Theoretical Upper and Lower Limits for Normalized Bandwidth of Digital Phase-Locked Loop in GNSS Receivers.

Determining the loop noise bandwidth and the coherent integration time is essential and important for the design of a reliable digital phase-locked loop (DPLL) in global navigation satellite system (GNSS) receivers. In general, designers set such parameters approximately by utilizing the well-known fact that the DPLL is stable if the normalized bandwidth, which is the product of the integration time and the noise bandwidth, is much less than one. However, actual limit points are not fixed at exactly one, and they vary with the loop filter order and implementation method. Furthermore, a lower limit on the normalized bandwidth may exist. This paper presents theoretical upper and lower limits for the normalized bandwidth of DPLL in GNSS receivers. The upper limit was obtained by examining the stability of DPLL with a special emphasis on the digital integration methods. The stability was investigated in terms of z-plane root loci with and without the consideration of the computational delay, which is a delay induced by the calculation of the discriminator and the loop filter. The lower limit was analyzed using the DPLL measurement error composed of the thermal noise, oscillator phase noise, and dynamic stress error. By utilizing the carrier-to-noise density ratio threshold which indicates the crossing point between the measurement error and the corresponding threshold, the lower limit of the normalized bandwidth is obtained.

Table-Based Adaptive Digital Phase-Locked Loop for GNSS Receivers Operating in Moon Exploration Missions.

An adaptive digital phase-locked loop (DPLL) continually adjusts the noise bandwidth of the loop filter in global navigation satellite system (GNSS) receivers to track signals by measuring the signal-to-noise ratio and/or dynamic stress. Such DPLLs have a relatively large amount of computational complexity compared with the conventional DPLL. A table-based adaptive DPLL is proposed that adjusts the noise bandwidth value by extracting it from the pre-generated table without additional calculations. The values of the noise bandwidth table are computed in an optimal manner in consideration of the thermal noise, oscillator phase noise, and dynamic stress error. The calculation method of the proper integration time to maintain the stability of the loop filter is presented. Additionally, the simulation is configured using the trajectory analysis results from the Moon exploration mission and shows that the proposed algorithm operates stably in harsh environments, while a conventional fixed bandwidth loop cannot. The proposed algorithm has a similar phase jitter performance to the existing adaptive DPLL algorithms and has an execution time that is approximately 2.4-5.4 times faster. It is verified that the proposed algorithm is computationally efficient while maintaining jitter performance.

Physicochemically modified high-resolution tin oxide thin film using soft imprinting.

We carried out nanoimprinting lithography on solution-processed tin oxide (SnO) film for use as a liquid crystal (LC) alignment layer, for which we used a parallel configuration. To transfer the nanostructures onto the SnO film, we conducted an experiment according to curing, from which fine nanostructures on the SnO film were obtained at a curing temperature of 200 °C. These acted as a guide for the arrangement of the LC molecules and induced geometric restriction which minimized elastic distortion energies, and so the LC molecules could be aligned in the direction of the nanostructures. The LC alignment state was investigated using polarized optical microscopy, and the pre-tilt angle was measured using a crystal rotation method. With high thermal endurance and drastically low power consumption, the nano patterned SnO was shown to be a promising candidate for LC applications. The nanopatterning process combined with nanoimprinting lithography and solution-processed inorganic materials exhibited the possibility of broadening the features of nanostructure-mounted applications, including LC devices.

A Comprehensive Evaluation of Possible RNSS Signals in the S-Band for the KPS.

Recently, the Korean government has announced a plan to develop a satellite-based navigation system called the Korean Positioning System (KPS). When designing a new Radio Navigation Satellite Service (RNSS) signal, the use of the S-band has emerged as an alternative to avoiding signal congestion in the L-bands, and South Korea is considering using the S-band with the L-bands. Therefore, this study proposed possible S-band signal candidates and evaluated their performance, such as the radio frequency (RF) compatibility, spectral efficiency, ranging performance, and receiver complexity. Several figures-of-merit (FoMs) were introduced for quantitative performance evaluation for each candidate. Each FoM was calculated using an analytical equation by considering the signal design parameters, such as the center frequency, modulation scheme, and chip rate. The results showed that the outstanding candidate signal was different depending on the signal performance of interest and the reception environments. Therefore, we discuss and summarize the signal performance analysis results considering the whole FoMs together. Under the assumptions given in this paper, the binary phase shift keying (BPSK)(1), sine-phased binary offset carrier (BOCs)(5,2), and BPSK signals were superior for the spectral efficiency, ranging performance, and receiver complexity, respectively.

