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Descemet&rsquo, s membrane detachment (DMD) is an uncommon, vision-threatening, ocular surface complication of cataract surgery. Among several treatment strategies, sulfur hexafluoride (SF6) descemetopexy is the standard of care. Herein, we report three cases of DMD after cataract surgery managed with SF6 descemetopexy, showing different outcomes. Anatomical success was achieved in cases 1 and 2 while intraocular pressure (IOP) was elevated in case 2. In case 3, despite SF6 descemetopexy, recurrent DMD was observed. Due to persistent corneal edema and possible corneal decompensation in case 3, Descemet&rsquo, s stripping automated endothelial keratoplasty was performed and a clear graft was found at the final visit. In conclusion, descemetopexy with 20 % SF6 is an effective and safe procedure for repairing DMD in most cases. Pupillary block with elevated IOP is another concern and prophylactic peripheral iridectomy is recommended. For recurrent DMDs, repeat descemetopexy could be considered. However, close monitoring is advocated since secondary management, such as endothelial keratoplasty, may be required.

Patients with cancer often confront the decision of whether to continue high-dose chemotherapy at the expense of cumulative toxicities. Reducing the dose of chemotherapy regimens while preserving efficacy is sorely needed to preserve the performance status of these vulnerable patients, yet has not been prioritized. Here, we introduce a dual pronged approach to modulate the microenvironment of desmoplastic pancreatic tumors and enable significant dose deescalation of the FDA-approved chemotherapeutic nanoliposomal irinotecan (nal-IRI) without compromising tumor control. We demonstrate that light-based photodynamic priming (PDP) coupled with vitamin D3 receptor (VDR) activation within fibroblasts increases intratumoral nal-IRI accumulation and suppresses protumorigenic CXCL12/CXCR7 crosstalk. Combined photodynamic and biochemical modulation of the tumor microenvironment enables a 75% dose reduction of nal-IRI while maintaining treatment efficacy, resulting in improved tolerability. Modifying the disease landscape to increase the susceptibility of cancer, via preferentially modulating fibroblasts, represents a promising and relatively underexplored strategy to enable dose deescalation. The approach presented here, using a combination of three clinically available therapies with nonoverlapping toxicities, can be rapidly translated with minimal modification to treatment workflow, and challenges the notion that significant improvements in chemotherapy efficacy can only be achieved at the expense of increased toxicity.

A key reason for the persistently grim statistics associated with metastatic ovarian cancer is resistance to conventional agents, including platinum-based chemotherapies. A major source of treatment failure is the high degree of genetic and molecular heterogeneity, which results from significant underlying genomic instability, as well as stromal and physical cues in the microenvironment. Ovarian cancer commonly disseminates via transcoelomic routes to distant sites, which is associated with the frequent production of malignant ascites, as well as the poorest prognosis. In addition to providing a cell and protein-rich environment for cancer growth and progression, ascitic fluid also confers physical stress on tumors. An understudied area in ovarian cancer research is the impact of fluid shear stress on treatment failure. Here, we investigate the effect of fluid shear stress on response to platinum-based chemotherapy and the modulation of molecular pathways associated with aggressive disease in a perfusion model for adherent 3D ovarian cancer nodules. Resistance to carboplatin is observed under flow with a concomitant increase in the expression and activation of the epidermal growth factor receptor (EGFR) as well as downstream signaling members mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK) and extracellular signal-regulated kinase (ERK). The uptake of platinum by the 3D ovarian cancer nodules was significantly higher in flow cultures compared to static cultures. A downregulation of phospho-focal adhesion kinase (p-FAK), vinculin, and phospho-paxillin was observed following carboplatin treatment in both flow and static cultures. Interestingly, low-dose anti-EGFR photoimmunotherapy (PIT), a targeted photochemical modality, was found to be equally effective in ovarian tumors grown under flow and static conditions. These findings highlight the need to further develop PIT-based combinations that target the EGFR, and sensitize ovarian cancers to chemotherapy in the context of flow-induced shear stress.

Longitudinal monitoring of tumor size in vivo can provide important biological information about disease progression and treatment efficacy that is not captured by other modes of quantification. Ultrasound enables high throughput evaluation of orthotopic mouse models via fast acquisition of three-dimensional tumor images and calculation of volume with a reasonable degree of accuracy. Herein, we compare orthotopic pancreatic tumor volume measurements determined by ultrasound with volume measured by calipers and tumor weight, and found strong correlations between the three modalities over a large range of tumor sizes, suggesting ultrasound can accurately quantify tumor volumes in this model. Furthermore, we demonstrate the unique ability of longitudinal treatment monitoring to reveal a tumor size-dependent dependent response to Benzoporphyrin Derivative photodynamic therapy (BPD-PDT) and irinotecan. Small tumors (5–35 mm(3)) were found to respond well to a single round of PDT, while large tumors (35–65 mm(3)) showed no response to the same treatment. These results highlight the role that tumor size can play in preclinical interpretation of treatment response, and more generally suggest that careful evaluation of subtle biological features such as this must be carefully considered in order to grant a more comprehensive understanding of disease biology in vivo.

Regulators and practitioners argue the relative merits of firm and partner rotation, while researchers report mixed results on the consequences of rotation. This study uses an experiment to examine the effect of an upcoming rotation on perceptions of auditor competence and independence and finds that participants appear to be indifferent to whether rotation is at the firm or partner level; they only react to concurrent changes in audit fees and the industry specialization status of the new auditor. Specifically, participants assess auditor competence and independence (specifically attention to detail, effort, and skeptical attitude) to be higher when fees increase rather than decrease significantly at the time of a rotation, and they assess auditor competence to be higher when rotation is to an industry specialist rather than a nonindustry specialist. These findings hold regardless of whether rotation is at the firm or partner level. JEL Classifications: M42. Data Availability: Data and the tasks used in this study are available on request.

