Interfacial reaction-induced roughening in polymer thin films.

Reactive blending of immiscible polymers is an important process for synthesizing polymer blends with superior properties. We use a phase-field model to understand reaction dynamics and morphology evolution by diffusive transport in layered films of incompatible, end-reactive polymers. We thoroughly investigate this phenomenon over a large parameter space of interface shapes, layer thicknesses, reaction rates specified by a Damkohler number (Daf), and Flory-Huggins interaction parameter (χ), under static conditions with no external fields. For films of the same thickness, the dynamics of the system is not significantly influenced by the length of the film or the initial shape of the interface. The interface between the polymers is observed to roughen, leading to the formation of a spontaneous emulsion. The reaction progresses slower and the interface roughens later for thicker films, and systems with higher χ. Increasing Daf increases the reaction rate and hastens the onset of roughening. The quasi-static interfacial tension decreases with the extent of reaction, but does not become vanishingly small or negative at the onset of roughening. Simulations with reversible reactions and systems where only a fraction of the homopolymers have reactive end groups show that a critical diblock (reaction product) concentration exists, below which interfacial roughening and spontaneous emulsification is not observed. We also demonstrate that thermal fluctuations accelerate the onset of interfacial roughening, and help sustain the system in an emulsified state.

Robust, 3-Dimensional Visualization of Human Colon Enteric Nervous System Without Tissue Sectioning.

BACKGROUND & AIMS: Small, 2-dimensional sections routinely used for human pathology analysis provide limited information about bowel innervation. We developed a technique to image human enteric nervous system (ENS) and other intramural cells in 3 dimensions. METHODS: Using mouse and human colon tissues, we developed a method that combines tissue clearing, immunohistochemistry, confocal microscopy, and quantitative analysis of full-thickness bowel without sectioning to quantify ENS and other intramural cells in 3 dimensions. RESULTS: We provided 280 adult human colon confocal Z-stacks from persons without known bowel motility disorders. Most of our images were of myenteric ganglia, captured using a 20× objective lens. Full-thickness colon images, viewed with a 10× objective lens, were as large as 4 × 5 mm2. Colon from 2 pediatric patients with Hirschsprung disease was used to show distal colon without enteric ganglia, as well as a transition zone and proximal pull-through resection margin where ENS was present. After testing a panel of antibodies with our method, we identified 16 antibodies that bind to molecules in neurons, glia, interstitial cells of Cajal, and muscularis macrophages. Quantitative analyses demonstrated myenteric plexus in 24.5% ± 2.4% of flattened colon Z-stack area. Myenteric ganglia occupied 34% ± 4% of myenteric plexus. Single myenteric ganglion volume averaged 3,527,678 ± 573,832 mm3 with 38,706 ± 5763 neuron/mm3 and 129,321 ± 25,356 glia/mm3. Images of large areas provided insight into why published values of ENS density vary up to 150-fold-ENS density varies greatly, across millimeters, so analyses of small numbers of thin sections from the same bowel region can produce varying results. Neuron subtype analysis revealed that approximately 56% of myenteric neurons stained with neuronal nitric oxide synthase antibody and approximately 33% of neurons produce and store acetylcholine. Transition zone regions from colon tissues of patients with Hirschsprung disease had ganglia in multiple layers and thick nerve fiber bundles without neurons. Submucosal neuron distribution varied among imaged colon regions. CONCLUSIONS: We developed a 3-dimensional imaging method for colon that provides more information about ENS structure than tissue sectioning. This approach could improve diagnosis for human bowel motility disorders and may be useful for other bowel diseases as well.

Muscularis macrophage development in the absence of an enteric nervous system.

