Palladium-Percolated Networks Enabled by Low Loadings of Branched Nanorods for Enhanced H2 Separations.

Nanoparticles (NPs) at high loadings are often used in mixed matrix membranes (MMMs) to improve gas separation properties, but they can lead to defects and poor processability that impede membrane fabrication. Herein, it is demonstrated that branched nanorods (NRs) with controlled aspect ratios can significantly reduce the required loading to achieve superior gas separation properties while maintaining excellent processability, as demonstrated by the dispersion of palladium (Pd) NRs in polybenzimidazole for H2 /CO2 separation. Increasing the aspect ratio from 1 for NPs to 40 for NRs decreases the percolation threshold volume fraction by a factor of 30, from 0.35 to 0.011. An MMM with percolated networks formed by Pd NRs at a volume fraction of 0.039 exhibits H2 permeability of 110 Barrer and H2 /CO2 selectivity of 31 when challenged with simulated syngas at 200 °C, surpassing Robeson's upper bound. This work highlights the advantage of NRs over NPs and nanowires and shows that right-sizing nanofillers in MMMs is critical to construct highly sieving pathways at minimal loadings. This work paves the way for this general feature to be applied across materials systems for a variety of chemical separations.

Potentiometric Biosensors Based on Molecular-Imprinted Self-Assembled Monolayer Films for Rapid Detection of Influenza A Virus and SARS-CoV-2 Spike Protein.

Rapid, yet accurate and sensitive testing has been shown to be critical in the control of spreading pandemic diseases such as COVID-19. Current methods which are highly sensitive and can differentiate different strains are slow and cannot be conveniently applied at the point of care. Rapid tests, meanwhile, require a high titer and are not sufficiently sensitive to discriminate between strains. Here, we report a rapid and facile potentiometric detection method based on nanoscale, three-dimensional molecular imprints of analytes on a self-assembled monolayer (SAM), which can deliver analyte-specific detection of both whole virions and isolated proteins in microliter amounts of bodily fluids within minutes. The detection substrate with nanoscale inverse surface patterns of analytes formed by a SAM identifies a target analyte by recognizing its surface nano- and molecular structures, which can be monitored by temporal measurement of the change in substrate open-circuit potential. The sensor unambiguously detected and differentiated H1N1 and H3N2 influenza A virions as well as the spike proteins of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and Middle-East respiratory syndrome (MERS) coronavirus in human saliva with limits of detection reaching 200 PFU/mL and 100 pg/mL for the viral particles and spike proteins, respectively. The demonstrated speed and specificity of detection, combined with a low required sample volume, high sensitivity, ease of potentiometric measurement, and simple sample collection and preparation, suggest that the technique can be used as a highly effective point-of-care diagnostic platform for a fast, accurate, and specific detection of various viral pathogens and their variants.

A fully integrated bacterial pathogen detection system based on count-on-a-cartridge platform for rapid, ultrasensitive, highly accurate and culture-free assay.

Rapid, sensitive and accurate point-of-care-testing (POCT) of bacterial load from a variety of samples can help prevent human infections caused by pathogenic bacteria and mitigate their spreading. However, there is an unmet demand for a POCT device that can detect extremely low concentrations of bacteria in raw samples. Herein, we introduce the 'count-on-a-cartridge' (COC) platform for quantitation of the food-borne pathogenic bacteria Staphylococcus aureus. The system comprised of magnetic concentrator, sensing cartridge and fluorescent image reader with a built-in counting algorithm facilitated fluorescent microscopic bacterial enumeration in user-convenient manner with high sensitivity and accuracy within a couple of hours. The analytical performance of this assay is comparable to that of a standard plate count. The COC assay shows a sensitivity of 92.9% and specificity of 100% performed according to global microbiological criteria for S. aureus which is acceptable below 100 CFU/g in the food matrix. This culture-independent, rapid, ultrasensitive and highly accurate COC assay has great potential for places where prompt bacteria surveillance is in high demand.

Fully Stretchable Capillary Microfluidics-Integrated Nanoporous Gold Electrochemical Sensor for Wearable Continuous Glucose Monitoring.