Two-dimensional (2D) transition metal dichalcogenides (TMDCs) have attracted great attention as alternatives to graphene with semiconducting band gaps. Mono- or few-layer TMDCs can be prepared by various methods, but regardless of the fabrication methods [such as mechanical exfoliation and chemical vapor deposition (CVD)], TMDCs contain many structural defects, which significantly affect their physical properties and limit their performance in applications. Metallophthalocyanines (MPcs) are organic semiconductors, and as dopants, they are capable of modulating the optical and electrical properties of other semiconducting materials. Here, we report that besides the ability to modulate the optoelectronic properties of 2D TMDCs, MPc molecules can be used to heal defects and improve the physicochemical properties of TMDCs. Doping of planar MPc molecules to TMDCs is achieved by a simple solution dip-coating method and results in a significant improvement in the optical properties and thermal responses of CVD-grown TMDCs, even comparable to those of mechanically exfoliated counterparts. Study of carrier dynamics shows that the adsorption of MPc on the TMDC surface leads to the complete suppression of the mid-gap defect-induced absorption in TMDCs. Furthermore, MPc molecules with a large lateral size are found to effectively reduce the point defects in mechanically exfoliated TMDCs introduced during the preparation process. Our results not only clarify the optoelectronic modulation mechanism of chemical doping but also offer a simple method to control the nanosized defects in 2D TMDCs.

The shortage of medical staffs has become a critical issue due to the rapid growth of aging population. Numerous attempts have been made on devising pervasive healthcare systems to precisely and continuously monitor patients’ health status. However, the transformation from prototypes in the laboratory to practical systems in a medical institution is still a challenging task. This paper reports our progress and lessons learned from designing and deploying a pervasive healthcare system that runs persistently for more than six months in a real-world smart ward of National Taiwan University Hospital, one of the most heavy-loaded hospitals with high occupancy rate in Taiwan. We describe techniques proposed to deal with three essential challenges: design for essential needs, design for user acceptance, and design for maintenance. The system is evaluated empirically by deploying two applications in the field. Based on the results of field interview and questionnaires, we believe that this work is a milestone of a persistently running pervasive healthcare system deployed in a hospital.

Contains fulltext : 283288.pdf (Publisher’s version ) (Open Access) Malaria transmission-blocking vaccines (TBVs) aim to elicit human antibodies that inhibit sporogonic development of Plasmodium falciparum in mosquitoes, thereby preventing onward transmission. Pfs48/45 is a leading clinical TBV candidate antigen and is recognized by the most potent transmission-blocking monoclonal antibody (mAb) yet described; still, clinical development of Pfs48/45 antigens has been hindered, largely by its poor biochemical characteristics. Here, we used structure-based computational approaches to design Pfs48/45 antigens stabilized in the conformation recognized by the most potently inhibitory mAb, achieving >25°C higher thermostability compared with the wild-type protein. Antibodies elicited in mice immunized with these engineered antigens displayed on liposome-based or protein nanoparticle-based vaccine platforms exhibited 1-2 orders of magnitude superior transmission-reducing activity, compared with immunogens bearing the wild-type antigen, driven by improved antibody quality. Our data provide the founding principles for using molecular stabilization solely from antibody structure-function information to drive improved immune responses against a parasitic vaccine target.

BackgroundInduction of CD8+ T cells that recognize immunogenic, mutated protein fragments in the context of major histocompatibility class I (MHC-I) is a pressing challenge for cancer vaccine development.MethodsUsing the commonly used murine renal adenocarcinoma RENCA cancer model, MHC-I restricted neoepitopes are predicted following next-generation sequencing. Candidate neoepitopes are screened in mice using a potent cancer vaccine adjuvant system that converts short peptides into immunogenic nanoparticles. An identified functional neoepitope vaccine is then tested in various therapeutic experimental tumor settings.ResultsConversion of 20 short MHC-I restricted neoepitope candidates into immunogenic nanoparticles results in antitumor responses with multivalent vaccination. Only a single neoepitope candidate, Nesprin-2 L4492R (Nes2LR), induced functional responses but still did so when included within 20-plex or 60-plex particles. Immunization with the short Nes2LR neoepitope with the immunogenic particle-inducing vaccine adjuvant prevented tumor growth at doses multiple orders of magnitude less than with other vaccine adjuvants, which were ineffective. Nes2LR vaccination inhibited or eradicated disease in subcutaneous, experimental lung metastasis and orthotopic tumor models, synergizing with immune checkpoint blockade.ConclusionThese findings establish the feasibility of using short, MHC-I-restricted neoepitopes for straightforward immunization with multivalent or validated neoepitopes to induce cytotoxic CD8+ T cells. Furthermore, the Nes2LR neoepitope could be useful for preclinical studies involving renal cell carcinoma immunotherapy.

Many approaches exist for stimuli-triggered cargo release from nanocarriers, but few can provide for on-demand release of multiple payloads, selectively. Here, we report the synthesis of purpurin-phospholipid (Pur-P), a lipid chromophore that has near-infrared absorbance red-shifted by 30 nm compared to a structurally similar pyropheophorbide-phospholipid (Pyr-P). Liposomes containing small amounts of either Pur-P or Pyr-P exhibited similar physical properties and fluorescence self-quenching. Loaded with distinct cargos, Pur-P and Pyr-P liposomes were mixed into a single colloidal suspension and selectively released cargo depending on irradiation wavelength. Spatiotemporal control of distinct cargo release was achieved by controlling multicolor laser placement. Using basic orange and doxorubicin anthraquinones, multidimensional cytotoxicity gradients were established to gauge efficacy against cancer cells using light-released drug. Wavelength selectivity of cargo release was maintained following intramuscular administration to mice.

