Altered pupil light and darkness reflex and eye-blink responses in late-life depression.

BACKGROUND: Late-life depression (LLD) is a prevalent neuropsychiatric disorder in the older population. While LLD exhibits high mortality rates, depressive symptoms in older adults are often masked by physical health conditions. In younger adults, depression is associated with deficits in pupil light reflex and eye blink rate, suggesting the potential use of these responses as biomarkers for LLD. METHODS: We conducted a study using video-based eye-tracking to investigate pupil and blink responses in LLD patients (n = 25), older (OLD) healthy controls (n = 29), and younger (YOUNG) healthy controls (n = 25). The aim was to determine whether there were alterations in pupil and blink responses in LLD compared to both OLD and YOUNG groups. RESULTS: LLD patients displayed significantly higher blink rates and dampened pupil constriction responses compared to OLD and YOUNG controls. While tonic pupil size in YOUNG differed from that of OLD, LLD patients did not exhibit a significant difference compared to OLD and YOUNG controls. GDS-15 scores in older adults correlated with light and darkness reflex response variability and blink rates. PHQ-15 scores showed a correlation with blink rates, while MoCA scores correlated with tonic pupil sizes. CONCLUSIONS: The findings demonstrate that LLD patients display altered pupil and blink behavior compared to OLD and YOUNG controls. These altered responses correlated differently with the severity of depressive, somatic, and cognitive symptoms, indicating their potential as objective biomarkers for LLD.

Functional and structural investigation of a broadly neutralizing SARS-CoV-2 antibody.

Since its emergence, SARS-CoV-2 has been continuously evolving, hampering the effectiveness of current vaccines against COVID-19. mAbs can be used to treat patients at risk of severe COVID-19. Thus, the development of broadly protective mAbs and an understanding of the underlying protective mechanisms are of great importance. Here, we isolated mAbs from donors with breakthrough infection with Omicron subvariants using a single-B cell screening platform. We identified a mAb, O5C2, which possesses broad-spectrum neutralization and antibody-dependent cell-mediated cytotoxic activities against SARS-CoV-2 variants, including EG.5.1. Single-particle analysis by cryo-electron microscopy revealed that O5C2 targeted an unusually large epitope within the receptor-binding domain of spike protein that overlapped with the angiotensin-converting enzyme 2 binding interface. Furthermore, O5C2 effectively protected against BA.5 Omicron infection in vivo by mediating changes in transcriptomes enriched in genes involved in apoptosis and interferon responses. Our findings provide insights into the development of pan-protective mAbs against SARS-CoV-2.

Linking tonic and phasic pupil responses to P300 amplitude in an emotional face-word Stroop task.

The locus coeruleus-norepinephrine (LC-NE) system, which regulates arousal levels, is important for cognitive control, including emotional conflict resolution. Additionally, the LC-NE system is implicated in P300 generation. If the P300 is mediated by the LC-NE system, and considering the established correlations between LC activity and pupil dilation, P300 amplitude should correlate with task-evoked (phasic) pupil dilation on a trial-by-trial basis. However, prior studies, predominantly utilizing oddball-type paradigms, have not demonstrated correlations between concurrently recorded task-evoked pupil dilation and P300 responses. Using a recently developed emotional face-word Stroop task that links pupil dilation to the LC-NE system, here, we examined both intra- and inter-individual correlations between task-evoked pupil dilation and P300 amplitude. We found that lower accuracy, slower reaction times, and larger task-evoked pupil dilation were obtained in the incongruent compared to the congruent condition. Furthermore, we observed intra-individual correlations between task-evoked pupil dilation and P300 amplitude, with larger pupil dilation correlating with a greater P300 amplitude. In contrast, pupil dilation did not exhibit consistent correlations with N450 and N170 amplitudes. Baseline (tonic) pupil size also showed correlations with P300 and N170 amplitudes, with smaller pupil size corresponding to larger amplitude. Moreover, inter-individual differences in task-evoked pupil dilation between the congruent and incongruent conditions correlated with differences in reaction time and P300 amplitude, though these effects only approached significance. To summarize, our study provides evidence for a connection between task-evoked pupil dilation and P300 amplitude at the single-trial level, suggesting the involvement of the LC-NE system in P300 generation.

Time-on-task effects on human pupillary and saccadic metrics after theta burst transcranial magnetic stimulation over the frontal eye field.

