The viable bioengineered allogeneic cellularized construct StrataGraft® synthesizes, deposits, and organizes human extracellular matrix proteins into tissue type-specific structures and secretes soluble factors associated with wound healing.

BACKGROUND: StrataGraft® (allogeneic cultured keratinocytes and dermal fibroblasts in murine collagen-dsat) is an FDA-approved viable bioengineered allogeneic cellularized construct for adult patients with deep partial-thickness burns requiring surgery. We characterized the structural and functional properties of StrataGraft to improve product understanding by evaluating extracellular matrix (ECM) molecule distribution and secreted protein factor expression in vitro. METHODS: ECM protein expression was determined using indirect immunofluorescence on construct cross sections using commercial antibodies against collagen III, IV, VI, laminin-332, and decorin. Human collagen I expression was verified by enzyme-linked immunosorbent assay (ELISA) for collagen I C-terminal propeptide. Soluble protein factor secretion was quantified by multiplex biomarker assays and singleplex ELISA in conditioned media from meshed constructs. RESULTS: StrataGraft cellular components produced collagen I, collagen III, collagen VI, and decorin in patterns indicating an organized ECM. Distributions of collagen IV and laminin-332 indicated formation of basement membranes and dermal-epidermal junctions. Soluble protein factors were observed in the pg/cm2/h range from 1 h to the experiment end at 168 h. CONCLUSIONS: The organization of the ECM proteins was like human skin and the viable cellular components provided sustained secretion of soluble wound healing factors, making StrataGraft an attractive option for treating severe burns.

Discovery of Ligands for TRIM58, a Novel Tissue-Selective E3 Ligase.

Redirecting E3 ligases to neo-substrates, leading to their proteasomal disassembly, known as targeted protein degradation (TPD), has emerged as a promising alternative to traditional, occupancy-driven pharmacology. Although the field has expanded tremendously over the past years, the choice of E3 ligases remains limited, with an almost exclusive focus on CRBN and VHL. Here, we report the discovery of novel ligands to the PRY-SPRY domain of TRIM58, a RING ligase that is specifically expressed in erythroid precursor cells. A DSF screen, followed by validation using additional biophysical methods, led to the identification of TRIM58 ligand TRIM-473. A basic SAR around the chemotype was established by utilizing a competitive binding assay employing a short FP peptide probe derived from an endogenous TRIM58 substrate. The X-ray co-crystal structure of TRIM58 in complex with TRIM-473 gave insights into the binding mode and potential exit vectors for bifunctional degrader design.

Strategies for implementing the Appointment-Based Model at a rural independent chain of community pharmacies.

BACKGROUND: The Appointment-Based Model (ABM) is a care model that helps community pharmacists streamline their medication dispensing workflow while simultaneously integrating patient care into the medication preparation process through medication synchronization. Implementation of the ABM has varied across community pharmacies. Further studies that identify tailored implementation approaches are needed to support broad adoption of the ABM in practice. OBJECTIVES: (1) To determine facilitators and barriers to ongoing adoption and implementation of the ABM at a small chain of rural independent pharmacies where adoption has stalled and (2) to identify implementation strategies to support further adoption of the ABM at these pharmacies METHODS: This project was an exploratory, mid-implementation study. Semistructured interviews were conducted with pharmacy staff who participated in the ongoing implementation and use of the ABM at the pharmacies. Interviews elicited stakeholder-centered perspectives on (1) experiences with the ABM to date, (2) processes and roles for the ABM, and (3) opinions on how implementation of the ABM could be improved at the pharmacies. Rapid qualitative assessment methodology was used for analysis to identify facilitators and barriers and to select implementation strategies. RESULTS: Thirty-one pharmacy personnel were interviewed: pharmacists (n = 10), pharmacy technicians (n = 7), and fill clerks (n = 14). The research team identified 6 facilitators and 4 barriers to the implementation of the ABM at the pharmacies. Five implementation strategies were selected based on the facilitators and barriers: (1) capture and share local knowledge across pharmacy sites, (2) conduct educational outreach visits, (3) conduct ongoing training, (4) prepare patients to be active participants in the ABM, and (5) organize clinician implementation team meetings. CONCLUSIONS: Development of a stakeholder-driven implementation approach may support further implementation and adoption of the ABM in practice.

A systematic review of community pharmacy initiatives to improve treatment of depression and pain: Focus on types of programs and patient-reported outcomes.

