CCR2-Mediated Uptake of Constitutively Produced CCL2: A Mechanism for Regulating Chemokine Levels in the Blood.

C-C chemokine receptor 2 (CCR2) is a key driver of monocyte/macrophage trafficking to sites of inflammation and has long been considered a target for intervention in autoimmune disease. However, systemic administration of CCR2 antagonists is associated with marked increases in CCL2, a CCR2 ligand, in the blood. This heretofore unexplained phenomenon complicates interpretation of in vivo responses to CCR2 antagonism. We report that CCL2 elevation after pharmacological CCR2 blockade is due to interruption in a balance between CCL2 secretion by a variety of cells and its uptake by constitutive internalization and recycling of CCR2. We observed this phenomenon in response to structurally diverse CCR2 antagonists in wild-type mice, and also found substantially higher CCL2 plasma levels in mice lacking the CCR2 gene. Our findings suggest that CCL2 is cleared from blood in a CCR2-dependent but G protein (Gαi, Gαs or Gαq/11)-independent manner. This constitutive internalization is rapid: on a given monocyte, the entire cell surface CCR2 population is turned over in <30 minutes. We also found that constitutive receptor internalization/recycling and ligand uptake are not universal across monocyte-expressed chemokine receptors. For example, CXCR4 does not internalize constitutively. In summary, we describe a mechanism that explains the numerous preclinical and clinical reports of increased CCL2 plasma levels following in vivo administration of CCR2 antagonists. These findings suggest that constitutive CCL2 secretion by monocytes and other cell types is counteracted by constant uptake and internalization by CCR2-expressing cells. The effectiveness of CCR2 antagonists in disease settings may be dependent upon this critical equilibrium.

Retinoblastoma in a 23-year-old adult treated with primary intra-arterial and intravitreal chemotherapy.

Intra-arterial chemotherapy (IAC) continues to provide a globe-sparing alternative as primary treatment for retinoblastoma with few adverse events. While there is growing evidence to highlight the utility of IAC in children with retinoblastoma, adult cases treated with primary IAC have not previously been characterized. We describe a rare case of Group D retinoblastoma in a 23-year-old adult treated successfully with IAC and intravitreal chemotherapy. This is a retrospective case report of a single patient. Subsequent to IAC and intravitreal chemotherapeutic treatments, at last follow-up 14 months following initial presentation and 8 months since last treatment, the retinoblastoma demonstrated complete regression into a partially calcified scar, with complete resolution of intravitreal and subretinal seeds and no evidence of tumor recurrence. Visual acuity improved to 20/30 in the left eye. There were no adverse events from therapy. Despite its rarity, it is important to consider retinoblastoma in the differential diagnosis of a white mass, even in an adult. Furthermore, this case highlights the utility of IAC for retinoblastoma, despite older patient age.

Efficacy of Chemokine Receptor Inhibition in Treating IL-36α-Induced Psoriasiform Inflammation.

Several types of psoriasiform dermatitis are associated with increased IL-36 cytokine activity in the skin. A rare, but severe, psoriasis-like disorder, generalized pustular psoriasis (GPP), is linked to loss-of-function mutations in the gene encoding IL-36RA, an important negative regulator of IL-36 signaling. To understand the effects of IL-36 dysregulation in a mouse model, we studied skin inflammation induced by intradermal injections of preactivated IL-36α. We found the immune cells infiltrating IL-36α-injected mouse skin to be of dramatically different composition than those infiltrating imiquimod-treated skin. The IL-36α-induced leukocyte population comprised nearly equal numbers of CD4+ αß T cells, neutrophils, and inflammatory dendritic cells, whereas the imiquimod-induced population comprised γδ T cells and neutrophils. Ligands for chemokine receptors CCR6 and CXCR2 are increased in both GPP and IL-36α-treated skin, which led us to test an optimized small-molecule antagonist (CCX624) targeting CCR6 and CXCR2 in the IL-36α model. CCX624 significantly reduced the T cell, neutrophil, and inflammatory dendritic cell infiltrates and was more effective than saturating levels of an anti-IL-17RA mAb at reducing inflammatory symptoms. These findings put CCR6 and CXCR2 forward as novel targets for a mechanistically distinct therapeutic approach for inflammatory skin diseases involving dysregulated IL-36 signaling, such as GPP.

