The Bantam microRNA Is Associated with Drosophila Fragile X Mental Retardation Protein and Regulates the Fate of Germline Stem Cells

Fragile X syndrome, a common form of inherited mental retardation, is caused by the loss of fragile X mental retardation protein (FMRP). We have previously demonstrated that dFmr1, the Drosophila ortholog of the fragile X mental retardation 1 gene, plays a role in the proper maintenance of germline stem cells in Drosophila ovary; however, the molecular mechanism behind this remains elusive. In this study, we used an immunoprecipitation assay to reveal that specific microRNAs (miRNAs), particularly the bantam miRNA (bantam), are physically associated with dFmrp in ovary. We show that, like dFmr1, bantam is not only required for repressing primordial germ cell differentiation, it also functions as an extrinsic factor for germline stem cell maintenance. Furthermore, we find that bantam genetically interacts with dFmr1 to regulate the fate of germline stem cells. Collectively, our results support the notion that the FMRP-mediated translation pathway functions through specific miRNAs to control stem cell regulation

Image-based non-photorealistic rendering for realtime virtual sculpting

This paper discusses a real-time virtual sculpting system in a non-photorealistic manner. Up till now, a majority of researchers pursuing virtual sculpting have highlighted the physical fidelity of simulation, with approaches for realizing photorealism being principal specialized contributions. We aim at capitalizing on the strengths of non-photorealistic rendering with respect to aesthetics, expression and computing expense. By means of touch-enabled manipulation, the haptic interaction is conducted to form the deformable surfaces, and the edge extraction is employed to stress boundaries. Afterwards, we utilize anisotropic diffusion to lessen unimportant details as well as a fresh inverse Gaussian bilateral filter for spot removal. Processed in a spatiotemporal style, the non-photorealistic rendering works to maintain coherence in time. Moreover, the parallel realization of the system put forward on graphics hardware (GPU) guarantees real-time behavior. © 2014 Springer Science+Business Media New York.This paper discusses a real-time virtual sculpting system in a non-photorealistic manner. Up till now, a majority of researchers pursuing virtual sculpting have highlighted the physical fidelity of simulation, with approaches for realizing photorealism being principal specialized contributions. We aim at capitalizing on the strengths of non-photorealistic rendering with respect to aesthetics, expression and computing expense. By means of touch-enabled manipulation, the haptic interaction is conducted to form the deformable surfaces, and the edge extraction is employed to stress boundaries. Afterwards, we utilize anisotropic diffusion to lessen unimportant details as well as a fresh inverse Gaussian bilateral filter for spot removal. Processed in a spatiotemporal style, the non-photorealistic rendering works to maintain coherence in time. Moreover, the parallel realization of the system put forward on graphics hardware (GPU) guarantees real-time behavior. © 2014 Springer Science+Business Media New York

Preparation of Cleaning Agent for Treatment of Abandoned Oil Pipeline

China crude oil pipeline industry commonly faces with abandonment problem because of the decreasing output of oilfield and pipeline integrity problem. China released its first standard on abandoned pipeline (SY/T 7413-2018) named specifications for the disposal of abandoned oil&gas pipelines and it is implemented on the April 1st 2019. Pipeline cleaning is very necessary to dispose the abandoned pipeline to eliminate the risk of environment and safety. There are many cleaning agents for abandoned pipeline in the market. This chemical cleaning agent is not suitable for china pipeline because the wax remained in china pipeline is different that of foreign pipeline. The wax content in residual of china pipeline is very high. So, preparation of cleaning agent suitable for china pipeline is very important to remove wax with low cost. In this study one kind of hydrophilic cleaning agent is prepared. The result of lab test result and cleaning project both show that the kind of cleaning have excellent performance for the abandoned pipeline

The bantam miRNA is required for GSC maintenance.

<p>A. A schematic diagram of a <i>Drosophila</i> germarium with different cell types labeled by different colors: GSCs, cystoblast (CB) and cysts, spectrosomes (SS), terminal filaments (TF), cap cells (CPC), inner germarium sheath cells (IGC), follicle cells (FC), SSC (somatic stem cells) and fusomes. B–G: Ovaries from wild-type (B), <i>ban</i> mutant flies at different ages (C–F), and <i>ban</i> mutants carrying transgene P<i>{banP-ban}</i> (G) were stained with anti-<i>Vasa</i> (Green) and anti-<i>Hts</i> (Red) antibodies. H. Quantitative analyses of the number of GSCs in <i>ban</i> mutant; the x-axis shows the day of examination post-eclosion, while the y-axis shows the average number of GSCs per germarium in <i>ban</i> mutants and WT. <i>P<0.001</i> when <i>ban</i> mutant was compared with WT at different time points. Arrows indicate GSCs.</p

<i>Bantam</i> genetically interacts with <i>dFmr1</i> in modulating GSCs.