Pupil size undergoes constant changes primarily influenced by ambient luminance. These changes are referred to as the pupillary light reflex (PLR), where the pupil transiently constricts in response to light. PLR kinematics provides valuable insights into autonomic nervous system function and have significant clinical applications. Recent research indicates that attention plays a role in modulating the PLR, and the circuit involving the frontal eye field (FEF) and superior colliculus is causally involved in controlling this pupillary modulation. However, there is limited research exploring the role of the human FEF in these pupillary responses, and its impact on PLR metrics remains unexplored. Additionally, although the protocol of continuous theta-burst stimulation (cTBS) is well-established, the period of disruption after cTBS is yet to be examined in pupillary responses. Our study aimed to investigate the effects of FEF cTBS on pupillary and saccadic metrics in relation to time spent performing a task (referred to as time-on-task). We presented a bright stimulus to induce the PLR in visual- and memory-delay saccade tasks following cTBS over the right FEF or vertex. FEF cTBS, compared to vertex cTBS, resulted in decreased baseline pupil size, peak constriction velocities, and amplitude. Furthermore, the time-on-task effects on baseline pupil size, peak amplitude, and peak time differed between the two stimulation conditions. In contrast, the time-on-task effects on saccadic metrics were less pronounced between the two conditions. In summary, our study provides the first evidence that FEF cTBS affects human PLR metrics and that these effects are modulated by time-on-task.

Mechanism of Antigen Presentation and Specificity of Antibody Cross-Reactivity Elicited by an Oligosaccharide-Conjugate Cancer Vaccine.

Polysaccharides have been successfully used as immunogens for the development of vaccines against bacterial infection; however, there are no oligosaccharide-based vaccines available to date and no previous studies of their processing and presentation. We reported here the intracellular enzymatic processing and antigen presentation of an oligosaccharide-conjugate cancer vaccine prepared from the glycan of Globo-H (GH), a globo-series glycosphingolipid (GSL). This oligosaccharide-conjugate vaccine was shown to elicit antibodies against the glycan moieties of all three globo-series GSLs that are exclusively expressed on many types of cancer and their stem cells. To understand the specificity and origin of cross-reactivity of the antibodies elicited by the vaccine, we found that the vaccine is first processed by fucosidase 1 in the early endosome of dendritic cells to generate a common glycan antigen of the GSLs along with GH for MHC class II presentation. This work represents the first study of oligosaccharide processing and presentation and is expected to facilitate the design and development of glycoconjugate vaccines based on oligosaccharide antigens.

Water-soluble graphene quantum dot-based polymer nanoparticles with internal donor/acceptor heterojunctions for efficient and selective detection of cancer cells.

We present a new, insightful donor-acceptor (D-A) energy transfer-based strategy for the preparation and development of water-soluble multifunctional pH-responsive heterojunction nanoparticles. Hydrophilic tertiary amine-grafted polythiophene (WPT) as a donor and blue fluorescent graphene quantum dots (GQD) as an acceptor spontaneously form co-assembled nanoparticles that function as a highly pH-sensitive and efficient biosensor appropriate for the detection of cancer cells. These WPT/GQD nanoparticles exhibit a number of unique physical characteristics-such as broad-range, tunable GQD-loading contents and particle sizes, extremely low cytotoxicity in normal and cancer cells, and highly sensitive pH-responsiveness and rapid acid-triggered fluorescent behavior under aqueous acidic conditions. We show these features are conferred by self-aggregation of the GQD within the nanoparticles and subsequent aggregation-induced fluorescence of GQD after disassembly of the nanoparticles and dissociation of the D-A interactions under acidic conditions. Importantly, in vitro fluorescence imaging experiments clearly demonstrated the WPT/GQD nanoparticles were gradually taken up into normal and cancer cells in vitro. Selective formation of GQD aggregates subsequently occurred in the acidic microenvironment of the cancer cells and the interior of the cancer cells exhibited strong blue fluorescence; these phenomena did not occur in normal cells. In contrast, pristine WPT and GQD did not exhibit cellular microenvironment-triggered fluorescence transitions in cancer or normal cell lines. Therefore, this newly discovered water-soluble heterojunction system may represent a strongly fluorescent highly pH-sensitive bioprobe for rapid detection of cancer cells.

Vaccination with SARS-CoV-2 spike protein lacking glycan shields elicits enhanced protective responses in animal models.