Optical Coherence Tomography and Management of a Retinal Granuloma in Presumed Ocular Sarcoidosis.

Purpose: To highlight the utility of EDI-OCT and periocular steroid administration for the treatment of a retinal granuloma due to presumed ocular sarcoidosis. Methods: Retrospective case-study of a single patient. Results: A 45-year-old African-American male with blurred vision in the right eye was found to have a macula-involving retinal granuloma. Laboratory, imaging, and clinical findings were consistent with a diagnosis of presumed ocular sarcoidosis. Optical coherence tomography (OCT) and fundus photographs were used to document granuloma evolution. Oral prednisone 60mg was initiated and tapered to 10mg within 2 months. Due to systemic side-effects and inadequate treatment response with prednisone, 5 sub-Tenon's triamcinolone acetonide (PSTA) injections were then administered over 7 months, and oral prednisone was discontinued at 6 months. Conclusion: This case demonstrated marked granuloma regression occurring with improvement in visual acuity, highlighting the utility of OCT and PSTA in managing retinal sarcoid granulomas.

CCR2 antagonism leads to marked reduction in proteinuria and glomerular injury in murine models of focal segmental glomerulosclerosis (FSGS).

Focal segmental glomerulosclerosis (FSGS) comprises a group of uncommon disorders that present with marked proteinuria, nephrotic syndrome, progressive renal failure and characteristic glomerular lesions on histopathology. The current standard of care for patients with FSGS include immunosuppressive drugs such as glucocorticoids followed by calcineurin inhibitors, if needed for intolerance or inadequate response to glucocorticoids. Renin-angiotensin-aldosterone (RAAS) blockers are also used to control proteinuria, an important signature of FSGS. Existing treatments, however, achieved only limited success. Despite best care, treatment failure is common and FSGS is causal in a significant proportion of end stage renal disease. Thus, an unmet need exists for novel disease modifying treatments for FSGS. We employed two widely-used murine models of FSGS to test the hypothesis that systemic inhibition of chemokine receptor CCR2 would have therapeutic benefit. Here we report that administration CCX872, a potent and selective small molecule antagonist of CCR2, achieved rapid and sustained attenuation of renal damage as determined by urine albumin excretion and improved histopathological outcome. Therapeutic benefit was present when CCX872 was used as a single therapy, and moreover, the combination of CCX872 and RAAS blockade was statistically more effective than RAAS blockade alone. In addition, the combination of CCR2 and RAAS blockade was equally as effective as endothelin receptor inhibition. We conclude that specific inhibition of CCR2 is effective in the Adriamycin-induced and 5/6 nephrectomy murine models of FSGS, and thus holds promise as a mechanistically distinct therapeutic addition to the treatment of human FSGS.

IL-17-Secreting γδ T Cells Are Completely Dependent upon CCR6 for Homing to Inflamed Skin.