Solution-Driven Imprinting Lithography of Sol-Gel ZnO Thin Films for Liquid Crystal Display.

We present a simple and economically convenient method to fabricate nanopatterned ZnO films by imprinting lithography and use them for the layer alignment of liquid crystal (LC) displays. First, a one-dimensional nanopattern was obtained by laser interference lithography on a silicon wafer, and the silicon mold replica was transferred onto a flexible polydimethylsiloxane (PDMS) sheet for conformal patterning. The so-obtained PDMS mold was then applied on a ZnO film spin-coated on a glass substrate. During the imprinting process, the temperature was controlled from 100 to 250 °C to observe the transferring morphologies of the ZnO film; the nanopattern was successfully transferred at annealing temperatures of 200 and 250 °C because the ZnO film at the sol state filled the cavities of the PDMS nanopattern and solidified, forming a negative replica of the nanopattern. The direction of the nanopatterned ZnO film served as a guide for aligning the LC molecules on the LC surface at the centimeter scale and, due to their elastic characteristics and group behavior, propagating their directional states in the LC bulk. The resulting LC cell exhibited an enhanced electro-optical performance and high thermal endurance above 180 °C. The geometry of the alignment layer increased the electric field on the ZnO film and showed reduced threshold voltage. In addition, since flexible devices are generally based on polyimide, which imidized at around 200 °C, the relatively low annealing temperatures of our fabricated nanopatterned ZnO film allow it to be mounted on such devices without any deterioration of the underlying thermoplastic substrate. Therefore, nanopatterned ZnO has a considerable potential for advanced LC displays.

Wg secreted by conventional Golgi transport diffuses and forms Wg gradient whereas Wg tethered to extracellular vesicles do not diffuse.

Wingless (Wg)/Wnt family proteins are essential for animal development and adult homeostasis. Drosophila Wg secreted from the dorsal-ventral (DV) midline in wing discs forms a concentration gradient that is shaped by diffusion rate and stability of Wg. To understand how the gradient of extracellular Wg is generated, we compared the secretion route of NRT-Wg, an artificial membrane-tethered form of Wg that is supposedly not secreted but still supports fly development, to that of wild-type Wg. We found that wild-type Wg is secreted by both conventional Golgi transport and via extracellular vesicles (EVs), and NRT-Wg can be also secreted via EVs. Furthermore, wild-type Wg secreted by Golgi transport diffused and formed Wg gradient but Wg-containing EVs did not diffuse at all. In case of Wg stability, Sol narae (Sona), a metalloprotease that cleaves Wg, contributes to generate a steep Wg gradient. Interestingly, Wg was also produced in the presumptive wing blade region, which indicates that NRT-Wg on EVs expressed in the blade allows the blade cells to proliferate and differentiate without Wg diffused from the DV midline. We propose that EV-associated Wg induces Wg signaling in autocrine and juxtaposed manners whereas Wg secreted by Golgi transport forms gradient and acts in the long-range signaling, and different organs differentially utilize these two types of Wg signaling for their own development.

Role of Armadillo repeat 2 and kinesin-II motor subunit Klp64D for wingless signaling in Drosophila.