A major challenge to end the pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is to develop a broadly protective vaccine that elicits long-term immunity. As the key immunogen, the viral surface spike (S) protein is frequently mutated, and conserved epitopes are shielded by glycans. Here, we revealed that S protein glycosylation has site-differential effects on viral infectivity. We found that S protein generated by lung epithelial cells has glycoforms associated with increased infectivity. Compared to the fully glycosylated S protein, immunization of S protein with N-glycans trimmed to the mono-GlcNAc-decorated state (SMG) elicited stronger immune responses and better protection for human angiotensin-converting enzyme 2 (hACE2) transgenic mice against variants of concern (VOCs). In addition, a broadly neutralizing monoclonal antibody was identified from SMG-immunized mice that could neutralize wild-type SARS-CoV-2 and VOCs with subpicomolar potency. Together, these results demonstrate that removal of glycan shields to better expose the conserved sequences has the potential to be an effective and simple approach for developing a broadly protective SARS-CoV-2 vaccine.

High UV-Vis-NIR Light-Induced Antibacterial Activity by Heterostructured TiO2-FeS2 Nanocomposites.

PURPOSE: Antibiotic resistance issues associated with microbial pathogenesis are considered to be one of the most serious current threats to health. Fortunately, TiO2, a photoactive semiconductor, was proven to have antibacterial activity and is being widely utilized. However, its use is limited to the short range of absorption wavelength. METHODS: In this work, heterostructured TiO2-FeS2 nanocomposites (NCs) were successfully prepared by a facile solution approach to enhance light-induced antibacterial activity over a broader absorption range. RESULTS: In TiO2-FeS2 NCs, FeS2 NPs, as light harvesters, can effectively increase light absorption from the visible (Vis) to near-infrared (NIR). Results of light-induced antibacterial activities indicated that TiO2-FeS2 NCs had better antibacterial activity than that of only TiO2 nanoparticles (NPs) or only FeS2 NPs. Reactive oxygen species (ROS) measurements also showed that TiO2-FeS2 NCs produced the highest relative ROS levels. Unlike TiO2 NPs, TiO2-FeS2 NCs, under light irradiation with a 515-nm filter, could absorb light wavelengths longer than 515 nm to generate ROS. In the mechanistic study, we found that TiO2 NPs in TiO2-FeS2 NCs could absorb ultraviolet (UV) light to generate photoinduced electrons and holes for ROS generation, including ⋅O2 - and ⋅OH; FeS2 NPs efficiently harvested Vis to NIR light to generate photoinduced electrons, which then were transferred to TiO2 NPs to facilitate ROS generation. CONCLUSION: TiO2-FeS2 NCs with superior light-induced antibacterial activity could be a promising antibacterial agent against bacterial infections.

CO2-Responsive Water-Soluble Conjugated Polymers for In Vitro and In Vivo Biological Imaging.

Water-soluble conjugated polymers (WCPs) composed of a hydrophobic polythiophene main chain with hydrophilic tertiary amine side-chains can directly self-assemble into sphere-like nano-objects in an aqueous solution due to phase separation between the hydrophilic and hydrophobic segments of the polymeric structure. Due to the presence of gas-responsive tertiary amine moieties in the spherical structure, the resulting polymers rapidly and reversibly tune their structural features, surface charge, and fluorescence performance in response to alternating carbon dioxide (CO2) and nitrogen (N2) bubbling, which leads to significantly enhanced fluorescence and surface charge switching properties and a stable cycle of on and off switching response. In vitro studies confirmed that the CO2-treated polymers exhibited extremely low cytotoxicity and enhanced cellular uptake ability in normal and tumor cells, and thus possess significantly improved fluorescence stability, distribution, and endocytic uptake efficiency within cellular organisms compared to the pristine polymer. More importantly, in vivo assays demonstrated that the CO2-treated polymers displayed excellent biocompatibility and high fluorescence enhancement in living zebrafish, whereas the fluorescence intensity and stability of zebrafish incubated with the pristine polymer decreased linearly over time. Thus, these CO2 and N2-responsive WCPs could potentially be applied as multifunctional fluorescent probes for in vivo biological imaging.

Plastic wrap combined with alcohol wiping is an effective method of preventing bacterial colonization on mobile phones.

INTRODUCTION: Using mobile phones for communication in emergency departments is a common practice; however, several studies have demonstrated that they may act as vectors for bacteria and viruses. This study evaluated the effectiveness of plastic wrapping in decreasing bacterial contamination on mobile phone surfaces. METHOD: We used culture dishes and a luminometer to detect bacterial colonies and contamination on the phone surfaces. RESULT: Our experiment showed that bacterial colonies exist on mobile phones before and after work. We found that wiping with 75% alcohol sanitizers effectively reduces the number of colonies on either a mobile phone or a temporary plastic covering. In addition, we found that bacterial colonies do not contaminate or adhere to plastic wrap any easier than to mobile phones. CONCLUSION: These results demonstrated the effectiveness of plastic wrap for protecting mobile phone surfaces against bacterial colonization. In addition, applying a layer of plastic wrap protects the phone from potential damage due to the alcohol.