The nervous system of the bowel regulates the inflammatory phenotype of tissue resident muscularis macrophages (MM), and in adult mice, enteric neurons are the main local source of colony stimulating factor 1 (CSF1), a protein required for MM survival. Surprisingly, we find that during development MM colonize the bowel before enteric neurons. This calls into question the requirement for neuron-derived CSF1 for MM colonization of the bowel. To determine if intestinal innervation is required for MM development, we analyzed MM of neonatal Ret-/- (Ret KO) mice that have no enteric nervous system in small bowel or colon. We found normal numbers of well-patterned MM in Ret KO bowel. Similarly, the abundance and distribution of MM in aganglionic human colon obtained from Hirschsprung disease patients was normal. We also identify endothelial cells and interstitial cells of Cajal as the main sources of CSF1 in the developing bowel. Additionally, MM from neonatal Ret KOs do not differ from controls in baseline activation status or cytokine-production in response to lipopolysaccharide. Unexpectedly, these data demonstrate that the enteric nervous system is dispensable for MM colonization and patterning in the bowel, and suggest that modulatory interactions between MM and the bowel nervous system are established postnatally.

Colloidal stability dictates drop breakup under electric fields.

Electric fields can deform drops of fluid from their equilibrium shape, and induce breakup at sufficiently large field strengths. In this work, the electric field induced breakup of a squalane drop containing a colloidal suspension of carbon black particles with polyisobutylene succinimide (OLOA 11000) surfactant is studied. The drop is suspended in silicone oil. The breakup mode of a drop containing carbon black depends strongly on the suspension stability. It is observed that a drop of a stable suspension of carbon black has the same breakup mode as a drop with surfactant alone, i.e., without added carbon black. At lower electric fields, these drops break by the formation of lobes at the two ends of the drop; and at higher fields the homogeneous lobes break in a non-axisymmetric manner. However, a drop of an unstable suspension shows a drastically different breakup mode, and undergoes breakup much faster compared to a drop with surfactant alone. These drops elongate and form asymmetric lobes that develop into fingers and eventually disintegrate in an inhomogeneous, three-dimensional fashion. As a basis for comparison, the breakup of a pure squalane drop, and a squalane drop with equivalent surfactant concentrations but no carbon black particles is examined. Axisymmetric boundary integral computations are used to elucidate the mechanism of breakup. Our work demonstrates the impact of colloidal stability on the breakup of drops under an electric field. Colloidal stability on the time scale of drop deformation leads to rich and unexplored breakup phenomena.

Dynamics of cracking in drying colloidal sheets.

Colloidal dispersions are known to display a fascinating network of cracks on drying. We probe the fracture mechanics of free-standing films of aqueous polymer-particle dispersions. Thin films of the dispersion are cast between a pair of plain steel wires and allowed to dry under ambient conditions. The strain induced on the particle network during drying is relieved by cracking. The stress which causes the films to crack has been calculated by measuring the deflection of the wires. The critical cracking stress varied inversely to the two-thirds' power of the film thickness. We also measure the velocity of the tip of a moving crack. The motion of a crack has been modeled as a competition between the release of the elastic energy stored in the particle network, the increase in surface energy as a result of the growth of a crack, the rate of viscous dissipation of the interstitial fluid and the kinetic energy associated with a moving crack. There is fair agreement between the measured crack velocities and predictions.

AACR Cancer Disparities Progress Report 2024: Achieving the Bold Vision of Health Equity.

Advances in cancer prevention, early detection, and treatments have led to unprecedented progress against cancer. However, these advances have not benefited everyone equally. Because of a long history of structural inequities and systemic injustices in the United States, many segments of the US population continue to shoulder a disproportionate burden of cancer. The American Association for Cancer Research (AACR) Cancer Disparities Progress Report 2024 (CancerDisparitiesProgressReport.org) outlines the recent progress against cancer disparities, the ongoing challenges faced by medically underserved populations, and emphasizes the vital need for further advances in cancer research and patient care to benefit all populations.

Mechanistic Insights into Propylparaben Sorption on Polyvinyl Chloride.

