Metabolization of Resolvin E4 by ω-Oxidation in Human Neutrophils: Synthesis and Biological Evaluation of 20-Hydroxy-Resolvin E4 (20-OH-RvE4).

Resolvin E4 (RvE4) belongs to the resolvin family of specialized pro-resolving mediators (SPMs). The resolvins are endogenously formed mediators with both potent pro-resolving and anti-inflammatory biological activities and have attracted considerable attention in both inflammation research and drug discovery. Hence, further metabolism of the resolvins is of interest. Gaining knowledge about the structure-function of further metabolites of the resolvins is important due to their interest in drug-discovery efforts. For the first time, the total synthesis and biological evaluations of the ω-20 hydroxylated metabolite of RvE4, named herein 20-OH-RvE4, are presented. RvE4 was converted to 20-OH-RvE4 by human polymorphonuclear leukocytes. LC-MS/MS analysis and UV spectrophotometry reveal that the synthetic 20-OH-RvE4 matched RvE4-converted product 20-OH-RvE4 by human neutrophils. Cellular studies have revealed that RvE4 is formed from eicosapentaenoic acid in physiologic hypoxia by human neutrophils and macrophages, and we herein established that 20-OH-RvE4 is a secondary metabolite formed by the ω-oxidation of RvE4 in human neutrophils. A direct comparison of the biological actions between RvE4 and its metabolic product suggested that 20-OH-RvE4 displayed reduced bioactions in stimulating the efferocytosis of human senescent erythrocytes by human M2-like macrophages. At concentrations down to 0.1 nM, RvE4 increased macrophage erythrophagocytosis, an important pro-resolving function that was diminished due to metabolic transformation. The results provided herein contribute to a novel molecular insight on the further local metabolization of RvE4, the newest member among the SPM superfamily.

Active entanglement enables stochastic, topological grasping.

Grasping, in both biological and engineered mechanisms, can be highly sensitive to the gripper and object morphology, as well as perception and motion planning. Here, we circumvent the need for feedback or precise planning by using an array of fluidically actuated slender hollow elastomeric filaments to actively entangle with objects that vary in geometric and topological complexity. The resulting stochastic interactions enable a unique soft and conformable grasping strategy across a range of target objects that vary in size, weight, and shape. We experimentally evaluate the grasping performance of our strategy and use a computational framework for the collective mechanics of flexible filaments in contact with complex objects to explain our findings. Overall, our study highlights how active collective entanglement of a filament array via an uncontrolled, spatially distributed scheme provides options for soft, adaptable grasping.

Combing a double helix.

Combing hair involves brushing away the topological tangles in a collective curl, defined as a bundle of interacting elastic filaments. Using a combination of experiment and computation, we study this problem that naturally links topology, geometry and mechanics. Observations show that the dominant interactions in hair are those of a two-body nature, corresponding to a braided homochiral double helix. This minimal model allows us to study the detangling of an elastic double helix driven by a single stiff tine that moves along it and leaves two untangled filaments in its wake. Our results quantify how the mechanics of detangling correlates with the dynamics of a topological quantity, the link density, that propagates ahead of the tine and flows out the free end as a link current. This in turn provides a measure of the maximum characteristic length of a single combing stroke in the many-body problem on a head of hair, producing an optimal combing strategy that balances trade-offs between comfort, efficiency and speed of combing in hair curls of varying geometrical and topological complexity.

Stereoselective Synthesis and Structural Confirmation of the Specialized Pro-Resolving Mediator Resolvin E4.

Herein, we report the stereoselective and convergent synthesis of resolvin E4, a newly identified specialized pro-resolving mediator. This synthesis proves the absolute configuration and exact olefin geometry. Key elements of the successful strategy include a highly stereoselective MacMillan organocatalytic oxyamination, a Midland Alpine borane reduction, and the use of a 1,4-pentadiyne unit as a linchpin building block. The application of reaction telescoping in several of the synthetic transformations enabled the preparation of the resolvin E4 methyl ester in 10% yield over 10 steps (longest linear sequence). The physical property (UV-Vis and LC-MS/MS) data of synthetic resolvin E4 matched those obtained from biologically produced material.