O-GlcNAcylation and its role in the immune system.

O-linked-N-acetylglucosaminylation (O-GlcNAcylation) is a type of glycosylation that occurs when a monosaccharide, O-GlcNAc, is added onto serine or threonine residues of nuclear or cytoplasmic proteins by O-GlcNAc transferase (OGT) and which can be reversibly removed by O-GlcNAcase (OGA). O-GlcNAcylation couples the processes of nutrient sensing, metabolism, signal transduction and transcription, and plays important roles in development, normal physiology and physiopathology. Cumulative studies have indicated that O-GlcNAcylation affects the functions of protein substrates in a number of ways, including protein cellular localization, protein stability and protein/protein interaction. Particularly, O-GlcNAcylation has been shown to have intricate crosstalk with phosphorylation as they both modify serine or threonine residues. Aberrant O-GlcNAcylation on various protein substrates has been implicated in many diseases, including neurodegenerative diseases, diabetes and cancers. However, the role of protein O-GlcNAcylation in immune cell lineages has been less explored. This review summarizes the current understanding of the fundamental biochemistry of O-GlcNAcylation, and discusses the molecular mechanisms by which O-GlcNAcylation regulates the development, maturation and functions of immune cells. In brief, O-GlcNAcylation promotes the development, proliferation, and activation of T and B cells. O-GlcNAcylation regulates inflammatory and antiviral responses of macrophages. O-GlcNAcylation promotes the function of activated neutrophils, but inhibits the activity of nature killer cells.

Effects of influenza vaccine and sun exposure time against laboratory-confirmed influenza hospitalizations among young children during the 2012-13 to 2015-16 influenza seasons.

BACKGROUND: Influenza is a major cause of acute respiratory infection burden worldwide, leading to many hospitalizations. An annual influenza vaccine is believed to be the best way to prevent influenza-related illnesses. We focused on the efficacies of other possible preventive measures such as increasing sun exposure time and dietary supplements to prevent these illnesses. METHODS: We conducted a matched-pair case-control study along with the Taiwan Pediatric Infectious Disease Alliance. We included influenza-related hospitalized patients with age ranging from 6 months to 5 years during the 2012-2013, 2013-2014, 2014-2015, and 2015-2016 influenza seasons. The controls were comparable to cases in age, sex, and residential area and had no influenza-related hospitalization records in the same season. We extracted data from vaccination histories and got the patients' guardians to complete questionnaires. Data were analyzed using conditional logistic regression. RESULTS: We enrolled 1514 children (421 influenza-infected cases and 1093 controls) in the study. We found seasonal influenza vaccination to be an independent protective factor against hospitalizations owing to influenza [p < 0.01; odds ratio (OR), 0.427; 95% confidence interval (CI), 0.306-0.594]. Children with mean sun exposure time of >7 h/week had a significantly lower risk of influenza-related hospitalizations than those with the mean sun exposure time of ≤7 h/week (p < 0.05; OR, 0.667; 95% CI, 0.491-0.906). CONCLUSIONS: Seasonal influenza vaccination effectively prevents influenza-related hospitalizations in children aged ≤5 years. Besides, >7 h of sun exposure/week may also be associated with lower risk of influenza-related hospitalizations in children.

High-Performance Sample Substrate of Gold Nanoparticle Multilayers for Surface-Assisted Laser Desorption/Ionization Mass Spectrometry.

The development of a sample substrate with superior performance for desorption and ionization of analyte is the key issue to ameliorate the quality of mass spectra for measurements of small molecules in surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). Herein, the homogeneous sample substrate of gold nanoparticle multilayers (AuNPs-ML) with hexagonal lattice was successfully prepared by self-assembly technique. With strong surface plasmon resonance absorption and superior photothermal effect, the sample substrate of AuNPs-ML exhibited high signal sensitivity and low background noise for the detection of model analyte of glucose without additional matrixes in SALDI-MS. Furthermore, compared to merchant matrixes of α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB), the sample substrate of AuNPs-ML was demonstrated to ameliorate the quality of mass spectra, including signal strength, background interference and signal/noise (S/N) ratio. The sucrose and tryptophan were also measured to show the extensive applications of AuNPs-ML sample substrate for the detections of small molecules in SALDI-MS. Most importantly, the remarkable reproducibility of glucose mass spectra with relative signal of 7.3% was obtained by the use of AuNPs-ML sample substrate for SALDI-MS. The homogeneous sample substrate of AuNPs-ML greatly improved the quality of mass spectra because of its strong absorption of laser energy, low specific heat, high heat conductivity and extraordinary homogeneity. We believe that AuNPs-ML could be a practical sample substrate for small molecule detection in SALDI-MS.

