Venom-inspired somatostatin receptor 4 (SSTR4) agonists as new drug leads for peripheral pain conditions.

Persistent pain affects one in five people worldwide, often with severely debilitating consequences. Current treatment options, which can be effective for mild or acute pain, are ill-suited for moderate-to-severe persistent pain, resulting in an urgent need for new therapeutics. In recent years, the somatostatin receptor 4 (SSTR 4 ), which is expressed in sensory neurons of the peripheral nervous system, has emerged as a promising target for pain relief. However, the presence of several closely related receptors with similar ligand-binding surfaces complicates the design of receptor-specific agonists. In this study, we report the discovery of a potent and selective SSTR 4 peptide, consomatin Fj1, derived from extensive venom gene datasets from marine cone snails. Consomatin Fj1 is a mimetic of the endogenous hormone somatostatin and contains a minimized binding motif that provides stability and drives peptide selectivity. Peripheral administration of synthetic consomatin Fj1 provided analgesia in mouse models of postoperative and neuropathic pain. Using structure-activity studies, we designed and functionally evaluated several Fj1 analogs, resulting in compounds with improved potency and selectivity. Our findings present a novel avenue for addressing persistent pain through the design of venom-inspired SSTR 4 -selective pain therapeutics. One Sentence Summary: Venom peptides from predatory marine mollusks provide new leads for treating peripheral pain conditions through a non-opioid target.

Comparative electron microscopy particle sizing of TiO2 pigments: sample preparation and measurement.

Titanium dioxide (TiO2) pigment is a non-toxic, particulate material in widespread use and found in everyone's daily life. The particle size of the anatase or rutile crystals are optimised to produce a pigment that provides the best possible whiteness and opacity. The average particle size is intentionally much larger than the 100 nm boundary of the EU nanomaterial definition, but the TiO2 pigment manufacturing processes results in a finite nanoscale content fraction. This optically inefficient nanoscale fraction needs to be quantified in line with EU regulations. In this paper, we describe the measurement procedures used for product quality assurance by three TiO2 manufacturing companies and present number-based primary particle size distributions (PSDs) obtained in a round-robin study performed on five anatase pigments fabricated by means of sulfate processes in different plants and commonly used worldwide in food, feed, pharmaceutical and cosmetic applications. The PSDs measured by the three titanium dioxide manufacturers based on electron micrographs are in excellent agreement with one another but differ significantly from those published elsewhere. Importantly, in some cases, the PSDs result in a different regulatory classification for some of the samples tested. The electron microscopy results published here are supported by results from other complementary methods including surface area measurements. It is the intention of this publication to contribute to an ongoing discussion on size measurements of TiO2 pigments and other particulate materials and advance the development of widely acceptable, precise, and reproducible measurement protocols for measuring the number-based PSDs of particulate products in the size range of TiO2 pigments.

Fungal Carbon: A Cost-Effective Tunable Network Template for Creating Supercapacitors.

Carbons form critical components in biogas purification and energy storage systems and are used to modify polymer matrices. The environmental impact of producing carbons has driven research interest in biomass-derived carbons, although these have yield, processing, and resource competition limitations. Naturally formed fungal filaments are investigated, which are abundantly available as food- and biotechnology-industry by-products and wastes as cost-effective and sustainable templates for carbon networks. Pyrolyzed Agaricus bisporus and Pleurotus eryngii filament networks are mesoporous and microscale with a size regime close to carbon fibers. Their BET surface areas of ≈282 m2 g-1 and ≈60 m2 g-1, respectively, greatly exceed values associated with carbon fibers and non-activated pyrolyzed bacterial cellulose and approximately on par with values for carbon black and CNTs in addition to pyrolyzed pinewood, rice husk, corn stover or olive mill waste. They also exhibit greater specific capacitance than both non-activated and activated pyrolyzed bacterial cellulose in addition to YP-50F (coconut shell based) commercial carbons. The high surface area and specific capacitance of fungal carbon coupled with the potential to tune these properties through species- and growth-environment-associated differences in network and filament morphology and inclusion of inorganic material through biomineralization makes them potentially useful in creating supercapacitors.

From Unregulated Networks to Designed Microstructures: Introducing Heterogeneity at Different Length Scales in Photopolymers for Additive Manufacturing.

