Computational Design of Phosphotriesterase Improves V-Agent Degradation Efficiency.

Organophosphates (OPs) are a class of neurotoxic acetylcholinesterase inhibitors including widely used pesticides as well as nerve agents such as VX and VR. Current treatment of these toxins relies on reactivating acetylcholinesterase, which remains ineffective. Enzymatic scavengers are of interest for their ability to degrade OPs systemically before they reach their target. Here we describe a library of computationally designed variants of phosphotriesterase (PTE), an enzyme that is known to break down OPs. The mutations G208D, F104A, K77A, A80V, H254G, and I274N broadly improve catalytic efficiency of VX and VR hydrolysis without impacting the structure of the enzyme. The mutation I106 A improves catalysis of VR and L271E abolishes activity, likely due to disruptions of PTE's structure. This study elucidates the importance of these residues and contributes to the design of enzymatic OP scavengers with improved efficiency.

Micro-DRIFTS for small area hyper-black spectroscopy.

We have developed a low-cost micro-diffuse reflectance infrared Fourier transform spectroscopic (micro-DRIFTS) setup for measuring the reflectance of small area diffuse samples. The system performance is characterized and then demonstrated on small area vertically aligned carbon nanotube (VACNT) samples. We find that our system can measure samples with a spatial resolution of approximately 140 µm with sensitivities of 10s of ppm in the 2 µm - 18 µm spectral window. Our uncertainty budget is presented along with how our measured reflectance can be equated to directional-hemispherical reflectance.

The growth of recovery capital in clients of recovery residences in Florida, USA: a quantitative pilot study of changes in REC-CAP profile scores.

BACKGROUND: There is a growing evidence base around predictors of retention and completion in a range of recovery residence models, particularly Oxford Houses and Sober Living Houses, and recovery housing is recognized as a clearly evidenced area of recovery intervention. The aim of the study was to quantitatively assess recovery capital in a sample of recovery residence clients. METHOD: The study used a repeated measures self-completion of a standardized recovery capital instrument (REC-CAP) for clients retained across various houses within one Level 2 recovery residence provider whose program was based on a 12-step approach. While 823 clients participated in the baseline assessment, a sample of 267 clients was achieved for six-month follow-up interview, based on those retained in the residence. A logistic regression model examined factors associated with retention and a repeated measures marginal mixed model evaluated the factors associated with changes in recovery capital between the baseline and the follow-up assessment. RESULTS: Members of the group that remained in recovery residences were more likely to be older with a record of high participation in recovery groups, with greater drop-out among younger residents, female residents and those with an identified housing need. For those retained to follow-up, greater recovery capital growth was associated with employment, higher levels of social support and more recovery group involvement, as well as age and a higher quality of life. The need for family support was shown to reduce levels of recovery capital. However, those younger people who were retained reported better recovery capital growth during the initial six months of residence. CONCLUSION: The key conclusion is that while recovery capital generally increases during a stay in a recovery residence, it does not do so consistently across the sample population. This has implications for how pathways to recovery group engagement are supported for women and young people and how social support (encompassing housing, employment and family issues) is provided to those populations during periods of residence. This suggests the potential need for training and guidance for house managers working with these groups.

siRNA high throughput screening identifies regulators of chloropicrin and hydrogen fluoride injury in human corneal epithelial cell models.

Corneal injuries induced by various toxicants result in similar clinical presentations such as corneal opacity and neovascularization. Many studies suggest that several weeks post-exposure a convergence of the molecular mechanisms drives these progressive pathologies. However, chemical agents vary in toxicological properties, and early molecular responses are anticipated to be somewhat dissimilar for different toxicants. We chose 3120 targets from the Dharmacon Human Druggable genome to screen for chloropicrin (CP) and hydrogen fluoride (HF) corneal injury as we hypothesized that targets identified in vitro may be effective as therapeutic targets in future studies. Human immortalized corneal epithelial cells (SV40-HCEC) were used for screening. Cell viability and IL-8 were analyzed to down-select hits into validation studies, where multiplex cytokine analysis and high content analysis were performed to understand toxicant effect and target function. Some endpoints were also evaluated in a second human immortalized corneal epithelial cell line, TCEpi. Over 20 targets entered validation studies for CP and HF; of these, only three targets were shared: NR3C1, RELA, and KMT5A. These findings suggest that early molecular responses to different toxicants may be somewhat distinctive and present dissimilar targets for possible early intervention.