Armadillo (Arm) is crucial for transducing Wingless (Wg) signaling. Previously, we have shown that Klp64D, a motor subunit of Drosophila kinesin-II, interacts with Arm for Wg signaling. Molecular basis for this interaction has remained unknown. Here we identify a critical Arm repeat (AR) required for binding Klp64D and Wg signaling. Arm/[Formula: see text]-catenin family proteins contain a conserved domain of 12 Arm repeats (ARs). Five of these ARs can interact with Klp64D, but only the second AR (AR2) binds to the cargo/tail domain of Klp64D. Overexpression of AR2 in wing imaginal disc is sufficient to cause notched wing margin. This phenotype by AR2 is enhanced or suppressed by reducing or increasing Klp64D expression, respectively. AR2 overexpression inhibits Wg signaling activity in TopFlash assay, consistent with its dominant-negative effects on Klp64D-dependent Wg signaling. Overexpression of the Klp64D cargo domain also results in dominant-negative wing notching. Genetic rescue data indicate that both AR2 and Klp64D cargo regions are required for the function of Arm and Klp64D, respectively. AR2 overexpression leads to an accumulation of Arm with GM130 Golgi marker in Klp64D knockdown. This study suggests that Wg signaling for wing development is regulated by specific interaction between AR2 and the cargo domain of Klp64D.

Formation of Wrinkle Structures on Styrene-b-isoprene-b-styrene Films Using One-Step Ion-Beam Irradiation.

We investigated the wrinkle formation on ion-beam (IB)-irradiated substrates coated with the thermoplastic elastomer styrene-b-isoprene-b-styrene (SIS) and demonstrate a relation of the wrinkle structure and the newly formed top layer induced by IB. IB irradiation led to polymer cross-linking on the surface, thereby forming a new skin layer, a finding which was supported by an X-ray photoelectron spectroscopy analysis, Young moduli calculated using force-distance curves, and time-of-flight secondary ion mass spectrometry depth profiling. The wrinkle wavelength increased according to the irradiation time, which indicates that the latter mainly increased the thickness of the cross-linking layer. The increase in the wrinkle wavelength varied from 420 to 670 nm by changing the IB irradiation time. In this paper, we present not only the expectation of wrinkle fabrication using our method but also the possibility of choosing diverse materials such as the thermoplastic elastomer SIS for fabrication of wrinkle structures.

Author Correction: An ADAMTS Sol narae is required for cell survival in Drosophila.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A novel population of extracellular vesicles smaller than exosomes promotes cell proliferation.

BACKGROUND: Extracellular vesicles (EVs) play important roles in intercellular communication by delivering RNA, lipid, and proteins to neighboring or distant cells. Identification and classification of EVs secreted from diverse cell types are essential for understanding their signaling properties. METHODS: In this study, EVs from the culture media were isolated by ultracentrifugation and analyzed by electron microscopy (EM) and nanoparticle tracking analyses. Conditioned media (CM) from HEK293 cells culture grown either in serum-free (SF) or 10% fetal bovine serum (FBS) containing media were centrifuged at 100,000×g to separate the SNΔ supernatant and the P100 pellet in which exosomes are enriched. Then, the SNΔ fraction was centrifuged at 200,000×g to yield the P200 pellet fraction containing novel EVs smaller than exosomes. The exosomal markers in the EV subgroups were examined by western blotting and immune-EM, and the functional analyses of EVs were conducted on HEK293 and THP-1 cell culture. RESULTS: We identified a new group of EVs in the P200 fraction that was smaller than exosomes in size. Typical exosome markers such as Hsp70, TSG101, and CD63 were found in both P100 exosomes and the P200 vesicles, but CD81 was highly enriched in exosomes but not in the P200 vesicles. Furthermore, chemicals that inhibit the major exosome production pathway did not decrease the level of P200 vesicles. Therefore, these small EVs indeed belong to a distinguished group of EVs. Exosomes and the P200 vesicles were found in CM of human cell lines as well as FBS. Addition of the exosomes and the P200 vesicles to human cell cultures enhanced exosome production and cell proliferation, respectively. CONCLUSIONS: Our study identifies a novel population of EVs present in the P200 fraction. This EV population is distinguished from exosomes in size, protein contents, and biogenesis pathway. Furthermore, exosomes promote their own production whereas the P200 vesicles support cell proliferation. In sum, we report a new group of EVs that are distinct physically, biologically and functionally from exosomes.

ADAMTS Sol narae cleaves extracellular Wingless to generate a novel active form that regulates cell proliferation in Drosophila.