Photopolymers have been optimized as protective and decorative coating materials for decades. However, with the rise of additive manufacturing technologies, vat photopolymerization has unlocked the use of photopolymers for three-dimensional objects with new material requirements. Thus, the originally highly cross-linked, amorphous architecture of photopolymers cannot match the expectations for modern materials anymore, revealing the largely unanswered question of how diverse properties can be achieved in photopolymers. Herein, we review how microstructural features in soft matter materials should be designed and implemented to obtain high performance materials. We then translate these findings into chemical design suggestions for enhanced printable photopolymers. Based on this analysis, we have found microstructural heterogenization to be the most powerful tool to tune photopolymer performance. By combining the chemical toolbox for photopolymerization and the analytical toolbox for microstructural characterization, we examine current strategies for physical heterogenization (fillers, inkjet printing) and chemical heterogenization (semicrystalline polymers, block copolymers, interpenetrating networks, photopolymerization induced phase separation) of photopolymers and put them into a material scientific context to develop a roadmap for improving and diversifying photopolymers' performance.

Characterization of PPS Piston and Packing Ring Materials for High-Pressure Hydrogen Applications.

The widespread adoption of renewable energy hinges on the efficient transportation of hydrogen. Reciprocating piston compressor technology in non-lubricated operation will play a key role, ensuring high flow rates and compression ratios. These systems rely on advanced high-strength sealing solutions for piston and rod packing rings utilizing advanced fiber-reinforced polymers. Polyphenylene sulfide (PPS) polymer matrix composites have seen use in tribological applications and promise high mechanical strength and wear resistance. The presented work describes carbon and glass fiber-reinforced PPS matrix polymers in comparison, which are characterized by complementary methods to investigate their properties and potential for application in reciprocating compressor under non-lubricated operation. Thermo-mechanical and tribological testing was supported by microstructure analysis utilizing advanced X-ray and electron imaging techniques. New insights in micromechanical deformation behavior in regard to fiber materials, interface strength and orientation in fiber-reinforced polymers are given. Conclusions on the suitability of different PPS matrix composites for high-pressure hydrogen compression applications were obtained.

Approaching Standardization: Mechanical Material Testing of Macroscopic Two-Photon Polymerized Specimens.

Two-photon polymerization (2PP) is becoming increasingly established as additive manufacturing technology for microfabrication due to its high-resolution and the feasibility of generating complex parts. Until now, the high resolution of 2PP is also its bottleneck, as it limited throughput and therefore restricted the application to the production of microparts. Thus, mechanical properties of 2PP materials can only be characterized using nonstandardized specialized microtesting methods. Due to recent advances in 2PP technology, it is now possible to produce parts in the size of several millimeters to even centimeters, finally permitting the fabrication of macrosized testing specimens. Besides suitable hardware systems, 2PP materials exhibiting favorable mechanical properties that allow printing of up-scaled parts are strongly demanded. In this work, the up-scalability of three different photopolymers is investigated using a high-throughput 2PP system and low numerical aperture optics. Testing specimens in the cm-range are produced and tested with common or even standardized material testing methods available in conventionally equipped polymer testing labs. Examples of the characterization of mechanical, thermo-mechanical, and fracture properties of 2PP processed materials are shown. Additionally, aspects such as postprocessing and aging are investigated. This lays a foundation for future expansion of the 2PP technology to broader industrial application.

microRNAs are differentially expressed in equine plasma of horses with osteoarthritis and osteochondritis dissecans versus control horses.