The mechanism of loss of propylparaben potency from formulations when in contact with polyvinyl chloride has been determined. It is caused by the adsorption of propylparaben onto polyvinyl chloride surfaces. The adsorption kinetics is best described using a pseudo-second order model based on non-linear fit. The rate of adsorption increases with increasing bulk concentration of propylparaben. Adsorption equilibrium isotherm was fitted to three isotherm models: Langmuir, Freundlich, and Temkin, using non-linear fit. The Freundlich and Temkin models show the best fit, indicating a multi-layer adsorption. Using this case study, we present a methodology to provide mechanistic insights into the compatibility data between pharmaceutical ingredients and product contact materials when sorption is involved.

Sequencing Reveals miRNAs Enriched in the Developing Mouse Enteric Nervous System.

The enteric nervous system (ENS) is an essential network of neurons and glia in the bowel wall. Defects in ENS development can result in Hirschsprung disease (HSCR), a life-threatening condition characterized by severe constipation, abdominal distention, bilious vomiting, and failure to thrive. A growing body of literature connects HSCR to alterations in miRNA expression, but there are limited data on the normal miRNA landscape in the developing ENS. We sequenced small RNAs (smRNA-seq) and messenger RNAs (mRNA-seq) from ENS precursor cells of mid-gestation Ednrb-EGFP mice and compared them to aggregated RNA from all other cells in the developing bowel. Our smRNA-seq results identified 73 miRNAs that were significantly enriched and highly expressed in the developing ENS, with miR-9, miR-27b, miR-124, miR-137, and miR-488 as our top 5 miRNAs that are conserved in humans. However, contrary to prior reports, our follow-up analyses of miR-137 showed that loss of Mir137 in Nestin-cre, Wnt1-cre, Sox10-cre, or Baf53b-cre lineage cells had no effect on mouse survival or ENS development. Our data provide important context for future studies of miRNAs in HSCR and other ENS diseases and highlight open questions about facility-specific factors in development.

Site-specific phosphorylation of CXCR4 is dynamically regulated by multiple kinases and results in differential modulation of CXCR4 signaling.

The chemokine receptor CXCR4 is a widely expressed G protein-coupled receptor that has been implicated in a number of diseases including human immunodeficiency virus, cancer, and WHIM syndrome, with the latter two involving dysregulation of CXCR4 signaling. To better understand the role of phosphorylation in regulating CXCR4 signaling, tandem mass spectrometry and phospho-specific antibodies were used to identify sites of agonist-promoted phosphorylation. These studies demonstrated that Ser-321, Ser-324, Ser-325, Ser-330, Ser-339, and two sites between Ser-346 and Ser-352 were phosphorylated in HEK293 cells. We show that Ser-324/5 was rapidly phosphorylated by protein kinase C and G protein-coupled receptor kinase 6 (GRK6) upon CXCL12 treatment, whereas Ser-339 was specifically and rapidly phosphorylated by GRK6. Ser-330 was also phosphorylated by GRK6, albeit with slower kinetics. Similar results were observed in human astroglia cells, where endogenous CXCR4 was rapidly phosphorylated on Ser-324/5 by protein kinase C after CXCL12 treatment, whereas Ser-330 was slowly phosphorylated. Analysis of CXCR4 signaling in HEK293 cells revealed that calcium mobilization was primarily negatively regulated by GRK2, GRK6, and arrestin3, whereas GRK3, GRK6, and arrestin2 played a primary role in positively regulating ERK1/2 activation. In contrast, GRK2 appeared to play a negative role in ERK1/2 activation. Finally, we show that arrestin association with CXCR4 is primarily driven by the phosphorylation of far C-terminal residues on the receptor. These studies reveal that site-specific phosphorylation of CXCR4 is dynamically regulated by multiple kinases resulting in both positive and negative modulation of CXCR4 signaling.

Cell-autonomous retinoic acid receptor signaling has stage-specific effects on mouse enteric nervous system.