Physiologic barriers to drug delivery and selection for drug resistance limit survival outcomes in cancer patients. In this study, we present preclinical evidence that a subtumoricidal photodynamic priming (PDP) strategy can relieve drug delivery barriers in the tumor microenvironment to safely widen the therapeutic window of a nanoformulated cytotoxic drug. In orthotopic xenograft models of pancreatic cancer, combining PDP with nanoliposomal irinotecan (nal-IRI) prevented tumor relapse, reduced metastasis, and increased both progression-free survival and 1-year disease-free survival. PDP enabled these durable improvements by targeting multiple tumor compartments to (i) increase intratumoral drug accumulation by >10-fold, (ii) increase the duration of drug exposure above a critical therapeutic threshold, and (iii) attenuate surges in CD44 and CXCR4 expression, which mediate chemoresistance often observed after multicycle chemotherapy. Overall, our results offer preclinical proof of concept for the effectiveness of PDP to minimize risks of tumor relapse, progression, and drug resistance and to extend patient survival. Significance: A biophysical priming approach overcomes key treatment barriers, significantly reduces metastases, and prolongs survival in orthotopic models of human pancreatic cancer. Cancer Res; 78(2); 558–71. ©2017 AACR.

Doxorubicin (Dox)-loaded stealth liposomes (similar to those in clinical use) can incorporate small amounts of porphyrin–phospholipid (PoP) to enable chemophototherapy (CPT). PoP is also an intrinsic and intrabilayer 64Cu chelator, although how radiolabeling impacts drug delivery has not yet been assessed. Here, we show that 64Cu can radiolabel the stable bilayer of preformed Dox-loaded PoP liposomes with inclusion of 1% ethanol without inducing drug leakage. Dox-PoP liposomes labeled with intrabilayer copper behaved nearly identically to unlabeled ones in vitro and in vivo with respect to physical parameters, pharmacokinetics, and CPT efficacy. Positron emission tomography and near-infrared fluorescence imaging visualized orthotopic mammary tumors in mice with passive liposome accumulation following administration. A single CPT treatment with 665 nm light (200 J/cm2) strongly inhibited primary tumor growth. Liposomes accumulated in lung metastases, based on NIR imaging. These results establish the feasib...

Systemic injection of therapeutic antibodies may cause serious adverse effects due to on-target toxicity to the antigens expressed in normal tissues. To improve the targeting selectivity to the region of disease sites, we developed protease-activated pro-antibodies by masking the binding sites of antibodies with inhibitory domains that can be removed by proteases that are highly expressed at the disease sites. The latency-associated peptide (LAP), C2b or CBa of complement factor 2/B were linked, through a substrate peptide of matrix metalloproteinase-2 (MMP-2), to an anti-epidermal growth factor receptor (EGFR) antibody and an anti-tumor necrosis factor-α (TNF-α) antibody. Results showed that all the inhibitory domains could be removed by MMP-2 to restore the binding activities of the antibodies. LAP substantially reduced (53.8%) the binding activity of the anti-EGFR antibody on EGFR-expressing cells, whereas C2b and CBa were ineffective (21% and 9.3% reduction, respectively). Similarly, LAP also blocked 53.9% of the binding activity of the anti-TNF-α antibody. Finally, molecular dynamic simulation showed that the masking efficiency of LAP, C2b and CBa was 33.7%, 10.3% and −5.4%, respectively, over the binding sites of the antibodies. This strategy may aid in designing new protease-activated pro-antibodies that attain high therapeutic potency yet reduced systemic on-target toxicity.

// Chun-Wei Wang 1, 2 , Chiung-Chiao Huang 1 , Pei-Hsin Chou 4 , Yu-Ping Chang 1 , Shouzuo Wei 5 , Frederick Peter Guengerich 6 , Yueh-Ching Chou 3, 7, 9 , Sheng-Fan Wang 1, 7 , Ping-Shan Lai 8 , Pavel Soucek 11 and Yune-Fang Ueng 1, 2, 3, 10 1 National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan, R.O.C 2 Institute of Biopharmaceutical Sciences, School of Pharmacy, National Yang-Ming University, Taipei, Taiwan, R.O.C 3 Department of Pharmacology, School of Medicine, National Yang-Ming University, Taipei, Taiwan, R.O.C 4 Department of Environmental Engineering, National Chung-Kung University, Tainan, Taiwan, R.O.C 5 Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN, USA 6 Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA 7 Department of Pharmacy, Taipei Veterans General Hospital, Taipei, Taiwan, R.O.C 8 Department of Chemistry, College of Science, National Chung-Hsin University, Taichung, Taiwan, R.O.C 9 Department of Pharmacy, Taipei Medical University, Taipei, Taiwan, R.O.C 10 Institute of Medical Sciences, Taipei Medical University, Taipei, Taiwan, R.O.C 11 Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic Correspondence to: Yune-Fang Ueng, email: ueng@nricm.edu.tw Keywords: 7-ketocholesterol, 27-hydroxycholesterol, doxorubicin, P-glycoprotein, estrogen receptor Received: March 13, 2017 Accepted: June 27, 2017 Published: August 02, 2017 ABSTRACT Hypercholesterolemia is one of the risk factors for poor outcome in breast cancer therapy. To elucidate the influence of the main circulating oxysterols, cholesterol oxidation products, on the cell-killing effect of doxorubicin, cells were exposed to oxysterols at a subtoxic concentration. When cells were exposed to oxysterols in fetal bovine serum-supplemented medium, 7-ketocholesterol (7-KC), but not 27-hydroxycholesterol (27-HC), decreased the cytotoxicity of doxorubicin in MCF-7 (high estrogen receptor (ER)α/ERβ ratio) cells and the decreased cytotoxicity was restored by the P-glycoprotein inhibitor verapamil. 7-KC stimulated the efflux function of P-glycoprotein and reduced intracellular doxorubicin accumulation in MCF-7 but not in ERα(-) MDA-MB-231 and the resistant MCF-7/ADR cells. In MCF-7 cells, 7-KC increased the mRNA and protein levels of P-glycoprotein. The 7-KC-suppressed doxorubicin accumulation was restored by the fluvestrant and ERα knockdown. In a yeast reporter assay, the ERα activation by 7-KC was more potent than 27-HC. 7-KC, but not 27-HC, stimulated the expression of an ER target, Trefoil factor 1 in MCF-7 cells. When charcoal-stripped fetal bovine serum was used, both 7-KC and 27-HC induced Trefoil factor 1 expression and reduced doxorubicin accumulation in MCF-7 cells. 7-KC-reduced doxorubicin accumulation could be reversed by inhibitors of phosphatidylinositol 3-kinase, Akt, and mammalian target of rapamycin (mTOR). These findings demonstrate that 7-KC decreases the cytotoxicity of doxorubicin through the up-regulation of P-glycoprotein in an ERα- and mTOR-dependent pathway. The 7-KC- and 27-HC-elicited estrogenic effects are crucial in the P-glycoprotein induction in breast cancer cells.

