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Firearm-related injuries in America have been under increasing scrutiny over the last several years. Few studies have examined the burden of these injuries in the pediatric population. The objective of this study was to describe the incidence of firearm-related injuries in hospitalized pediatric patients in the United States and identify the risk factors associated with readmission in this young population.The Nationwide Readmission Database was examined from 2010 to 2017. Pediatric patients (aged ≤18 y) who survived their index hospitalization for any firearm injury were analyzed to determine incidence rate, case fatality rate, risk factors for 30-d readmission, and financial health care burden.There were 35,753 pediatric firearm injuries (86.8% male) with an overall incidence rate of 10.49 (95% confidence interval [CI]: 9.26-11.71) per 100,000 pediatric hospitalizations. Adolescents aged12 y had the highest incidence rate (60.51, 95% CI: 55.19-65.84). In-hospital mortality occurred in 1948 cases (5.5%), with higher case fatality rates in males. There were 1616 (5.7%) unplanned 30-d readmissions. Multivariate analysis showed abdominal firearm injuries (hazard ratio: 1.13, 95% CI: 1.03-1.24; P = 0.006) and longer length of stay (hazard ratio: 1.27, 95% CI: 1.04-1.55; P = 0.016) were associated with a greater risk of 30-d readmission. The median health care cost for firearm-related injuries was $36,535 (interquartile range: $19,802-$66,443), 22% of which was due to readmissions. Cost associated with 30-d readmissions was $7978 (interquartile range: $4305-$15,202).Firearm-related injury is a major contributor to pediatric morbidity, mortality, and health care costs. Males are disproportionately affected by firearm injury, but females are more likely to require unplanned 30-d readmissions. Interventions should target female sex, injuries of suicidal intent, psychiatric comorbidities, prolonged index hospitalization, and abdominal injuries.

Introduction: Time is an essential element in outcomes of trauma patients. The relationship of time to treatment in management of noncompressible torso hemorrhage (NCTH) with resuscitative endovascular balloon occlusion of the aorta (REBOA) or resuscitative thoracotomy (RT) has not been previously described. We hypothesized that shorter times to intervention would reduce mortality. Methods: A review of the Aortic Occlusion for Resuscitation in Trauma and Acute Care Surgery registry from 2013 to 2022 was performed to identify patients who underwent emergency department aortic occlusion (AO). Multivariate logistic regression was used to examine the impact of time to treatment on mortality. Results: A total of 1,853 patients (1,245 [67%] RT, 608 [33%] REBOA) were included. Most patients were male (82%) with a median age of 34 years (interquartile range, 30). Median time from injury to admission and admission to successful AO were 31 versus 11 minutes, respectively. Patients who died had shorter median times from injury to successful AO (44 vs. 72 minutes, Plt; 0.001) and admission to successful AO (10 vs. 22 minutes, Plt; 0.001). Multivariate logistic regression demonstrated that receiving RT was the strongest predictor of mortality (odds ratio [OR], 6.6; 95% confidence interval [CI], 4.4-9.9; Plt; 0.001). Time from injury to admission and admission to successful AO were not significant. This finding was consistent in subgroup analysis of RT-only and REBOA-only populations. Conclusions: Despite expedited interventions, time to aortic occlusion did not significantly impact mortality. This may suggest that rapid in-hospital intervention was often insufficient to compensate for severe exsanguination and hypovolemia that had already occurred before emergency department presentation. Selective prehospital advanced resuscitative care closer to the point of injury with "scoop and control" efforts including hemostatic resuscitation warrants special consideration.

Gun violence, particularly in the form of mass shooting events (MSE), is a growing, significant public health crisis in the US. Whether stricter gun laws decrease MSE is not known. We hypothesized that stronger state gun laws would be associated with lower MSE incidence.Mass shooting events, defined as at least 4 people injured in a single event, and state gun law grade data for years 2014 through 2021 were obtained from the Gun Violence Archive and Giffords Law Center, respectively. An A grade indicated strictest gun control laws, and F indicated the weakest. US 2020 Census data were used to estimate MSE per million per state. The number of MSE per million was examined for association with gun law grades.From 2014 through 2020, there were a total of 2,736 recorded MSE, with at least a 2-fold increase in incidence from 272 in 2014 to 626 in 2020. Concomitantly, the number of F grade states decreased from 27 to 21 (22%). The MSE mean (SD) per F state increased from 4.0 (5.1) in 2014 to 9.7 (10.3) in 2020 (p = 0.03). No differences were found in unadjusted number of MSE per year by gun law grade for any study year examined (p = 0.67). After adjusting for population, this finding of no difference persisted.Strength of state gun law grades does not affect MSE incidence, even after correction for population size. This suggests that legislation by itself is not an effective prevention measure and other broader and meaningful primary gun violence interventions are needed.