Extreme laser pulse-energy measurements by means of photon momentum.

Extreme lasers capable of short, high-energy pulses are probing the frontiers of science and advancing practical technology. The utility of such lasers increases with their average power delivery, which enables faster data acquisition, higher flux of laser-driven particle and radiation sources and more efficient material processing. However, the same extreme energies and electric field strengths of these lasers are currently preventing their direct and high accuracy measurement for these experimental applications. To overcome this limitation, we use the momentum of the laser pulses as a measurement proxy for their energy. When light reflects from an ideal mirror, its momentum is transferred to the mirror, but its energy is reflected. We demonstrate here a force-sensing mirror configuration to measure laser pulse energies up to 100 J/pulse (10 ns duration, 10 Hz repetition rate) from a kilowatt-level average power multi-slab laser operated at the HiLASE facility of the Czech Academy of Sciences. We combine a radiation-pressure power meter with a charge integrator photodiode to form what we refer to as a Radiation Pressure Energy Meter. To our knowledge, this is the first demonstration of a high-accuracy, non-absorbing, SI traceable primary standard measurement of both single and average pulse energies of a 1-kW-average-power pulsed laser source. With this, we demonstrate a practical method for in-situ calibration of the traditional thermal instruments (pyroelectric detectors) currently used for indirect measurements of energy and power of such extreme lasers.

Chk1 and the Host Cell DNA Damage Response as a Potential Antiviral Target in BK Polyomavirus Infection.

The human BK polyomavirus (BKPyV) is latent in the kidneys of most adults, but can be reactivated in immunosuppressed states, such as following renal transplantation. If left unchecked, BK polyomavirus nephropathy (PyVAN) and possible graft loss may result from viral destruction of tubular epithelial cells and interstitial fibrosis. When coupled with regular post-transplant screening, immunosuppression reduction has been effective in limiting BKPyV viremia and the development of PyVAN. Antiviral drugs that are safe and effective in combating BKPyV have not been identified but would be a benefit in complementing or replacing immunosuppression reduction. The present study explores inhibition of the host DNA damage response (DDR) as an antiviral strategy. Immunohistochemical and immunofluorescent analyses of PyVAN biopsies provide evidence for stimulation of a DDR in vivo. DDR pathways were also stimulated in vitro following BKPyV infection of low-passage human renal proximal tubule epithelial cells. The role of Chk1, a protein kinase known to be involved in the replication stress-induced DDR, was examined by inhibition with the small molecule LY2603618 and by siRNA-mediated knockdown. Inhibition of Chk1 resulted in decreased replication of BKPyV DNA and viral spread. Activation of mitotic pathways was associated with the reduction in BKPyV replication. Chk1 inhibitors that are found to be safe and effective in clinical trials for cancer should also be evaluated for antiviral activity against BKPyV.

High-accuracy room temperature planar absolute radiometer based on vertically aligned carbon nanotubes.

We have developed a planar absolute radiometer for room temperature (PARRoT) that will replace the legacy C-series calorimeter as the free-space continuous-wave laser power detector standard at the National Institute of Standards and Technology (NIST). This instrument will lower the combined relative expanded measurement uncertainty (k = 2) from 0.84 % to 0.13 %. PARRoT's performance was validated by comparing its response against a transfer standard silicon trap detector traceable to NIST's primary standard laser optimized cryogenic radiometer (LOCR) and against the C-series calorimeter. On average, these comparisons agreed to better than 0.008 % and 0.05 %, respectively.

High amplification laser-pressure optic enables ultra-low uncertainty measurements of the optical laser power at kilowatt levels.

