The glymphatic system and cerebral small vessel disease.

OBJECTIVES: Cerebral small vessel disease is a group of pathologies in which alterations of the brain's blood vessels contribute to stroke and neurocognitive changes. Recently, a neurotoxic waste clearance system composed of perivascular spaces abutting the brain's blood vessels, termed the glymphatic system, has been identified as a key player in brain homeostasis. Given that small vessel disease and the glymphatic system share anatomical structures, this review aims to reexamine small vessel disease in the context of the glymphatic system and highlight novel aspects of small vessel disease physiology. MATERIALS AND METHODS: This review was conducted with an emphasis on studies that examined aspects of small vessel disease and on works characterizing the glymphatic system. We searched PubMed for relevant articles using the following keywords: glymphatics, cerebral small vessel disease, arterial pulsatility, hypertension, blood-brain barrier, endothelial dysfunction, stroke, diabetes. RESULTS: Cerebral small vessel disease and glymphatic dysfunction are anatomically connected and significant risk factors are shared between the two. These include hypertension, type 2 diabetes, advanced age, poor sleep, obesity, and neuroinflammation. There is clear evidence that CSVD hinders the effective functioning of glymphatic system. CONCLUSION: These shared risk factors, as well as the model of cerebral amyloid angiopathy pathogenesis, hint at the possibility that glymphatic dysfunction could independently contribute to the pathogenesis of cerebral small vessel disease. However, the current evidence supports a model of cascading dysfunction, wherein concurrent small vessel and glymphatic injury hinder glymphatic-mediated recovery and promote the progression of subclinical to clinical disease.

Nano-sandwich composite by kinetic trapping assembly from protein and nucleic acid.

Design and preparation of layered composite materials alternating between nucleic acids and proteins has been elusive due to limitations in occurrence and geometry of interaction sites in natural biomolecules. We report the design and kinetically controlled stepwise synthesis of a nano-sandwich composite by programmed noncovalent association of protein, DNA and RNA modules. A homo-tetramer protein core was introduced to control the self-assembly and precise positioning of two RNA-DNA hybrid nanotriangles in a co-parallel sandwich arrangement. Kinetically favored self-assembly of the circularly closed nanostructures at the protein was driven by the intrinsic fast folding ability of RNA corner modules which were added to precursor complex of DNA bound to the protein. The 3D architecture of this first synthetic protein-RNA-DNA complex was confirmed by fluorescence labeling and cryo-electron microscopy studies. The synthesis strategy for the nano-sandwich composite provides a general blueprint for controlled noncovalent assembly of complex supramolecular architectures from protein, DNA and RNA components, which expand the design repertoire for bottom-up preparation of layered biomaterials.

Olfaction in pregnancy: systematic review and meta-analysis.

Little attention has been paid to olfactory changes during pregnancy with contemporary studies limited in number and sample size. We examined whether pregnancy is associated with differences in olfactory performance and if there were any specific gestational ages at which these differences occur through a comprehensive systematic review and meta-analysis of the current literature. An initial electronic database search identified 234 citations, which were screened at the abstract level. Twenty-three citations were germane for full-text review, and 13 met criteria for inclusion. Our review assessed 5 olfactory measures of interest: odor identification (n = 11 articles), threshold (n = 8), discrimination (n = 5), hedonics (n = 6), and intensity (n = 5). Nine of these 13 studies contained sufficient data for meta-analysis, and these studies included a total of 523 pregnant women and 365 non-pregnant controls. Despite previous subjective and objective reports of odor intolerances and odor hypersensitivity, we did not find any significant differences between pregnant and non-pregnant women in odor discrimination, thresholds, or hedonics. However, meta-analysis of 506 cases and 333 controls showed worse odor identification in pregnant women compared to controls in a random-effects model. Thus, we demonstrate worse performance at odor identification during pregnancy. In this review, we discuss the current evidence (and lack thereof) regarding olfaction in pregnancy as well as highlight current knowledge gaps in this field.

Impact of social determinants of health on access to rhinology care and patient outcomes: A pilot study.