The findings that nitrosamine impurities, mutagenic carcinogens, have been detected in several classes of medicines, including valsartan, have raised safety concerns; regulatory authorities responded by strengthening surveillance, creating specific guidance and risk assessment strategies. At the core of these responses are a number of highly sensitive analytical methodologies to detect and quantitate trace amounts of nitrosamines in pharmaceutical products. However, many of the issued methods rely on high resolution or other advanced mass spectrometers which may not be readily available or ideally suited for routine analysis of large sample sets for risk assessment. Here we describe our efforts to develop an alternative approach based on a LC method coupled with a single quadrupole mass spectrometer. Six nitrosamines, N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine (NDEA), N-nitrosodiisopropylamine (NDIPA), N-nitrosoethylisopropylamine (NEIPA), N-nitrosodibutylamine (NDBA), and N-nitrosomethylaminobutyric acid (NMBA) were investigated and a LC/MS method was developed for the detection and quantitation of the six nitrosamine impurities in valsartan. Two commonly used techniques, large-volume injection (LVI) and matrix precipitation for sample preparation, were incorporated to enhance sensitivity. The developed single quadrupole LC/MS method has the capacity for detection and quantitation of the six nitrosamines listed with a LOQ of 0.05 ppm relative to valsartan. Furthermore, the method was validated within a wide dynamic range (0.05 – 3.6 ppm) and applied to commercial valsartan samples.

Sulconazole has been reported to degrade into sulconazole sulfoxide via sulfur oxidation; however, structural characterization data was lacking and the potential formation of an N-oxide or sulfone could not be excluded. To clarify the degradation pathways and incorporate the impurity profile of sulconazole into the United States Pharmacopeia–National Formulary (USP–NF) monographs, a multifaceted approach was utilized to confirm the identity of the degradant. The approach combines stress testing of sulconazole nitrate, chemical synthesis of the degradant via a hydrogen peroxide-mediated oxidation reaction, semi-preparative HPLC purification, and structural elucidation by LC–MS/MS and NMR spectroscopy. Structural determination was primarily based on the comparison of spectroscopic data of sulconazole and the oxidative degradant. The mass spectrometric data have revealed a McLafferty-type rearrangement as the characteristic fragmentation pathway for alkyl sulfoxides with a β-hydrogen atom, and was used to distinguish the sulfoxide from N-oxide or sulfone derivatives. Moreover, the generated sulconazole sulfoxide was utilized as reference material for compendial procedure development and validation, which provides support for USP monograph modernization. Keywords: Sulconazole nitrate, Sulconazole sulfoxide, Forced degradation, Structural characterization, LC–MS/MS

The analysis of protein glycosylation by mass spectrometry (MS) has been a challenging technical problem. Quantification by HPLC of N-linked glycans can be executed by the use of peptide-N-glycosidase F to release them from the protein, followed by attachment of a fluorescent label and subsequent fluorescence detection. Similar quantification of O-linked glycans is not possible, as a result of the lack of a universal deglycosylation enzyme. Site-specific analyses by MS, such as the use of proteases to digest the glycoprotein, are difficult to use for quantification of glycans, as a result of the presence of miscleavages. Here, we present a new application of a digestion method for native proteins using resin-bound, thermally stabilized proteases. The use of this enzymatic treatment eliminates miscleavages around the site of glycosylation, thereby allowing site-specific relative quantification of glycans on glycoproteins. A native, intact human mAb was digested using a thermally stable, resin-bound trypsin to produce glycopeptides from the Fc region using a single-step protocol. A 1 mg sample was treated with 60 µg trypsin for 3 h at 70°C. After digestion, acetonitrile was added, and the mixture was centrifuged to remove the resin before analysis. Liquid chromatography (LC)/MS with hydrophilic interaction chromatography was used to analyze the glycopeptides produced. All of the glycopeptides found resulted from a single peptide (EEQYNSTYR). The LC/MS analysis of the glycopeptides is compared with that of fluorescently labeled glycans. Quantitative analysis produced a correlation coefficient of 0.87 for the linear fit between the glycopeptide and released glycan methods.

A revolutionary impact on the performance of many optical systems and components can come from the integrative design of multifunctional coatings. Such coatings should be mechanically robust, and combine user-defined optical and wetting functions with scalable fabrication formulations. By taking cues from the properties of some natural biological structures, we report here the formation of low-refractive index antireflective glass films that embody omni-directional optical properties over a wide range of wavelengths, while also possessing specific wetting capabilities. The coatings comprise an interconnected network of nanoscale pores surrounded by a nanostructured silica framework. These structures result from a novel fabrication method that utilizes metastable spinodal phase separation in glass-based materials. The approach not only enables design of surface microstructures with graded-index antireflection characteristics, where the surface reflection is suppressed through optical impedance matching between interfaces, but also facilitates self-cleaning ability through modification of the surface chemistry. Based on near complete elimination of Fresnel reflections (yielding >95% transmission through a single-side coated glass) and corresponding increase in broadband transmission, the fabricated nanostructured surfaces are found to promote a general and an invaluable ∼3–7% relative increase in current output of multiple direct/indirect bandgap photovoltaic cells. Moreover, these antireflective surfaces also demonstrate superior resistance against mechanical wear and abrasion. Unlike conventional counterparts, the present antireflective coatings are essentially monolithic, enabling simultaneous realization of graded index anti-reflectivity, self-cleaning capability, and mechanical stability within the same surface. The concept represents a fundamental basis for development of advanced coated optical quality products, especially where environmental exposure is required.