We present the first measurements of kilowatt laser power with an uncertainty less than 1 %. These represent progress toward the most accurate measurements of laser power above 1 kW at 1070 nm wavelength and establish a more precise link between force metrology and laser power metrology. Radiation pressure, or photon momentum, is a relatively new method of non-destructively measuring laser power. We demonstrate how a multiple reflection optical system amplifies the pressure of a kilowatt class laser incoherently to improve the signal to noise ratio in a radiation pressure-based measurement. With 14 incoherent reflections of the laser, we measure a total uncertainty of 0.26 % for an input power of 10 kW and 0.46 % for an input power of 1 kW at the 95 % confidence level. These measurements of absolute power are traceable to the SI kilogram and mark a state-of-the-art improvement in measurement precision by a factor of four.

Feedback Control of a Nonlinear Electrostatic Force Transducer.

We document a feedback controller design for a nonlinear electrostatic transducer that exhibits a strong unloaded resonance. Challenging features of this type of transducer include the presence of multiple fixed points (some of which are unstable), nonlinear force-to-deflection transfer, effective spring-constant softening due to electrostatic loading and associated resonance frequency shift. Furthermore, due to the utilization of lowpass filters in the electronic readout circuitry, a significant amount of transport delay is introduced in the feedback loop. To stabilize this electro-mechanical system, we employ an active disturbance-rejecting controller with nonlinear force mapping and delay synchronization. As demonstrated by numerical simulations, the combination of these three control techniques stabilizes the system over a wide range of electrode deflections. The proposed controller shows good setpoint tracking and disturbance rejection, and improved settling time, compared to the sensor alone.

Simplified kilogram traceability for high-power laser measurement using photon momentum radiometers.

Photon momentum radiometers measure the force imparted by a reflected laser beam to determine the laser's optical power. This requires high-accuracy calibration of the force sensors using milligram and microgram mass artifacts. Calibrated test masses can therefore be used to provide traceability of these radiometers to the International System of Units, but low-noise calibration at these mass levels is difficult. Here, we present the improvement in calibration capability that we have gained from implementing a robotic mass delivery system. We quantify this in terms of the specific nuances of force measurements as implemented for laser power metrology.

Miniature force sensor for absolute laser power measurements via radiation pressure at hundreds of watts.

We present a small power meter that detects the radiation pressure of an incident high-power laser. Given its small package and non-destructive interaction with the laser, this power meter is well suited to realizing a robust real-time, high-accuracy power measurement in laser-based manufacturing environments. The incident laser power is determined through interferometric measurement of displacement of a 20 mm diameter high reflectivity mirror, mounted at the center of a dual element spiral flexure. This device can measure laser power from 25 W to 400 W with a 260 m W/H z noise floor and ≤ 3.2% expanded uncertainty. We validate our device against a calibrated thermopile with simultaneous measurements of an unpolarized 1070 nm laser and report good agreement between the two systems. Finally, by referencing to an identical mechanical spring that does not see the incident laser, we suppress vibration noise in the power measurement by 14.8 dB over a 600 Hz measured bandwidth. This is an improvement over other radiation pressure based power meters that have previously been demonstrated.

Development of mouse models for the study of chloropicrin and hydrogen fluoride ocular injury.

The possibility of chemical terrorism within the United States is a rising concern, with the eye being one of the most sensitive tissues to toxicant exposure. We sought to develop mouse models of toxicant-induced ocular injury for the purpose of evaluating potential therapeutics. Chloropicrin (CP) and hydrogen fluoride (HF) were selected for the study owing to their reportedly high potential to induce ocular injury. Eyes of female BALB/c mice were exposed to CP or HF vapor in order to produce a moderate injury, as defined by acute corneal epithelial loss followed by progressive corneal pathology with the absence of injury to deeper eye structures. Clinical injury progression was evaluated up to 12 weeks postexposure, where a significant dose-dependent induction of corneal neovascularization was measured. Histopathology noted epithelial necrosis and stromal edema as early as 24 h after exposure but was resolved by 12 weeks. A significant increase in inflammatory cytokine concentrations was measured in the cornea 24 h after exposure and returned to baseline by day 14. The ocular injury models we developed here for CP and HF exposure should serve as a valuable tool for the future evaluation of novel therapeutics and the molecular mechanisms of injury.

Calibration of free-space and fiber-coupled single-photon detectors.