Objective: This novel pilot study constructs a social deprivation index (SDI) and utilizes an area deprivation index (ADI) to evaluate the link between social determinants of health and rhinology patient experiences. Methods: Adult patients undergoing outpatient care of chronic rhinitis and chronic rhinosinusitis at a tertiary academic medical center were recruited to participate in a telephone survey assessing symptoms, social/emotional consequences of disease, and barriers to care on a 5-point Likert scale. Sociodemographic characteristics were utilized to rate SDI on an 8-point scale. ADI was obtained by area code of residence. Ordered logistic regression was used to examine associations between the SDI/ADI and perceptions of rhinology care. Results: Fifty patients were included. Individuals with higher SDI scores (i.e., more socially deprived) experienced more severe nasal congestion (p = .007). Furthermore, higher national ADI correlated with increased severity of smell changes (p = .050) and facial pressure (p = .067). No association was seen between either deprivation index and global/psychiatric symptoms. While no correlations were found between higher SDI and difficulties with the costs of prescriptions, rhinologist's visits, or saline, higher SDI was correlated with decreased difficulty with surgery costs (p = .029), and individuals with higher national ADI percentile had increased difficulties obtaining nasal saline (p = .029). Conclusion: Worse social deprivation is associated with difficulties obtaining saline rinses and increased severity of nasal/sinus symptoms in an urban, underserved, majority-Black population. These findings suggest social factors affect access to and quality of rhinology care in a complex and nuanced way and highlight the need for a specific SDI to further study social determinants of health in rhinology. Level of Evidence: 2c.

Metalated Nucleic Acid Nanostructures.

Nucleic acid nanotechnology takes advantage of the self-assembling property of nucleic acids to form a variety of shapes and structures. The incorporation of metal ions into these structures introduces functionality for sensor and molecular electronic applications. Here, we describe a protocol for the incorporation of silver ions into polygonal nanoshapes that self-assemble from RNA and DNA modules.

Duration and Magnitude of Opioid Use After Minimally Invasive Sacroiliac Joint Fusion.

STUDY DESIGN: Retrospective cohort study performed in a nationwide insurance claims database. OBJECTIVE: To evaluate the duration and magnitude of post-operative opioid prescriptions after minimally invasive surgical sacroiliac joint fusion (MIS SIJF) as compared to other common spine surgeries. SUMMARY OF BACKGROUND DATA: MIS SIJF has been reported to significantly improve quality of life and reduce pain. However, there is a paucity of reported data on post-operative opioid use in patients undergoing MIS SIJF for sacroiliac joint dysfunction. METHODS: A nationwide insurance claims database was queried to identify 4,666 patients who underwent MIS SIJF. Patients were stratified by pre-operative opioid use: Opioid naïve, sporadic use, or chronic use were respectively defined as 0,1, or≥2 opioid prescriptions filled within 6 months prior to surgery. Duration of opioid use was defined by the time between MIS SIJF and last opioid prescription filled while magnitude of opioid use was determined by milligram morphine equivalents filled by 30 days post-operation. This opioid use data was compared to that of other common spine surgeries. RESULTS: Patients undergoing MIS SIJF continued to fill opioid prescriptions 1-year post-operatively at significantly higher proportions than those undergoing other common spine procedures assessed by prior literature within each of the pre-operative opioid use cohorts (chronic: 73% vs. 49-62%; P <0.0001, sporadic: 39% vs. 23-28%; P <0.0001, opioid naïve: 22% vs. 15-18%; P <0.0001). Chronic users filled the highest opiate dosages during the 30-day post-operative period, filling on average 64.75 MME/d compared to 19.75 MME/d and 24.25 MME/d by the opioid naïve and sporadic users, respectively. CONCLUSION: After MIS SIJF, opioid naïve patients use fewer opioids and for a shorter period of time compared to patients with sporadic or chronic pre-operative opioid use. MIS SIJF may result in less effective pain reduction when compared to other common spine surgeries evaluated via identical methodology.

Corticosteroid injection of the knee within one month prior to meniscus repair increases the risk of repair failure requiring meniscectomy.

