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TAX1BP1 is a selective macroautophagy/autophagy receptor that inhibits NFKB and RIGI-like receptor (RLR) signaling to prevent excessive inflammation and maintain homeostasis. Selective autophagy receptors such as SQSTM1/p62 and OPTN are phosphorylated by the kinase TBK1 to stimulate their selective autophagy function. However, it is unknown if TAX1BP1 is regulated by TBK1 or other kinases under basal conditions or during RNA virus infection. Here, we found that TBK1 and IKBKE/IKKi function redundantly to phosphorylate TAX1BP1 and regulate its autophagic turnover through canonical macroautophagy. TAX1BP1 phosphorylation promotes its localization to lysosomes resulting in its degradation. Additionally, we found that during vesicular stomatitis virus infection, TAX1BP1 is targeted to lysosomes in an Atg8-family protein-independent manner. Furthermore, TAX1BP1 plays a critical role in the clearance of MAVS aggregates, and phosphorylation of TAX1BP1 controls its MAVS aggrephagy function. Together, our data support a model whereby TBK1 and IKBKE license TAX1BP1-selective autophagy function to inhibit MAVS and RLR signaling. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • The chitosan-carbon nanotube hybrid was prepared by homogenizer easily. • And chitosan-carbon nanotube hybrid was applied for sensing the glucose electrochemically. • The electrochemical performance of glucose was characterized by cyclic voltammetry. Abstract In glucose sensors, the direct electron transfer (DET) method delivers electrons from glucose oxidase (GOx) to an electrode without the intervention of a mediator. Many researchers have developed the DET method using carbon nanotube (CNT)-based composites for the permeation into the enzyme co-factor. We synthesized chitosan (Chit) core-shelled CNTs that were hydrophilic, biocompatible, and easily prepared. Electrodes with three different Chit-CNT weight ratios were prepared, and their morphological and physicochemical properties were characterized by Fourier-transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), x-ray diffraction (XRD), scanning electron microscopy (SEM), and transition electron microscopy (TEM). The Chit-CNT85 composite (85 wt% Chit) performed optimally by cyclic voltammetry (CV), and was covalently modified with GOx via a routine coupling protocol to form the GOx-Chit-CNT85/ITO electrode. This electrode was electrochemically characterized by impedance spectroscopy (EIS). The GOx loading and pH were optimized against the CV redox potential. The electrode exhibited a fast electron-transfer constant (k s) of 8.2 s−1. The cathodic peak of GOx decreased in response to increasing glucose concentrations, but no changes were observed in the presence of physiological interfering substances such as ascorbic acid and uric acid. Therefore, our GOx-Chit-CNT85/ITO electrode has the potential for use in glucose sensing directly without a mediator. [ABSTRACT FROM AUTHOR]

Human herpesvirus 8 (HHV-8) is causally related to human malignancies. HHV-8 latent viral FLICE-inhibitory protein (vFLIP) is a viral oncoprotein that is linked to pathogenesis, but how its expression is regulated is largely unknown. In an attempt to understand the role of the mitochondrial antiviral signaling (MAVS) adaptor in HHV-8 infection, we discovered that vFLIP expression was post-translationally up-regulated by the MAVS signaling complex on peroxisomes. Furthermore, we demonstrated that vFLIP could be targeted to the peroxisomes, where it was oncogenically active, in a PEX19-dependent manner. Targeted disruption of vFLIP and MAVS interaction resulted in a decrease in vFLIP expression and selectively promoted death of latently HHV-8-infected cells, providing therapeutic potential for treating HHV-8 diseases. Collectively, our experimental results suggest novel involvement of peroxisomes and MAVS in the stabilization of vFLIP and thereby in the establishment or maintenance of HHV-8 latency and associated pathogenesis. [ABSTRACT FROM AUTHOR]

