Activation-free printed carbon nanotube field emitters.

When a carbon nanotube paste is formulated based on highly functional hyperbranched polymers such as dipentaerythritol hexaacrylate, the volume shrinkage during thermal curing builds up internal stress that generates microcrack patterns on the printed surface. The nanotubes exposed in the cracks emit electrons successfully at such an extremely low electric field as 0.5 V µm( - 1), and reach 25.5 mA cm( - 2) of current density at 2 Vµm( - 1) from an optimized paste concerning mainly the size and spatial uniformity of the crack. In addition to the superior field emission properties with low manufacturing cost, this activation-free technology can provide a minimized nanohazard in the device fabrication process, compared to those conventional activation technologies developing serious nanoflakes by using destructive methods.

The immunoglobulin G response to Malassezia pachydermatis extracts in atopic and non-atopic dogs.

IgG immunoreactivity to Malassezia pachydermatis was compared in atopic and non-atopic dogs. Malassezia pachydermatis proteins with a molecular weight of 98 kDa were recognized at a significantly higher frequency in the sera of atopic dogs. Most of the atopic dogs with Malassezia dermatitis had a greater IgG response than did normal dogs.

Bunched specifically regulates alpha/beta mushroom body neuronal cell proliferation during metamorphosis.

In Drosophila, mushroom bodies are centers for higher order behavior. Mushroom body neurons consist of three distinct types of neuronal cells, alpha, alpha'/beta', and alpha/beta, which are all generated by the same neuroblasts. The mechanism by which a single neuroblast generates three different types of mushroom body neurons is a compelling area of research. Here, we report that bunched (bun) is expressed only in alpha/beta-type mushroom body neurons and that mutation of the bun gene only affects the development of alpha/beta neurons. Reduced bun expression causes decreased and premature arrest of neuroblast cell division, which results in reduced numbers of alpha/beta neurons and thin axon bundled formation. We propose that bun acts as a specific factor in regulating neuroblast mitotic activity during the development of alpha/beta neurons.

The MED-7 transcriptional mediator encoded by let-49 is required for gonad and germ cell development in Caenorhabditis elegans.

Transcription mediators are evolutionarily conserved from yeast to human. We previously reported the specific in vivo roles of mediators during development. Transcriptional mediators including med-6, med-7, and med-10 were shown to be involved in the regulated transcription of specific genes, but not in the transcription of ubiquitous genes. In this report we have identified and characterized the Caenorhabditis elegans med-7 gene. A genetic mutation in the med-7 gene was identified by comparing genetic and physical maps and determining the molecular lesion. let-49 was found to have a nonsense mutation in the coding region of the med-7 gene. The identification of let-49 as the med-7 gene was confirmed by rescue experiments. The phenotype of the let-49 mutation indicated that the med-7 gene is required for normal postembryonic development. RNAi experiments showed that med-7 is also involved in embryogenesis and the gonad and germ cell development.

Isolation of true Rbp9 null alleles by imprecise P element excisions.

The P element has been widely used as a mutagen because of its convenience in locating the site of mutagenesis. However, P element-induced mutations often result in varied mutant phenotypes, making it difficult to identify the null phenotype. Previously, three Rbp9 alleles were isolated using P element mutagenesis. Although the coding regions of Rbp9 were disrupted by P elements in all three cases, they showed different degrees of defects. In order to characterize the null phenotype of Rbp9, Rbp9 alleles with chromosomal deletions were created by inducing imprecise excisions of the P elements. All Rbp9 alleles generated from imprecise excisions showed the same mutant phenotype: female flies were sterile and cystocyte differentiation was blocked. This result reveals that the primary function of Rbp9 resides in the regulation of cystocyte differentiation. In addition, this result shows that a P element does not always completely inactivate gene activity, even when it is incorporated into the coding region.

Requirement of Rbp9 in the maintenance of Drosophila germline sexual identity.

Drosophila germline sex determination is controlled by a group of genes expressed at early stages of oogenesis (ovo, otu, bam, and Sxl, etc.). Mutations in these genes cause not only sex transformation of female germ cells, but also ovarian tumors. Although mutations at the Rbp9 locus also cause an ovarian tumor phenotype, Rbp9 has been shown to function during later developmental stages than do other ovarian tumor-causing genes. To test whether Rbp9 is also required for germline sex determination, we examined the sex transformation process of female germ cells in Rbp9 mutant flies. The detection of Sxl male transcripts and other male germline markers in Rbp9 mutant ovaries revealed that the Rbp9 mutation caused a partial germline sex transformation. Therefore, sex determination signals that persist throughout oogenesis appear to be required for proper maintenance of germline sexual identity.

Requirement of RBP9, a Drosophila Hu homolog, for regulation of cystocyte differentiation and oocyte determination during oogenesis.