Quantitative landslide susceptibility assessment is necessary for mitigating casualties, property damage, and economic loss. Identification of landslides and preparation of landslide susceptibility maps are crucial steps in landslide susceptibility assessment. Therefore, an optimal landslide susceptibility model is presented that is capable of producing accurate landslide susceptibility maps and assessing landslide susceptibility. To construct the optimal landslide susceptibility model, the effectiveness of the improved self-organizing linear output map (ISOLO), support vector machines (SVM) with four kernel functions (LN-SVM, PL-SVM, RBF-SVM, and SIG-SVM) and logistic regression (LR) was compared. Twelve landslide causative factors (namely, slope, slope aspect, elevation, curvature, profile curvature, plan curvature, slope length, topographic wetness index, distance to river, distance to road, distance to fault and annual maximum 24- and 48-h rainfalls) were used in this landslide susceptibility analysis. These models were applied to the Kaoping River basin in Southwestern Taiwan to assess its performance. Landslide inventory maps from 2008 to 2011 were collected. Data from the first three-year period were used for training and the remaining data was used for testing. The performance of the models was compared using accuracy and the area under the receiver operating characteristic curve as criteria. The results show that the RBF-SVM model outperformed the logistic regression in the study area. Using the RBF-SVM model, the landslide susceptibility under the annual 48-h maximum rainfall of various return periods were analyzed to assist local administrations and decision makers in disaster planning.

This article proposes an interleaved soft-switching converter with high voltage conversion ratio as an alternative to existing hard-switching converters, such that the switching loss can be reduced. This proposed soft-switching converter not merely boosts the output voltage of a fuel cell to a high level, that is applied to a high voltage DC link or a DC load, but also reduces the current stress in main switches, the input current and the output voltage ripples. A resonant auxiliary branch is connected in parallel with either main switch employed in a conventional hard-switching converter, such that the voltage drop across the main switch falls to zero before the main switch is triggered. In doing so, the aim of zero voltage switching (ZVS) is achieved, leading to a reduced switching loss, promoted conversion efficiency and lowered switching stress. In the end, this work is applied to fuel cells and validated by a good agreement between simulations and experiments.

Governments in many developing countries, such as China, are investing a lot of human, financial and material resources to ensure that as much of municipal solid waste (MSW) as possible is collected for centralized harmless treatment and disposal. This is regarded as an effective way to alleviate the “waste siege” problem in governance caused by the continuous enormous increase in MSW quantity. Therefore, how to improve the efficiency of MSW collection service system is an important governance issue because the inputs that can be utilized to manage MSW problem are limited due to budget and resource constraints. However, studies on MSW collection efficiency in developing countries like China are under-represented in the existing literature. This paper applies a stochastic frontier analysis (SFA) approach to evaluate the efficiency of MSW collection services in 30 provinces of China from 2008 to 2017, and explore the relative importance of several factors that might influence collection efficiency. The results indicate that there exists considerable room (62.8%) to enhance efficiency since its current efficiency value is merely 0.372. With regard to influencing factors, the proportion of population aged 15–64 appears to have the greatest positive impact on efficiency, along with per capita GDP, added value of tertiary industry and education level. This paper also explores spatial variations of MSW collection efficiency across the eastern, central and western regions. These findings have policy implications and can inform the related government departments how to formulate proper policies to improve collection efficiency.

Additional file 1. Additional Figures S1–S4 and Tables S1–S7.

Additional file 1. This Additional file contains Figures S1-S16, Tables S1-S7, and Additional Method S1.

Photothermal therapy (PTT) is an externally activated approach that generates heat to damage, repair, or modulate biological targets. Gold nanoparticles (GNPs) combined with PTT provide unique opportunities for biomedical applications, where light activation can trigger both direct heat-mediated cytotoxicity and the release of drug payloads from nanoparticles. The combination of PTT and other cancer therapies, based on interactive mechanisms that target multiple nonoverlapping growth and survival pathways, is key to improving treatment outcomes. With advances in the development of GNPs and miniaturized light delivery systems, PTT can now access diseases at hard-to-reach anatomical sites in our body. This section introduces engineering aspects interfacing PTT and GNPs, as well as their preclinical and clinical applications in therapy, surgery, and diagnosis.