Osteoarthritis (OA) is a leading cause of lameness in horses with no effective disease-modifying treatment and challenging early diagnosis. OA is considered a disease of the joint involving the articular cartilage, subchondral bone, synovial membrane, and ligaments. Osteochondritis dissecans (OCD) is a joint disease consisting of focal defects in the osteochondral unit which may progress to OA later in life. MicroRNAs (miRNAs) have been recognized as small non-coding RNAs that regulate a variety of biological processes and have been detected in biological fluids. MiRNAs are currently investigated for their utility as biomarkers and druggable targets for a variety of diseases. The current study hypothesizes that miRNA profiles can be used to actively monitor joint health and differences in miRNA profiles will be found in healthy vs diseased joints and that differences will be detectable in blood plasma of tested horses. Five horses with OA, OCD, and 4 controls (C) had blood plasma and synovial fluid collected. Total RNA, including miRNA was isolated before generating miRNA libraries from the plasma of the horses. Libraries were sequenced at the Schroeder Arthritis Institute (Toronto). Differential expression analysis was done using DESeq2 and validated using ddPCR. KEGG pathway analysis was done using mirPath v.3 (Diana Tools). 57 differentially expressed miRNAs were identified in OA vs C plasma, 45 differentially expressed miRNAs in OC vs C plasma, and 21 differentially expressed miRNAs in OA vs OCD plasma. Notably, miR-140-5p expression was observed to be elevated in OA synovial fluid suggesting that miR-140-5p may serve as a protective marker early on to attenuate OA progression. KEGG pathway analysis of differentially expressed plasma miRNAs showed relationships with glycan degradation, glycosaminoglycan degradation, and hippo signaling pathway. Interestingly, ddPCR was unable to validate the NGS data suggesting that isomiRs may play an integral role in miRNA expression when assessed using NGS technologies.

One Health: Circadian Medicine Benefits Both Non-human Animals and Humans Alike.

Circadian biology's impact on human physical health and its role in disease development and progression is widely recognized. The forefront of circadian rhythm research now focuses on translational applications to clinical medicine, aiming to enhance disease diagnosis, prognosis, and treatment responses. However, the field of circadian medicine has predominantly concentrated on human healthcare, neglecting its potential for transformative applications in veterinary medicine, thereby overlooking opportunities to improve non-human animal health and welfare. This review consists of three main sections. The first section focuses on the translational potential of circadian medicine into current industry practices of agricultural animals, with a particular emphasis on horses, broiler chickens, and laying hens. The second section delves into the potential applications of circadian medicine in small animal veterinary care, primarily focusing on our companion animals, namely dogs and cats. The final section explores emerging frontiers in circadian medicine, encompassing aquaculture, veterinary hospital care, and non-human animal welfare and concludes with the integration of One Health principles. In summary, circadian medicine represents a highly promising field of medicine that holds the potential to significantly enhance the clinical care and overall health of all animals, extending its impact beyond human healthcare.

The consequences of firm scope and scale on patient access to healthcare.

OBJECTIVE: The aim was to quantify changes in the market structure of primary care physicians and examine its relationship with access to care. DATA SOURCES AND STUDY SETTING: We created measures of market structure from a 5% sample of Medicare fee-for-service claims and examined access to care using nationally representative data from the Medical Expenditure Panel Survey (MEPS). Our study spanned from 2008 to 2019. STUDY DESIGN: We used a linear probability model to estimate the relationship between access to care and two measures of market structure: concentration, measured by the Herfindahl-Hirschman Index (HHI), and vertical integration, measured by the market share of multispecialty firms. Our model controlled for year and ZIP code fixed effects, respondents' demographics and health status, and other measures of market structure. DATA COLLECTION/EXTRACTION METHODS: All adult respondents in the MEPS were included. PRINCIPAL FINDINGS: The percentage of people living in concentrated ZIP codes (HHI above 1500) increased from 37% in 2008 to 53% in 2019. During the same period, the median market share of multispecialty firms rose from 30% to 48%. Respondents in highly concentrated ZIP codes (HHI over 2500) were 5.9 percentage points (95% CI: -1.4 to -10.4) less likely to report having access to immediate care than respondents in unconcentrated ZIP codes. The association was largest among Medicaid beneficiaries, a 17.3 percentage point reduction (95% CI: -5.1 to -29.4). When we applied a model that was robust to biases from treatments with staggered timing, the estimated association remained negative but was not statistically significant. We found no association between HHI and indicators for having a usual source of care and annual checkups. The multispecialty market share was negatively associated with checkups, but not other measures of access. CONCLUSIONS: Increases in concentration may reduce some types of access to healthcare. These effects appear most pronounced among Medicaid beneficiaries.

Bioprocess development for cord blood mesenchymal stromal cells on microcarriers in Vertical-Wheel bioreactors.