Applying a nanocomposite to increase the conductivity of an electrode can facilitate electrochemical analysis. In this regard, multi-walled carbon nanotubes (MWCNTs) evenly dispersed in hydrophilic solution can play an important role in electrochemical bio-sensing due to their unique properties, such as their high electrical conductivity and ability to conjugate with hydrophilic enzymes. Herein, we report the simple ultrasonic synthesis of a highly dispersible, enzyme-binding nanocomposite, poly(acrylamide)- co -poly(vinyl imidazole) (7:1 mol ratio)-MWCNTs (PAA-PVI@MWCNTs). This material, having a zeta potential of 36.6 ± 0.53 mV, was applied as a film to an electrode surface and stably bound with glucose oxidase to transfer an electron between the enzyme and electrode in the presence of glucose. The PAA-PVI@MWCNTs composite, which was readily dispersed in deionized water, can be used as a biocompatible material for applications such as bio-sensing, point-of-care testing (POCT), and other health care functions. [ABSTRACT FROM AUTHOR]

Recently, viruses have been shown to regulate selective autophagy for productive infections. For instance, human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma-associated herpesvirus (KSHV), activates selective autophagy of mitochondria, termed mitophagy, thereby inhibiting antiviral innate immune responses during lytic infection in host cells. We previously demonstrated that HHV-8 viral interferon regulatory factor 1 (vIRF-1) plays a crucial role in lytic replication-activated mitophagy by interacting with cellular mitophagic proteins, including NIX and TUFM. However, the precise molecular mechanisms by which these interactions lead to mitophagy activation remain to be determined. Here, we show that vIRF-1 binds directly to mammalian autophagy-related gene 8 (ATG8) proteins, preferentially GABARAPL1 in infected cells, in an LC3-interacting region (LIR)-independent manner. Accordingly, we identified key residues in vIRF-1 and GABARAPL1 required for mutual interaction and demonstrated that the interaction is essential for mitophagy activation and HHV-8 productive replication. Interestingly, the mitophagy receptor NIX promotes vIRF-1-GABARAPL1 interaction, and NIX/vIRF-1-induced mitophagy is significantly inhibited in GABARAPL1-deficient cells. Moreover, a vIRF-1 variant defective in GABARAPL1 binding substantially loses the ability to induce vIRF-1/NIX-induced mitophagy. These results suggest that NIX supports vIRF-1 activity as a mitophagy mediator. In addition, we found that NIX promotes vIRF-1 aggregation and stabilizes aggregated vIRF-1. Together, these findings indicate that vIRF-1 plays a role as a viral mitophagy mediator that can be activated by a cellular mitophagy receptor. Author summary: In addition to their role in energy metabolism, mitochondria act as platforms for antiviral signaling that leads to apoptosis and type I interferon expression in response to virus infection. However, for their benefit, viruses have evolved strategies to attenuate mitochondria-mediated antiviral signaling, including selective autophagy of mitochondria (termed mitophagy) that eliminates dysfunctional or superfluous mitochondria. We previously demonstrated that mitochondria-localized viral interferon regulatory factor 1 (vIRF-1) plays a role in the activation of mitophagy during human herpesvirus 8 (HHV-8) via interaction with the mitophagy proteins NIX and TUFM, thereby inhibiting antiviral responses and contributing to productive replication. However, it remains to elucidate the precise molecular mechanisms and regulation of vIRF-1-mediated mitophagy, particularly interactions with the core autophagy machinery. In this report, we discover that vIRF-1 interacts selectively with GABARAPL1 among ATG8 proteins in a non-canonical manner in lytically HHV-8-infected cells, and the mitophagy receptor NIX promotes vIRF-1 oligomerization and interaction with the ATG8 protein. Our results reveal the presence of a functional complex of vIRF-1/NIX/GABARAPL1 for the promotion of mitophagy and also provide the first evidence of the post-translational regulation of viral mitophagic protein by cellular mitophagy receptor. [ABSTRACT FROM AUTHOR]