Bio-inspired superhydrophobic surfaces were fabricated based on fossilized silica fresh water diatomaceous earth (DE) particles. These nanostructured silicified diatom frustules of cylindrical and circular structures were fluorinated to impart them with superhydrophobic properties. Substrates coated with superhydrophobic DE structures of varying size and shape were found to have water contact angles of approximately 170° and sliding angles of approximately 3°. The substrates were subjected to significant abrasion forces using a standard surface abrader. The ability to retain their superhydrophobic properties was observed to depend on the geometry and average size of the DE particles. The wettability of the abraded coatings was determined by their surface topology, and a transition from a non-wetted state to a partially wetted state was observed to occur and was dependent on the surface roughness. The proposed coatings are scalable, cost-effective, and can be applied on a variety of surfaces on critical infrastructures requiring protection from water saturation, ice formation and water based corrosion.

Over the past few years, engineers have been exploring the utilization of various types of substrates like fabrics, organic, and even papers substrate to fabricate microwave components. Use of such flexible substrates renders simplified fabrication process, and can easily lead to low cost mass production and fast turn around. But they generally have a serious problem that is water absorption that would significantly degrade their performance. In this paper, we utilize a super-hydrophobic material to cover these circuits to minimize water absorption while sustaining their RF performance. The results show very consistent performance before and after applying the super-hydrophobic coating on the antenna surface and minimal degradation if any.

In this paper, the icephobic properties of superhydrophobic surfaces are investigated under dynamic flow conditions using a closed-loop low-temperature wind tunnel. Superhydrophobic surfaces were prepared by coating aluminum and steel substrate plates with nano-structured hydrophobic particles. The superhydrophobic plates, along with uncoated controls, were exposed to a wind tunnel air flow of 12 m/s and −7 °C with deviations of ±1 m/s and ±2.5 °C, respectively, containing micrometer-sized (∼50 μm in diameter) water droplets. The ice formation and accretion were observed by CCD cameras. Results show that the superhydrophobic coatings significantly delay ice formation and accretion even under the dynamic flow condition of highly energetic impingement of accelerated supercooled water droplets. It is found that there is a time scale for this phenomenon (delay in ice formation) which has a clear correlation with contact angle hysteresis and the length scale of the surface roughness of the superhydrophobic surface samples, being the highest for the plate with the lowest contact angle hysteresis and finest surface roughness. The results suggest that the key for designing icephobic surfaces under the hydrodynamic pressure of impinging droplets is to retain a non-wetting superhydrophobic state with low contact angle hysteresis, rather than to only have a high apparent contact angle (conventionally referred to as a “static” contact angle).

To date, most of the proteomic analyses on lung cancer tissue samples have been performed using surgical specimens, which are obtained after a diagnosis is made. To determine if a proteomic signature obtained from bronchoscopic biopsy samples could be found to assist with diagnosis, 50 lung cancer bronchoscopic biopsy samples and 13 adjacent normal lung tissue samples were analyzed using histology-directed, matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). Lung tissue samples were cryosectioned, and sinapinic acid was robotically deposited on areas of each tissue section enriched in epithelial cells, either tumor or normal. Mass spectra were acquired using a MALDI-time of flight instrument. Small cell lung cancers (SCLCs) demonstrated clearly different protein profiles from normal lung tissue and from non-small cell lung cancers (NSCLCs). Calcyclin (m/z= 10,094.7) was identified to be underexpressed in small cell lung cancers, as compared with non-small cell lung cancers and normal lung tissue. An immunohistochemistry study using 152 NSCLCs and 21 SCLCs confirmed significantly reduced calcyclin stain in SCLCs. Thus, protein profiles obtained from bronchoscopic biopsy samples via MALDI MS distinguish cancerous epithelium from normal lung tissue and between NSCLCs and SCLCs.

Over the past few years, the scientific community, as well as the world' s coatings industry have seen the introduction of oxide/polymer-based hydrophobic surfaces and coatings with exceptional water repelling properties. This paper addresses the RF behavior of antennas coated with a recently introduced super-hydrophobic coating that could fundamentally change how water interacts with ink-jet printed antennas on fabrics, papers, or any other surface, in addition to its low loss at high frequencies. There are numerous possibilities of printing sensors on any surface for various applications like body area networks, checking athletes' vitals, and elderly healthcare monitoring applications.

We report a novel method of producing ordered arrays of glass nanocones with precisely controlled height, lattice constant and aspect ratio. As with nanochannel glass, fibre drawing, bundling and redrawing are used to produce structured glass composite material. The surface of the composite is etched to form nanocones through a differential etching process. The lattice constant of the arrays ranges from 40 µm to 1.6 µm, while the aspect ratio of the nanocones is varied from 0.4 to 13 by simple changes in the chemistry of the hydrofluoric acid etching solution.

The recent observation of optical gain from silicon nanocrystals embedded in SiO2 opens an opportunity to develop a nanoscale silicon-based laser. However, the challenge remains to design and develop a laser architecture using CMOS-compatible materials. In this paper we present two designs for a waveguide laser in which silicon nanocrystals embedded in SiO2 are used as the optical gain media. One design employs a SiO2 membrane containing encapsulated Si nanocrystals. Preliminary calculations given here show that a highly resonant laser cavity can be produced in a SiO2 membrane using sub-wavelength structures. This photonic crystal architecture, used to guide and contain the light, can be combined with a gain medium of optically active Si nanocrystals synthesized in the SiO2 membrane using ion implantation/thermal annealing to produce a Si-based laser. The laser cavity dimensions can be matched to the near-infrared wavelengths where optical gain has been observed from Si nanocrystals. The second design utilizes silicon nanocrystals embedded in a distributed-feedback laser cavity fabricated in SiO2. Lasing action over a broad wavelength range centered at ∼770 nm should be possible in both of these configurations.