mAbs that neutralize IL-17 or its receptor have proven efficacious in treating moderate-to-severe psoriasis, confirming IL-17 as an important driver of this disease. In mice, a rare population of T cells, γδT17 cells, appears to be a dominant source of IL-17 in experimental psoriasis. These cells traffic between lymph nodes and the skin, and are identified by their coexpression of the TCR variable regions γ4 and δ4. These cells are homologous to the Vγ9Vδ2 T cell population identified in human psoriatic plaques. In this study we report that a potent and specific small molecule antagonist of the CCR6 chemokine receptor, CCX2553, was efficacious in reducing multiple aspects of psoriasis in two different murine models of the disease. Administration of CCX2553 ameliorated skin inflammation in both the IL-23-induced ear swelling model and the topical imiquimod model, and significantly reduced the number of γδT17 cells in inflamed skin. γδT17 cells were greatly reduced in imiquimod-treated skin of CCR6-/- mice, but adoptively transferred wild-type (CCR6+/+) γδT17 cells homed normally to the skin of imiquimod-treated CCR6-/- mice. Our data suggest that γδT17 cells are completely dependent on CCR6 for homing to psoriasiform skin. Thus, CCR6 may constitute a novel target for a mechanistically distinct therapeutic approach to treating psoriasis.

Coupled wave equations theory of surface-enhanced femtosecond stimulated Raman scattering.

We present a coupled wave semiclassical theory to describe plasmonic enhancement effects in surface-enhanced femtosecond stimulated Raman scattering (SE-FSRS). A key result is that the plasmon enhanced fields which drive the vibrational equation of motion for each normal mode results in dispersive lineshapes in the SE-FSRS spectrum. This result, which reproduces experimental lineshapes, demonstrates that plasmon-enhanced stimulated Raman methods provide unique sensitivity to a plasmonic response. Our derived SE-FSRS theory shows a plasmonic enhancement of |gpu|(2)ImχR(ω)gst (2)/ImχR(ω), where |gpu|(2) is the absolute square of the plasmonic enhancement from the Raman pump, χR(ω) is the Raman susceptibility, and gst is the plasmonic enhancement of the Stokes field in SE-FSRS. We conclude with a discussion on potential future experimental and theoretical directions for the field of plasmonically enhanced coherent Raman scattering.

Retinal metastasis from systemic cancer in 8 cases.

IMPORTANCE: Metastatic tumors of the retina are rare, simulate retinitis, and are associated with poor patient survival. OBJECTIVE: To describe the clinical features and outcomes of patients with retinal metastasis from systemic cancer. DESIGN, SETTING, AND PARTICIPANTS: Retrospective case series of 8 patients with retinal metastasis from cutaneous melanoma (n = 4), breast cancer (n =2), esophageal cancer (n =1), and lung cancer (n = 1). At presentation, the mean patient age was 62 years and all were white. INTERVENTION: Treatment included plaque radiotherapy (n = 1) for localized disease or enucleation (n =3) for extensive tumor hemorrhage (n = 1), total retinal detachment (n = 1), or pain (n = 1). For 4 preterminal patients, observation was preferred. MAIN OUTCOMES AND MEASURES: Clinical features and systemic outcomes. RESULTS: The mean interval from primary cancer diagnosis to retinal metastasis was 63 months. Initial misdiagnosis as retinitis (n = 5), hemangioma (n = 1), choroidal neovascular membrane (n = 1), or nerve fiber layer infarction (n = 1) for a mean interval of 5 months was recorded. Visual acuity in the affected eye was 20/40 to 20/60 (n = 5) or 20/400 to light perception (n = 3). The tumors were unilateral (n = 7), involved the macula (n = 3), and had a mean distance to the foveola of 6 mm. In one case, dense vitreous blood precluded fundus visualization. The mean tumor basal dimension was 7.4 mm, and the mean thickness was 2.3 mm. The tumors appeared white (n = 2), yellow (n = 4), or brown (n = 1); were located in the inner retina (n = 6) or full-thickness retina (n = 1); and had vitreous seeds (n = 3), vitreous hemorrhage (n = 2), retinal hemorrhage (n = 4), subretinal fluid (n = 4), and/or intraretinal exudation (n = 1). Fluorescein angiography disclosed early retinal hypofluorescence and late hyperfluorescence with staining. Fine-needle aspiration biopsy confirmed the diagnoses (n = 4). Metastasis-related death occurred in 5 patients within 1 month in each case. Of the remaining 3 patients, 2 were alive at 4 and 17 months and 1 was too sick to return. CONCLUSIONS AND RELEVANCE: Retinal metastases resemble retinitis, often with delay in diagnosis and poor life prognosis.