The human T-cell leukemia virus type 1 (HTLV-1) Tax protein hijacks the host ubiquitin machinery to activate IκB kinases (IKKs) and NF-κB and promote cell survival; however, the key ubiquitinated factors downstream of Tax involved in cell transformation are unknown. Using mass spectrometry, we undertook an unbiased proteome-wide quantitative survey of cellular proteins modified by ubiquitin in the presence of Tax or a Tax mutant impaired in IKK activation. Tax induced the ubiquitination of 22 cellular proteins, including the anti-apoptotic BCL-2 family member MCL-1, in an IKK-dependent manner. Tax was found to promote the nondegradative lysine 63 (K63)-linked polyubiquitination of MCL-1 that was dependent on the E3 ubiquitin ligase TRAF6 and the IKK complex. Tax interacted with and activated TRAF6, and triggered its mitochondrial localization, where it conjugated four carboxyl-terminal lysine residues of MCL-1 with K63-linked polyubiquitin chains, which stabilized and protected MCL-1 from genotoxic stress-induced degradation. TRAF6 and MCL-1 played essential roles in the survival of HTLV-1 transformed cells and the immortalization of primary T cells by HTLV-1. Therefore, K63-linked polyubiquitination represents a novel regulatory mechanism controlling MCL-1 stability that has been usurped by a viral oncogene to precipitate cell survival and transformation. [ABSTRACT FROM AUTHOR]

A voltammetric enzyme electrode was developed based on nicotinamide-independent trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), which catalyses the oxidation of trimethylamine (TMA) to dimethylamine and formaldehyde. A quaternized osmium hydrogel polymer, poly(vinylimidazole-[Os(4,4′-dimethyl-2,2′-bipyridine)Cl]) with ethylamine (PVI-Os-EA), was prepared as a potential redox mediator in an electrochemical biosensor. TMA was detected using TMADH that was co-immobilized with an osmium hydrogel polymer on electrodeposited gold nanoparticles (Au-NPs) on screen-printed carbon electrodes (SPCEs). The Au-NPs deposited onto SPCEs provided about a three times higher electrochemical response compared to that of a planar gold electrode. As TMA was catalyzed by wired TMADH, the electrical signal was monitored at 0.3 V versus Ag/AgCl by cyclic voltammetry and chronoamperometry. The anode currents increased linearly in proportion to the TMA concentration over the 0 ∼ 2.5 mM range with a detection limit of 1 μM ( R = 0.9972). [ABSTRACT FROM AUTHOR]

Abstract: Human herpesvirus 8 (HHV-8) is associated with Kaposi''s sarcoma (KS), an endothelial cell lesion believed to be initiated and driven primarily by cytokine dysregulation. Among the viral proteins suspected as contributing to viral pathogenesis is the lytically expressed viral G protein-coupled receptor (vGPCR), which can induce various cellular cytokines. CC ligand-2 (CCL2/MCP-1) is a vGPCR-regulated angiogenic chemokine found at elevated levels in KS lesions and induced by HHV-8 infection of endothelial cells. Here we show that vGPCR induces CCL2 in endothelial cells via activation of C/EBPβ and that vGPCR and C/EBPβ are critical components of CCL2 induction by HHV-8 infection of endothelial cultures. To our knowledge, this is the first report of vGPCR-mediated cytokine induction, and its characterization, in the context of virus infection. Our results identify a mechanism by which vGPCR can contribute, in a host cell shutoff-independent manner, to viral pathogenesis. [Copyright &y& Elsevier]

Urinary hippuric acid (HA), of molecular weight 180 Da, is one of the major metabolites in toluene-exposed humans and is a major biological indicator. Simple and ubiquitous monitoring of exposure to toluene is very important in occupational health care, and a microfluidic chip-based electrochemical immunoassay for rapid and quantitative detection of HA in human urine is proposed in this paper. The system employs a conjugate of ferrocene (Fc) and hippuric acid (HA). The competition between hippuric acid (HA) and the ferrocene-hippuric acid complex (Fc-Lys-HA) to bind with a HA antibody coated onto polybeads generated electrical signals proportional to the HA concentration in the range of 0–40 mg mL−1. All the complicated HA detection processes were integrated on the single microfluidic platform. The quantitative advantages of our HA detection chip are as follows: (1) the total chip size was reduced to 3.0 × 2.0 × 0.5 cm and is small enough to be portable, (2) the assay time took 1 min, and is shorter than that of conventional electrochemical HA immunoassay systems (about 20 min) and (3) 40 μL of the sample solution was enough to detect HA in the range of 0–40 mg mL−1, which is enough range to be used for the point-of-care system. In addition, we suggest the improved chip-based HA assay method by the combination of electrochemical and enzymatic amplification processes for the detection of greater electrical signals. The sensitivity of the combined method was increased about three times compared to that of the non-enzymatic process. [ABSTRACT FROM AUTHOR]