INTRODUCTION: Depression and pain are common, disabling, mutually exacerbating conditions. Many patients living with these conditions present to community pharmacies on a regular schedule to purchase both prescribed and over-the-counter medications. Community-pharmacy based programs have been developed to improve depression and pain outcomes. METHODS: The PRISMA guidelines were utilized to answer the following question: In patients with depression and/or pain, what is the effect of the existing community pharmacy programs on depression and/or pain outcomes. Queried databases included Pubmed, EMBASE, and PsychINFO. DistillerSR was used to organize the screening, abstraction, and review of data. All potential articles were evaluated by two authors, and conflicts were discussed to achieve resolution. In addition to primary outcomes, sources of potential bias and quality indicators were abstracted for every article. RESULTS: Three thousand nine hundred and twenty articles were reviewed, and 13 studies met eligibility criteria (n = 7 for depression; n = 6 for pain). Most studies demonstrated improvement in measures of depression or pain. However, compared to usual care or other control conditions, most of the depression and pain-specific interventions did not provide additional symptomatic benefit. The community pharmacy-based interventions were superior for other outcomes including medication adherence, reducing stigma, improvement in self-efficacy, and improvement in general management of disease. CONCLUSION: Community pharmacies may be uniquely positioned to deliver interventions that improve outcomes associated with successful depression and pain treatment outcomes. However, the benefits of published community pharmacy-based treatments for actually improving depression and pain severity has not yet been established. Innovative interventions and additional research may be needed to achieve clinical success for pharmacy interventions for depression and pain.

Improving Effective Mental Health Consultation for Rural Older Adults Living With Depression and Pain: Learning From the Experiences of Rural Primary Care Physicians.

OBJECTIVE: Novel approaches are needed to assist rural primary care physicians (PCPs) in caring for older patients living with depression and pain who are at an elevated suicide risk. To refine and improve a model of care (PREDICTOR: Pharmacy Identification and Primary Care Intervention of Older Adults at Risk for Suicide), we conducted qualitative interviews with rural PCPs about (1) caring for seniors with depression, pain, and suicidality and (2) their favored procedures for working with psychiatric consultants and the professional characteristics desired in an effective consultant. METHODS: The study utilized a best-practice approach (including double coding) for qualitative interviews with 10 PCPs practicing in rural Pennsylvania. PCPs were interviewed about 3 themes related to caring for older adults with depression, pain, and suicidal ideation and working with psychiatric consultants. The study was conducted from January 2019 to May 2019. RESULTS: Four primary themes emerged from the interviews. (1) Rural PCPs become comfortable managing depression in older adults out of necessity, but desire collaboration on more complex mental health care. (2) Comorbid depression and pain are universally described as related through a vicious cycle in older adults. (3) Rural PCPs experience varying comfort with prescribing opioids for pain management in older patients, but most prefer not to prescribe opioids, and some refuse to do so. (4) PCPs endorsed the PREDICTOR remote consultation model as potentially beneficial to themselves and their older patients, but strongly desired that the consultant work with them as collaborators and for a collegial professional relationship with the mental health specialist. CONCLUSIONS: Rural PCPs are comfortable with remote consultation for older patients living with depression but desire collegial relationships with these consultants, supporting a collaborative approach. We describe explicit plans for implementing these findings as we refine PREDICTOR, in efforts to promote PCP practice change.

On the bandwidth dependent performance of split transmitter-receiver optical fiber nonlinearity compensation.

The Gaussian noise model is used to estimate the performance of three digital nonlinearity compensation (NLC) algorithms in C-band, long-haul, optical fiber transmission, when the span length and NLC bandwidth are independently varied. The algorithms are receiver-side digital backpropagation (DBP), transmitter-side DBP (digital precompensation), and Split NLC (an equal division of DBP between transmitter and receiver). For transmission over 100×100 km spans, the model predicts a 0.2 dB increase in SNR when applying Split NLC (versus DBP) to a single 32 GBd channel (from 0.4 dB to 0.6 dB), monotonically increasing with NLC bandwidth up to 1.6 dB for full-field NLC. The underlying assumptions of this model and the practical considerations for implementation of Split NLC are discussed. This work demonstrates, theoretically, that, regardless of the transmission scenario, it is always beneficial to divide NLC between transmitter and receiver, and identifies the transmission regimes where Split NLC is particularly advantageous.

Reduction Dynamics of Doped Ceria, Nickel Oxide, and Cermet Composites Probed Using In Situ Raman Spectroscopy.