Quantum Monte Carlo Benchmark of Exchange-Correlation Functionals for Bulk Water.

The accurate description of the thermodynamic and dynamical properties of liquid water from first-principles is a very important challenge to the theoretical community. This represents not only a critical test of the predictive capabilities of first-principles methods, but it will also shed light into the microscopic properties of such an important substance. Density Functional Theory, the main workhorse in the field of first-principles methods, has been so far unable to properly describe water and its unusual properties in the liquid state. With the recent introduction of exact exchange and an improved description of dispersion interaction, the possibility of an accurate description of the liquid is finally within reach. Unfortunately, there is still no way to systematically improve exchange-correlation functionals, and the number of available functionals is very large. In this article we use highly accurate quantum Monte Carlo calculations to benchmark a selection of exchange-correlation functionals typically used in Density Functional Theory simulations of bulk water. This allows us to test the predictive capabilities of these functionals in water, giving us a way to choose optimal functionals for first-principles simulations. We compare and contrast the importance of different features of functionals, including the hybrid component, the vdW component, and their importance within different aspects of the PES. In addition, in order to correct the inaccuracies in the description of short-range interactions in the liquid, we test a recently introduced scheme that combines Density Functional Theory with Coupled Cluster calculations through a Many-Body expansion of the energy.

Flow diversion versus traditional aneurysm embolization strategies: analysis of fluoroscopy and procedure times.

BACKGROUND AND OBJECTIVE: Flow diverters are increasingly used for treatment of complex intracranial aneurysms. The purpose of this study was to compare the pipeline embolization device (PED) and traditional embolization strategies in terms of fluoroscopy and procedure time. MATERIAL AND METHODS: Fluoroscopy and procedure times (in minutes) were retrospectively analyzed and compared between 127 patients treated with the PED, 86 patients treated with single stage stent assisted coiling (SAC), and 16 patients treated with Onyx HD 500 at our institution. A multivariate logistic regression analysis was performed to determine independent predictors of fluoroscopy and procedure time. RESULTS: The three groups were comparable with respect to patient age, gender, and ruptured/unruptured aneurysm status. Aneurysms treated with the PED were significantly larger than stent coiled aneurysms, and aneurysm location distribution differed significantly between the three groups. Mean fluoroscopy time was significantly increased in the SAC (55±31 min, p<0.001) and Onyx HD 500 (91±36 min, p<0.001) groups relative to the PED group (34±23 min). Likewise, mean procedure time was significantly longer in SAC (155±50 min, p<0.001) and Onyx HD 500 (176±65 min, p<0.001) patients compared with PED patients (131±36 min). In multivariate analysis, SAC/Onyx HD 500 versus PED independently predicted longer procedure and fluoroscopy times. CONCLUSIONS: PED treatment requires significantly shorter fluoroscopy and procedure times compared with SAC and Onyx HD 500. The results of this study may be used by advocates of flow diverters as an additional argument for using this treatment modality to treat intracranial aneurysms.

Comparison of flow diversion and coiling in large unruptured intracranial saccular aneurysms.