Topology, Geometry, and Mechanics of Strongly Stretched and Twisted Filaments: Solenoids, Plectonemes, and Artificial Muscle Fibers.

Soft elastic filaments that can be stretched, bent, and twisted exhibit a range of topologically and geometrically complex morphologies. Recently, a number of experiments have shown how to use these building blocks to create filament-based artificial muscles that use the conversion of writhe to extension or contraction, exposing the connection between topology, geometry, and mechanics. Here, we combine numerical simulations of soft elastic filaments that account for geometric nonlinearities and self-contact to map out the basic structures underlying artificial muscle fibers in a phase diagram that is a function of the extension and twist density. We then use ideas from computational topology to track the interconversion of link, twist, and writhe in these geometrically complex physical structures to explain the physical principles underlying artificial muscle fibers and provide guidelines for their design.

Author Correction: Blunting neuroinflammation with resolvin D1 prevents early pathology in a rat model of Parkinson's disease.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Blunting neuroinflammation with resolvin D1 prevents early pathology in a rat model of Parkinson's disease.

Neuroinflammation is one of the hallmarks of Parkinson's disease (PD) and may contribute to midbrain dopamine (DA) neuron degeneration. Recent studies link chronic inflammation with failure to resolve early inflammation, a process operated by specialized pro-resolving mediators, including resolvins. However, the effects of stimulating the resolution of inflammation in PD - to modulate disease progression - still remain unexplored. Here we show that rats overexpressing human α-synuclein (Syn) display altered DA neuron properties, reduced striatal DA outflow and motor deficits prior to nigral degeneration. These early alterations are coupled with microglia activation and perturbations of inflammatory and pro-resolving mediators, namely IFN-γ and resolvin D1 (RvD1). Chronic and early RvD1 administration in Syn rats prevents central and peripheral inflammation, as well as neuronal dysfunction and motor deficits. We also show that endogenous RvD1 is decreased in human patients with early-PD. Our results suggest there is an imbalance between neuroinflammatory and pro-resolving processes in PD.

Resolvin D5 Inhibits Neuropathic and Inflammatory Pain in Male But Not Female Mice: Distinct Actions of D-Series Resolvins in Chemotherapy-Induced Peripheral Neuropathy.

Earlier studies have demonstrated that essential fatty acid-derived specialized pro-resolving mediators (SPMs) promote the resolution of inflammation and pain. However, the potential analgesic actions of SPMs in chemotherapy-induced peripheral neuropathy (CIPN) are not known. Recent results also showed sex dimorphism in immune cell signaling in neuropathic pain. Here, we evaluated the analgesic actions of D-series resolvins (RvD1, RvD2, RvD3, RvD4, and RvD5) on a CIPN in male and female mice. Paclitaxel (PTX, 2 mg/kg), given on days 0, 2, 4, and 6, produced robust mechanical allodynia in both sexes at 2 weeks. Intrathecal injection of RvD1 and RvD2 (100 ng, i.t.) at 2 weeks reversed PTX-induced mechanical allodynia in both sexes, whereas RvD3 and RvD4 (100 ng, i.t.) had no apparent effects on either sex. Interestingly, RvD5 (100 ng, i.t.) only reduced mechanical allodynia in male mice but not in female mice. Notably, PTX-induced mechanical allodynia was fully developed in Trpv1 or Trpa1 knockout mice, showing no sex differences. Also, intrathecal RvD5 reduced mechanical allodynia in male mice lacking Trpv1 or Trpa1, whereas female mice with Trpv1 or Trpa1 deficiency had no response to RvD5. Finally, RvD5-induced male-specific analgesia was also confirmed in an inflammatory pain condition. Formalin-induced second phase pain (licking and flinching) was reduced by intrathecal RvD5 in male but not female mice. These findings identified RvD5 as the first SPM that shows sex dimorphism in pain regulation. Moreover, these results suggest that specific resolvins may be used to treat CIPN, a rising health concern in cancer survivors.