The purpose of this study was to control the size and shape of fabricated glass microcone arrays for use in optical surgery instruments by varying the fiber drawing and etching methods. The spacing of the cones is controlled via the drawing process while the etching process controls the cone height-to-base ratio. The device viability was tested by imprinting, and subsequent staining, of low-density polyethylene and porcine corneas, resulting in a consistent patterned structure of micron-sized perforations. After imprint, the device was examined and no evidence of microcone fracture or over-penetration was present during the course of these experiments. This research promises to lead to advances in optical surgery for the treatment of recurrent corneal erosions, providing quicker, safer, and more cost-effective procedures with decreased risk of vision loss and scarring associated with current procedures such as anterior stromal puncture. The ease of procedure and micron sized incisions could potentially replace current techniques and provide a viable treatment alternative for recurrent corneal erosions in the visual axis.

We describe the formation and properties of atomically bonded, optical quality, nanostructured thin glass film coatings on glass plates, utilizing phase separation by spinodal decomposition in a sodium borosilicate glass system. Following deposition via magnetron sputtering, thermal processing and differential etching, these coatings are structurally superhydrophilic (i.e., display anti-fogging functionality) and demonstrate robust mechanical properties and superior abrasion resistance. After appropriate chemical surface modification, the surfaces display a stable, non-wetting Cassie-Baxter state and exhibit exceptional superhydrophobic performance, with water droplet contact angles as large as 172°. As an added benefit, in both superhydrophobic and superhydrophilic states these nanostructured surfaces can block ultraviolet radiation and can be engineered to be anti-reflective with broadband and omnidirectional transparency. Thus, the present approach could be tailored toward distinct coatings for numerous markets, such as residential windows, windshields, specialty optics, goggles, electronic and photovoltaic cover glasses, and optical components used throughout the US military.

Ferroelectric(FE) films exhibit interesting electro-optic (EO) properties and are utilized in devices used for second harmonic generation, spatial light modulators and optical switches. These films typically yield large values of linear and quadratic electro-optic coefficients. Solgel derived FE films namely PZT 53 47 and PLZT 28/0/100 were prepared on conductive glass substrates. 0.5M precursor solutions based on the appropriate stoichiometric amounts of lead acetate, La nitrate and Ti Zr alkoxides were refluxed for 1 hour and later spincoated on the substrates. They were the fired to 600C to crystallize them into single phase perovskite films. Top Au Pd electrodes were deposited to form the top contacts for the capacitors. The refractive indices, extinction coefficients, linear and quadratic coefficients of the films were obtained using multiangle reflection ellipsometry. These parameters were then used to calculate the linear and quadratic electro-optic coefficients of the films. The quadratic EO coefficients of PLZT 28/0/100 and PZT 53 47 films were measured to be 0.07 and 0.38 × 10−16 m2/V2 respectively while the linear electrooptic coefficients of PLZT 28/0/100 and PZT 53 47 were found to be 59 and 315 pm/V respectively, with the reff value for PZT 53 47 the highest among reported ferroelectric films in literature.

Ferroelectric (FE) films constitute an important component of electro-optic devices; such films have been used for second harmonic generation, spatial light modulators and optical switches. FE films typically yield large values of linear and quadratic electro-optic coefficients. Most electric field induced birefringence measurements have been made with transverse electrodes, i. e. with light beam between poled electrodes. In the present study, an Ultimate Ellipsometer was used; a HeNe laser was directed perpendicularly through FE capacitors with applied voltages and the phase change monitored. The layout of this device is discussed in details. Sol-gel derived PZT 53/47 and PLZT 28/0/100 films were prepared on conductive glass substrates. 0·5M precursor solutions based on the stoichiometric amounts of Pb acetate, La nitrate and Ti/Zr alkoxides were refluxed for 1 hour and spincoated on the substrates under clean room conditions. The films were then fired to 550C to crystallize them to single phase...

To date, most of the proteomic analyses on lung cancer tissue samples have been performed using surgical specimens, which are obtained after a diagnosis is made. To determine if a proteomic signature obtained from bronchoscopic biopsy samples could be found to assist with diagnosis, 50 lung cancer bronchoscopic biopsy samples and 13 adjacent normal lung tissue samples were analyzed using histology-directed, matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS). Lung tissue samples were cryosectioned, and sinapinic acid was robotically deposited on areas of each tissue section enriched in epithelial cells, either tumor or normal. Mass spectra were acquired using a MALDI-time of flight instrument. Small cell lung cancers (SCLCs) demonstrated clearly different protein profiles from normal lung tissue and from non-small cell lung cancers (NSCLCs). Calcyclin (m/z= 10,094.7) was identified to be underexpressed in small cell lung cancers, as compared with non-small cell lung cancers and normal lung tissue. An immunohistochemistry study using 152 NSCLCs and 21 SCLCs confirmed significantly reduced calcyclin stain in SCLCs. Thus, protein profiles obtained from bronchoscopic biopsy samples via MALDI MS distinguish cancerous epithelium from normal lung tissue and between NSCLCs and SCLCs.