BACKGROUND AND PURPOSE: Flow diversion has emerged as an important tool for the management of intracranial aneurysms. The purpose of this study was to compare flow diversion and traditional embolization strategies in terms of safety, efficacy, and clinical outcomes in patients with unruptured, large saccular aneurysms (≥10 mm). METHODS: Forty patients treated with the Pipeline Embolization Device (PED) were matched in a 1:3 fashion with 120 patients treated with coiling based on patient age and aneurysm size. Fusiform and anterior communicating artery aneurysms were eliminated from the analysis. Procedural complications, angiographic results, and clinical outcomes were analyzed and compared. RESULTS: There were no differences between the 2 groups in terms of patient age, sex, aneurysm size, and aneurysm location. The rate of procedure-related complications did not differ between the PED (7.5%) and the coil group (7.5%; P=1). At the latest follow-up, a significantly higher proportion of aneurysms treated with PED (86%) achieved complete obliteration compared with coiled aneurysms (41%; P<0.001). In multivariable analysis, coiling was an independent predictor of nonocclusion. Retreatment was necessary in fewer patients in the PED group (2.8%) than the coil group (37%; P<0.001). A similar proportion of patients attained a favorable outcome (modified Rankin Scale, 0-2) in the PED group (92%) and in the coil group (94%; P=0.8). CONCLUSIONS: The PED provides higher aneurysm occlusion rates than coiling, with no additional morbidity and similar clinical outcomes. These findings suggest that the PED might be a preferred treatment option for large unruptured saccular aneurysms.

Nuclear quantum effects and nonlocal exchange-correlation functionals applied to liquid hydrogen at high pressure.

Using first-principles molecular dynamics, we study the influence of nuclear quantum effects (NQEs) and nonlocal exchange-correlation density functionals (DFs) near molecular dissociation in liquid hydrogen. NQEs strongly influence intramolecular properties, such as bond stability, and are thus an essential part of the dissociation process. Moreover, by including DFs that account for either the self-interaction error or dispersion interactions, we find a much better description of molecular dissociation and metallization than previous studies based on classical protons and/or local or semilocal DFs. We obtain excellent agreement with experimentally measured optical properties along Hugoniot curves for precompressed states, and while we still find a first-order liquid-liquid transition at low temperatures, transition pressures are increased by more than 100 GPa.

Plasmonics in the ultraviolet with the poor metals Al, Ga, In, Sn, Tl, Pb, and Bi.

We discuss how the poor metals Al, Ga, In, Sn, Tl, Pb, and Bi can be used for plasmonics in the near to far ultraviolet (UV) range, similar to the noble metals Ag and Au in the visible (Vis) range. We first discuss the empirical dielectric functions of the poor metals, contrasting them with Ag and Au, and also fitting them to a Drude and multiple Lorentz oscillator form. Using Mie theory, we then compare the optical responses of spherical poor metal nanoparticles to noble metal ones. Finally, nanoparticle dimers are studied using a vectorial finite element method. We show how the poor metals exhibit large electric field enhancements in the UV, comparable to Au in the Vis, which makes them particularly attractive for sensing applications, such as surface enhanced Raman spectroscopy.

Pathogenesis of NOD diabetes is initiated by reactivity to the insulin B chain 9-23 epitope and involves functional epitope spreading.

Type 1 diabetes (T1D) is mediated by destruction of pancreatic ß-cells by CD4 and CD8 T cells specific for epitopes on numerous diabetogenic autoantigens resulting in loss of glucose homeostasis. Employing antigen-specific tolerance induced by i.v. administration of syngeneic splenocytes ECDI cross-linked to various diabetogenic antigens/epitopes (Ag-SP), we show that epitope spreading plays a functional role in the pathogenesis of T1D in NOD mice. Specifically, Ag-SP coupled with intact insulin, Ins B(9-23) or Ins B(15-23), but not GAD65(509-528), GAD65(524-543) or IGRP(206-214), protected 4-6 week old NOD mice from the eventual development of clinical disease; infiltration of immune cells to the pancreatic islets; and blocked the induction of DTH responses in a Treg-dependent, antigen-specific manner. However, tolerance induction in 19-21 week old NOD mice was effectively accomplished only by Ins-SP, suggesting Ins B(9-23) is a dominant initiating epitope, but autoimmune responses to insulin epitope(s) distinct from Ins B(9-23) emerge during disease progression.

Ground-state structures of atomic metallic hydrogen.