Equine mesenchymal stromal cells (MSCs) have been found to be beneficial for the treatment of many ailments, including orthopedic injuries, due to their superior differentiation potential and immunomodulating properties. Cell therapies require large cell numbers, which are not efficiently generated using conventional static expansion methods. Expansion of equine cord blood-derived MSCs (eCB-MSCs) in bioreactors, using microcarriers as an attachment surface, has the potential to generate large numbers of cells with increased reproducibility and homogeneity compared with static T-flask expansion. This study investigated the development of an expansion process using Vertical-Wheel (VW) bioreactors, a single-use bioreactor technology that incorporates a wheel instead of an impeller. Initially, microcarriers were screened at small scale to assess eCB-MSC attachment and growth and then in bioreactors to assess cell expansion and harvesting. The effect of different donors, serial passaging, and batch versus fed batch were all examined in 0.1 L VW bioreactors. The use of VW bioreactors with an appropriate microcarrier was shown to be able to produce cell densities of up to 1E6 cells/mL, while maintaining cell phenotype and functionality, thus demonstrating great potential for the use of these bioreactors to produce large cell numbers for cell therapies.

Towards the validation of quantitative contrast sensitivity as a clinical endpoint: correlations with vision-related quality of life in bilateral AMD.

AIM: To investigate if active learning of contrast sensitivity (CS) in bilateral age-related macular degeneration (AMD) correlates better than visual acuity (VA) with vision-related quality of life (VRQoL) using factor analysis-calibrated National Eye Institute Visual Function Questionnaire-25 (NEI VFQ-25). METHODS: Prospective cross-sectional observational study in 93 patients (186 eyes) with bilateral AMD. CS was measured in one eye at a time with the quantitative CS function (qCSF) method (Adaptive Sensory Technology). Same-day VRQoL was assessed with factor analysis-calibrated NEI VFQ-25 visual function and socioemotional scales. Mixed-effects multiple linear regression analyses evaluated the associations of the qCSF outcomes and VA with the NEI VFQ-25 scales. A subgroup analysis on patients with AMD with VA more than 20/25 in both eyes was performed. RESULTS: Compared with VA, CS outcomes were associated with larger effect on both visual function scale (standardised beta coefficients (ß*) for area under the logarithm of CSF (AULCSF) curve and CS thresholds at 1.5, 3 and 6 cycles per degree (cpd): ß*=0.50, 0.48, 0.52, 0.46, all p<0.001, respectively, vs ß*=-0.45 for VA, all p<0.001) and socioemotional scale (ß* for AULCSF and CS threshold at 6 cpd: ß*=0.44, 0.44 vs ß*=-0.42 for VA, all p<0.001). In patients with AMD with VA more than 20/25 in both eyes (N=20), both VFQ-25 scales and all CS outcomes were significantly reduced. CONCLUSIONS: qCSF-measured CS strongly correlates with patient-reported VRQoL in bilateral AMD, even stronger than VA does. This study further validates qCSF-measured CS as a promising functional endpoint for future clinical trials in AMD.

Comparative study of widefield swept-source optical coherence tomography angiography in eyes with concomitant age-related macular degeneration and diabetic retinopathy.

BACKGROUND/AIMS: We sought to evaluate widefield swept-source optical coherence tomography angiography (WF SS-OCTA) among eyes with concomitant age-related macular degeneration (AMD) and diabetes mellitus or diabetic retinopathy (DM/DR). METHODS: This cross-sectional, comparative study consisted of three study groups: eyes with (1) AMD and DM/DR, (2) AMD alone and (3) DM/DR alone. WF SS-OCTA (3×3, 6×6 and 12×12 mm) images were captured. Vascular metrics included foveal avascular zone (FAZ), vessel density (VD) and vessel skeletonised density (VSD). Mixed-effects multivariable regression models adjusted for age were performed by cohort and subgroup based on AMD and DR stages. RESULTS: Our cohort included 287 eyes from 186 patients with an average age of 64±14.0 years old. Results revealed significantly reduced vascular metrics in concomitant AMD and DM/DR eyes (N=68) compared with AMD-only eyes (N=71) on all angiograms but not compared with DM/DR-only eyes (N=148). For example, when compared with AMD-only eyes, AMD and DM/DR eyes had significantly reduced VD (ß=-0.03, p=0.016) and VSD (ß=-1.09, p=0.022) on 12×12 mm angiograms, increased FAZ perimeter (ß=0.51, p=0.025) and FAZ area (ß=0.11, p=0.015) on 6×6 mm angiogram, and reductions in all VD and VSD metrics on 3×3 and 6×6 mm angiograms. However, only 3×3 mm angiogram FAZ metrics were significantly different when comparing DM/DR eyes with concomitant AMD and DM/DR eyes. CONCLUSION: WF SS-OCTA revealed significant reductions in retinal microvasculature metrics in AMD and DM/DR eyes compared with AMD-only eyes but not compared with DM/DR-only eyes.