Entrapment of an adenine derivative by a photo-irradiated uracil-functionalized micelle confers controlled self-assembly behavior.

HYPOTHESIS: Invoking cooperative assembly of the uracil-functionalized supramolecular polymer BU-PPG [uracil end-capped poly(propylene glycol)] upon association with the nucleobase adenine derivative A-MA [methyl 3-(6-amino-9H-purin-9-yl)propanoate] as a model drug provides a new concept to control and tune the properties of supramolecular complexes and holds significant potential for the development of safer, more effective drug delivery systems. EXPERIMENTS: BU-PPG and A-MA were successfully developed and exhibited specific recognition and high affinity, which enabled reversible complementary adenine-uracil (A-U) hydrogen bonding-induced formation of spherical micelles in aqueous solution. The self-assembly and controllable A-MA release behavior of BU-PPG/A-MA micelles were studied using morphological analysis and optical and light scattering techniques to investigate the effect of photoirradiation and temperature on the complementary hydrogen bond interactions between BU-PPG and A-MA. FINDINGS: The resulting micelles possess unusual physical properties, including controlled photoreactivity kinetics, controllable self-assembled morphology and low cytotoxicity in vitro, as well as reversible temperature-responsive behavior. Importantly, irradiated micelles exhibited excellent long-term structural stability under normal physiological conditions and serum disturbance. Increasing the temperature triggered rapid release of A-MA by disrupting A-U complexes. These findings represent an entirely new, promising strategy for the development of multi-controlled release drug delivery nanocarriers based on complementary hydrogen bonding interactions.

Osteoporosis risk assessment using multilayered gold-nanoparticle thin film via SALDI-MS measurement.

A powerful technique to detect bone biomarkers has been developed for assessment of osteoporosis at the early stage. Two-dimensional multilayered gold-nanoparticle thin film (MTF-AuNPs) was demonstrated as a promising test platform for detection of bone biomarker, hydroxyproline (HYP), measured by surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). With strong surface plasmon resonance and excellent homogeneity, facilely prepared, highly ordered, and large-scale MTF-AuNPs revealed high sensitivity of HYP in the SALDI-MS measurement without additional matrixes, such as α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB). Furthermore, the mass spectrum of HYP with MTF-AuNPs was significantly improved in signal intensity enhancement, background noise reduction, and signal-to-noise ratio amplification. The excellent reproducibility of HYP spectra with only 9.3% relative signal variation could be attributed to MTF-AuNPs' high absorbance at a wavelength of 337 nm, low heat capacity, superior thermal conductivity, and outstanding homogeneity. The calibration curve showed high linear correlation between mass spectrum intensity and HYP concentration in the range of 1 to 100 µM, covering the whole level in healthy people and osteoporosis patients. In particular, the serum sample was directly deposited onto the MTF-AuNP sample substrate without any pretreatment and its HYP concentration was then successfully determined. We believe that the combination of SALDI-MS and MTF-AuNP sample substrates would be a potential approach for bone biomarker detection in the osteoporosis risk assessment. Graphical abstract.

Guidelines for DNA recombination and repair studies: Mechanistic assays of DNA repair processes.

Genomes are constantly in flux, undergoing changes due to recombination, repair and mutagenesis. In vivo, many of such changes are studies using reporters for specific types of changes, or through cytological studies that detect changes at the single-cell level. Single molecule assays, which are reviewed here, can detect transient intermediates and dynamics of events. Biochemical assays allow detailed investigation of the DNA and protein activities of each step in a repair, recombination or mutagenesis event. Each type of assay is a powerful tool but each comes with its particular advantages and limitations. Here the most commonly used assays are reviewed, discussed, and presented as the guidelines for future studies.

Supramolecular Polymer Network-Mediated Structural Phase Transitions within Polymeric Micelles in Aliphatic Alcohols.