Biosensor systems for wearable continuous monitoring are desired to be developed into conformal patch platforms. However, developing such patches is very challenging owing to the difficulty of imparting materials and components with both high stretchability and high performance. Herein, we report a fully stretchable microfluidics-integrated glucose sensor patch comprised of an omnidirectionally stretchable nanoporous gold (NPG) electrochemical biosensor and a stretchable passive microfluidic device. A highly electrocatalytic NPG electrode was formed on a stress-absorbing 3D micropatterned polydimethylsiloxane (PDMS) substrate to confer mechanical stretchability, high sensitivity, and durability in non-enzymatic glucose detection. A thin, stretchable, and tough microfluidic device was made by embedding stretchable cotton fabric as a capillary into a thin polyurethane nanofiber-reinforced PDMS channel, enabling collection and passive, accurate delivery of sweat from skin to the electrode surface, with excellent replacement capability. The integrated glucose sensor patch demonstrated excellent ability to continuously and accurately monitor the sweat glucose level.

A smartphone fluorescence imaging-based mobile biosensing system integrated with a passive fluidic control cartridge for minimal user intervention and high accuracy.

A key challenge for realizing mobile device-based on-the-spot environmental biodetection is that a biosensor integrated with a fluid handling sensor cartridge must have acceptable accuracy comparable to that of conventional standard analytical methods. Furthermore, the user interface must be easy to operate, technologically plausible, and concise. Herein, we introduced an advanced smartphone imaging-based fluorescence microscope designed for Hg2+ monitoring by utilizing a biosensor cartridge that reduced user intervention via time-sequenced passive fluid handling. The cartridge also employed a metal-nanostructured plastic substrate for complementing the fluorescence signal output; this helped the realization of high-accuracy detection, in which a ratiometric dual-wavelength detection method was applied. Using 30 samples of Hg2+-spiked wastewater, we showed that our device, which has a detection limit of ∼1 pM, can perform analytical assays accurately. The detection results from our method were in good linearity and agreement with those of conventional standard methods. We conclude that the integration of a simple-to-use biosensor cartridge, fluorescence signal-enhancing substrate, dual-wavelength detection, and quantitative image data processing on a smartphone has great potential to make any population accessible to small-molecule detection, which has been performed in centralized laboratories for environmental monitoring.

MicroRNA expression profiling of adult hippocampal neural stem cells upon cell death reveals an autophagic cell death-like pattern.

Adult hippocampal neural (HCN) stem cells promptly undergo irreversible autophagic cell death (ACD) if deprived of insulin in culture. Small, non-coding microRNAs (miRNA) play an important role in regulating biological processes, including proliferation and cell death. However, there have been no reports thus far regarding miRNA involvement in the induction of adult HCN stem cell death under insulin-deprived conditions, for which we performed a microarray-based analysis to examine the expression signature of miRNAs in adult rat HCN stem cells. Three independent specimens per culture condition either with or without insulin were prepared and a miRNA microarray analysis carried out. A total of 12 exhibited significantly altered expression levels upon cell death due to the absence of insulin when compared to HCN stem cells cultured with insulin present (cut-off limit; p < 0.05 and fold-change >1.3) The resulting volcano plot showed that, among these miRNAs, seven were upregulated and five were downregulated. The upregulated miRNAs were capable of modulating HCN stem cell death. Caspase-3 activity analysis, LC3 conversion, and TEM of autophagosome formation consistently suggested that ACD, not apoptosis, was most likely the mechanism affecting HCN cell death. As such, we have come to term these miRNAs, "HCN stem cell-specific autophagic cell death regulators." Taken together, our data suggest that the miRNA expression profile of HCN stem cells is altered during ACD occurring due to insulin deprivation and that differentially expressed miRNAs are involved in HCN stem cell viability. Detailed explorations of the underlying mechanisms regarding HCN stem cell viability modulation by these miRNAs would be beneficial in further understanding the physiological features of adult HCN stem cells and are currently being investigated.

Culture-free, highly sensitive, quantitative detection of bacteria from minimally processed samples using fluorescence imaging by smartphone.

A critical unmet need in the diagnosis of bacterial infections, which remain a major cause of human morbidity and mortality, is the detection of scarce bacterial pathogens in a variety of samples in a rapid and quantitative manner. Herein, we demonstrate smartphone-based detection of Staphylococcus aureus in a culture-free, rapid, quantitative manner from minimally processed liquid samples using aptamer-functionalized fluorescent magnetic nanoparticles. The tagged S. aureus cells were magnetically captured in a detection cassette, and then fluorescence was imaged using a smartphone camera with a light-emitting diode as the excitation source. Our results showed quantitative detection capability with a minimum detectable concentration as low as 10 cfu/ml by counting individual bacteria cells, efficiently capturing S. aureus cells directly from a peanut milk sample within 10 min. When the selectivity of detection was investigated using samples spiked with other pathogenic bacteria, no significant non-specific detection occurred. Furthermore, strains of S. aureus from various origins showed comparable results, ensuring that the approach can be widely adopted. Therefore, the quantitative fluorescence imaging platform on a smartphone could allow on-site detection of bacteria, providing great potential assistance during major infectious disease outbreaks in remote and resource-limited settings.