Resolving Inflammation: Synthesis, Configurational Assignment, and Biological Evaluations of RvD1n-3 DPA.

New drugs that can resolve inflammation without immunosuppressive effects are at the medicinal chemistry frontier. Pro-resolving endogenously formed small molecules, that is, the resolvins, are excellent candidates displaying such bioactions. The first total synthesis of the specialized pro-resolving mediator RvD1n-3 DPA has been achieved using the underutilized sp3 -sp3 Negishi cross coupling reaction and an alkyne hydrosilylation-protodesilylation protocol. Biological evaluations revealed that this novel mediator displays low nanomolar pro-resolving properties and potently activates the human DRV1/GPR32 receptor. As such, this endogenous natural product is a lead compound for the development of novel immunoresolvents.

Development of global health education at Johns Hopkins University School of Medicine: a student-driven initiative.

Global health is increasingly present in the formal educational curricula of medical schools across North America. In 2008, students at Johns Hopkins University School of Medicine (JHUSOM) perceived a lack of structured global health education in the existing curriculum and began working with the administration to enhance global health learning opportunities, particularly in resource-poor settings. Key events in the development of global health education have included the introduction of a global health intersession mandatory for all first-year students; required pre-departure ethics training for students before all international electives; and the development of a clinical global health elective (Global Health Leadership Program, GHLP). The main challenges to improving global health education for medical students have included securing funding, obtaining institutional support, and developing an interprofessional program that benefits from the resources of the Schools of Medicine, Public Health, and Nursing. Strategies used included objectively demonstrating the need for and barriers to more structured global health experiences; obtaining guidance and modifying existing resources from other institutions and relevant educational websites; and harnessing institution-specific strengths including the large Johns Hopkins global research footprint and existing interprofessional collaborations across the three schools. The Johns Hopkins experience demonstrates that with a supportive administration, students can play an important and effective role in improving global health educational opportunities. The strategies we used may be informative for other students and educators looking to implement global health programs at their own institutions.

Total synthesis of the anti-inflammatory and pro-resolving lipid mediator MaR1n-3 DPA utilizing an sp(3) -sp(3) Negishi cross-coupling reaction.

The first total synthesis of the lipid mediator MaR1n-3 DPA (5) has been achieved in 12 % overall yield over 11 steps. The stereoselective preparation of 5 was based on a Pd-catalyzed sp(3) -sp(3) Negishi cross-coupling reaction and a stereocontrolled Evans-Nagao acetate aldol reaction. LC-MS/MS results with synthetic material matched the biologically produced 5. This novel lipid mediator displayed potent pro-resolving properties stimulating macrophage efferocytosis of apoptotic neutrophils.

Impaired proinsulin secretion before and during oral glucose stimulation in HIV-infected patients who display fat redistribution.

The beta-cell function of HIV-infected patients on highly active antiretroviral therapy who display lipodystrophy may be impaired. An early defect in beta-cell function may be characterized by an increase in secretion of 32-33 split proinsulin (SP) and intact proinsulin (IP). To address this issue, the secretion patterns of SP and IP of 16 HIV-infected men with lipodystrophy (LIPO) and 15 HIV-infected men without lipodystrophy (NONLIPO) were studied during an oral glucose tolerance test (OGTT). All patients received highly active antiretroviral therapy. Insulin secretion rates were determined by deconvolution of plasma C-peptide concentrations. More LIPO than NONLIPO patients displayed diabetes mellitus and impaired glucose tolerance than normal glucose tolerance (LIPO 2/8/6 vs NONLIPO 1/2/12, P = .05). LIPO patients had increased fasting levels of SP and IP, ratio of SP/IP, and area under the curve of SP and IP during the early phase (0, 10, and 20 minutes) and during the late phase (45, 75, and 105 minutes) of the OGTT compared with NONLIPO patients (Ps < .05). LIPO patients exhibited significantly increased fasting SP/IP ratio, fasting SP/insulin ratio, and total proinsulin to C-peptide ratio during the OGTT. LIPO patients displayed increased incremental secretion of IP during the first 10 minutes of the OGTT (P < .05), although the incremental insulin secretion during this period did not differ between LIPO and NONLIPO patients. These data suggest that HIV-infected patients with lipodystrophy display major perturbations of proinsulin secretion in the fasting state and during an OGTT, which is compatible with the notion of a beta-cell dysfunction of such patients.