The redox properties of gadolinium doped ceria (CGO) and nickel oxide (NiO) composite cermets underpin the operation of solid oxide electrochemical cells. Although these systems have been widely studied, a full comprehension of the reaction dynamics at the interface of these materials is lacking. Here, in situ Raman spectroscopic monitoring of the redox cycle is used to investigate the interplay between the dynamic and competing processes of hydrogen spillover and water dissociation on the doped ceria surface. In order to elucidate these mechanisms, the redox process in pure CGO and NiO is studied when exposed to wet and dry hydrogen and is compared to the cermet behavior. In dry hydrogen, CGO reduces relatively rapidly via a series of intermediate phases, while NiO reduces via a single-step process. In wet reducing atmospheres, however, the oxidation state of pure CGO is initially stabilized due to the dissociation of water by reduced Ce(III) and subsequent incorporation of oxygen into the structure. In the reduction process involving the composite cermet, the close proximity of the NiO improves the efficiency and speed of the composite reduction process. Although NiO is already incorporated into working cells, these observations suggest direct routes to further improve cell performance.

Increasing the information rates of optical communications via coded modulation: a study of transceiver performance.

Optical fibre underpins the global communications infrastructure and has experienced an astonishing evolution over the past four decades, with current commercial systems transmitting data rates in excess of 10 Tb/s over a single fibre core. The continuation of this dramatic growth in throughput has become constrained due to a power dependent nonlinear distortion arising from a phenomenon known as the Kerr effect. The mitigation of fibre nonlinearities is an area of intense research. However, even in the absence of nonlinear distortion, the practical limit on the transmission throughput of a single fibre core is dominated by the finite signal-to-noise ratio (SNR) afforded by current state-of-the-art coherent optical transceivers. Therefore, the key to maximising the number of information bits that can be reliably transmitted over a fibre channel hinges on the simultaneous optimisation of the modulation format and code rate, based on the SNR achieved at the receiver. In this work, we use an information theoretic approach based on the mutual information and the generalised mutual information to characterise a state-of-the-art dual polarisation m-ary quadrature amplitude modulation transceiver and subsequently apply this methodology to a 15-carrier super-channel to achieve the highest throughput (1.125 Tb/s) ever recorded using a single coherent receiver.

Maximizing the optical network capacity.

Most of the digital data transmitted are carried by optical fibres, forming the great part of the national and international communication infrastructure. The information-carrying capacity of these networks has increased vastly over the past decades through the introduction of wavelength division multiplexing, advanced modulation formats, digital signal processing and improved optical fibre and amplifier technology. These developments sparked the communication revolution and the growth of the Internet, and have created an illusion of infinite capacity being available. But as the volume of data continues to increase, is there a limit to the capacity of an optical fibre communication channel? The optical fibre channel is nonlinear, and the intensity-dependent Kerr nonlinearity limit has been suggested as a fundamental limit to optical fibre capacity. Current research is focused on whether this is the case, and on linear and nonlinear techniques, both optical and electronic, to understand, unlock and maximize the capacity of optical communications in the nonlinear regime. This paper describes some of them and discusses future prospects for success in the quest for capacity.

Detection of T790M, the Acquired Resistance EGFR Mutation, by Tumor Biopsy versus Noninvasive Blood-Based Analyses.

PURPOSE: The T790M gatekeeper mutation in the EGFR is acquired by some EGFR-mutant non-small cell lung cancers (NSCLC) as they become resistant to selective tyrosine kinase inhibitors (TKI). As third-generation EGFR TKIs that overcome T790M-associated resistance become available, noninvasive approaches to T790M detection will become critical to guide management. EXPERIMENTAL DESIGN: As part of a multi-institutional Stand-Up-To-Cancer collaboration, we performed an exploratory analysis of 40 patients with EGFR-mutant tumors progressing on EGFR TKI therapy. We compared the T790M genotype from tumor biopsies with analysis of simultaneously collected circulating tumor cells (CTC) and circulating tumor DNA (ctDNA). RESULTS: T790M genotypes were successfully obtained in 30 (75%) tumor biopsies, 28 (70%) CTC samples, and 32 (80%) ctDNA samples. The resistance-associated mutation was detected in 47% to 50% of patients using each of the genotyping assays, with concordance among them ranging from 57% to 74%. Although CTC- and ctDNA-based genotyping were each unsuccessful in 20% to 30% of cases, the two assays together enabled genotyping in all patients with an available blood sample, and they identified the T790M mutation in 14 (35%) patients in whom the concurrent biopsy was negative or indeterminate. CONCLUSIONS: Discordant genotypes between tumor biopsy and blood-based analyses may result from technological differences, as well as sampling different tumor cell populations. The use of complementary approaches may provide the most complete assessment of each patient's cancer, which should be validated in predicting response to T790M-targeted inhibitors.

High spectral density transmission emulation using amplified spontaneous emission noise.