A smartphone imaging-based label-free and dual-wavelength fluorescent biosensor with high sensitivity and accuracy.

The accuracy of a bioassay based on smartphone-integrated fluorescent biosensors has been limited due to the occurrence of false signals from non-specific reactions as well as a high background and low signal-to-noise ratios for complementary metal oxide semiconductor image sensors. To overcome this problem, we demonstrate dual-wavelength fluorescent detection of biomolecules with high accuracy. Fluorescent intensity can be quantified using dual wavelengths simultaneously, where one decreases and the other increases, as the target analytes bind to the split capture and detection aptamer probes. To do this, we performed smartphone imaging-based fluorescence microscopy using a microarray platform on a substrate with metal-enhanced fluorescence (MEF) using Ag film and Al2O3 nano-spacer. The results showed that the sensitivity and specificity of the dual-wavelength fluorescent quantitative assay for the target biomolecule 17-ß-estradiol in water were significantly increased through the elimination of false signals. The detection limit was 1pg/mL and the area under the receiver operating characteristic curve of the proposed assay (0.922) was comparable to that of an enzyme-linked immunosorbent assay (0.956) from statistical accuracy tests using spiked wastewater samples. This novel method has great potential as an accurate point-of-care testing technology based on mobile platforms for clinical diagnostics and environmental monitoring.

Seesawed fluorescence nano-aptasensor based on highly vertical ZnO nanorods and three-dimensional quantitative fluorescence imaging for enhanced detection accuracy of ATP.

Probe-mediated fluorescence biosensing methods based on spectrophotometry still have limitations such as detection inaccuracy caused by the occurrence of false signals and lack of simultaneous qualitative and quantitative read-outs with an ultra-low detection limit. Herein, we describe a novel seesawed fluorescence detection strategy based on dual-colour imaging-based quantitation in which the green fluorescence of the capture aptamer decreases and the red fluorescence of the detection aptamer increases simultaneously upon their respective interactions with the target biomolecule. This approach enhances detection accuracy through facilitating identification of probable false-positives in biological samples. Furthermore, combining the seesawed detection scheme with three-dimensional imaging of fluorescence signal enhanced by highly vertical ZnO nanorods increases signal-to-noise ratio, which addresses the limited performance of digital cameras and, in turn, enhances sensitivity and dynamic range. This simple, robust, scalable, imaging-based and label-free fluorescence method allows highly specific and sensitive quantification of biomolecules with excellent reliability.

Effect of Cervical Interlaminar Epidural Steroid Injection: Analysis According to the Neck Pain Patterns and MRI Findings.

BACKGROUND: It is widely accepted that cervical interlaminar steroid injection (CIESI) is more effective in treating radicular pain than axial neck pain, but without direct comparison. And the differences of effect after CIESI according to MRI findings are inconsistent. In this retrospective study, we evaluated the therapeutic response of CIESI according to pain sites, durations, MRI findings, and other predictive factors altogether, unlike previous studies, which evaluated them separately. METHODS: The medical records of 128 patients who received fluoroscopy guided CIESI were analyzed. We evaluated the therapeutic response (more than a 50% reduction on the visual analog scale [VAS] by their second visit) after CIESI by (1) pain site; neck pain without radicular pain/radicular pain with or without neck pain, (2) pain duration; acute/chronic (more than 6 month), and (3) findings of MRI; herniated intervertebral disc (HIVD)/spinal stenosis, respectively and altogether. RESULTS: Eighty-eight patients (68%) responded to CIESI, and there were no significant differences in demographic data, initial VAS score, or laboratory findings. And there were no significant differences in the response rate relating to pain site, pain duration, or MRI findings, respectively. In additional analysis, acute radicular pain with HIVD patients showed significantly better response than chronic neck pain with spinal stenosis (P = 0.04). CONCLUSIONS: We cannot find any sole predictive factor of therapeutic response to the CIESI. But the patients having acute radicular pain with HIVD showed the best response, and those having other chronic neck pain showed the worst response to CIESI.

Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

Combined antitumor gene therapy with herpes simplex virus-thymidine kinase and short hairpin RNA specific for mammalian target of rapamycin.

A proof-of-concept study is presented using dual gene therapy that employed a small hairpin RNA (shRNA) specific for mammalian target of rapamycin (mTOR) and a herpes simplex virus-thymidine kinase (HSV-TK) gene to inhibit the growth of tumors. Recombinant adeno-associated virus (rAAV) vectors containing a mutant TK gene (sc39TK) were transduced into HeLa cells, and the prodrug ganciclovir (GCV) was administered to establish a suicide gene-therapy strategy. Additionally, rAAV vectors expressing an mTOR-targeted shRNA were employed to suppress mTOR-dependent tumor growth. GCV selectively induced death in tumor cells expressing TK, and the mTOR-targeted shRNA altered the cell cycle to impair tumor growth. Combining the TK-GCV system with mTOR inhibition suppressed tumor growth to a greater extent than that achieved with either treatment alone. Furthermore, HSV-TK expression and mTOR inhibition did not mutually interfere with each other. In conclusion, gene therapy that combines the TK-GCV system and mTOR inhibition shows promise as a novel strategy for cancer therapy.

Ultrarapid and ultrasensitive electrical detection of proteins in a three-dimensional biosensor with high capture efficiency.

The realization of a high-throughput biosensor platform with ultrarapid detection of biomolecular interactions and an ultralow limit of detection in the femtomolar (fM) range or below has been retarded due to sluggish binding kinetics caused by the scarcity of probe molecules on the nanostructures and/or limited mass transport. Here, as a new method for the highly efficient capture of biomolecules at extremely low concentration, we tested a three-dimensional (3D) platform of a bioelectronic field-effect transistor (bio-FET) with vertically aligned and highly dense one-dimensional (1D) ZnO nanorods (NRs) as a sensing surface capped by an ultrathin TiO2 layer for improved electrolytic stability on a chemical-vapor-deposited graphene (Gr) channel. The ultrarapid detection capability with a very fast response time (∼1 min) at the fM level of proteins in the proposed 3D bio-FET is primarily attributed to the fast binding kinetics of the probe-target proteins due to the small diffusion length of the target molecules to reach the sensor surface and the substantial number of probe molecules available on the largely increased surface area of the vertical ZnO NRs. This new 3D electrical biosensor platform can be easily extended to other electrochemical nanobiosensors and has great potential for practical applications in miniaturized biosensor integrated systems.

Correction of target-controlled infusion following wrong selection of emulsion concentrations of propofol.

BACKGROUND: We investigated the correction methods following wrong-settings of emulsion concentrations of propofol as a countermeasure against erroneous target-controlled infusions (TCI). METHODS: TCIs were started with targeting 4.0 µg/ml of effect-site concentration (Ceff) of propofol, and the emulsion concentrations were selected for 2.0% instead of 1.0% (FALSE1-2, n = 24), or 1.0% instead of 2.0% (FALSE2-1, n = 24). These wrong TCIs were corrected at 3 min after infusion start. During FALSE1-2, the deficit was filled up while injecting after equilibrium (n = 12), or while overriding (n = 12). During FALSE2-1, the overdose was evacuated while targeting Ceff (n = 12) or targeting plasma concentration (Cp) (n = 12). The gravimetrical measurements of TCI reproduced the Cp and Ceff using simulations. The reproduced Ceff at 3 min (Ceff-3min) and the time to be normalized within ± 5% of target Ceff (T±5%), were compared between the correction methods. RESULTS: During the wrong TCI, Ceff-3min was 1.98 ± 0.01 µg/ml in FALSE1-2, and 7.99 ± 0.05 µg/ml in FALSE2-1. In FALSE1-2, T±5% was significantly shorter when corrected while overriding (3.9 ± 0.25 min), than corrected after equilibrium (6.9 ± 0.05 min) (P < 0.001). In FALSE2-1, T±5% was significantly shorter during targeting Cp (3.6 ± 0.04 min) than targeting Ceff (6.7 ± 0.15 min) (P < 0.001). CONCLUSIONS: The correction methods, based on the pharmacokinetic and pharmacodynamic characteristics, could effectively and rapidly normalize the wrong TCI following erroneously selections of the emulsion concentration of propofol.

Administration of four different doses of gabapentin reduces awakening from breakthrough pain and adverse effects in outpatients with neuropathic pain during the initial titration.