Liposomes hold great potential as gene and drug delivery vehicles due to their biocompatibility and modular properties, coupled with the major advantage of attenuating the risk of systemic toxicity from the encapsulated therapeutic agent. Decades of research have been dedicated to studying and optimizing liposomal formulations for a variety of medical applications, ranging from cancer therapeutics to analgesics. Some effort has also been made to elucidate the toxicities and immune responses that these drug formulations may elicit. Notably, intravenously injected liposomes can interact with plasma proteins, leading to opsonization, thereby altering the healthy cells they come into contact with during circulation and removal. Additionally, due to the pharmacokinetics of liposomes in circulation, drugs can end up sequestered in organs of the mononuclear phagocyte system, affecting liver and spleen function. Importantly, liposomal agents can also stimulate or suppress the immune system depending on their physiochemical properties, such as size, lipid composition, pegylation, and surface charge. Despite the surge in the clinical use of liposomal agents since 1995, there are still several drawbacks that limit their range of applications. This review presents a focused analysis of these limitations, with an emphasis on toxicity to healthy tissues and unfavorable immune responses, to shed light on key considerations that should be factored into the design and clinical use of liposomal formulations.

BackgroundThe endothelial cell–cell junctions of the blood–brain barrier (BBB) play a pivotal role in the barrier’s function. Altered cell–cell junctions can lead to barrier dysfunction and have been implicated in several diseases. Despite this, the driving forces regulating junctional protein presentation remain relatively understudied, largely due to the lack of efficient techniques to quantify their presentation at sites of cell–cell adhesion. Here, we used our novel Junction Analyzer Program (JAnaP) to quantify junction phenotype (i.e., continuous, punctate, or perpendicular) in response to various substrate compositions, cell culture times, and cAMP treatments in human brain microvascular endothelial cells (HBMECs). We then quantitatively correlated junction presentation with barrier permeability on both a “global” and “local” scale.MethodsWe cultured HBMECs on collagen I, fibronectin, collagen IV, laminin, fibronectin/collagen IV/laminin, or hyaluronic acid/gelatin for 2, 4, and 7 days with varying cAMP treatment schedules. Images of immunostained ZO-1, VE-cadherin, and claudin-5 were analyzed using the JAnaP to calculate the percent of the cell perimeter presenting continuous, punctate, or perpendicular junctions. Transwell permeability assays and resistance measurements were used to measure bulk (“global”) barrier properties, and a “local” permeability assay was used to correlate junction presentation proximal to permeable monolayer regions.ResultsSubstrate composition was found to play little role in junction presentation, while cAMP supplements significantly increased the continuous junction architecture. Increased culture time required increased cAMP treatment time to reach similar ZO-1 and VE-cadherin coverage observed with shorter culture, though longer cultures were required for claudin-5 presentation. Prolonged cAMP treatment (6 days) disrupted junction integrity for all three junction proteins. Transwell permeability and TEER assays showed no correlation with junction phenotype, but a local permeability assay revealed a correlation between the number of discontinuous and no junction regions with barrier penetration.ConclusionsThese results suggest that cAMP signaling influences HBMEC junction architecture more than matrix composition. Our studies emphasized the need for local barrier measurement to mechanistically understand the role of junction phenotype and supported previous results that continuous junctions are indicative of a more mature/stable endothelial barrier. Understanding what conditions influence junction presentations, and how they, in turn, affect barrier integrity, could lead to the development of therapeutics for diseases associated with BBB dysfunction.

A range of cellular, architectural, and physical cues in the tumor microenvironment influence the intrinsic and acquired resistance mechanisms that lead to treatment failure. Strategies that leverage photodynamic therapy (PDT), a photochemistry-based biophysical treatment modality, to regionally target and prime stubborn tumor populations may be essential to realizing durable improvements in cancer management while minimizing toxicity from traditional agents. Capturing these attributes in rationally-designed combinations has shown promise by synergistically reducing tumor area in 3D models, and durably controlling tumor burden in vivo. Among the areas that remain understudied is the influence of mechanical forces, such as hydrodynamic shear stress, on resistance, and the development of 3D tumor models and in vivo models that account for physical stress. To evaluate and optimize PDT regimens, and PDT-based combinations, designed to overcome resistance to conventional therapies due to physical stress, a multi-faceted approach is needed. Here the impact of hydrodynamic stress is evaluated in bioengineered 3D tumor models in the context of ovarian cancer. The potential value of using biologically inspired in vitro models to guide customized, rationally-designed PDT-based combination regimens will be presented.

Photosensitizing biomolecules (PSBM) represent a new generation of light-absorbing compounds with improved optical and physicochemical properties for biomedical applications. Despite numerous advances in lipid-, polymer-, and protein-based PSBMs, their effective use requires a fundamental understanding of how macromolecular structure influences the physicochemical and biological properties of the photosensitizer. Here, we prepared and characterized three well-defined PSBMs based on a clinically used photosensitizer, benzoporphyrin derivative (BPD). The PSBMs include 16:0 lysophosphocholine-BPD (16:0 Lyso PC-BPD), distearoyl-phosphoethanolamine-polyethylene-glycol-BPD (DSPE-PEG-BPD), and anti-EGFR cetuximab-BPD (Cet-BPD). In two glioma cell lines, DSPE-PEG-BPD exhibited the highest singlet oxygen yield but was the least phototoxic due to low cellular uptake. The 16:0 Lyso PC-BPD was most efficient in promoting cellular uptake but redirected BPD&rsquo, s subcellular localization from mitochondria to lysosomes. At 24 h after incubation, proteolyzed Cet-BPD was localized to mitochondria and effectively disrupted the mitochondrial membrane potential upon light activation. Our results revealed the variable trafficking and end effects of PSBMs, providing valuable insights into methods of PSBM evaluation, as well as strategies to select PSBMs based on subcellular targets and cytotoxic mechanisms. We demonstrated that biologically informed combinations of PSBMs to target lysosomes and mitochondria, concurrently, may lead to enhanced therapeutic effects against gliomas.