Wnt/ Wingless (Wg) is essential for embryonic development and adult homeostasis in all metazoans, but the mechanisms by which secreted Wnt/Wg is processed remain largely unknown. A Drosophila Sol narae (Sona) is a member of A Disintegrin And Metalloprotease with ThromboSpondin motif (ADAMTS) family, and positively regulates Wg signaling by promoting Wg secretion. Here we report that Sona and Wg are secreted by both conventional Golgi and exosomal transports, and Sona cleaves extracellular Wg at the two specific sites, leading to the generation of N-terminal domain (NTD) and C-terminal domain (CTD) fragments. The cleaved forms of extracellular Wg were detected in the extracellular region of fly wing discs, and its level was substantially reduced in sona mutants. Transient overexpression of Wg-CTD increased wing size while prolonged overexpression caused lethality and developmental defects. In contrast, Wg-NTD did not induce any phenotype. Moreover, the wing defects and lethality induced by sona RNAi were considerably rescued by Wg-CTD, indicating that a main function of extracellular Sona is the generation of Wg-CTD. Wg-CTD stabilized cytoplasmic Armadillo (Arm) and had genetic interactions with components of canonical Wg signaling. Wg-CTD also induced Wg downstream targets such as Distal-less (Dll) and Vestigial (Vg). Most importantly, Cyclin D (Cyc D) was induced by Wg-CTD but not by full-length Wg. Because Sona also induces Cyc D in a cell non-autonomous manner, Wg-CTD generated by Sona in the extracellular region activates a subset of Wg signaling whose major function is the regulation of cell proliferation.

One-dimensional surface wrinkling for twisted nematic liquid crystal display based on ultraviolet nanoimprint lithography.

Surface wrinkling method is used to fabricate a 1-dimensional nanostructure. The structure is transferred to an ultraviolet cured polymer which is used as an alignment layer. The anisotropic geometry serves as a guide for aligning liquid crystal molecules uniformly without defects. The TN-LC cell showed a successful LC switching, with a response time of 20.5 ms, and a threshold voltage of 2.00 V. It also exhibited high thermal stability above 180°C. The proposed UV-cured polymers with 1-D nano wrinkle geometry can be a candidate for alternative alignment techniques, for advanced liquid crystal devices with high thermal budgets.

Physicochemical Modification Effect on Homogeneously Aligned Liquid Crystals Based on the Nickel Oxide Thin Film.

We demonstrated homogeneous liquid crystal (LC) alignment on Nickel Oxide (NiO) films subjected to ion beam (IB) irradiation. Uniform LC alignment was achieved at high IB intensity values of 1200 and 1800 eV. To determine the mechanism of LC alignment following IB irradiation, physicochemical analysis was performed using atomic force microscopy and X-ray photoelectron spectroscopy. IB irradiation with high intensity increases uniformity of the surface, and IB irradiation induces the formation of oxygen vacancies and increases the NiO (Ni2+) phase components. Hence, both of the smooth surface and the strong van der Waals interactions between the NiO film and the LC molecules provides the LC molecules with a stable anchor on the surface, leading to uniform LC alignment on NiO films after IB irradiation. IB-irradiated NiO exhibited a high transparency of 85% in the visible light range as compared with the 83% average transmittance of conventional polyimide, which makes the IB-irradiated NiO alignment layer attractive in display devices.

An ADAMTS Sol narae is required for cell survival in Drosophila.

Cell survival is essential for all living organisms to cope against multiple environmental insults. Intercellular signaling between dying and surviving cells plays an important role to ensure compensatory proliferation, preventing tissue loss after environmental stresses. Here, we show that Sol narae (Sona), a Disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) in Drosophila is required for cell survival. sona exhibited a positive genetic interaction with Death-associated inhibitor of apoptosis 1 (Diap1), and a negative genetic interaction with reaper (rpr). Transcription patterns of sona, Diap1, and rpr genes in the pouch region of wing discs were coordinately changed after irradiation. Interestingly, there was a negative correlation in the expression levels of Sona and DIAP1, and both cell types, one with high Sona level and the other with high Diap1 level, were resistant to irradiation-induced cell death. The sona-expressing cells rarely entered into cell cycle themselves but promoted the nearby cells to proliferate in irradiation conditions. We found that these sona-expressing cells are able to upregulate Cyclin D (Cyc D) and increase tissue size. Furthermore, transient Sona overexpression increased survival rate and promoted development of flies in irradiation conditions. We propose that the two types of radiation-resistant cells, one with high Sona level and the other with high Diap1 level, communicate with dying cells and between each other for cell survival and proliferation in response to irradiation.