We measure the detection efficiency of single-photon detectors at wavelengths near 851 nm and 1533.6 nm. We investigate the spatial uniformity of one free-space-coupled single-photon avalanche diode and present a comparison between fusion-spliced and connectorized fiber-coupled single-photon detectors. We find that our expanded relative uncertainty for a single measurement of the detection efficiency is as low as 0.70% for fiber-coupled measurements at 1533.6 nm and as high as 1.78% for our free-space characterization at 851.7 nm. The detection-efficiency determination includes corrections for afterpulsing, dark count, and count-rate effects of the single-photon detector with the detection efficiency interpolated to operation at a specified detected count rate.

Inline laser power measurement by photon momentum.

We present a novel measurement scheme and instrumentation for quantifying laser power by means of photon momentum. The optical design is optimized such that the incoming laser beam is minimally perturbed and is available for other purposes along the incoming beam axis. Additionally, the geometry of the instrument gives access to the small but measurable transmittance between two passive mirrors that face the force sensor. The force sensor is based on a commercially available weighing instrument ("scale") that has a temporal response of approximately 5 s and a readability of approximately 1 µg (∼2 W). Our measurement results demonstrate beam profile and power for 500 W, but the mirror and mass (or force) calibration are suitable for very high power up to 50 kW and beyond. The optics are based on commercially available, off-the-shelf mirrors optimized for the angle of incidence and maximum reflectance at the wavelength of 1070 nm. The size of the complete instrument has an input aperture of Ø75 mm, but this constraint is only a matter of optimizing the beam path and box geometry.

Spectral, spatial, and survivability evaluation of a flash-dried plasma-etched nanotube spray coating.

We demonstrate improved manufacturability of spectrally flat detectors for visible to mid-infrared wavelengths by characterizing a carbon nanotube spray coating compatible with lithium tantalate and other thermal sensors. Compared against previous spray coatings, it demonstrated the highest responsivity yet attained due to both higher absorptivity and thermal diffusivity, while also being matured to a commercially available product. It demonstrated spectral nonuniformity from 300 nm to 12 µm less than 1% with uncertainty (k=2) under 0.4%. The spatial nonuniformity of the assembled sensor was less than 0.5% over the central half (4 mm) of the absorber. As with previous developments employing isotropic carbon nanotube coatings, the absorber surface was sufficiently robust to withstand cleaning by compressed air blast and survived repeated vacuum cycling without measurable impact upon responsivity.

High Throughput SiRNA Screening for Chloropicrin and Hydrogen Fluoride-Induced Cornea Epithelial Cell Injury.

Toxicant-induced ocular injury is a true ocular emergency because chemicals have the potential to rapidly inflict significant tissue damage. Treatments for toxicant-induced corneal injury are generally supportive as no specific therapeutics exist to treat these injuries. In the efforts to develop treatments and therapeutics to care for exposure, it can be important to understand the molecular and cellular mechanisms of these injuries. We propose that utilization of high throughput small inhibitory RNA (siRNA) screening can be an important tool that could help to more rapidly elucidate the molecular mechanisms of chemical cornea epithelial injury. siRNA are double stranded RNA molecules that are 19-25 nucleotides long and utilize the post-transcriptional gene silencing pathway to degrade mRNA which have homology to the siRNA. The resulting reduction of expression of the specific gene can then be studied in toxicant exposed cells to ascertain the function of that gene in the cellular response to the toxicant. The development and validation of in vitro exposure models and methods for the high throughput screening (HTS) of hydrogen fluoride- (HF) and chloropicrin- (CP) induced ocular injury are presented in this article. Although we selected these two toxicants, our methods are applicable to the study of other toxicants with minor modifications to the toxicant exposure protocol. The SV40 large T antigen immortalized human corneal epithelial cell line SV40-HCEC was selected for study. Cell viability and IL-8 production were selected as endpoints in the screening protocol. Several challenges associated with the development of toxicant exposure and cell culture methods suitable for HTS studies are presented. The establishment of HTS models for these toxicants allows for further studies to better understand the mechanism of injury and to screen for potential therapeutics for chemical ocular injury.

Synthesis of multi-omic data and community metabolic models reveals insights into the role of hydrogen sulfide in colon cancer.