PELP1 (proline-, glutamic acid-, and leucine-rich protein 1) has been recognized as a coactivator of estrogen receptor (ER)-recruiting p300/CREB-binding protein histone acetyltransferase to the target chromosome. The present study shows that PELP1 does indeed coactivate ER-mediated transcription but also serves as a corepressor of other nuclear hormone receptors (NR)- and non-NR sequence-specific transcription factors tested, including GR, Nut77, AP1, NF-κB, and TCF/SRF. PELP1 expression also retarded the proliferation of mouse fibroblast cell lines in which there was no detectable ER. This was due, at least in part, to the suppressed activation of serum-response genes such as c-fos that in turn resulted from the blocked histone hyperacetylation of nucleosomes containing the c-fos promoter region. The N-terminal leucine-abundant region of PELP1 was observed to interact with HDAC2 and exhibited repressive activity when tethered to the chromatin. In addition, the C-terminal glutamic acid-abundant region bound to the hypoacetylated histones H3 and H4 and prevented them from becoming substrates of histone acetyltransferase. Thus PELP1 promotes and maintains the hypoacetylated state of histones at the target genomic site, and ER binding reverses its role to hyperacetylate histones through an as yet unidentified mechanism. [ABSTRACT FROM AUTHOR]

Biofuel cells (BFCs) offer an eco‐friendly route to convert biochemical energy into electricity. However, their performance is hindered by insufficient enzyme immobilization as well as limited electron transfer within the enzymatic electrode. While the incorporation of redox mediators (RMs) into enzyme layers has been shown to improve BFC performance through enhanced electron transfer, progress has plateaued in the last decade. Herein, a major breakthrough is presented realized by a novel strategy that exploits electron transfer‐intensified RM layers. Metal nanoparticles covalently bridged between neighboring RMs facilitate electron transfer ubiquitously. Electron transfer characteristics are enhanced not only within the RM layers themselves, but also at the glucose oxidase (GOx)/host electrode and GOx/GOx interfaces. This leads to a remarkable performance boost in the enzymatic anode. A hybrid BFC constructed with innovative anode and Pt‐based cathode exhibits a striking combination of high power output (2.3 and 8.5 mW cm−2 at 10 and 300 mmol L−1 glucose, respectively) and exceptional operational stability (≈80% and 47% power retention after 10 days and 1 month, respectively), outperforming all previously reported BFCs by a significant margin. [ABSTRACT FROM AUTHOR]

Selective autophagy has emerged as a key mechanism of quality and quantity control responsible for the autophagic degradation of specific subcellular organelles and materials. In addition, a specific type of selective autophagy (xenophagy) is also activated as a line of defense against invading intracellular pathogens, such as viruses. However, viruses have evolved strategies to counteract the host's antiviral defense and even to activate some proviral types of selective autophagy, such as mitophagy, for their successful infection and replication. This review discusses the current knowledge on the regulation of selective autophagy by human herpesviruses. [ABSTRACT FROM AUTHOR]

A study was conducted to evaluate the effectiveness and safety of a topical emollient in treating mild-to-moderate atopic dermatitis (AD) in adolescents and adults. The study involved 100 participants and lasted for 24 weeks. The results showed that the group using the emollient had significant improvements in various measures compared to the placebo group. No safety concerns were reported. However, the study had limitations and further research is needed to explore other aspects of AD and the effects of the emollient. Overall, the emollient shows promise as a therapeutic option for long-term care of AD patients. [Extracted from the article]