In this paper, the iceophobic properties of superhydrophobic surfaces are compared to those of uncoated aluminum and steel plate surfaces as investigated under dynamic flow conditions by using a closed loop low-temperature wind tunnel. Superhydrophobic surfaces were prepared at the Oak Ridge National Laboratory by coating aluminum and steel plates with nano-structured hydrophobic particles. The contact angle and contact angle hysteresis measured for these surfaces ranged from 165–170° and 1–8°, respectively. The superhydrophobic plates along with uncoated control ones were exposed to an air flow of 12 m/s and 20°F with micron-sized water droplets in the icing wind tunnel and the ice formation and accretion were probed by using high speed cameras for 90 seconds. Results show that the developed superhydrophobic coatings significantly delay the ice formation and accretion even with the impingement of accelerated super-cooled water droplets, but there is a time scale for this phenomenon which has a clear relation with contact angle hysteresis of the samples. Among the different superhydrophobic coating samples, the plate having the lowest contact angle hysteresis showed the most pronounced iceophobic effects, while the correlation between static contact angles and the iceophobic effects was not evident. The results suggest that the key parameter for designing iceophobic surfaces is to retain a low contact angle hysteresis, rather than to have only a low contact angle, which can result in more efficient anti-icing properties in dynamic flow conditions.Copyright © 2011 by ASME

In this paper, we report the characterization of large-scale superhydrophobic surfaces for hydrodynamic drag reduction in boundary layer flows using a high-speed towing tank system. For making superhydrophobic surfaces, flat aluminum plates (4 ft × 2 ft × 3/8 in, with sharpened leading/trailing edges) were prepared and coated with nano-structured hydrophobic particles. The static and dynamic contact angle measurements indicate that the coated surfaces correspond to a de-wetting (Cassie) state with air retained on the nano-structured surfaces. Hydrodynamic drag of the large-area superhydrophobic plates was measured to cover turbulent flows (water flow speeds up to 30 ft/s, Reynolds number in the range of 105 −107 ) and compared with that of an uncoated bare aluminum control plate. Results show that an acceptable drag reduction was obtained up to ∼30% in the early stage of the turbulent regime which is due to reduced shear forces on the plates because of the lubricating air layer on the surface. However, in a fully developed turbulent flow regime, an increase in drag was measured which is mainly attributed to the amplified surface roughness due to the protrusions of air bubbles formed on the surface. Meanwhile, a qualitative observation suggests that the air bubbles are prone to be depleted during several runs of the high shear-rate flows, as revealed by streak lines of depleted air bubbles. This suggests that the superhydrophobic coating is unstable in maintaining the de-wetted state under dynamic flow conditions and that the increased drag results from the inherent surface roughness of the coating layer where the de-wetted state collapses to a wetted (Wenzel) state due to the depletion of air bubbles. However, it was also observed that the air bubbles would reform on the surface, with the same properties as a dry surface immersed in water, while the plate was kept statically immersed in water for 12 hours, suggesting that the superhydrophobic coating retains static stability for a de-wetted state. The experimental results illustrate that drag reduction is not solely dependent on the superhydrophobicity of a surface (e.g., contact angle and air fraction), but the morphology and stability of the surface air layer are also critical for the design and use of superhydrophobic surfaces for large-scale hydrodynamic drag reduction, especially in turbulent flow regimes.Copyright © 2011 by ASME

Arachidonylethanolamide (anandamide), a candidate endogenous cannabinoid ligand, has recently been isolated from porcine brain and displayed cannabinoid-like binding activity to synaptosomal membrane preparations and mimicked cannabinoid-induced inhibition of the twitch response in isolated murine vas deferens. In this study, anandamide and several congeners were evaluated as cannabinoid agonists by examining their ability to bind to the cloned cannabinoid receptor, inhibit forskolin-stimulated cAMP accumulation, inhibit N-type calcium channels, and stimulate one or more functional second messenger responses. Synthetic anandamide, and all but one congener, competed for [3H]CP55,940 binding to plasma membranes prepared from L cells expressing the rat cannabinoid receptor. The ability of anandamide to activate receptor-mediated signal transduction was evaluated in Chinese hamster ovary (CHO) cells expressing the human cannabinoid receptor (HCR, termed CHO-HCR cells) and compared to control CHO cells expressing the muscarinic m5 receptor (CHOm5 cells). Anandamide inhibited forskolin-stimulated cAMP accumulation in CHO-HCR cells, but not in CHOm5 cells, and this response was blocked with pertussis toxin. N-type calcium channels were inhibited by anandamide and several active congeners in N18 neuroblastoma cells. Anandamide stimulated arachidonic acid and intracellular calcium release in both CHOm5 and CHO-HCR cells and had no effect on the release of inositol phosphates or phosphatidylethanol, generated after activation of phospholipase C and D, respectively. Anandamide appears to exhibit the essential criteria required to be classified as a cannabinoid/anandamide receptor agonist and shares similar nonreceptor effects on arachidonic acid and intracellular calcium release as other cannabinoid agonists.