Multi-omic data and genome-scale microbial metabolic models have allowed us to examine microbial communities, community function, and interactions in ways that were not available to us historically. Now, one of our biggest challenges is determining how to integrate data and maximize data potential. Our study demonstrates one way in which to test a hypothesis by combining multi-omic data and community metabolic models. Specifically, we assess hydrogen sulfide production in colorectal cancer based on stool, mucosa, and tissue samples collected on and off the tumor site within the same individuals. 16S rRNA microbial community and abundance data were used to select and inform the metabolic models. We then used MICOM, an open source platform, to track the metabolic flux of hydrogen sulfide through a defined microbial community that either represented on-tumor or off-tumor sample communities. We also performed targeted and untargeted metabolomics, and used the former to quantitatively evaluate our model predictions. A deeper look at the models identified several unexpected but feasible reactions, microbes, and microbial interactions involved in hydrogen sulfide production for which our 16S and metabolomic data could not account. These results will guide future in vitro, in vivo, and in silico tests to establish why hydrogen sulfide production is increased in tumor tissue.

Time-Resolved Absorptance and Melt Pool Dynamics during Intense Laser Irradiation of a Metal.

High-irradiance lasers incident on metal surfaces create a complex, dynamic process through which the metal can rapidly change from highly reflective to strongly absorbing. Absolute knowledge of this process underpins important industrial laser processes such as laser welding, cutting, and metal additive manufacturing. Determining the time-dependent absorptance of the laser light by a material is important, not only for gaining a fundamental understanding of the light-matter interaction but also for improving process design in manufacturing. Measurements of the dynamic optical absorptance are notoriously difficult due to the rapidly changing nature of the absorbing medium. These data are also of vital importance to process modelers, whose complex simulations need reliable, accurate input data; yet, there are very few available. In this work, we measure the time-dependent, reflected light during a 10-ms laser spot weld using an integrating-sphere apparatus. From this, we calculate the dynamic absorptance for 1070-nm-wavelength light incident on 316L stainless steel. The time resolution of our experiment (less than 1 µs) allows the determination of the precise conditions under which several important physical phenomena occur, such as melt and keyhole formation. The average absorptances determined optically are compared with calorimetrically determined values, and it is found that the calorimeter severely underestimates the absorbed energy due to mass lost during the spot weld. Weld-nugget cross sections are also presented to verify our interpretation of the optical results, as well as to provide experimental data for weld-model validation.

Onsite multikilowatt laser power meter calibration using radiation pressure.

We have demonstrated the calibration of a thermal power meter against a radiation pressure power meter in the range of 20 kW in a manufacturing test environment. The results were compared to a traditional calorimeter-based laboratory calibration undertaken at the National Institute of Standards and Technology. The results are reported, and the effects of nonideal conditions typical of measurements in low-stability environments are discussed.

Recovery capital pathways: Modelling the components of recovery wellbeing.

BACKGROUND: In recent years, there has been recognition that recovery is a journey that involves the growth of recovery capital. Thus, recovery capital has become a commonly used term in addiction treatment and research yet its operationalization and measurement has been limited. Due to these limitations, there is little understanding of long-term recovery pathways and their clinical application. METHODS: We used the data of 546 participants from eight different recovery residences spread across Florida, USA. We calculated internal consistency for recovery capital and wellbeing, then assessed their factor structure via confirmatory factor analysis. The relationships between time, recovery barriers and strengths, wellbeing and recovery capital, as well as the moderating effect of gender, were estimated using structural equations modelling. RESULTS: The proposed model obtained an acceptable fit (χ2 (141, N=546)=533.642, p<0.001; CMIN/DF=3.785; CFI=0.915; TLI=0.896; RMSEA=0.071). Findings indicate a pathway to recovery capital that involves greater time in residence ('retention'), linked to an increase in meaningful activities and a reduction in barriers to recovery and unmet needs that, in turn, promote recovery capital and positive wellbeing. Gender differences were observed. CONCLUSIONS: We tested the pathways to recovery for residents in the recovery housing population. Our results have implications not only for retention as a predictor of sustained recovery and wellbeing but also for the importance of meaningful activities in promoting recovery capital and wellbeing.