Human T-cell leukemia virus type 1 (HTLV-1) is the etiological agent of adult T-cell leukemia/lymphoma (ATLL), and the neurological disease HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The HTLV-1 Tax protein persistently activates the NF-κB pathway to enhance the proliferation and survival of HTLV-1 infected T cells. Lysine 63 (K63)-linked polyubiquitination of Tax provides an important regulatory mechanism that promotes Tax-mediated interaction with the IKK complex and activation of NF-κB; however, the host proteins regulating Tax ubiquitination are largely unknown. To identify new Tax interacting proteins that may regulate its ubiquitination we conducted a yeast two-hybrid screen using Tax as bait. This screen yielded the E3/E4 ubiquitin conjugation factor UBE4B as a novel binding partner for Tax. Here, we confirmed the interaction between Tax and UBE4B in mammalian cells by co-immunoprecipitation assays and demonstrated colocalization by proximity ligation assay and confocal microscopy. Overexpression of UBE4B specifically enhanced Tax-induced NF-κB activation, whereas knockdown of UBE4B impaired Tax-induced NF-κB activation and the induction of NF-κB target genes in T cells and ATLL cell lines. Furthermore, depletion of UBE4B with shRNA resulted in apoptotic cell death and diminished the proliferation of ATLL cell lines. Finally, overexpression of UBE4B enhanced Tax polyubiquitination, and knockdown or CRISPR/Cas9-mediated knockout of UBE4B attenuated both K48- and K63-linked polyubiquitination of Tax. Collectively, these results implicate UBE4B in HTLV-1 Tax polyubiquitination and downstream NF-κB activation. Author summary: Infection with the retrovirus HTLV-1 leads to the development of either CD4+CD25+ leukemia/lymphoma (ATLL) or a demyelinating neuroinflammatory disease (HAM/TSP) in a subset of infected individuals. The HTLV-1 Tax protein is a regulatory protein which regulates viral gene expression and persistently activates cellular signaling pathways such as NF-κB to drive the clonal expansion and longevity of HTLV-1 infected CD4+ T cells. Polyubiquitination of Tax is a key mechanism of NF-κB activation by assembling and activating IκB kinase (IKK) signaling complexes; however, the host factors regulating Tax ubiquitination have remained elusive. Here, we have identified the E3/E4 ubiquitin conjugation factor UBE4B as a novel Tax binding protein that promotes both K48- and K63-linked polyubiquitination of Tax. Knockdown or knockout of UBE4B impairs Tax-induced NF-κB activation and triggers apoptosis of HTLV-1-transformed cells. Therefore, UBE4B is an integral host factor that supports HTLV-1 Tax polyubiquitination, NF-κB activation and cell survival. [ABSTRACT FROM AUTHOR]

High performance with stability, easy-handling electrodes, and biofluid-flow controllable system with mechanical strength of the biofuel cell can be considered as the critical issues for future human body implant. These three challenges are sufficiently considered by using the effective platform regarding the high surface area from multi-walled carbon nanotube-conducting polymer with poly(3,4-ethylenedioxythiophene), and size/shape dependent flexible yarn electrodes for the implantation of biofuel cell. High power biofuel cell of mW cm −2 range in physiological condition (low glucose-containing phosphate buffered saline solution and human blood serum) controlling the stirring degree is also first demonstrated for future implantation in this study. Biofuel cells for future implantation in human body vitally require long-term stability and high power outputs. We have demonstrated that a high-surface area yarn-based biofuel cell retained over 70% of its initial power output after an extended 20 days period of continuous operation in human blood serum, while delivering a power density of ∼1.0 mW cm −2 . Subsequently, our enhanced enzymatic biofuel cell system would be potentially used as an innovative power source for the next generation implantable electronics. [ABSTRACT FROM AUTHOR]