We demonstrate the use of spectrally shaped amplified spontaneous emission (SS-ASE) noise for wideband channel loading in the investigation of nonlinear transmission limits in wavelength-division multiplexing transmission experiments using Nyquist-spaced channels. The validity of this approach is explored through statistical analysis and experimental transmission of Nyquist-spaced 10 GBaud polarization-division multiplexing (PDM) quadrature phase-shift keying and PDM-16-ary quadrature amplitude modulation (QAM) channels, co-propagated with SS-ASE over single mode fiber. It is shown that this technique, which is simpler to implement than a fully modulated comb of channels, is valid for distances exceeding 240 km for PDM-16QAM with dispersion of 16 ps/nm/km, yields a good agreement with theory, and provides a conservative measure of system performance.

Unrepeatered Nyquist PDM-16QAM transmission over 364 km using Raman amplification and multi-channel digital back-propagation.

Transmission of a net 467-Gb/s PDM-16QAM Nyquist-spaced superchannel is reported with an intra-superchannel net spectral efficiency (SE) of 6.6 (b/s)/Hz, over 364-km SMF-28 ULL ultra-low loss optical fiber, enabled by bi-directional second-order Raman amplification and digital nonlinearity compensation. Multi-channel digital back-propagation (MC-DBP) was applied to compensate for nonlinear interference; an improvement of 2 dB in Q(2) factor was achieved when 70-GHz DBP bandwidth was applied, allowing an increase in span length of 37 km.

Spectrally shaped DP-16QAM super-channel transmission with multi-channel digital back-propagation.

The achievable transmission capacity of conventional optical fibre communication systems is limited by nonlinear distortions due to the Kerr effect and the difficulty in modulating the optical field to effectively use the available fibre bandwidth. In order to achieve a high information spectral density (ISD), while simultaneously maintaining transmission reach, multi-channel fibre nonlinearity compensation and spectrally efficient data encoding must be utilised. In this work, we use a single coherent super-receiver to simultaneously receive a DP-16QAM super-channel, consisting of seven spectrally shaped 10GBd sub-carriers spaced at the Nyquist frequency. Effective nonlinearity mitigation is achieved using multi-channel digital back-propagation (MC-DBP) and this technique is combined with an optimised forward error correction implementation to demonstrate a record gain in transmission reach of 85%; increasing the maximum transmission distance from 3190 km to 5890 km, with an ISD of 6.60 b/s/Hz. In addition, this report outlines for the first time, the sensitivity of MC-DBP gain to linear transmission line impairments and defines a trade-off between performance and complexity.

Printing in three dimensions with graphene.

Responsive graphene oxide sheets form non-covalent networks with optimum rheological properties for 3D printing. These networks have shear thinning behavior and sufficiently high elastic shear modulus (G') to build self-supporting 3D structures by direct write assembly. Drying and thermal reduction leads to ultra-light graphene-only structures with restored conductivity and elastomeric behavior.

The Drosophila Huntington's disease gene ortholog dhtt influences chromatin regulation during development.

Huntington's disease is an autosomal dominant neurodegenerative disorder caused by a CAG expansion mutation in HTT, the gene encoding huntingtin. Evidence from both human genotype-phenotype relationships and mouse model systems suggests that the mutation acts by dysregulating some normal activity of huntingtin. Recent work in the mouse has revealed a role for huntingtin in epigenetic regulation during development. Here, we examine the role of the Drosophila huntingtin ortholog (dhtt) in chromatin regulation in the development of the fly. Although null dhtt mutants display no overt phenotype, we found that dhtt acts as a suppressor of position-effect variegation (PEV), suggesting that it influences chromatin organization. We demonstrate that dhtt affects heterochromatin spreading in a PEV model by modulating histone H3K9 methylation levels at the heterochromatin-euchromatin boundary. To gain mechanistic insights into how dhtt influences chromatin function, we conducted a candidate genetic screen using RNAi lines targeting known PEV modifier genes. We found that dhtt modifies phenotypes caused by knockdown of a number of key epigenetic regulators, including chromatin-associated proteins, histone demethylases (HDMs) and methyltransferases. Notably, dhtt strongly modifies phenotypes resulting from loss of the HDM dLsd1, in both the ovary and wing, and we demonstrate that dhtt appears to act as a facilitator of dLsd1 function in regulating global histone H3K4 methylation levels. These findings suggest that a fundamental aspect of huntingtin function in heterochromatin/euchromatin organization is evolutionarily conserved across phyla.

Mesoscale assembly of chemically modified graphene into complex cellular networks.