Sol narae (Sona) is a Drosophila ADAMTS involved in Wg signaling.

ADAMTS (a disintegrin and metalloproteases with thrombospondin motif) family consists of secreted proteases, and is shown to cleave extracellular matrix proteins. Their malfunctions result in cancers and disorders in connective tissues. We report here that a Drosophila ADAMTS named Sol narae (Sona) promotes Wnt/Wingless (Wg) signaling. sona loss-of-function mutants are lethal and rare escapers had malformed appendages, indicating that sona is essential for fly development and survival. sona exhibited positive genetic interaction with wntless (wls) that encodes a cargo protein for Wg. Loss of sona decreased the level of extracellular Wg, and also reduced the expression level of Wg effector proteins such as Senseless (Sens), Distalless (Dll) and Vestigial (Vg). Sona and Wg colocalized in Golgi and endosomal vesicles, and were in the same protein complex. Furthermore, co-expression of Wg and Sona generated ectopic wing margin bristles. This study suggests that Sona is involved in Wg signaling by regulating the level of extracellular Wg.

Cell type-specific responses to wingless, hedgehog and decapentaplegic are essential for patterning early eye-antenna disc in Drosophila.

The Drosophila eye-antenna imaginal disc (ead) is a flattened sac of two-layered epithelia, from which most head structures are derived. Secreted morphogens like Wingless (Wg), Hedgehog (Hh), and Decapentaplegic (Dpp) are important for early patterning of ead, but the underlying mechanisms are still largely unknown. To understand how these morphogens function in the ead of early larval stages, we used wg-LacZ and dpp-Gal4 markers for the examination of wild-type and mutant eads. We found that the ead immediately after hatching was crescent-shaped with the Bolwig's nerve at the ventral edge, suggesting that it consists of dorsal domain. In a subsequent step, transcriptional induction of dpp in the cells along the Bolwig's nerve was followed by rapid growth of the ventral domain. Both Wg and Hh were required for the formation of the ventral domain. Wg was crucial for the growth of the entire ead, but Hh was essential for cell division only in the dorsal domain. In the ventral domain, Hh regulated dpp transcription. Based on these data, we propose that signaling among distinct groups of cells expressing Wg, Dpp, or Hh in the ead of the first-instar larvae are critical for coordinated growth and patterning of ead.

Identification of Heat Shock Protein 60 as a Regulator of Neutral Sphingomyelinase 2 and Its Role in Dopamine Uptake.

Activation of sphingomyelinase (SMase) by extracellular stimuli is the major pathway for cellular production of ceramide, a bioactive lipid mediator acting through sphingomyelin (SM) hydrolysis. Previously, we reported the existence of six forms of neutral pH-optimum and Mg(2+)-dependent SMase (N-SMase) in the membrane fractions of bovine brain. Here, we focus on N-SMase ε from salt-extracted membranes. After extensive purification by 12,780-fold with a yield of 1.3%, this enzyme was eventually characterized as N-SMase2. The major single band of 60-kDa molecular mass in the active fractions of the final purification step was identified as heat shock protein 60 (Hsp60) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. Proximity ligation assay and immunoprecipitation study showed that Hsp60 interacted with N-SMase2, prompting us to examine the effect of Hsp60 on N-SMase2 and ceramide production. Interestingly, Hsp60 siRNA treatment significantly increased the protein level of N-SMase2 in N-SMase2-overexpressed HEK293 cells. Furthermore, transfection of Hsp60 siRNA into PC12 cells effectively increased both N-SMase activity and ceramide production and increased dopamine re-uptake with paralleled increase. Taken together, these results show that Hsp60 may serve as a negative regulator in N-SMase2-induced dopamine re-uptake by decreasing the protein level of N-SMase2.