The human skin sebum suggests that it (along with other epidermal surface lipids) plays a role in skin barrier formation, the moderation of cutaneous inflammation, and antimicrobial defense. Various methods have been developed for collecting and measuring skin sebum. We tested methods of detection using "color intensity", by staining the skin casual sebum. This process was conducted in three steps; first, the selection of materials for sebum collection; second, staining the collected sebum; third, the development of a device that can measure the level of stained sebum. A plastic film was used to effectively collect sebum that increased with the replacement time of the sebum. In addition, the collected sebum was stained with Oil Red O (ORO) and checked with RGB; as a result, the R2 value was higher than 0.9. It was also confirmed that the correlation value was higher than 0.9 in the comparison result with Sebumeter®, which is a common standard technology. Finally, it was confirmed that the R2 value was higher than 0.9 in the detection value using the sensor. In conclusion, we have proven the proof of concept (PoC) for this method, and we would like to introduce an effective sebum measurement method that differs from the existing method. [ABSTRACT FROM AUTHOR]

The electrochemical-based detection of glucose is widely used for diagnostic purposes and is mediated by enzyme-mediated signal transduction mechanisms. For such applications, recent attention has focused on utilizing the oxygen-insensitive glucose dehydrogenase (GDH) enzyme in place of the glucose oxidase (GOx) enzyme, which is sensitive to oxygen levels. Currently used Ru-based redox mediators mainly work with GOx, while Ru(dmo–bpy)2Cl2 has been proposed as a promising mediator that works with GDH. However, there remains an outstanding need to improve Ru(dmo–bpy)2Cl2 attachment to electrode surfaces. Herein, we report the use of polydopamine-functionalized multi-walled carbon nanotubes (PDA-MWCNTs) to effectively attach Ru(dmo–bpy)2Cl2 and GDH onto screen-printed carbon electrodes (SPCEs) without requiring a cross–linker. PDA-MWCNTs were characterized by Fourier transform infrared (FT–IR) spectroscopy, Raman spectroscopy, and thermal gravimetric analysis (TGA), while the fabrication and optimization of Ru(dmo–bpy)2Cl2/PDA-MWCNT/SPCEs were characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) measurements. The experimental results demonstrate a wide linear range of glucose-concentration-dependent responses and the multi-potential step (MPS) technique facilitated the selective detection of glucose in the presence of physiologically relevant interfering species, as well as in biological fluids (e.g., serum). The ease of device fabrication and high detection performance demonstrate a viable pathway to develop glucose sensors based on the GDH enzyme and Ru(dmo–bpy)2Cl2 redox mediator and the sensing strategy is potentially extendable to other bioanalytes as well. [ABSTRACT FROM AUTHOR]

Expression of Kaposi's sarcoma herpesvirus vFLIP, a potent activator of NFkB signaling, promotes latency. Inhibition of NFkB signaling promotes lytic reactivation. We previously reported that lytic inducer, RTA, inhibits vFLIP induced NFkB signaling by inducing the degradation of vFLIP via the proteasome. Here we report that the cellular ubiquitin ligase, Itch, is required for RTA induced degradation of vFLIP. Expression of either Itch targeting shRNA or a dominant negative mutant of the ubiquitin ligase both increased the stability of vFLIP in the presence of RTA. Itch potently ubiquitinated vFLIP in vivo and in vitro . We provide evidence for interaction between RTA, vFLIP and Itch and we identified an RTA resistant mutant of vFLIP that is unable to interact with Itch. These observations contribute to our understanding of how RTA counteracts the activities of vFLIP. [ABSTRACT FROM AUTHOR]