Determining the molecular weight of nanoparticles can be challenging. The molecular weight characterization of dendrimers, for example, with varying covalent and noncovalent modifications is critical to their use as therapeutics. As such, we describe in this chapter a protocol for the analysis of these molecules by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Determining the molecular weight of nanoparticles can be challenging. The molecular weight characterization of dendrimers, for example, with varying covalent and noncovalent modifications is critical to their use as therapeutics. As such, we describe in this chapter a protocol for the analysis of these molecules by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Since its introduction as an ionization technique in mass spectrometry, matrix-assisted laser desorption ionization (MALDI) has been applied to a wide range of applications. Quantitative small molecule analysis by MALDI, however, is limited due to the presence of intense signals from the matrix coupled with non-homogeneous surfaces. The surface used in nano-structured laser desorption ionization (NALDI) eliminates the need for a matrix and the resulting interferences, and allows for quantitative analysis of small molecules. This study was designed to analyze and quantitate phospholipid components of liposomes. Here we have developed an assay to quantitate the DPPC and DC(8,9)PC in liposomes by NALDI following various treatments. To test our method we chose to analyze a liposome system composed of DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and DC(8,9)PC (1,2-bis(tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine), as DC(8,9)PC is known to undergo cross-linking upon treatment with UV (254 nm) and this reaction converts the monomer into a polymer. First, calibration curves for pure lipids (DPPC and DC(8,9)PC) were created using DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) as an internal standard. The calibration curve for both DPPC and DC(8,9)PC showed an R(2) of 0.992, obtained using the intensity ratio of analyte and internal standard. Next, DPPC:DC(8,9)PC liposomes were treated with UV radiation (254 nm). Following this treatment, lipids were extracted from the liposomes and analyzed. The analysis of the lipids before and after UV exposure confirmed a decrease in the signal of DC(8,9)PC of about 90%. In contrast, there was no reduction in DPPC signal.

In this study, a closed loop low-temperature wind tunnel was custom-built and uniquely used to investigate the anti-icing mechanism of superhydrophobic surfaces in regulated flow velocities, temperatures, humidity, and water moisture particle sizes. Silica nanoparticle-based hydrophobic coatings were tested as superhydrophobic surface models. During tests, images of ice formation were captured by a camera and used for analysis of ice morphology. Prior to and after wind tunnel testing, apparent contact angles of water sessile droplets on samples were measured by a contact angle meter to check degradation of surface superhydrophobicity. A simple peel test was also performed to estimate adhesion of ice on the surfaces. When compared to an untreated sample, superhydrophobic surfaces inhibited initial ice formation. After a period of time, random droplet strikes attached to the superhydrophobic surfaces and started to coalesce with previously deposited ice droplets. These sites appear as mounds of accreted ice across the surface. The appearance of the ice formations on the superhydrophobic samples is white rather than transparent, and is due to trapped air. These ice formations resemble soft rime ice rather than the transparent glaze ice seen on the untreated sample. Compared to untreated surfaces, the icing film formed on superhydrophobic surfaces was easy to peel off by shear flows.Copyright © 2010 by ASME

In 2007, The Association of Biomolecular Resource Facilities (ABRF) Survey Committee surveyed the ABRF membership and scientists at-large concerning the current state of funding in service-oriented laboratories. Questions pertained to services offered, cost recovery, capital equipment funding, and future outlook. The web-based survey, available for 3 weeks, achieved participation from 209 respondents in 13 countries, 77% of which represented academic laboratories. Most respondents (75%) directed their laboratories. Laboratories depend largely on institutional support and customer recharges to fund operations, but National Institutes of Health and National Science Foundation Shared Instrumentation Grant programs are considered critical to meeting future needs. Source allocations supporting capital equipment acquisitions, operations, and laboratory director salary are presented.

Carbohydrate-protein conjugates are utilized extensively in basic research and as immunogens in a variety of bacterial vaccines and cancer vaccines. As a result, there have been significant efforts to develop simple and reliable methods for the construction of these conjugates. While direct coupling via reductive amination is an appealing approach, the reaction is typically very inefficient. In this paper, we report improved reaction conditions providing an approximately 500% increase in yield. In addition to optimizing a series of standard reaction parameters, we found that addition of 500 mM sodium sulfate improves the coupling efficiency. To illustrate the utility of these conditions, a series of high mannose BSA conjugates were produced and incorporated into a carbohydrate microarray. Ligand binding to ConA could be observed and apparent affinity constants ( K ds) measured using the array were in good agreement with values reported by surface plasmon resonance. The results show that the conditions are suitable for microgram-scale reactions, are compatible with complex carbohydrates, and produce biologically active conjugates.

The mutant beta1,4-galactosyltransferase (beta4Gal-T1), beta4Gal-T1-Y289L, in contrast to wild-type beta4Gal-T1, can transfer GalNAc from the sugar donor UDP-GalNAc to the acceptor, GlcNAc, with efficiency as good as that of galactose from UDP-Gal. Furthermore, the mutant can also transfer a modified sugar, C2 keto galactose, from its UDP derivative to O-GlcNAc modification on proteins that provided a functional handle for developing a highly sensitive chemoenzymatic method for detecting O-GlcNAc post-translational modification on proteins. We report herein that the modified sugar, C2 keto galactose, can be transferred to free GlcNAc residues on N-linked glycoproteins, such as ovalbumin or asialo-agalacto IgG1. The transfer is strictly dependent on the presence of both the mutant enzyme and the ketone derivative of the galactose. Moreover, the PNGase F treatment of the glycoproteins, which cleaves the N-linked oligosaccharide chain, shows that the modified sugar has been transferred to the N-glycan chains of the glycoproteins and not to the protein portion. The application of the mutant galactosyltransferase, beta4Gal-T1-Y289L, to produce glycoconjugates carrying sugar moieties with reactive groups, is demonstrated. We envision a broad potential for this technology such as the possibilities to link cargo molecules to glycoproteins, such as monoclonal antibodies, via glycan chains, thereby assisting in the glycotargeting of drugs to the site of action or used as biological probes.