OBJECTIVE: Meniscal tears are common knee injuries with limited endogenous healing capacity. This study aimed to investigate the association between the timing and administration of preoperative intra-articular corticosteroid injections (CSIs) and the risk of subsequent meniscectomy following meniscus repair. METHODS: Using a national insurance claims database, patients aged 18-40 years undergoing meniscus repair within six months of tear diagnosis were studied. Patients were categorized based on whether they received preoperative CSIs within three intervals prior to repair. Multivariable logistic regression was used to analyze the risk of follow-up meniscectomy while controlling for various patient-related variables. RESULTS: Among 5,390 patients meeting inclusion criteria, 201 received preoperative CSIs. The CSI group was older and had higher rates of diabetes, obesity, and knee osteoarthritis. The overall rate of follow-up meniscectomy did not differ between groups. However, CSIs performed within one month prior to repair were associated with significantly higher odds of subsequent meniscectomy compared to CSIs performed between three and six months prior. Obesity, tobacco use, and knee osteoarthritis were also independently associated with higher risk, while increasing age was associated with lower risk. CONCLUSION: The study highlights an increased risk of repair failure requiring follow-up meniscectomy for patients receiving intra-articular CSIs within one month prior to meniscus repair. These findings suggest caution when considering CSIs as a treatment option for patients scheduled for meniscus repair. Further research is needed to establish optimal timing guidelines for CSIs in relation to meniscus repair and to understand the underlying mechanisms.

Versatile kit of robust nanoshapes self-assembling from RNA and DNA modules.

DNA and RNA have emerged as a material for nanotechnology applications that take advantage of the nucleic acids' ability to encode folding and programmable self-assembly through mainly base pairing. The two types of nucleic acid have rarely been used in combination to enhance structural diversity or for partitioning of functional and architectural roles. Here, we report a design and screening strategy to integrate combinations of RNA motifs as architectural joints and DNA building blocks as functional modules for programmable self-assembly of a versatile toolkit of polygonal nucleic acid nanoshapes. Clean incorporation of diverse DNA modules with various topologies attest to the extraordinary robustness of the RNA-DNA hybrid framework. The design and screening strategy enables systematic development of RNA-DNA hybrid nanoshapes as programmable platforms for applications in molecular recognition, sensor and catalyst development as well as protein interaction studies.

Synthetic Biomaterials to Rival Nature's Complexity-a Path Forward with Combinatorics, High-Throughput Discovery, and High-Content Analysis.

Cells in tissue receive a host of soluble and insoluble signals in a context-dependent fashion, where integration of these cues through a complex network of signal transduction cascades will define a particular outcome. Biomaterials scientists and engineers are tasked with designing materials that can at least partially recreate this complex signaling milieu towards new materials for biomedical applications. In this progress report, recent advances in high throughput techniques and high content imaging approaches that are facilitating the discovery of efficacious biomaterials are described. From microarrays of synthetic polymers, peptides and full-length proteins, to designer cell culture systems that present multiple biophysical and biochemical cues in tandem, it is discussed how the integration of combinatorics with high content imaging and analysis is essential to extracting biologically meaningful information from large scale cellular screens to inform the design of next generation biomaterials.

Combinatorial Discovery of Defined Substrates That Promote a Stem Cell State in Malignant Melanoma.

The tumor microenvironment is implicated in orchestrating cancer cell transformation and metastasis. However, specific cell-ligand interactions between cancer cells and the extracellular matrix are difficult to decipher due to a dynamic and multivariate presentation of many signaling molecules. Here we report a versatile peptide microarray platform that is capable of screening for cancer cell phenotypic changes in response to ligand-receptor interactions. Using a screen of 78 peptide combinations derived from proteins present in the melanoma microenvironment, we identify a proteoglycan binding and bone morphogenic protein 7 (BMP7) derived sequence that selectively promotes the expression of several putative melanoma initiating cell markers. We characterize signaling associated with each of these peptides in the activation of melanoma pro-tumorigenic signaling and reveal a role for proteoglycan mediated adhesion and signaling through Smad 2/3. A defined substratum that controls the state of malignant melanoma may prove useful in spatially normalizing a heterogeneous population of tumor cells for discovery of therapeutics that target a specific state and for identifying new drug targets and reagents for intervention.