A toxin-based approach to neuropeptide and peptide hormone discovery.

Peptide hormones and neuropeptides form a diverse class of bioactive secreted molecules that control essential processes in animals. Despite breakthroughs in peptide discovery, many signaling peptides remain undiscovered. Recently, we demonstrated the use of somatostatin-mimicking toxins from cone snails to identify the invertebrate ortholog of somatostatin. Here, we show that this toxin-based approach can be systematically applied to discover other unknown secretory peptides that are likely to have signaling function. Using large sequencing datasets, we searched for homologies between cone snail toxins and secreted proteins from the snails' prey. We identified and confirmed expression of five toxin families that share strong similarities with unknown secretory peptides from mollusks and annelids and in one case also from ecdysozoans. Based on several lines of evidence we propose that these peptides likely act as signaling peptides that serve important physiological functions. Indeed, we confirmed that one of the identified peptides belongs to the family of crustacean hyperglycemic hormone, a peptide not previously observed in Spiralia. We propose that this discovery pipeline can be broadly applied to other systems in which one organism has evolved molecules to manipulate the physiology of another.

Overview of Equine Stem Cells: Sources, Practices, and Potential Safety Concerns.

Over the past 2 decades, equine veterinarians are turning increasingly to stem cell therapies to repair damaged tissues or to promote healing through modulation of the immune system. Research is ongoing into optimizing practices associated with stem cell product transport, dosage, and administration. Culture-expanded equine mesenchymal stem cell therapies seem safe, even when used allogeneically, but various safety concerns should be considered. Stem cells and cellular reprogramming tools hold great promise for future equine therapies.

MicroRNAs as Prognostic Markers for Chondrogenic Differentiation Potential of Equine Mesenchymal Stromal Cells.

Mesenchymal stromal cells (MSCs) are a promising cell source for cartilage tissue regeneration in animals and humans but with large interdonor variation in their in vitro chondrogenic differentiation potential. Underlying molecular mechanisms responsible for culture-expanded MSC heterogeneity remain poorly understood. In this study, we sought to identify variations in microRNA (miRNA) signatures associated with cultured equine MSC chondrogenic differentiation potential from different donors. Neocartilage tissue generated from equine cord blood-derived MSCs was categorized as having either high or low chondrogenic potential (LCP) based on their histological appearance and quantification of glycosaminoglycan deposition. Using next-generation sequencing, we identified 30 differentially expressed miRNAs among undifferentiated MSC cultures that corresponded with their chondrogenic potential. Of note, MSCs with LCP upregulated miR-146a and miR-487b-3p, which was also observed by quantitative real-time polymerase chain reaction. Our findings suggest that miRNA profiling of equine MSC cultures may have prognostic value in selecting MSC donors with regard to their chondrogenic differentiation potential.

A previously unrecognized superfamily of macro-conotoxins includes an inhibitor of the sensory neuron calcium channel Cav2.3.

Animal venom peptides represent valuable compounds for biomedical exploration. The venoms of marine cone snails constitute a particularly rich source of peptide toxins, known as conotoxins. Here, we identify the sequence of an unusually large conotoxin, Mu8.1, which defines a new class of conotoxins evolutionarily related to the well-known con-ikot-ikots and 2 additional conotoxin classes not previously described. The crystal structure of recombinant Mu8.1 displays a saposin-like fold and shows structural similarity with con-ikot-ikot. Functional studies demonstrate that Mu8.1 curtails calcium influx in defined classes of murine somatosensory dorsal root ganglion (DRG) neurons. When tested on a variety of recombinantly expressed voltage-gated ion channels, Mu8.1 displayed the highest potency against the R-type (Cav2.3) calcium channel. Ca2+ signals from Mu8.1-sensitive DRG neurons were also inhibited by SNX-482, a known spider peptide modulator of Cav2.3 and voltage-gated K+ (Kv4) channels. Our findings highlight the potential of Mu8.1 as a molecular tool to identify and study neuronal subclasses expressing Cav2.3. Importantly, this multidisciplinary study showcases the potential of uncovering novel structures and bioactivities within the largely unexplored group of macro-conotoxins.