Over the past few years, the scientific community, as well as the world's coatings industry has seen the introduction of oxide/polymer-based superhydrophobic surfaces and coatings with exceptional water repellency. Online videos have caught the public's imagination by showing people walking through mud puddles without getting their tennis shoes wet or muddy, and water literally flying off coated surfaces. This article attempts to explain the basics of this behavior and to discuss and explain the latest superhydrophobic technological breakthroughs. Since superhydrophobic surfaces and coatings can fundamentally change how water interacts with surfaces, and the fact that earth is a water world, it can legitimately be said that this technology has the potential to literally change the world.

The dramatic progress in mass spectrometry-based methods of protein identification has triggered a new quest for disease-associated biomarkers. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and its variant surface-enhanced laser desorption/ionization mass spectrometry, provide effective means to explore the less studied information slice of the human serum proteome -- low-molecular-weight proteins and peptides. These low-molecular-weight proteins and peptides are promising for the detection of important biomarkers. Due to the significant experimental problems imposed by high-abundance and high-molecular-weight proteins, it is important to effectively remove these species prior to mass spectrometry analysis of the low-molecular-weight serum and plasma proteomes. In this review, the advantages afforded by recently introduced methods for prefractionation of serum, as they pertain to the detection and identification of biomarkers, will be discussed.

Spatial heterodyning is an interferometric technique that allows a full complex optical wavefront to be recorded and quickly reconstructed with a single image capture. Oak Ridge National Laboratory (ORNL) has combined a high-speed, image capture technique with a Fourier reconstruction algorithm to produce a method for recovery of both the phase and magnitude of the optical wavefront. Single frame spatial heterodyne interferometry (SHI) enables high-speed inspection applications such as those needed in the semiconductor industry. While the wide range of materials on wafers make literal interpretation of surface topology difficult, the wafers contain multiple copies of the same die and die-to-die comparisons are used to locate defects in high-aspect-ratio structures such as contacts, vias, and trenches that are difficult to detect with other optical techniques. Metrology with SHI has also been investigated by ORNL, in particular the use of SHI to perform metrology of line widths and heights on photolithographic masks for semiconductor wafer production. Several types of masks are currently in use with phase shifting techniques being employed to extend the wafer printing resolution. With the ability to measure the phase of the wavefront, SHI allows a more complete inspection and measurement of the phase shifting regions.

A method to extract peptides and low molecular weight proteins from serum under denaturing conditions using acetonitrile containing 0.1% trifluoroacetic acid has been developed. The extraction procedure precipitates large, abundant proteins to simplify subsequent mass spectral analysis. This sample preparation method provides an efficient way to extract serum peptides, enabling them to be compared and identified using different mass spectrometry approaches. Surface-enhanced laser desorption/ionization-time of flight mass spectrometry analysis of mouse blood serum samples prepared by this method allowed detection of two markers which were significantly reduced in mice with B cell lymphoma tumor. One of these markers has been identified as apolipoprotein A-II.

Cyanovirin-N (CV-N) exerts a potent human immunodeficiency virus (HIV)-inactivating activity against diverse strains of HIV by binding to the viral surface envelope glycoprotein gp120 and blocking its essential interactions with cellular receptors. Based on previous thermodynamic analyses, it has been speculated that discrete protein-protein interactions might play an important ancillary role in the CV-N/gp120 binding event, in addition to the interactions of CV-N with specific oligosaccharides present on gp120. Here, we report the identification and characterization of CV-N-binding peptides, which were isolated by screening of M13 phage-displayed peptide libraries. After performing three rounds of biopanning of the libraries against biotinylated CV-N, a CV-N-binding motif, X3CX6(W/F)(Y/F)CX2(Y/F), was evident. A vector was designed to express CV-N-binding peptides as a fusion with thioredoxin (Trx) containing a penta-His affinity tag. The CV-N-binding peptides fused with His-tagged Trx inhibited binding of the corresponding peptide-bearing phages to CV-N, confirming that the peptides possessed CV-N-binding activity. Optical biosensor binding studies showed that the one of the CV-N-binding peptide, TN10-1, bound to CV-N with a KD value of 1.9 microM. The results of alanine scanning mutagenesis of the peptide showed that aromatic residues at positions 11, 12, and 16, as well as the conformational structure of the peptide secured by a disulfide bond, were important for the binding interactions. A series of competitive binding assays confirmed that gp120 inhibited CV-N binding of the corresponding peptide-bearing phages, and suggested that TN10-1 peptides were mimicking the protein component of gp120 rather than mimicking specific oligosaccharides present on gp120.