BACKGROUND: Gabapentin is a safe and well-tolerated anticonvulsant with a wide therapeutic index, and it is used for neuropathic pain. The aim of this study was to compare previous dosing methods with the administration of four different doses of gabapentin while maintaining the same maximum daily dose for the safe administration of high doses of the medication. METHODS: THE SUBJECTS WERE OUTPATIENTS WITH VARIOUS NEUROPATHIC PAIN SYNDROMES, WITH AT LEAST TWO OF THE FOLLOWING SYMPTOMS: allodynia, burning pain, shooting pain, or hyperalgesia. The TID group received equal doses of gabapentin 3 times per day, while the QID group received 4 different doses of gabapentin per day. The pain score, frequency of breakthrough pain (BTP), severity and the duration of pain, sleep disturbance due to nocturnal pain, and adverse effects were recorded each day. RESULTS: The average daily pain score and sleep disturbance were significantly reduced in the QID group between days 3 and 10 of the experiment. The adverse effects of the medication were also reduced in the QID group. However, the frequency of BTP and severity and duration of pain were not significantly different between two groups. CONCLUSIONS: Administration of 4 different doses of gabapentin during the initial titration in outpatients with neuropathic pain resulted in a significant reduction in awakening from breakthrough pain and a reduction in the adverse effects of the medication.

Efficacy of stellate ganglion block in cholinergic urticaria with acquired generalized hypohidrosis.

Cholinergic urticaria with acquired generalized hypohidrosis, and its pathophysiology is not well known. Autoimmunity to sweat glands or to acetylcholine receptors on sweat glands has been mentioned as one of the possible etiologies. Systemic steroid therapy, antihistamines, anticholinergics, and avoidance of the stimulatory situations are recommended for treatment. We experienced a case of cholinergic urticaria with acquired generalized hypohidrosis in a patient who had no other associated disease, and the symptoms eased after repeated bilateral stellate ganglion block. Stellate ganglion block normalized the elevated sympathetic tone and may relieve symptoms in patients with this condition.

Gastroprotective and safety effects of WIN-34B, a novel treatment for osteoarthritis, compared to NSAIDs.

ETHNOPHARMACOLOGICAL RELEVANCE: The dried flowers of Lonicera japonica, also known as Japanese honeysuckle, and the dried root of Anemarrhena asphodeloides, the component herbs of WIN-34B, are traditionally used in Eastern medicine to treat various inflammatory conditions including arthritis. OBJECTIVE: To study the acute and chronic toxicities of WIN-34B and to compare its effects on gastric mucosa with those of diclofenac, a widely used NSAID, and celecoxib, a selective COX-2 inhibitor. MATERIALS AND METHODS: To investigate acute toxicity, we orally administered a single dose of 5,000 mg/kg WIN-34B to rats. To investigate chronic toxicity, we orally administered 500, 1000 or 2,000 mg/kg WIN-34B to rats daily for 13 weeks. To assess its effects on gastric mucosa, rats received either a single dose or repeated doses of WIN-34B (400, 1000, or 2,000 mg/kg), diclofenac (10, 40, or 80 mg/kg), celecoxib (100 or 1,000 mg/kg), or vehicle, after which samples of gastric mucosa were assessed grossly and histologically. We also measured tissue activity of myeloperoxidase and synthesis of eicosanoids, including prostaglandin E(2) (PGE(2)) and leukotriene B(4) (LTB(4)). To further assess its effects, we administered WIN-34B to rats either intraperitoneally or orally, measured gastric injury scores using a rat model of diclofenac-induced gastric injury, and measured eicosanoid synthesis. RESULTS: WIN-34B showed no signs of acute or chronic toxicity in terms of general behavior, gross appearance of the internal organs, blood chemistry, or mortality. WIN-34B did not cause significant gastric mucosal damage after single or repeated doses. In contrast, diclofenac and celecoxib both caused gastric damage. In terms of eicosanoid synthesis, WIN-34B significantly suppressed LTB(4) synthesis while both diclofenac and celecoxib increased LTB(4) synthesis. WIN-34B slightly reduced PGE(2) production, while both diclofenac and celecoxib significantly reduced PGE(2) production. In a rat model of diclofenac-induced gastric injury, WIN-34B significantly suppressed LTB(4) synthesis and restored PGE(2) release. CONCLUSIONS: These results demonstrate that WIN-34B did not cause acute or chronic toxicity in male or female rats. In addition, WIN-34B did not cause significant gastric mucosal damage, instead appearing to protect the mucosa from diclofenac-induced gastric damage through the regulation of PGE(2) and LTB(4).