The widespread technological introduction of graphene beyond electronics rests on our ability to assemble this two-dimensional building block into three-dimensional structures for practical devices. To achieve this goal we need fabrication approaches that are able to provide an accurate control of chemistry and architecture from nano to macroscopic levels. Here, we describe a versatile technique to build ultralight (density ≥1 mg cm(-3)) cellular networks based on the use of soft templates and the controlled segregation of chemically modified graphene to liquid interfaces. These novel structures can be tuned for excellent conductivity; versatile mechanical response (elastic-brittle to elastomeric, reversible deformation, high energy absorption) and organic absorption capabilities (above 600 g per gram of material). The approach can be used to uncover the basic principles that will guide the design of practical devices that by combining unique mechanical and functional performance will generate new technological opportunities.

Clinical consequences of polypharmacy in elderly.

INTRODUCTION: Polypharmacy, defined as the use of multiple drugs or more than are medically necessary, is a growing concern for older adults. MEDLINE and EMBASE databases were searched from January 1, 1986 to June 30, 2013) to identify relevant articles in people aged > 65 years. AREAS COVERED: We present information about: i) prevalence of polypharmacy and unnecessary medication use; ii) negative consequences of polypharmacy; and iii) interventions to improve polypharmacy. EXPERT OPINION: International research shows that polypharmacy is common in older adults with the highest number of drugs taken by those residing in nursing homes. Nearly 50% of older adults take one or more medications that are not medically necessary. Research has clearly established a strong relationship between polypharmacy and negative clinical consequences. Moreover, well-designed interprofessional (often including clinical pharmacist) intervention studies that focus on enrolling high-risk older patients with polypharmacy have shown that they can be effective in reducing aspects of unnecessary prescribing with mixed results on distal health outcomes.

Gain-switched multicarrier transmitter in a long-reach UDWDM PON with a digital coherent receiver.

The authors report on the downlink performance of a 10 Gb/s long-reach and ultra-dense wavelength-division multiplexed passive optical network, based on a multicarrier transmitter realized by using an externally injected gain-switched distributed-feedback laser diode. Each of the comb channels, spaced by 10 GHz, is modulated with a 3 Gbaud dual polarization quadrature phase shift keying signal that included a 20% overhead for forward error correction. Frequency selectivity and enhanced receiver sensitivity is achieved by employing a digital coherent receiver to receive the signal. Experimental results achieved in a back-to-back and 100 km transmission scenarios show an excellent worst case receiver sensitivity of -44 dBm.

Genetic and functional studies implicate synaptic overgrowth and ring gland cAMP/PKA signaling defects in the Drosophila melanogaster neurofibromatosis-1 growth deficiency.

Neurofibromatosis type 1 (NF1), a genetic disease that affects 1 in 3,000, is caused by loss of a large evolutionary conserved protein that serves as a GTPase Activating Protein (GAP) for Ras. Among Drosophila melanogaster Nf1 (dNf1) null mutant phenotypes, learning/memory deficits and reduced overall growth resemble human NF1 symptoms. These and other dNf1 defects are relatively insensitive to manipulations that reduce Ras signaling strength but are suppressed by increasing signaling through the 3'-5' cyclic adenosine monophosphate (cAMP) dependent Protein Kinase A (PKA) pathway, or phenocopied by inhibiting this pathway. However, whether dNf1 affects cAMP/PKA signaling directly or indirectly remains controversial. To shed light on this issue we screened 486 1(st) and 2(nd) chromosome deficiencies that uncover >80% of annotated genes for dominant modifiers of the dNf1 pupal size defect, identifying responsible genes in crosses with mutant alleles or by tissue-specific RNA interference (RNAi) knockdown. Validating the screen, identified suppressors include the previously implicated dAlk tyrosine kinase, its activating ligand jelly belly (jeb), two other genes involved in Ras/ERK signal transduction and several involved in cAMP/PKA signaling. Novel modifiers that implicate synaptic defects in the dNf1 growth deficiency include the intersectin-related synaptic scaffold protein Dap160 and the cholecystokinin receptor-related CCKLR-17D1 drosulfakinin receptor. Providing mechanistic clues, we show that dAlk, jeb and CCKLR-17D1 are among mutants that also suppress a recently identified dNf1 neuromuscular junction (NMJ) overgrowth phenotype and that manipulations that increase cAMP/PKA signaling in adipokinetic hormone (AKH)-producing cells at the base of the neuroendocrine ring gland restore the dNf1 growth deficiency. Finally, supporting our previous contention that ALK might be a therapeutic target in NF1, we report that human ALK is expressed in cells that give rise to NF1 tumors and that NF1 regulated ALK/RAS/ERK signaling appears conserved in man.