Outer surface protein A (OspA) is a Borrelia lipoprotein and an established Lyme disease vaccine target. Admixing non-lipidated, recombinant B. burgdorferi OspA with liposomes containing cobalt porphyrin-phospholipid (CoPoP) resulted in rapid, particulate surface display of the conformationally intact antigen. Particleization was serum-stable and led to enhanced antigen uptake in murine macrophages in vitro. Mouse immunization using CoPoP liposomes that also contained a synthetic monophosphoryl lipid A (PHAD) elicited a Th1-biased OspA antibody response with higher IgG production compared to other vaccine adjuvants. Antibodies were reactive with intact B. burgdorferi spirochetes and Borrelia lysates, and induced complement-mediated borreliacidal activity in vitro. One year after initial immunization, mice maintained high levels of circulating borreliacidal antibodies capable of blocking B. burgdorferi transmission from infected ticks to human blood in a feeding chamber.

Surfactant-stripped micelles are formed from a commercially available cyanine fluroalkylphosphate salt dye (CyFaP) and used for high contrast photoacoustic imaging in the second near infrared window (NIR-II). The co-loading of Coenzyme Q10 into surfactant-stripped CyFaP (ss-CyFaP) micelles improves yield, storage stability and results in a peak absorption wavelength in the NIR-II window close to the 1064 nm output of Nd-YAG lasers used for photoacoustic imaging. Aqueous ss-CyFaP dispersions exhibit intense NIR-II optical absorption, calculated to be greater than 500 at 1064 nm. ss-CyFaP is detected through 12 cm of chicken breast tissue with PAI. In preclinical animal models, ss-CyFaP is visualized in draining lymph nodes of rats through 3.1 cm of overlaid chicken breast tissue. Following intravenous administration, ss-CyFaP accumulates in neoplastic tissues of mice and rats bearing orthotopic mammary tumors without observation of acute toxic side effects. ss-CyFaP is imaged through whole compressed human breasts in three female volunteers at depths of 2.6 to 5.1 cm. Taken together, these data show that ss-CyFaP is an accessible contrast agent for deep tissue photoacoustic imaging in the NIR-II window.

Simple Summary Accumulation of excess fluid in the abdomen typically indicates abnormal function or disease, such as cancer, in the underlying tissues. This accumulation of fluid, or ascites, occurs more frequently in patients with advanced-stage ovarian cancer than any other type of cancer. The presence of ascites indicates the poorest outcomes for patients with advanced stage ovarian cancer, but little is known about the reasons for these dismal outcomes. This review discusses the current understanding of ascites, starting with an overview of ovarian cancer and ascites, followed by a description of the tools used to analyze the components of ascites and how these components modulate ovarian cancer biology. A perspective on the mechanical effects of ascites and the impact of mechanical stress on treatment resistance is provided. Lastly, treatment options for ascites and opportunities to develop new therapeutic strategies to improve outcomes are discussed. Abstract Ascites refers to the abnormal accumulation of fluid in the peritoneum resulting from an underlying pathology, such as metastatic cancer. Among all cancers, advanced-stage epithelial ovarian cancer is most frequently associated with the production of malignant ascites and is the leading cause of death from gynecologic malignancies. Despite decades of evidence showing that the accumulation of peritoneal fluid portends the poorest outcomes for cancer patients, the role of malignant ascites in promoting metastasis and therapy resistance remains poorly understood. This review summarizes the current understanding of malignant ascites, with a focus on ovarian cancer. The first section provides an overview of heterogeneity in ovarian cancer and the pathophysiology of malignant ascites. Next, analytical methods used to characterize the cellular and acellular components of malignant ascites, as well the role of these components in modulating cell biology, are discussed. The review then provides a perspective on the pressures and forces that tumors are subjected to in the presence of malignant ascites and the impact of physical stress on therapy resistance. Treatment options for malignant ascites, including surgical, pharmacological and photochemical interventions are then discussed to highlight challenges and opportunities at the interface of drug discovery, device development and physical sciences in oncology.

Amphiphilic poly(acrylic acid) terminated with tetraphenylthiophene (TP) was prepared by atom transfer radical polymerization (ATRP) of t -butyl acrylate monomer, initiated by amino-functionalized TP, and the following by acid-catalyzed hydrolysis. With the hydrophobic TP terminal, a fluorophore with aggregation-induced emission (AIE) property, and the hydrophilic polymeric chain, the resulting polymer of TP-PAA is amphiphilic and self-assembled, through preferable hydrogen bond interactions among the carboxylic acid pedants, in water to form aggregates with strong AIE-related emission. The convenient dispersion of TP-PAA aggregates in water facilitated its applications as fluorescent sensor for pH value and for the natural protein of bovine serum albumin (BSA). Fluorescence variations in term of aggregation level of the TP terminals in different environments will be discussed in this study.

FPGA-realization of a rotor position estimation algorithm based on a novel sliding mode observer (SMO) for permanent magnet synchronous motor (PMSM) drives is adopted in this paper. Comparing to the conventional SMO, the estimator based on the novel SMO is presented for reducing the chattering phenomenon and improving the estimated precision of the rotor position. However, a hybrid sensorless control system for PMSM drives using I-f startup method then a smooth transition to the novel SMO-based sensorless field oriented control (FOC) are also considered in this paper. In realization, a very high speed IC hardware description language (VHDL) is firstly introduced to describe the behavior of the adopted estimation and control algorithm which is called the sensorless control intellectual property (IP). Then, to verify the correctness of the designed VHDL code for the sensorless control IP, it will be downloaded to the FPGA, and an experimental system with a sensorless PMSM drive is set up and some experimental results are demonstrated.