Ab initio random structure searching using density functional theory is used to determine the ground-state structures of atomic metallic hydrogen from 500 GPa to 5 TPa. Including proton zero-point motion within the harmonic approximation, we estimate that molecular hydrogen dissociates into a monatomic body-centered tetragonal structure near 500 GPa (r(s)=1.23) that remains stable to 1 TPa (r(s)=1.11). At higher pressures, hydrogen stabilizes in an …ABCABC… planar structure that is similar to the ground state of lithium, but with a different stacking sequence. With increasing pressure, this structure compresses to the face-centered cubic lattice near 3.5 TPa (r(s)=0.92).

Structure-activity relationships in gold nanoparticle dimers and trimers for surface-enhanced Raman spectroscopy.

Understanding the detailed relationship between nanoparticle structure and activity remains a significant challenge for the field of surface-enhanced Raman spectroscopy. To this end, the structural and optical properties of individual plasmonic nanoantennas comprised of Au nanoparticle assemblies that are coated with organic reporter molecules and encapsulated by a SiO(2) shell have been determined using correlated transmission electron microscopy (TEM), dark-field Rayleigh scattering microscopy, surface-enhanced Raman scattering (SERS) microscopy, and finite element method (FEM) calculations. The distribution of SERS enhancement factors (EFs) for a structurally and optically diverse set of nanoantennas is remarkably narrow. For a collection of 30 individual nanoantennas ranging from dimers to heptamers, the EFs vary by less than 2 orders of magnitude. Furthermore, the EFs for the hot-spot-containing nanoparticles are uncorrelated to aggregation state and localized surface plasmon resonance (LSPR) wavelength but are crucially dependent on the size of the interparticle gap. This study demonstrates that the creation of hot spots, where two particles are in subnanometer proximity or have coalesced to form crevices, is paramount to achieving maximum SERS enhancements.

Optical properties of nanowire dimers with a spatially nonlocal dielectric function.

We study the optical spectra and electromagnetic field enhancements around cylindrical and triangular Ag nanowire dimers, allowing for a spatially nonlocal dielectric function that partially accounts for quantum mechanical effects. For the triangular structures, we pay particular attention to how these properties depend on the sharpness of the nanowire's tips. We demonstrate that significant differences exist from classical electrodynamics that employs a more common, spatially local dielectric function. These differences are shown to arise from the optical excitation of volume plasmons inside of the structures, analogous to one-particle quantum mechanical states, which lead to complex and striking patterns of material polarization. These results are important for further understanding the optical properties of structures at the nanoscale and have implications for numerous physical processes, such as surface-enhanced Raman scattering.

Reversing the size-dependence of surface plasmon resonances.

The size-dependence of surface plasmon resonances (SPRs) is poorly understood in the small particle limit due to complex physical/chemical effects and uncertainties in experimental samples. In this article, we report an approach for synthesizing an ideal class of colloidal Ag nanoparticles with highly uniform morphologies and narrow size distributions. Optical measurements and theoretical analyses for particle diameters in the d approximately 2-20 nm range are presented. The SPR absorption band exhibits an exceptional behavior: As size decreases from d approximately 20 nm it blue-shifts but then turns over near d approximately 12 nm and strongly red-shifts. A multilayer Mie theory model agrees well with the observations, indicating that lowered electron conductivity in the outermost atomic layer, due to chemical interactions, is the cause of the red-shift. We corroborate this picture by experimentally demonstrating precise chemical control of the SPR peak positions via ligand exchange.

Nonlocal optical response of metal nanostructures with arbitrary shape.

We present an implementation of Maxwell's equations that incorporates the spatially nonlocal response of materials, an effect necessary to describe the optical properties of structures with features less than 10 nm. For the first time it is possible to investigate the nonlocal optical response of structures without spherical or planar shape, and outside of the electrostatic limit. As an illustration, we calculate the optical properties of Au nanowires and show that nonlocal effects are particularly important in structures with apex features, even for arbitrarily large sizes.