygonal cellular pattern contracted independently as observed in the glycerinated fibroblasts. Contraction of the glycerinated pigmented epithelial cells was inhibited by N-ethylmaleimide but not by cytochalasin B. ITP and UTP also effected the contraction of the glycerinated cells, but GTP and ADP did not. Ca2+ was not required. This contractile model of pigmented epithelium provides a useful experimental system for analyzing the function of actin in cellular morphogenesis.

The A5 antigen, a candidate for the neuronal recognition molecule, has homologies to complement components and coagulation factors.

The A5 antigen is a neuronal cell surface protein of Xenopus presumed to be involved in the neuronal recognition between the optic nerve fibers and the visual centers. Analyses of cDNA clones revealed that the A5 antigen is a class I membrane protein containing two different internal repeats in the extracellular segment. The first repeat bears homology to domain III of complement components C1r and C1s, and the second repeat is homologous to the C1 and C2 domains of coagulation factors V and VIII. The mRNA for the A5 antigen was present in retinal ganglion cells and visual center neurons. Nonneuronal cells in the peripheral and central nervous systems did not express the mRNA for the A5 antigen.

Gene regulation and differentiation in vertebrate ocular tissues.

Molecular biological techniques have contributed greatly to the study of vertebrate ocular tissues. The specification of ocular tissues has been shown to be closely related to the expression of transcription factors encoded by genes such as Pax6 and microphthalmia. Lens-specific expression of the delta 1-crystallin gene is controlled by factors, such as delta EF1, binding to its enhancer sequences. Retinal activity of the glucocorticoid hormone receptor is regulated by its binding with another transcription factor. Degeneration of photoreceptors in a retinal disease, retinitis pigmentosa, can be caused by the introduction of a mutated opsin gene into mice. In addition, the process of transdifferentiation in ocular tissues has been described at the level of gene expression.

Nycthemeral Rhythm of Phlebotominae (Diptera: Psychodidae) in a Craggy Region, Transitioning Between the Wetland and the Plateau, Brazil.

Recording the nycthemeral rhythm of sand flies allows the evaluation of the daily activity in different ecotypes, the period of greatest activity, and their degree of anthropophily. We investigated the fauna and the rhythm of sand fly activity in an ecotourism region in Mato Grosso do Sul (MS) state, Brazil. Sand flies were captured monthly, using a Shannon trap for 24 h periods between July 2012 and June 2014. We collected 1,815 sand flies, in which Lutzomyia whitmani (=Nyssomyia whitmani, sensu Galati) and Lutzomyia longipalpis were the most abundant species during the dry season, with activity from 5 p.m.-7 a.m. and 6 p.m.-5 a.m., respectively. Both species require particular attention as vectors of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum in several regions of Brazil, including MS. However, Lutzomyia dispar was more anthropophilic, and was most active between January and March, from 5 p.m. to 5 a.m. Lutzomyia misionensis (=Pintomyia misionensis, sensu Galati) was present throughout both years, active from 4 p.m. to 5 a.m. Other species were active from 5 p.m. to 6 a.m. Due to intense tourism in the months that coincide with a high number of vectors for leishmaniases in Piraputanga, it is essential to determine vector-monitoring strategies in the area by investigating sand fly rhythm while not neglecting other periods of the year when the insects are present.

Studies on aspartase. IV. Reversible denaturation of Escherichia coli aspartase.

Aspartase (L-aspartate ammonia lyase, EC 4.3.1.1) of Escherichia coli, denatured in 4 M guanidine-HCl, was renatured in vitro by simple dilution with a concomitant restoration of the activity. While the native enzyme exhibited a marked negative Cotton effect centered at 233 +/- 1 nm in optical rotatory dispersion, the enzyme denatured in 4 M guanidine-HCl retained little optical activity. Upon dilution of the denatured enzyme, however, more than 90% of the ordered structure was recovered in 1 min, while the restoration of the activity proceeded much more slowly. Estimation of molecular weights by gel permeation chromatography indicated that the tetrameric enzyme is subject to reversible dissociation into monomeric subunits under the experimental conditions. Various environmental factors such as temperature, pH and protein concentration exhibited profound influence on the rate and extent of the reactivation. In order to examine the correlation between the restoration of the activity and the quaternary structure, electron microscopic inspection of the kinetic processes of reversible denaturation was attempted. Upon dilution of the denatured enzyme at 4 degrees C, neither the activity nor tetrameric images were detected over several min. Upon the temperature shift up to 25 degrees C, however, the activity regain was rapidly proceeded concomitant with the appearance of tetrameric molecules. These results are compatible with the possibility that the subunit assembly is an essential prerequisite, thought not sufficient, for enzyme activity.

Disappearance of calcium-induced phase separation in phosphatidylserine-phosphatidylcholine membranes caused by protonation and by electric current.