Impact of glycoprotein B genotype and naturally occurring ORF UL56 polymorphisms upon susceptibility of clinical human cytomegalovirus isolates to letermovir.

Letermovir is a novel anti-HCMV drug in Phase III development that targets the UL56 subunit of the viral terminase complex. In immunocompromised patients four major glycoprotein B (gB) subtypes are known and may influence pathogenesis and thus disease outcomes. Using a panel of 74 letermovir-naïve, low-passage, clinical HCMV isolates, we examined the potential impact of i) gB genotype and ii) naturally occurring UL56 sequence variations upon susceptibility to letermovir. Our data show that letermovir's potency is independent of gB subtype and show that naturally-occurring letermovir-resistance is rare or possibly absent.

Patterned porous silicon photonic crystals with modular surface chemistry for spatial control of neural stem cell differentiation.

We present a strategy to spatially define regions of gold and nanostructured silicon photonics, each with materials-specific surface chemistry, for azide-alkyne cycloaddition of different bioactive peptides. Neural stem cells are spatially directed to undergo neurogenesis and astrogenesis as a function of both surface properties and peptide identity.

Cell shape and the presentation of adhesion ligands guide smooth muscle myogenesis.

The reliable generation of smooth muscle cells is important for a number of tissue engineering applications. Human mesenchymal stem cells (MSCs) are a promising progenitor of smooth muscle, with high expression of smooth muscle markers observed in a fraction of isolated cells, which can be increased by introduction of soluble supplements that direct differentiation. Here we demonstrate a new micropatterning technique, where peptides of different ligand affinity can be microcontact printed onto an inert background, to explore MSC differentiation to smooth muscle through controlled biochemical and biophysical cues alone. Using copper-catalyzed alkyne-azide cycloaddition (CuAAC), we patterned our surfaces with RGD peptide ligands-both a linear peptide with low integrin affinity and a cyclic version with high integrin affinity-for the culture of MSCs in shapes with various aspect ratios. At low aspect ratio, ligand affinity is a prime determinant for smooth muscle differentiation, while at high aspect ratio, ligand affinity has less of an effect. Pathway analysis reveals a role for focal adhesion turnover, Rac1, RhoA/ROCK, and calpain during smooth muscle differentiation of MSCs in response to cell shape and the affinity of the cell adhesion interface. Controlling integrin-ligand affinity at the biomaterials interface is important for mediating adhesion but may also prove useful for directing smooth muscle myogenesis. Peptide patterning enables the systematic investigation of single to multiple peptides derived from any protein, at different densities across a biomaterials surface, which has the potential to direct multiple MSC differentiation outcomes without the need for soluble supplements.

Epidermal mechano-acoustic sensing electronics for cardiovascular diagnostics and human-machine interfaces.

Physiological mechano-acoustic signals, often with frequencies and intensities that are beyond those associated with the audible range, provide information of great clinical utility. Stethoscopes and digital accelerometers in conventional packages can capture some relevant data, but neither is suitable for use in a continuous, wearable mode, and both have shortcomings associated with mechanical transduction of signals through the skin. We report a soft, conformal class of device configured specifically for mechano-acoustic recording from the skin, capable of being used on nearly any part of the body, in forms that maximize detectable signals and allow for multimodal operation, such as electrophysiological recording. Experimental and computational studies highlight the key roles of low effective modulus and low areal mass density for effective operation in this type of measurement mode on the skin. Demonstrations involving seismocardiography and heart murmur detection in a series of cardiac patients illustrate utility in advanced clinical diagnostics. Monitoring of pump thrombosis in ventricular assist devices provides an example in characterization of mechanical implants. Speech recognition and human-machine interfaces represent additional demonstrated applications. These and other possibilities suggest broad-ranging uses for soft, skin-integrated digital technologies that can capture human body acoustics.

Rapid 3D Extrusion of Synthetic Tumor Microenvironments.