ORNL is developing a high-speed, full-duplex all weather communications link for ranges up to 5 kilometers. To accomplish this project, we have constructed an RF-driven waveguide CO2 laser and a dielectric-waveguide Stark modulator. The 10-micron wavelength was selected for its ability to penetrate smoke, fog, and rain. The modulator is based on the Stark shift of NH2D (deuterated ammonia). The laser is driven by a 60 MHz RF amplifier at a power level of approximately 50 watts. The resonator cavity of the laser is formed by a 2.4 mm internal diameter ceramic waveguide with external optics. The RF electrodes are formed from aluminum heatsink extrusions that also provide cooling for the discharge. Details of the laser design will be presented.

Nanocrystalline silicon (n-Si) is formed in a silicon dioxide thin-film matrix by ion implantation followed by thermal annealing in forming gas at 1100 °C for 1 hour. The ion implantation is performed using multiple implants with different implantation energies and doses to create a quasi-flat concentration of silicon atoms throughout the silicon dioxide film. These samples are then analyzed using spectroscopic ellipsometry to characterize their linear optical properties. Implantations with small doses (5 × 1020 Si atoms/cm3) increase the refractive index by a small amount (δn∼0.006 at 600nm), while implantations with moderate dose (5 × 1021 Si atoms/cm3) have a larger increase in refractive index and exhibit optical absorption above ∼1.9 eV (650 nm).

By exploiting phase-separation in oxide materials, we present a simple and potentially low-cost approach to create exceptional superhydrophobicity in thin-film based coatings. By selecting the TiO2–Cu2O system and depositing through magnetron sputtering onto single crystal and metal templates, we demonstrate growth of nanostructured, chemically phase-segregated composite films. These coatings, after appropriate chemical surface modification, demonstrate a robust, non-wetting Cassie–Baxter state and yield an exceptional superhydrophobic performance, with water droplet contact angles reaching to ~172° and sliding angles

This paper gives optical and magneto-optical (MO) characterization results (index of refraction, absorption coefficient, thickness, Faraday rotation, and coercivity) obtained from bismuth-doped dysprosium iron garnet samples. These samples were prepared under differing sol-gel processing conditions and were characterized using the Ultimate Ellipsometer. After first describing the design and measurement capabilities of the Ultimate Ellipsometer (Simpson, 1995), we then describe how various processing parameters, such as spin speed and annealing temperatures, affect the bismuth iron garnet material's optical and MO properties.© (1995) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

In previous studies from our laboratory we found that the CCl4 reactive metabolites produced during enzymatic in vitro or in vivo CCl4 biotransformation covalently bind to DNA. Further, chemically produced •CCl3 produce many adducts of unknown structure with the four DNA bases when the reaction proceeds in model systems. In the present work, we describe our attempt to elucidate by GLC/MS the structures of the adducts resulting when chemically generated •CCl3 interact with thymine. The following reaction products were identified: (i) 5-hydroxymethyl uracil; (ii) thymineglycol; (iii) 5-trichloroethyl uracil (tentative) and (iv) two isomeric 5,6-monochloro monohydroxy adducts of thymine (tentative). Reaction products found do not involve thymine positions directly participating in base-pairing processes. However, alterations in thymine structure reported if they occurred in DNA from livers of CCl4 poisoned animals, might potentially have biological significance that remains to be established.

Loose multi-wall carbon nanotubes (MWNTs) were processed into a bundle of 19,600 individual channels with an individual channel diameter of 0.4 microm using a fiber drawing process. First, a powder of sodium silicate solution containing purified MWNTs was created. A glass capillary tube was filled with the powder and drawn into fibers. The fibers were cut into segments, bundled and redrawn multiple times to create fibers with multiple channels containing MWNTs. This processing approach created thousands of uniformly ordered channels containing dispersed MWNTs in a glass matrix while simultaneously aligning the MWNTs. The bulk resistivity of the MWNT-silicate channel has been improved by 38% after two consecutive draws as a result of the increased MWNT fraction.

The authors investigate the properties of water drops on uniform arrays of vertically aligned nanocones. A simple model predicts a variety of behaviors depending on the nanocone aspect ratio. At low aspect ratios, surface tension pulls the water into the nanocone array, resulting in a wetted drop with high contact angle hysteresis. At high aspect ratios, the effect of the surface tension is reversed, resulting in little liquid-solid contact and drops which easily slide off the surface. Experiments show that the transition occurs as a gradual change in drop contact angles, but an abrupt change in sliding angles.

The HIV-1 nucleocapsid (NC) protein is a small, basic protein containing two retroviral zinc fingers. It is a highly active nucleic acid chaperone; because of this activity, it plays a crucial role in virus replication as a cofactor during reverse transcription, and is probably important in other steps of the replication cycle as well. We previously reported that NC binds with high-affinity to the repeating sequence d(TG)n. We have now analyzed the interaction between NC and d(TG)4 in considerable detail, using surface plasmon resonance (SPR), tryptophan fluorescence quenching (TFQ), fluorescence anisotropy (FA), isothermal titration calorimetry (ITC) and electrospray ionization Fourier transform mass spectrometry (ESI-FTMS). Our results show that the interactions between these two molecules are surprisngly complex: while the K(d) for binding of a single d(TG)4 molecule to NC is only approximately 5 nM in 150 mM NaCl, a single NC molecule is capable of interacting with more than one d(TG)4 molecule, and conversely, more than one NC molecule can bind to a single d(TG)4 molecule. The strengths of these additional binding reactions are quantitated. The implications of this multivalency for the functions of NC in virus replication are discussed.