Self-complementary supramolecular polymers (SCSPs), an efficient combination of sextuple hydrogen-bonded dimer moieties and a temperature-responsive polymer, can promote the construction of stable supramolecular polymer networks (SPNs) that enable the formation of well-defined nanospherical micelles in aliphatic alcohols. These micelles undergo tailorable, thermoresponsive phase transitions at the upper critical solution temperature (UCST) and have a desirable spherical morphology and size ranges, thus, are potential candidates for applications in interfacial engineering and biomedical fields. Moreover, concentration-dependent UCST measurements and variable-temperature experiments indicated that the hydrogen-bonded complexes are strong enough to form stable intermolecularly entangled SPNs within the micelles, even above the UCST or at low concentrations in solution, which enables the micelles to undergo reversible temperature-dependent conformational changes between insoluble and soluble globules without significant changes in particle size or size distribution. Thus, this newly discovered system offers a new approach toward the development of next-generation temperature-responsive SCSPs with the desired structural stability that undergoes UCST transitions.

Facile synthesis of two-dimensional Ruddlesden-Popper perovskite quantum dots with fine-tunable optical properties.

In hybrid organic-inorganic and all-inorganic metal halide perovskite nanomaterials, two-dimensional (2D) Ruddlesden-Popper (RP) perovskites have become one of the most interesting materials because of tunable bandgaps varied with the layer thickness, effective modulation of the electron-hole confinement, and high stability. Here, we report a one-pot synthesis of 2D RP perovskite (BA)2(MA)n - 1PbnX3n + 1 (BA = 1-butylammonium, MA = methylammonium, X = Br or I) quantum dots (QDs) with an average size of 10 nm at room temperature. The (BA)2(MA)n - 1PbnBr3n + 1 (Br series) QDs and (BA)2(MA)n - 1PbnI3n + 1 (I series) QDs exhibited tunable emitting spectrum in the range of 410-523 nm and 527-761 nm, respectively, with full width at half maximum (FWHM) of 12-75 nm. The emission color was tuned by the ratio of MA and halide. The photoluminescence quantum yield of 2D perovskite QDs reached 48.6% with more thermodynamic stability in comparison with 3D MAPbX3 QDs. Overall results indicated that developing a solution synthesis for 2D RP perovskite QDs with great optical properties paves the way toward future optoelectronic devices and perovskite quantum dot photovoltaics.

Highly oriented Langmuir-Blodgett film of silver cuboctahedra as an effective matrix-free sample plate for surface-assisted laser desorption/ionization mass spectrometry.

The design of a homogeneous sample plate to solve the sweet heating spot issues is the key step to expand the applicability of surface-assisted laser desorption/ionization mass spectrometry (SALDI MS). Herein, large-scale and highly oriented Langmuir-Blodgett (LB) films of uniform silver nanocrystals have been fabricated as a highly efficient and matrix-free sample plate for SALDI MS. Three individual silver nanocrystals (cubes, cuboctahedra and octahedra) assembled LB films have been applied as the sample plates for glucose detection by SALDI MS without an additional matrix. The results show that the signal intensity, background noise, signal-to-noise ratio and reproducibility have been significantly improved using LB films as the sample plate in comparison with commercial matrixes of CHCA and DHB. In particular, a relative signal of 5.7% was obtained for LB films of silver cuboctahedra. The significant improvement in the SALDI MS measurement could be attributed to the homogenous dissipation of laser irradiation energy to create a large area of the sweet heating spot on well-oriented silver cuboctahedra-based LB film. This ready-to-use sample plate has the potential for widespread commercial applications in SALDI MS.

In vivo inhibition of influenza A virus replication by RNA interference targeting the PB2 subunit via intratracheal delivery.

BACKGROUND: Influenza virus infection is a major threat to human health. Small interfering RNA (siRNA) is a promising approach for the prevention and treatment of viral infections. In this study, we constructed a series of DNA vector-based short hairpin RNAs (shRNAs) that target various genes of the influenza A virus using the polymerase III U6-RNA promoter to prevent influenza virus infection in vitro and in a mouse model. RESULTS: Three sets of DNA vector-based shRNA, two targeting genes encoding the polymerase acidic protein (PA) and one targeting polymerase basic protein 2 (PB2), efficiently inhibited the replication of influenza virus A/WSN/33(H1N1) in vitro. We also successfully prevented influenza virus A/WSN/33(H1N1) infection in a C57BL/6 mouse model by intratracheal delivery of anti-PB2 shRNA. CONCLUSIONS: Our findings suggest that the PB2-targeting shRNA plasmid showed potential for use as an RNAi-based therapeutic for influenza virus infection.