Acquisition of selective antitumoral effects of recombinant adeno-associated virus by genetically inserting tumor-targeting peptides into capsid proteins.

Recombinant adeno-associated virus serotype 5 (rAAV5) is considered to be a promising gene transfer vehicle. However, preferential gene delivery to the tumor remains a requirement for cancer treatment. We generated rAAV5 mutants bearing tumor marker-binding peptides and analyzed their properties as viral vectors, as well as their transduction efficiencies and preferential antitumoral potencies. All of the mutants were successfully produced. Transduction analyses showed that rAAV5 mutants harboring tumor-homing peptides, including RGD and TnC, transduced human cancer cells expressing corresponding receptors on their surfaces. RGDS peptides and TnC antibodies significantly suppressed transduction by rAAV5-RGD and rAAV5-TnC. Cytotoxicity was evident upon transfer of HSV-TK to cells by re-targeted rAAV5. These results provide evidence that rAAV5 vectors, genetically armed with tumor-targeting ligands, preferentially infect human cancer cells harboring the corresponding receptors, thereby inducing antitumoral effects. Further optimization of rAAV5 mutant viruses should thus facilitate practical exploitation of these vectors for gene-based cancer treatment.

Functional study of mucus secretion of the eustachian tube in Guinea pigs.

OBJECTIVES: Our aim was to verify neural regulation of submucous gland mucus secretions in the Eustachian tubes of guinea pigs. STUDY DESIGN: Prospective animal study. METHODS: Eustachian tubes harvested from 12 guinea pigs were used for this study. For real-time resolution of pure glandular secretion, we used a modified method of single-gland optical measurement. Secretory monitoring was undertaken after each preparation with phenylephrine, isoproterenol, forskolin, and substance P. To confirm the viability of each tissue, we examined glandular secretion after treatment with carbachol. Secretory effects of each agonist were evaluated by comparing with basal secretion using a Student's t test (p < 0.01). RESULTS: The Ca-elevating agonists carbachol and substance P showed greater effects on submucous gland secretions of the Eustachian tube than the cyclic adenosine monophosphate (cAMP)-elevating agonists forskolin and isoproterenol. However, phenylephrine, although it belongs to the Ca-elevating agonist group, did not show any significant secretory effect. CONCLUSION: The optical measurement method used in this study had the merit of real-time resolution of submucous glandular secretion. Submucous glandular secretion in the Eustachian tube was regulated by both Ca- and cAMP-elevating agonists, and Ca-elevating agonists seemed to be more potent than cAMP-elevating agonists except phenylephrine. Our results suggest that not only the autonomic nerve system but also the neuropeptides such as substance P are closely related to glandular secretion in the Eustachian tube, and beta-adrenergic receptors seem to be more related to submucous glandular secretion of the Eustachian tube in guinea pig than alpha-adrenergic receptors.

Effective relief of neuropathic pain by adeno-associated virus-mediated expression of a small hairpin RNA against GTP cyclohydrolase 1.

BACKGROUND: Recent studies show that transcriptional activation of GTP cyclohydrolase I (GCH1) in dorsal root ganglia (DRG) is significantly involved in the development and persistency of pain symptoms. We thus hypothesize that neuropathic pain may be attenuated by down-regulation of GCH1 expression, and propose a gene silencing system for this purpose. RESULTS: To interrupt GCH1 synthesis, we designed a bidirectional recombinant adeno-associated virus encoding both a small hairpin RNA against GCH1 and a GFP reporter gene (rAAV-shGCH1). After rAAV-shGCH1 was introduced into the sciatic nerve prior to or following pain-inducing surgery, therapeutic efficacy and the underlying mechanisms were subsequently validated in animal models. The GFP expression data indicates that rAAV effectively delivered transgenes to DRG. Subsequently reduced GCH1 expression was evident from immunohistochemistry and western-blotting analysis. Along with the down-regulation of GCH1, the von Frey test correspondingly indicated a sharp decline in pain symptoms upon both pre- and post-treatment with rAAV-shGCH1. Interestingly, GCH1 down-regulation additionally led to decreased microglial activation in the dorsal horn, implying an association between pain attenuation and reduced inflammation. CONCLUSION: Therefore, the data suggests that GCH1 levels can be reduced by introducing rAAV-shGCH1, leading to pain relief. Based on the results, we propose that GCH1 modulation may be developed as a clinically applicable gene therapy strategy to treat neuropathic pain.