Direct electron transfer (DET) glucose sensors are third-generation biosensors, which allow the enzyme to directly transfer electrons by glucose oxidation without a mediator. In previous studies, many researchers have focused on the glucose oxidase (GOx)-based DET glucose sensor; however, an efficient DET glucose sensor using flavin adenine dinucleotide (FAD)-glucose dehydrogenase (GDH) has not been developed. In the present study, we bound FAD-GDH and multi-walled carbon nanotubes (MWCNT), using chitosan (CS) compounds that support hydrogen bonding, van der Waals forces, and 3D structural adsorption. The GDH/CS-MWCNT-5 composite, which had a GDH concentration of 75 wt%, was physically adsorbed on screen-printed carbon electrodes (SPCEs), and cyclic voltammetry indicated that its oxidation and reduction peaks were at − 0.422 V and − 0.543 V (vs Ag/AgCl), respectively. In addition, the electrochemical results confirmed that the prepared GDH/CS-MWCNT/SPCEs were not affected by other interfering substances or oxygen at pH of 7. The GDH/CS-MWCNT/SPCEs displayed oxidation catalytic currents, which increased according to glucose concentrations across a range of 0–5.5 mM. Finally, the short-term stability of glucose, assessed for 10 days, was maintained at 80% of the GDH enzyme activity for 6 days, and it reduced to 50% of the initial activity for the remaining 4 days. Here, we illustrate the potential utility of the FAD-GDH-based DET method in continuous glucose monitoring sensors. Diagram of the GDH/CS-MWCNT fabrication process and glucose sensing [Display omitted] • CS-MWCNTs were conjugated with FAD-GDH by hydrogen bonding. • CS-MWCNTs were optimized with GDH for DET glucose sensing. • GDH/CS-MWCNT showed good response with glucose under interfering substances. [ABSTRACT FROM AUTHOR]

Abstract: We have screened 356 libraries of Korean herbal plant extracts to find potential anti-obesity drugs. We employed the recently developed fluorescence polarization high throughput screening (FP HTS) assays of human neuropeptide FF (NPFF) receptors in 384-well microtiter plates. The primary hits were cherry-picked from the libraries and further analyzed by secondary displacement curve assays, in vitro GTPγS binding assays and cell-based CRE luciferase reporter assays. Agonists of NPFF receptors showed biphasic affinity curves while the antagonist, BIBP 3226, gave a monophasic affinity curve in competitive binding assays. We isolated and characterized two agonists of human NPFF2 receptor, PC 314 with K i of 1.42μM, and PC 315 with K i of 2.17μM from Schizandra chinensis. PC 314 and PC 315 have been characterized as benzoylgomisin Q (M.W. 552) and gomisin G (M.W. 536). We report that PC 314 and PC 315 are the first non-peptide, natural compounds, which bind to human NPFF2 receptors with good affinity. PC 314 and PC 315 inhibit forskolin-stimulated luciferase expression when CHO cells are co-transfected with NPFF2 receptor and CRE reporter vector. They possess the pharmacological and functional profiles of full agonists. The FP HTS system provides a specific, sensitive and reproducible methodology for studying and screening NPFF receptor ligands. [Copyright &y& Elsevier]

Protein kinase C δ (PKCδ) plays an important role in the regulation of apoptosis in response to diverse anticancer agents. PKCδ is cleaved irreversibly to a catalytically active fragment in response to apoptotic stimuli; however, little information is available about the regulation of PKCδ gene expression. In this study, we found that the amount of steady-state PKCδ mRNA and protein was increased by etoposide in mouse L1210 leukemia cells. The transcriptional rate of the PKCδ gene and the stability of PKCδ mRNA were increased by treatment with etoposide, resulting in the accumulation of PKCδ protein. Rottlerin inhibited etoposide-induced PKCδ gene expression significantly, while Go6976, LY294002 and PD98059 had no effect. Further, both stable and adenovirus-mediated expression of a dominant negative PKCδ(KR) abrogated etoposide-induced PKCδ expression. Etoposide-stimulated PKCδ transcription but not PKCδ mRNA stability was blocked completely by pretreatment with rottlerin. Our data reveal a novel mechanism whereby PKCδ gene is regulated at the transcriptional and post-transcriptional level in the L1210 leukemia cell line. [Copyright &y& Elsevier]