A randomized trial of supine vs. prone positioning in patients undergoing escalated dose conformal radiotherapy for prostate cancer.

BACKGROUND AND PURPOSE: The optimal treatment position for patients receiving radical radiation therapy for prostate cancer has been a source of controversy. To resolve this issue, we conducted a randomized trial to evaluate the effects of supine and prone positioning on organ motion, positioning errors, and dose to critical organs during escalated dose conformal irradiation for localized prostate cancer and patient and therapist satisfaction with setup technique. PATIENTS AND METHODS: Twenty eight patients were randomized to commence treatment immobilized in the supine or prone position and were subsequently changed to the alternate positioning for the latter half of their treatment. Patients underwent CT simulation and conformal radiotherapy planning and treatment in both positions. The clinical target volume encompassed the prostate gland. Alternate day lateral port films were compared to corresponding simulator radiographs to measure the isocentre positioning errors (IPE). Prostate motion (PM) and total positioning error (TPE) were measured from the same films by the displacements of three implanted fiducial markers. Dose volume histograms (DVHs) for the two treatment positions were compared at the 95, 80 and 50% dose (D%) levels. The patients and radiation therapists completed weekly questionnaires regarding patient comfort and ease of setup. RESULTS: Seven patients, who started in the supine position, subsequently refused prone position and received their whole treatment supine. Small bowel in the treatment volume, not present in the supine position, prevented one patient from being treated prone. PM in anterior posterior direction was statistically significantly less in the supine position (P<0.05). There was no significant difference in superior inferior PM for the two treatment positions. No statistically significant difference between supine and prone positioning was observed in isocentre positioning error (IPE) or total positioning error (TPE) due to a policy of daily pre-treatment correction. However, more pre-treatment corrections were required for patients in the prone position. The DVH analysis demonstrated larger volumes of the bladder wall, rectal wall and small bowel within the D95, D80 and D50% when comparing the planning target volumes (PTVs) actually treated for prone positioning. When the prone PTV was expanded to account for the greater PM encountered in that position, a statistically significant difference (P<0.007) was observed in favour of the supine position at all dose levels. In the prone position, four patients had small bowel within the 60 Gray (Gy) isodose and in the supine position, no patients had small bowel in the 60 or 38Gy volumes. Supine position was significantly more comfortable for the patients and setup was significantly easier for the radiation therapists. The median patient comfort score was 0.79 (Standard deviation (SD) 0.03) supine and 0.45 (SD 0.05) prone (P<0.001) The therapist convenience of setup was 0.80 (SD 0.016) supine and 0.54 (SD 0.025) prone (P<0.005). No statistically significant difference was seen for the other parameters studied. CONCLUSIONS: We demonstrated significantly less PM in the supine treatment position. There was no difference for either treatment position in IPE or TPE, however, more pre-treatment corrections were required in the prone position. Prone position required a larger PTV with resulting increased dose to critical organs. There were statistically significant improvements at all dose levels for small bowel, rectal wall and bladder wall doses in the supine position once corrections were made for differences in organ motion. Linear analogue scores of patient comfort and radiation therapist convenience demonstrated statistically significant improvement in favour of the supine position. Supine positioning has been adopted as the standard for conformal prostatic irradiation at our centre.