The Drosophila RNA binding protein RBP9 and its Drosophila and human homologs, ELAV and the Hu family of proteins, respectively, are highly expressed in the nuclei of neuronal cells. However, biochemical studies suggest that the Hu proteins function in the regulation of mRNA stability, which occurs in the cytoplasm. In this paper, we show that RBP9 is expressed not only in the nuclei of neuronal cells but also in the cytoplasm of cystocytes during oogenesis. Despite the predominant expression of RBP9 in nerve cells, mutational analysis revealed a female sterility phenotype rather than neuronal defects for Rbp9 mutants. The female sterility phenotype of the Rbp9 mutants resulted from defects in oogenesis; the lack of Rbp9 activity caused the germarium region of the mutants to be filled with undifferentiated cystocytes. RBP9 appears to stimulate cystocyte differentiation by regulating the expression of bag-of-marbles (bam) mRNA, which encodes a developmental regulator of germ cells. RBP9 protein bound specifically to bam mRNA in vitro, which is required for cystocyte proliferation, and the number of cells that expressed BAM protein was increased 5- to 10-fold in the germarium regions of Rbp9 mutants. These results suggest that RBP9 protein binds to bam mRNA to down regulate BAM protein expression, which is essential for the initiation of cystocyte differentiation into functional egg chambers. In hypomorphic Rbp9 mutants, cystocytes differentiated into egg chambers; however, oocyte determination and positioning were perturbed. Therefore, the concentrated localization of RBP9 protein in the oocyte of the early egg chambers may be required for proper oocyte determination or positioning.

Down regulation of extramacrochaetae mRNA by a Drosophila neural RNA binding protein Rbp9 which is homologous to human Hu proteins.

Rbp9 is an RNA binding protein expressed mainly in the central nervous system of adult Drosophilamelanogaster. Rbp9 shares a high degree of sequence similarity with human neural proteins referred to as Hu antigens. Hu antigens bind to U-rich mRNA destabilizing elements with a high affinity and, thus, have been implicated as regulators of mRNA stability. Using in vitro RNA binding assays, we found that Rbp9 binds strongly to poly U sequences. We then employed a Selex system to identify a consensus Rbp9 binding site (UUUXUUUU). Information obtained from the Selex results allowed the detection of two repeats of the Rbp9 consensus binding sequence in the 3' untranslated region of extramacrochaetae mRNA. UV crosslinking experiments demonstrated that Rbp9 interactsspecifically with emc mRNA. The requirement of Rbp9 protein in the down regulation of emc mRNA was confirmed by northern (RNA) analysis, which revealed that the level of emc mRNA increased 10-fold in rbp9 mutant flies. Taken together with the in vitro RNA binding results, the genetic evidence obtained strongly supports the hypothesis that Rbp9 functions as a regulator of RNA stability.

Translational regulation of oskar mRNA by bruno, an ovarian RNA-binding protein, is essential.

Oskar (osk) protein directs the deployment of nanos (nos), the posterior body-patterning morphogen in Drosophila. To avoid inappropriate activation of nos, osk activity must appear only at the posterior pole of the oocyte, where the osk mRNA becomes localized during oogenesis. Here, we show that translation of osk mRNA is, and must be, repressed prior to its localization; absence of repression allows osk protein to accumulate throughout the oocyte, specifying posterior body patterning throughout the embryo. Translational repression is mediated by an ovarian protein, bruno, that binds specifically to bruno response elements (BREs), present in multiple copies in the osk mRNA 3'UTR. Addition of BREs to a heterologous mRNA renders it sensitive to translational repression in the ovary.

Multiple RNA regulatory elements mediate distinct steps in localization of oskar mRNA.

Pattern formation in the early development of many organisms relies on localized cytoplasmic proteins, which can be prelocalized as mRNAs. The Drosophila oskar gene, required both for posterior body patterning and germ cell determination, encodes one such mRNA. Localization of oskar mRNA is an elaborate process involving movement of the transcript first into the oocyte from adjacent interconnected nurse cells and then across the length of the oocyte to its posterior pole. We have mapped RNA regulatory elements that direct this localization. Using a hybrid lacZ/oskar mRNA, we identify several elements within the oskar 3' untranslated region that affect different steps in the process: the early movement into the oocyte, accumulation at the anterior margin of the oocyte and finally localization to the posterior pole. This use of multiple cis-acting elements suggests that localization may be orchestrated in a combinatorial fashion, thereby allowing localized mRNAs with ultimately different destinations to employ common mechanisms for shared intermediate steps.

oskar mRNA is localized to the posterior pole of the Drosophila oocyte.

Mutants of the maternal posterior-group genes of Drosophila lack posterior body pattern elements and germ cells, both of which form through the action of determinants localized to the posterior pole of the oocyte. We report that transcripts of one of these genes, oskar, become localized to the posterior pole of oocytes shortly after the oocyte begins to differentiate visibly. Analysis of various posterior-group mutants reveals that localization of oskar mRNA is an early step in the posterior localization pathway. In addition, we find that nonsense oskar mutations disrupt osk mRNA localization, while missense oskar mutations do not.