The electrochemical-based detection of uric acid (UA) is widely used for diagnostic purposes. However, various interfering species such as ascorbic acid, dopamine, and glucose can affect electrochemical signals, and hence there is an outstanding need to develop improved sensing platforms to detect UA with high selectivity. Herein, we report a pentagonal mediator-based non-enzymatic electrochemical biosensing platform to selectively measure UA in the presence of interfering species. The working electrode was fabricated by electrodepositing polymerized 1-vinylimidazole (PVI), which has an imidazole ligand, onto indium tin oxide (ITO), and then conjugating nickel ions to the PVI-coated ITO electrode. Electrode performance was characterized by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) measurements and integrated together with pentacyanoammineferrate, which can bind to the amine groups of UA and function as an electron transferring mediator. The experimental results showed a wide linear range of UA concentration-dependent responses and the multi-potential step (MPS) technique facilitated selective detection of UA in the presence of physiologically relevant interfering species. Altogether, these findings support that pentacyanoammineferrate-based non-enzymatic electrodes are suitable biosensing platforms for the selective measurement of UA, and such approaches could potentially be extended to other bioanalytes as well. [ABSTRACT FROM AUTHOR]

(A) Schematic illustration and SEM and TEM images of CS-shell/MWCNT-core nanohybrids structure. (B) Photograph of commercial alumina-based humidity sensor electrode. (C) Photograph of the humidity sensor electrode coated by CS-MWCNT nanohybrids and SEM image showing electrodes interconnected by CS-MWCNT nanohybrids. • Fabrication of chitosan-MWCNT core–shell-structured nanohybrid (CS-MWCNT) on a gold electrode as an electrochemical humidity sensor. • CS-MWCNT nanohybrid fibers exhibiting low resistance without moisture. • Remarkable results in the humidity response with an efficient humidity sensing range of 30–90%. Multi-walled carbon nanotubes (MWCNTs) were fabricated on a gold electrode to develop a resistance-sensing humidity sensor with low resistance. Core-shell-structured nanohybrids were prepared using Chitosan-MWCNT to obtain good hydrophilicity, resulting in a highly responsive humidity sensor. We also measured resistance, which changes due to hydrogen bonding between the amine group of chitosan and H 2 O. The structure of the fabricated chitosan-MWCNT core–shell-structured nanohybrid was measured. Furthermore, we determined the highest response and linearity of the CS-MWCNT25 nanohybrids based on the number of coats on the electrode and the composition ratio of chitosan and MWCNTs. The fabricated CS-MWCNT25 nanohybrid humidity sensor exhibited good electrical efficiency without voltage or frequency dependence and a low-temperature dependency with a temperature coefficient (RH/°C) of 0.187%. Finally, a fast recovery time of 40 s between 30% and 100% RH, a maximum humidity hysteresis of less than 0.300 ± 0.001%, and long-term stability for 2 months indicated that this is an excellent humidity sensor. [ABSTRACT FROM AUTHOR]

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

A glucose-reactive enzyme-based biofuel cell system (EBFC) was recently introduced in the scientific community for biomedical applications, such as implantable artificial organs and biosensors for drug delivery. Upon direct contact with tissues or organs, an implanted EBFC can exert effects that damage or stimulate intact tissue due to its byproducts or generated electrical cues, which have not been investigated in detail. Here, we perform a fundamental cell culture study using a glucose dehydrogenase (GDH) as an anode enzyme and bilirubin oxidase (BOD) as a cathode enzyme. The fabricated EBFC had power densities of 15.26 to 38.33 nW/cm2 depending on the enzyme concentration in media supplemented with 25 mM glucose. Despite the low power density, the GDH-based EBFC showed increases in cell viability (~150%) and cell migration (~90%) with a relatively low inflammatory response. However, glucose oxidase (GOD), which has been used as an EBFC anode enzyme, revealed extreme cytotoxicity (~10%) due to the lethal concentration of H2O2 byproducts (~1500 µM). Therefore, with its cytocompatibility and cell-stimulating effects, the GDH-based EBFC is considered a promising implantable tool for generating electricity for biomedical applications. Finally, the GDH-based EBFC can be used for introducing electricity during cell culture and the fabrication of organs on a chip and a power source for implantable devices such as biosensors, biopatches, and artificial organs. [ABSTRACT FROM AUTHOR]