Retinoic acid (RA) signaling is essential for enteric nervous system (ENS) development, since vitamin A deficiency or mutations in RA signaling profoundly reduce bowel colonization by ENS precursors. These RA effects could occur because of RA activity within the ENS lineage or via RA activity in other cell types. To define cell-autonomous roles for retinoid signaling within the ENS lineage at distinct developmental time points, we activated a potent floxed dominant-negative RA receptor α (RarαDN) in the ENS using diverse CRE recombinase-expressing mouse lines. This strategy enabled us to block RA signaling at premigratory, migratory, and postmigratory stages for ENS precursors. We found that cell-autonomous loss of RA receptor (RAR) signaling dramatically affected ENS development. CRE activation of RarαDN expression at premigratory or migratory stages caused severe intestinal aganglionosis, but at later stages, RarαDN induced a broad range of phenotypes including hypoganglionosis, submucosal plexus loss, and abnormal neural differentiation. RNA sequencing highlighted distinct RA-regulated gene sets at different developmental stages. These studies show complicated context-dependent RA-mediated regulation of ENS development.

Priority COVID-19 Vaccination for Patients with Cancer while Vaccine Supply Is Limited.

Published series on COVID-19 support the notion that patients with cancer are a particularly vulnerable population. There is a confluence of risk factors between cancer and COVID-19, and cancer care and treatments increase exposure to the virus and may dampen natural immune responses. The available evidence supports the conclusion that patients with cancer, in particular with hematologic malignancies, should be considered among the very high-risk groups for priority COVID-19 vaccination.

Morphine increases brain levels of ferritin heavy chain leading to inhibition of CXCR4-mediated survival signaling in neurons.

This study focuses on the effect of mu-opioid receptor agonists on CXCR4 signaling in neurons and the mechanisms involved in regulation of neuronal CXCR4 by opiates. The data show that CXCR4 is negatively modulated by long-term morphine treatments both in vitro and in vivo; CXCR4 inhibition is caused by direct stimulation of mu-opioid receptors in neurons, leading to alterations of ligand-induced CXCR4 phosphorylation and upregulation of protein ferritin heavy chain (FHC), a negative intracellular regulator of CXCR4. Reduced coupling of CXCR4 to G-proteins was found in the brain of morphine-treated rats, primarily cortex and hippocampus. CXCR4-induced G alpha(i)/G betagamma activities were suppressed after 24 h treatment of cortical neurons with morphine or the selective mu-opioid agonist DAMGO (D-Ala2-N-Me-Phe(4)-glycol(5)-enkephalin), as shown by analysis of downstream targets of CXCR4 (i.e., cAMP, Akt, and ERK1/2). These agonists also prevented CXCL12-induced phosphorylation of CXCR4, indicating a deficit of CXCR4 activation in these conditions. Indeed, morphine (or DAMGO) inhibited prosurvival signaling in neurons. These effects are not attributable to a reduction in CXCR4 expression or surface levels but rather to upregulation of FHC by opioids. The crucial role of FHC in inhibition of neuronal CXCR4 was confirmed by in vitro and in vivo RNA interference studies. Overall, these findings suggest that opiates interfere with normal CXCR4 function in the brain. By this mechanism, opiates could reduce the neuroprotective functions of CXCR4 and exacerbate neuropathology in opiate abusers who are affected by neuroinflammatory/infectious disorders, including neuroAIDS.

Alterations of CXCR4 function in µ-opioid receptor-deficient glia.