Prospects for Recyclable Multilayer Packaging: A Case Study.

Food preservation is an essential application for polymers, particularly in packaging. Complex multilayer films, such as those used for modified atmosphere packaging (MAP), extend the shelf life of sensitive foods. These mostly contain various polymers to achieve the necessary combination of mechanic, optic, and barrier properties that limit their recyclability. As the European Union's Circular Economy Action Plan calls for sustainable products and business models, including waste prevention policies and recycling quotas, with plastic packaging being a high priority, solutions towards more sustainable multilayer packaging are urgently needed. This study evaluated and compared the recycling potential of functionally equivalent PET (polyethylene terephthalate) and PP (polypropylene) post-consumer MAP through structure analysis and recycling simulation. The structure analysis revealed that both types of MAP contained functional (stability) and barrier layers (oxygen and moisture). The recycling simulation showed that the PP-based packaging was recyclable 10 times, maintaining its mechanical properties and functionality. At the same time, the PET-based MAP resulted in a highly brittle material that was unsuitable for reprocessing into similar economic value products. The secondary material from the PP-based MAP was successfully manufactured into films, demonstrating the functional possibility of closed-loop recycling. The transition from a linear to a circular economy for MAP is currently still limited by safety concerns due to a lack of sufficient and efficient purification methods, but the proper design of multilayers for recyclability is a first step towards circularity.

Association Between Contrast Sensitivity and Central Subfield Thickness in Center-Involving Diabetic Macular Edema.

Purpose: To evaluate the association between contrast sensitivity (CS) and central subfield thickness (CST) in diabetic macular edema (DME). Methods: This prospectively recruited, cross-sectional study included eyes with DME evaluated from November 2018 to March 2021. CST was measured using spectral-domain optical coherence tomography on the same day as CS testing. Only eyes with center-involving DME (CST >305 µm for women; >320 µm for men) were included. CS was evaluated using the quantitative CS function (qCSF) test. Outcomes included visual acuity (VA) and the following qCSF metrics: area under the log CS function, contrast acuity (CA), and CS thresholds at 1 to 18 cycles per degree (cpd). Pearson correlation and mixed-effects regression analyses were performed. Results: The cohort included 52 eyes of 43 patients. Pearson correlation analysis showed a stronger association between CST and CS thresholds at 6 cpd (r = -0.422, P = 0.002) than CST and VA (r = 0.293, P = 0.035). Mixed-effects univariate and multivariate regression analyses showed significant associations between CST and CA (ß = -0.001, P = .030), CS at 6 cpd (ß = -0.002, P = .008), and CS at 12 cpd (ß = -0.001, P = .049) but no significant associations between CST and VA. Among the visual function metrics, the effect size of CST was largest on CS at 6 cpd (ßStandardized = -0.37, P = .008). Conclusions: In patients with DME, CS may be more strongly associated with CST than VA. Including CS as an adjunct visual function outcome measure in eyes with DME may prove clinically valuable.

Hourly forecasting of traffic flow rates using spatial temporal graph neural networks.

Traffic congestion forms a large problem in many major metropolitan regions around the world, leading to delays and societal costs. As people resume travel upon relaxation of COVID-19 restrictions and personal mobility returns to levels prior to the pandemic, policy makers need tools to understand new patterns in the daily transportation system. In this paper we use a Spatial Temporal Graph Neural Network (STGNN) to train data collected by 34 traffic sensors around Amsterdam, in order to forecast traffic flow rates on an hourly aggregation level for a quarter. Our results show that STGNN did not outperform a baseline seasonal naive model overall, however for sensors that are located closer to each other in the road network, the STGNN model did indeed perform better.