Disappearance of Ca2+-induced phase separation in phosphatidylserine-phosphatidylcholine membrane has been studied under several conditions by monitoring electron spin resonance spectrum of spin-labeled phosphatidylcholine. The membranes were prepared in Millipore filters. Electron micrographs of the pre parations showed formation of multilayered structures lined on the pore surface. The phase separation was disappeared when the membrane was soaked in non-buffered salt solution (100 ml KCl, pH 5.5). It was markedly contrasting that when the bathing salt solution was buffered no disappearance was observed. Disappearance of the phase separation was also observed when the Ca2+-treated membrane was transferred to acidic salt solutions (less than or equal to pH 2.5) or to low ionic strength media (less than or equal to mM) buffered at pH 5.5, and then to the buffered salt solution (100 mM KCl, pH 5.5). These are due to replacement of Ca2+ by proton, proton-induced separation, followed by disappearance of the phase separation in the buffered salt solution. Biological significance of the competition between Ca2+ and proton for the phase separation or domain formation in the membranes was emphasized.

Studies on aspartase. VII. Subunit arrangement of Escherichia coli aspartase.

Aspartase (L-aspartate ammonia-lyase, EC 4.3.1.1) of Escherichia coli is composed of four subunits of seemingly identical molecular weight (Suzuki, S., Yamaguchi, J. And Tokushige, M.(1973) Biochim. Biophys. Acta 321, 369-381). The subunit arrangement of the enzyme was studied by two distinct methods, cross-linking of subunits with a bifunctional reagent, dimethyl suberimidate, and statistical classification of negatively stained electron microscopic images. In the former method, the densitometric patterns of the cross-linked aspartase were analyzed quantitatively after separating each component by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and the results were compared with the theoretical distribution. In the latter method, a number of electron microscopic images were classified into several groups according to their characteristic appearance. The results obtained by these two methods are compatible with the possibility that the enzyme has a tetrameric structure consisting of two pairs of dimers, in which the two pairs of rod-shape subunits meet perpendicularly, being typical of D2 symmetry.

Expression and distribution of regeneration-responsive molecule during normal development of the newt, Cynops pyrrhogaster.

We have investigated the expression and distribution of the regeneration-responsive molecule, 2NI-36, the loss of which is responsible for initiation of dedifferentiation of dorsal marginal iris pigmented epithelial cells to regenerate a lens. In the process of the normal development of the newt, the expression of 2NI-36 could not be detected in embryos at the early developmental stages, i.e., cleavages, gastrulation and neurulation, nor through later developmental stages to tail-bud, even though organogenesis was occurring. 2NI-36 was not detectable in any tissues until embryos reached developmental stage 40 (before hatching). In hatched larvae around developmental stage 46, strong expression of 2NI-36 was observed in several tissues including the vascular endothelium, the pigmented epithelium and the inner layer of skin epidermis. Moreover, 2NI-36 was present on the cell surface of these tissue cells. In conclusion, when the embryos hatch out to become swimming larvae that can feed by themselves, 2NI-36 begins to be expressed in some kinds of differentiated tissues. These results suggest that the function of 2NI-36 might be related to the completion of morphogenesis in development and also to the stabilization of the differentiated state of newly formed tissue cells.

The loss of gap junctional cell-to-cell communication is coupled with dedifferentiation of retinal pigmented epithelial cells in the course of transdifferentiation into the lens.

Retinal pigmented epithelial cells (PECs) of the chick embryo can be cultured as a monolayer of melanized hexagonal cells. Modifications of the culture condition make the cells lose most of the phenotypes and further transdifferentiate into lentoid bodies within a few weeks. Ultrastructural observations showed that PECs and the lentoids have gap junctions with distinct morphology. Diffusion of a fluorescent dye confirmed the presence of gap junctions in both phenotypes. However, cells in the intermediate stage of transdifferentiation, which show neither the phenotype of the PEC nor that of the lentoid and are called dedifferentiated PECs here, have almost no gap junction structure. We propose the possibility that the dedifferentiation of PECs and the loss of cell-to-cell communication are tightly coupled events. This cell culture system is a suitable material for further studying this relationship by cellular and molecular approaches.

Characterization of terminally differentiated cell state by categorizing cDNA clones derived from chicken lens fibers.

To characterize a terminally differentiated state of cells at the gene expression level, a cDNA library of chicken lens fibers was analyzed. The major population of the library consisted of cDNAs encoding delta-crystallin (about 35% of the recombinants) and other crystallins (alpha A-, alpha B-, beta A3/A1-, beta B1-, beta B2-), as well as cytoskeletal proteins (CP49, CP95), and membrane protein (MP28). These cDNA clones representing lens structural proteins known, accounted for about 60% of the library. When 96 clones were randomly selected from this library, 55 clones corresponded to the above-mentioned major class proteins. Analyses of the remaining clones indicated that many of them were expressed in a lens-specific manner at very low levels. The partial nucleotide sequence analysis of these clones revealed that two cDNAs corresponded to the genes encoding lens-type connexin, three cDNAs to the genes encoding housekeeping proteins, and some cDNAs to the genes encoding regulatory proteins. The mRNA composition in the chicken lens fiber cells indicated rather simple organization of mRNA species of this cell type, and gave scope to the possibility of full description of differentiated lens fiber cells at gene activity level.