With the rapid development of economy and urbanization, municipal solid waste (MSW) generation has also increased dramatically. MSW poses great threat to the ecological environment and the survival of humankind, and forms a bottleneck restricting the healthy and sustainable development of an economy. Research on the influence factors of MSW could provide useful information for formulation of pollution control policies. This article develops a multiple regression model to assess the impact of these influencing factors. We find that MSW generations are strongly and positively related to urban population, GDP, the quantity of solid waste transport vehicles, collection area of solid waste, and the per capita disposable income of urban residents. However, we find that MSW generations have strong and negative correlations with the inflation rate and the solid waste treatment rate. Finally, based on the empirical findings, we put forward policy recommendations to help government agencies to design measures that can effectively address the increasing trend of waste generation. © 2016 American Institute of Chemical Engineers Environ Prog, 2016

To ensure the optimal infectivity on contact with host cells, pathogenic Pseudomonas syringae has evolved a complex mechanism to control the expression and construction of the functional type III secretion system (T3SS) that serves as a dominant pathogenicity factor. In this study, we showed that the hrpF gene of P. syringae pv. averrhoi, which is located upstream of hrpG, encodes a T3SS-dependent secreted/translocated protein. Mutation of hrpF leads to the loss of bacterial ability on elicitation of disease symptoms in the host and a hypersensitive response in non-host plants, and the secretion or translocation of the tested T3SS substrates into the bacterial milieu or plant cells. Moreover, overexpression of hrpF in the wild-type results in delayed HR and reduced t3ss expression. The results of protein-protein interactions demonstrate that HrpF interacts directly with HrpG and HrpA in vitro and in vivo, and protein stability assays reveal that HrpF assists HrpA stability in the bacterial cytoplasm, which is reduced by a single amino acid substitution at the 67th lysine residue of HrpF with alanine. Taken together, the data presented here suggest that HrpF has two roles in the assembly of a functional T3SS: one by acting as a negative regulator, possibly involved in the HrpSVG regulation circuit via binding to HrpG, and the other by stabilizing HrpA in the bacterial cytoplasm via HrpF-HrpA interaction prior to the secretion and formation of Hrp pilus on the bacterial surface.

In this letter, a 75–110-GHz W-band high-linearity traveling-wave single-pole-double-throw switch in 90-nm CMOS GUTM 1P9M is presented. Compared with the traditional traveling-wave switch with negative body-biasing technique, the proposed design applies the negative biasing to the gate terminals simultaneously. Therefore, the third-order intermodulation product is significantly reduced. The measured results show that the insertion loss is less than 3.4 dB, return loss is better than 10 dB, and Tx-to-Rx isolation is higher than 40 dB, respectively. For the linearity of the switch, the measured IP1dB is higher than 15 dBm and IIP3 is 37.1 dBm at 94 GHz, respectively. The proposed switch presents good performance of insertion loss, isolation, IP1dB, and IIP3, among the compared millimeter-wave CMOS switches.

In article number 2005637, Jonathan F Lovell and co-workers show that the SARS-CoV-2 RBD surface protein becomes a potent immunogen when presented in nanoparticle format Using a vaccine adjuvant that spontaneously converts soluble recombinant antigens into stable particles, immunization studies in mice and rabbits shows that the particle-based RBD elicits strong immune responses and potent antibodies capable of neutralizing the virus

Thermostability and decreased component costs are desirable features for adjuvanted, recombinant vaccines. We previously showed that a model malaria transmission-blocking vaccine candidate antigen, Pfs25, can be rendered more immunogenic when mixed with liposomes containing cobalt porphyrin–phospholipid (CoPoP) and a synthetic monophosphoryl lipid A (MPLA) variant. CoPoP can induce stable particle formation of recombinant antigens based on interaction with their polyhistidine tag. In the present work, different synthetic MPLA variants and concentrations were assessed in CoPoP liposomes. Long-term biophysical stability and immunogenicity were not adversely impacted by a 60% reduction in MPLA content. When admixed with Pfs25, the adjuvant formulations effectively induced functional antibodies in immunized mice and rabbits. Lyophilized, antigen-bound liposomes were formed using sucrose and trehalose cryoprotectants, which improved vaccine reconstitution for a variety of model antigens. Compared to liquid storage, the lyophilized Pfs25 and CoPoP liposomes exhibited thermostability with respect to size, biochemical integrity, binding capacity, protein folding and immunogenicity. Following 6 weeks of storage at 60 °C, the most extended storage period assessed, the lyophilized formulation induced functional antibodies in mice with immunization.

The receptor‐binding domain (RBD) of the SARS‐CoV‐2 spike protein is a candidate vaccine antigen that binds angiotensin‐converting enzyme 2 (ACE2), leading to virus entry. Here, it is shown that rapid conversion of recombinant RBD into particulate form via admixing with liposomes containing cobalt‐porphyrin‐phospholipid (CoPoP) potently enhances the functional antibody response. Antigen binding via His‐tag insertion into the CoPoP bilayer results in a serum‐stable and conformationally intact display of the RBD on the liposome surface. Compared to other vaccine formulations, immunization using CoPoP liposomes admixed with recombinant RBD induces multiple orders of magnitude higher levels of antibody titers in mice that neutralize pseudovirus cell entry, block RBD interaction with ACE2, and inhibit live virus replication. Enhanced immunogenicity can be accounted for by greater RBD uptake into antigen‐presenting cells in particulate form and improved immune cell infiltration in draining lymph nodes. QS‐21 inclusion in the liposomes results in an enhanced antigen‐specific polyfunctional T cell response. In mice, high dose immunization results in minimal local reactogenicity, is well‐tolerated, and does not elevate serum cobalt levels. Taken together, these results confirm that particulate presentation strategies for the RBD immunogen should be considered for inducing strongly neutralizing antibody responses against SARS‐CoV‐2., The receptor‐binding domain (RBD) of SARS‐CoV‐2 is a target for COVID‐19 vaccine development. Using a next‐generation vaccine adjuvant system that rapidly converts soluble antigens into stable particles, the neutralizing antibody response induced by the RBD is greatly enhanced, demonstrating merit for particle‐based approaches toward RBD‐based vaccines.