<p>(A) The number of GSCs per germarium was measured from <i>dfmr1</i> heterozygotes, <i>ban</i> heterozygotes, and <i>dfmr1</i>, <i>ban</i> double heterozygotes at day 2 and day 12 post-eclosion, respectively. <i>P<0.01</i> when the percentage of type II and III germaria of the trans-heterozygotes were compared with those of either WT or single heterozygotes. (B) Ovaries from the genotypes above were stained with anti-<i>Vasa</i> (Green) and anti-<i>Hts</i> (Red); germaria carrying two GSCs, one GSC, and no GSCs were referred to as type I, II, III-a and III-b, respectively.</p

The bantam miRNA plays a non-autonomous role in GSC maintenance.

<p>GSC clones were induced by heat-shock treatment in adult female flies. Ovaries from FRT control flies (A and B) and FRT, <i>ban</i> flies (C and D) were dissected at day 2 and day 12 following heat-shock treatment; GSC clones were identified by the lack of GFP expression. Scale bar represents 10 µm. (E) Relative percentages of negatively GFP-marked GSC clones in FRT control and two FRT; <i>ban</i> null alleles at days 2, 7, 12, and 15 AHST are shown. <i>P>0.5</i>.</p

Generation and characterization of new <i>bantam</i> mutant alleles.

<p>Imprecise mobilization of the P-element from <i>ban^EP3622</i> was carried out to generate stronger alleles for the <i>ban</i> gene. One sterile line, <i>ban²⁰</i>, and one lethal line, <i>ban¹²</i>, were isolated. The breakpoints of two mutant lines are shown, with the adjacent genes indicated. Mature bantam miRNA is indicated in green. The chromosomal position and sequence region is based on <i>Drosophila melanogaster</i> (R5.13).</p

The bantam miRNA is required for repressing primordial germ cell (PGC) differentiation.

<p>Gonads from the third-instar larvae of <i>w¹¹¹⁸</i> (A) and <i>ban¹²</i> mutants (B, C, and C inset) were stained with anti-<i>Vasa</i> and anti-<i>Hts</i> antibodies. Anti-<i>Hts</i> (Red) was used to outline gonads and morphology of fusomes, while anti-<i>Vasa</i> (green) was used to visualize all germ cells. PGCs carrying a single round fusome are indicated by arrows, while differentiated germ cells are indicated by arrowheads. Scale bar represents 10 µm.</p

Specific miRNAs associated with <i>dFmr1</i> protein in <i>Drosophila</i> ovary.

<p>(A) Western blot shows that <i>dFmr1</i> protein (dFmrp) was immunoprecipitated from wild-type <i>Drosophila</i> ovary. A <i>dFmr1</i> null mutant (<i>dfmr1³</i>) was used as a negative control. (B) miRNA TaqMan assays of 72 known <i>Drosophila</i> miRNAs were performed in triplicate using both input and IP RNAs from both wild-type and <i>dfmr1³</i> mutants. The miRNAs that were enriched are shown in progressively brighter shades of red, and the miRNAs that were reduced in IP are shown in progressively brighter shades of green. The miRNAs shown in black were not changed. The fold of the change is indicated on both sides of the scale bar. The miRNAs that are specifically enriched in IP from wild-type ovary are shown. The data represent the average of two biological replicates (two independent immunoprecipitation experiments). (C) TaqMan assays of the bantam miRNA were performed in triplicate, and the enrichment of the bantam miRNA in dFmrp-IP (independent IP experiments from those presented in panel B) from wild-type ovary is shown. (D) Northern blot shows that the bantam miRNA is associated with dFmrp. Northern blots detecting the sense and anti-sense strands of the bantam miRNA in both input and IP RNAs from WT and <i>dfmr1³</i> mutants are shown.</p

Relative fertility of different mutant flies.

<p>Individual female was crossed with wildtype (<i>w¹¹¹⁸</i>) male flies and the number of fertile eggs was quantified.</p>*<p><i>P<0.01</i> when the trans-heterozygotes were compared with either WT or single heterozygotes.</p