Basic fibroblast growth factor as one of the essential factors regulating lens transdifferentiation of pigmented epithelial cells.

In vitro transdifferentiation of retinal pigmented epithelial cells of the chick embryo into lens cells can be markedly enhanced by culture in the presence of testicular hyaluronidase and phenylthiourea. Since the commercial preparations of hyaluronidase that had previously been used were very crude, a search for the actual effective molecule(s) enhancing lens transdifferentiation was conducted. First, we purified the enzyme and tested the effect of the purified hyaluronidase. Highly purified hyaluronidase itself did not enhance lens transdifferentiation. The crude hyaluronidase was then separated according to affinity with heparin, considering the possibility that the fibroblast growth factor (FGF) is contained in the crude hyaluronidase. Transdifferentiation-enhancing activity was detected in the fraction which was bound to heparin and eluted with 2 M NaCl, where no hyaluronate-degrading activity existed. Analysis of the fraction by SDS-PAGE revealed the existence of an 18 kDa protein whose NH2-terminal sequence was identical to that of basic FGF. The basic FGF derived from bovine brain also enhanced lens transdifferentiation of pigmented epithelial cells. These findings suggest that basic FGF must play a major role in enhancing transdifferentiation of pigmented epithelial cells to lens cells.

Analysis of a unique molecule responsible for regeneration and stabilization of differentiated state of tissue cells.

In Wolffian regeneration in the newt, a functional lens can be regenerated through cellular transdifferentiation of the pigmented epithelium of the mid-dorsal marginal iris. A novel monoclonal antibody, 2NI-36 mAb, generated in our laboratory has been utilized as a highly useful probe to study newt lens regeneration. The antigen molecule against this 2NI-36 mAb (2NI-36) became temporarily undetectable only at the site of lens regeneration. Moreover, the ventral iris pieces expressed the ability to differentiate a lens when pretreated with this monoclonal antibody and implanted in lentectomized eyes (Eguchi, Cell Differ. Dev. 25, Suppl., 1988). We have investigated the distribution of 2NI-36 in newt tissues. 2NI-36 was not specific to iris pigmented epithelium and distributed in many different kinds of mesodermal tissues, including dermis, blood vessel, mesonephros and so forth. 2NI-36 was also detected in either cell surface or intercellular spaces of cultured pigmented epithelial cells when they organized an epithelial cell sheet. Western blot analysis showed that 2NI-36 had the molecular weight of 50-200kD and was completely digested by trypsin, suggesting that 2NI-36 was a glycoprotein with many carbohydrate chains. It was also revealed by Western blot analysis that all the tissues in which 2NI-36 could be detected expressed this molecule similar to that in the iris epithelium. We expect that 2NI-36 is a glycoprotein expressed by various newt tissues and is functional to stabilize the differentiated state of each tissue cell in the same way as observed in the iris pigmented epithelial cells.

A unique aged human retinal pigmented epithelial cell line useful for studying lens differentiation in vitro.

Lens regeneration occurs in some urodeles and fish throughout their adult life. Such an event is possible by the transdifferentiation of the pigment epithelial cells (PECs) from the dorsal iris. Studies of this event at the cellular level have been facilitated owing to the ability of PECs to become lens cells even when they are placed in culture, outside of the eye. In fact, PECs possess the capacity for transdifferentiation regardless of the origin of species or age. However, studies at the molecular level are still hindered by the intrinsic problems of primary cultures, namely storage, reproducibility and genetic manipulation. In an attempt to establish an ideal model system for lens transdifferentiation, we have analyzed the ability of a human dedifferentiated PEC line to differentiate into lens. We have found that this cell line can indeed be induced to synthesize crystallin and morphologically differentiate to three-dimensional structures resembling lentoids under controlled treatment in vitro. Gene expression studies also provided important insights into the role of key genes. This human cell line can be used for detailed genetic studies in order to identify the key factors involved in lens transdifferentiation from PECs.

Isolation of a novel chick homolog of Serrate and its coexpression with C-Notch-1 in chick development.

Intercellular signaling mediated by the transmembrane proteins, Notch as receptor and its ligands, Delta and Serrate, plays essential roles in the developmental fate decision of many cell types in Drosophila. The Notch genes are highly conserved both in invertebrates and in vertebrates, suggesting that Notch pathway regulates cell fate decisions during vertebrates development. Notch, Delta and Serrate homologs in chicken have been cloned (Henrique et al., Nature 375: 787-790, 1995; Myat et al., Dev. Biol. 174: 233-247, 1996). We isolated a novel chick homolog of Drosophila Serrate, named C-Serrate-2, and examined its expression patterns during the early chick development using whole-mount in situ hybridization. C-Serrate-2 transcripts were detected in several tissues including the forebrain, the myotome and the apical ectodermal ridge (AER) of the limb bud of a 4-day-old chick embryo. In most of the regions where C-Serrate-2 was expressed, C-Notch-1 was also expressed. Our observations suggest that Serrate-2-Notch-1 signaling plays a role in a variety of morphogeneses during the chick development.