The spatial positioning of 3D tumor-mimetic microenvironments, containing multiple cell types, is controlled with a simple concentric flow device in a single step. A range of geometric architectures are demonstrated, and the migration of segregated tumor cells and macrophages is explored using drugs that inhibit heterotypic interactions.

Peptide microarrays for the discovery of bioactive surfaces that guide cellular processes: a single step azide-alkyne "click" chemistry approach.

Cell behavior in vivo is guided by a complex microenvironment containing many different molecules including extracellular matrix (ECM) proteins, growth factors, and proteoglycans. Controlling the interaction between these various components at the cell-material interface will be invaluable in developing new materials for biomedical devices and tissue engineering applications. We report a single step approach to forming mixed peptide conjugated self-assembled monolayers on gold using copper-catalyzed azide-alkyne cycloaddition chemistry to study the combinatorial effects of different peptide ligands on cellular processes. We synthesized ECM adhesion peptides (YIGSR, GRGDS), a bone morphogenetic protein 7 (BMP-7) derived peptide (KPSSAPTQLN), and a heparin binding peptide (KRSR), and arrayed them, alone and in combination, onto gold coated coverslips. SAMs were characterized by X-ray photoelectron spectroscopy (XPS) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry, and arrayed peptide combinations were seen to differentially bind to adipose derived stem cells (ADSCs) and mouse embryonic fibroblasts (MEFs). We further investigated the osteogenesis of ADSCs on SAMs containing combinations of adhesion peptide and BMP-7 peptide in both standard culture and osteogenic differentiation media. We demonstrate enhanced expression of osteogenic markers Runx2 and osteopontin when ADSCs are adherent to BMP-7 derived peptide alone or in combination with ECM adhesion peptides. The platform presented here enables immobilization of multiple peptides in a single step using a commercially available microarray spotter which will prove useful in fabricating biomolecule interfaces for cell biology studies and biochemical assays.

The effect of mesenchymal stem cell shape on the maintenance of multipotency.

Human mesenchymal stem cells (MSCs) have broad therapeutic potential due to their ability to differentiate into multiple cell types. However, when cultured ex vivo MSCs will spontaneously differentiate and have been shown to lose multipotency after prolonged passaging. Cell culture conditions that promote maintenance of multipotency during in vitro expansion are a critical need to fully realize the therapeutic potential of MSCs. Here we show that by confining MSCs to small islands, we can restrict inappropriate lineage specification and enhance the expression of mesenchymal stem cell markers Stro-1 and Endoglin. Even when released from the islands and reseeded, cells previously cultured in patterns maintain higher expression of MSC markers compared to cells cultured on plastic, while maintaining their ability to differentiate into adipocytes and osteoblasts. Exposure of non-patterned cells to inhibitors of myosin and Rho-associated protein kinase (ROCK) leads to increased expression of stem cell markers. Our findings suggest that maintenance of MSC "stemness" requires a low state of actomyosin contractility. This work will prove useful in the development of culture conditions for the maintenance of multipotent MSCs in vitro and for the design of niche-mimetic biomaterials.

Directing stem cell fate on hydrogel substrates by controlling cell geometry, matrix mechanics and adhesion ligand composition.

There is a dynamic relationship between physical and biochemical signals presented in the stem cell microenvironment to guide cell fate determination. Model systems that modulate cell geometry, substrate stiffness or matrix composition have proved useful in exploring how these signals influence stem cell fate. However, the interplay between these physical and biochemical cues during differentiation remains unclear. Here, we demonstrate a microengineering strategy to vary single cell geometry and the composition of adhesion ligands - on substrates that approximate the mechanical properties of soft tissues - to study adipogenesis and neurogenesis in adherent mesenchymal stem cells. Cells cultured in small circular islands show elevated expression of adipogenesis markers while cells that spread in anisotropic geometries tend to express elevated neurogenic markers. Arraying different combinations of matrix protein in a myriad of 2D and pseudo-3D geometries reveals optimal microenvironments for controlling the differentiation of stem cells to these "soft" lineages without the use of media supplements.