SUMMARY We examine whether increases in co-working experience between the lead and concurring audit partners affect engagement audit quality and audit efficiency. We define co-working experience as the lead and concurring audit partners having worked together in these roles on previous audit engagements for clients other than the focal client. A priori, increases in co-working experience could increase or decrease audit quality, but are expected to increase audit efficiency. Using data from Taiwan, where the identities of lead and concurring audit partners are known, we find that co-working experience is positively associated with audit quality and audit efficiency. Further, the effects of co-working experience on audit quality and audit efficiency are more pronounced when co-working experience is more intensive, the two partners are more accessible to each other, the audit firm is less experienced with the client or less knowledgeable about the client's industry, or client audit risks are higher. Data Availability: Data are available from the sources cited in the text. JEL Classifications: M4; M42.

Considering the various ways of Municipal Solid Waste (MSW) disposal that are currently in practice, this paper proposes a bundle pricing model of MSW disposal in Beijing, China. The bundle pricing model is set according to per capita annual disposable income, per capita annual consumption expenditure and MSW disposal cost based on the cost reimbursement principle and complementarity principle. This paper first estimates the unit costs of landfill and incineration respectively, and then designs three bundling packages according to three different income levels of households in Beijing (low-income households, middle-income households and high-income households). Specifically, based on our bundle pricing model, we propose to charge low-income households (of the average size of 3 persons) MSW fees 5.64 RMB per month, middle-income households 10.16 RMB, and high-income households 20.70 RMB per month. We also discuss and explain how the proposed mechanism can be expected to not only mitigate the capital shortage problem, but also reduce MSW quantity and improve the MSW incineration disposal technology.

Drug resistance to conventional therapies remains a major cause of treatment failure, tumor recurrence and dismal survival rates for patients with advanced stage cancers. Photodynamic therapy (PDT) provides an opportunity to exploit photochemically-triggered death mechanisms via targeting of sub-cellular, cellular and stromal compartments to overcome treatment resistance in unresponsive populations of stubborn disease. The informed design of mechanism-based combinations is emerging as increasingly important to targeting resistance and improving the efficacy of conventional treatments, while minimizing toxicity. PDT has been shown to synergize with conventional agents and to overcome the evasion pathways that cause resistance. Increasing evidence shows that PDT-based combinations cooperate mechanistically with, and improve the therapeutic index of, traditional chemotherapies. These and other findings emphasize the importance of including PDT as part of comprehensive treatment plans for cancer, particularly in complex disease sites. Identifying effective combinations requires a multi-faceted approach that includes the development of bioengineered cancer models and corresponding image analysis tools. The presentation will focus on the molecular and phenotypic basis of verteporfin PDT-based enhancement of chemotherapeutic efficacy and predictability in complex 3D models and in vivo models, with a particular emphasis on ovarian and pancreatic cancer.

Artificial superlattices, based on van der Waals heterostructures of two-dimensional atomic crystals such as graphene or molybdenum disulfide, offer technological opportunities beyond the reach of existing materials. Typical strategies for creating such artificial superlattices rely on arduous layer-by-layer exfoliation and restacking, with limited yield and reproducibility. The bottom-up approach of using chemical-vapour deposition produces high-quality heterostructures but becomes increasingly difficult for high-order superlattices. The intercalation of selected two-dimensional atomic crystals with alkali metal ions offers an alternative way to superlattice structures, but these usually have poor stability and seriously altered electronic properties. Here we report an electrochemical molecular intercalation approach to a new class of stable superlattices in which monolayer atomic crystals alternate with molecular layers. Using black phosphorus as a model system, we show that intercalation with cetyl-trimethylammonium bromide produces monolayer phosphorene molecular superlattices in which the interlayer distance is more than double that in black phosphorus, effectively isolating the phosphorene monolayers. Electrical transport studies of transistors fabricated from the monolayer phosphorene molecular superlattice show an on/off current ratio exceeding 10^7, along with excellent mobility and superior stability. We further show that several different two-dimensional atomic crystals, such as molybdenum disulfide and tungsten diselenide, can be intercalated with quaternary ammonium molecules of varying sizes and symmetries to produce a broad class of superlattices with tailored molecular structures, interlayer distances, phase compositions, electronic and optical properties. These studies define a versatile material platform for fundamental studies and potential technological applications.

The objectives of this paper are to provide a descriptive analysis of the Chinese stock market gyrations in 2015 and to assess the effectiveness of Chinese government’s attempts to rescue the stock market from collapse. This paper views the efforts taken up by China’s policy makers in 2015 to stabilize the stock market as a case study of public crisis management. We first briefly chronicle the boom and bust of China’s stock market in the recent past seven months, and analyze the underlying causes and background of these gyrations. Next, we describe various policy actions implemented by the government in response to this public crisis. Finally, we provide some assessments and projections of the effectiveness of these intervention measures. We conclude that while it is understandable and seems necessary for the Chinese authority to intervene to prevent the crisis from exploding further, it has unfortunately damaged the credibility of the current leadership’s management capability and resolve to “confine the power inside the cage”.

In this study of the amphiphilic polypeptide TP-PPLG-g-MEO2 containing a hydrophobic tetraphenylthiophene (TP) terminal, with aggregation-induced emission (AIE) properties, and hydrophilic ether side groups, it was found that secondary structures (α-helix, β-sheet and random coil) of the peptide chains affected the AIE-related emission of the TP terminal. The amphiphilic TP-PPLG-g-MEO2, prepared from a ring-opening polymerization and the following click reaction, contained a secondary structure mostly composed of α-helical chains. The main α-helical chain nevertheless can be converted into a β-sheet structure by adding NaCl into the aqueous solution of TP-PPLG-g-MEO2 and by doing so, the solution emission can be enhanced. In addition, the coil chain generated in the alkaline solution was found to emit efficiently with the highest emission intensity among all three conformations. The relationship between the AIE property and the secondary structure of the peptide chains can therefore be evaluated. All three secondary structures were also used as luminescent sensors to test their sensitivity for bovine serum albumin (BSA).