The chemokine receptor CXCR4 and the µ-opioid receptor (MOR) are G-protein-coupled receptors that are essential for normal function of the nervous and immune systems. Several studies have suggested that MOR is a key regulator of CXCR4 in the brain; however, the molecular basis of the opioid-chemokine interaction is not fully understood, and it may involve different mechanisms in neuronal and glial cells. Our previous studies demonstrated that MOR stimulation specifically upregulates the protein ferritin heavy chain - an inhibitor of CXCR4 - in neurons, and suggested that additional mechanisms could be operative in glia. In this study, we investigated CXCR4 function in brains and astroglial cultures deprived of MOR. Reduced coupling of CXCR4 to G-proteins was found in brain slices and tissue homogenates of MOR(-/-) mice as compared with wild-type controls. CXCR4-induced signaling was also reduced in glial cultures from MOR(-/-) mice, as shown by analysis of CXCR4 downstream targets (Akt and ERK1/2). Pharmacological studies with δ-opioid receptor (DOR)-specific ligands suggested that DOR-CXCR4 interactions are implicated in the inhibition of CXCR4 in MOR-deficient cells both in vitro and in vivo. Moreover, increased CXCR4/DOR co-immunoprecipitation was found in brain tissue and cultured glia from MOR(-/-) mice. Importantly, CXCR4 function was restored by pretreatment with a DOR antagonist. Overall, these findings indicate that DOR plays a crucial role in the regulation of CXCR4 in glia, probably via silent receptor heterodimers. The data also suggest that the opiate system interferes with normal CXCR4 function in different ways, depending on receptor subtypes.

Dynamic interfacial tension measurement under electric fields allows detection of charge carriers in nonpolar liquids.

HYPOTHESIS: Electric fields enhance surfactant transport to oil-water interfaces when the surfactant forms charged aggregates in the oil phase. Hence, transport under electric fields could be used to detect charged surfactant aggregates in nonpolar media. EXPERIMENTS: Two surfactants with different architecture were dispersed in Isopar-M. The transport of surfactants to an oil-water interface under a constant electric field was quantified using a custom-built electrified microtensiometer platform. Electrical conductivity of the oil with surfactant concentration was also measured to determine the presence of charge carriers. FINDINGS: The charging mechanism of the oil phase, and field-enhanced transport was different for the two surfactants. At low concentrations where the electrical conductivity of the surfactants is indistinguishable, dynamic interfacial tension measurements under electric fields can ascertain the presence of charge carriers in Isopar-M. The transport of ionic surfactants in the aqueous phase was unaffected by the field, confirming that the field-enhanced transport of oil-phase surfactants is due to electrophoresis of charge carriers. Moreover, the equilibrium interfacial tension was not found to change under an electric field, suggesting the adsorption isotherm is independent of the field strength. We demonstrate that dynamic interfacial tension measurements under electric fields is a sensitive technique to detect charge carriers in nonpolar fluids.

Disparities in Cancer Prevention in the COVID-19 Era.

Screening for cancer is a proven and recommended approach to prevent deaths from cancer; screening can locate precursor lesions and/or cancer at early stages when it is potentially curable. Racial and ethnic minorities and other medically underserved populations exhibit lower uptake of cancer screening than nonminorities in the United States. The COVID-19 pandemic, which disproportionately affected minority communities, has curtailed preventive services including cancer screening to preserve personal protective equipment and prevent spread of infection. While there is evidence for a rebound from the pandemic-driven reduction in cancer screening nationally, the return may not be even across all populations, with minority population screening that was already behind becoming further behind as a result of the community ravages from COVID-19. Fear of contracting COVID-19, limited access to safety-net clinics, and personal factors like, financial, employment, and transportation issues are concerns that are intensified in medically underserved communities. Prolonged delays in cancer screening will increase cancer in the overall population from pre-COVID-19 trajectories, and elevate the cancer disparity in minority populations. Knowing the overall benefit of cancer screening versus the risk of acquiring COVID-19, utilizing at-home screening tests and keeping the COVID-19-induced delay in screening to a minimum might slow the growth of disparity.

Electric fields enable tunable surfactant transport to microscale fluid interfaces.

The transport dynamics of oil-soluble surfactants to oil-water interfaces are quantified using a custom-built electrified capillary microtensiometer platform. Dynamic interfacial tension measurements reveal that surfactant transport is enhanced under a dc electric field, due to electro-migration of charge carriers in the oil toward the interface. Notably, this enhancement can be precisely tuned by altering the field strength and temporal scheduling. We demonstrate electric fields as a new parameter to manipulate surfactant transport to microscale fluid-fluid interfaces.