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Aims/Introduction The relationship between glycated hemoglobin (HbA1c) and cardiovascular events in older adults was investigated using a Japanese administrative medical database. Materials and Methods Anonymized medical data on patients with diabetes mellitus aged ≥65 years for the period from January 2010 to December 2019 were extracted from the EBM Provider database. The primary end‐point was a composite of cardiovascular events, whereas the other end‐points included severe hypoglycemia and fracture. The association between cardiovascular events and HbA1c at the index date (i.e., approximately 10 months after initial diabetes mellitus diagnosis) was evaluated using the Cox proportional hazards model. Results Among the 3,186,751 patients in the database, 3,946 older adults with diabetes mellitus were eligible for inclusion and were subsequently grouped according to HbA1c quartiles at the index date. Cardiovascular events occurred in 142 patients. Patients with HbA1c in the highest quartile had significantly higher risk of hospitalization for cardiovascular disease than those with HbA1c in the lowest quartile (hazard ratio 1.948; 95% confidence interval 1.252–3.031, P = 0.003). However, the events risk was similar across subgroups with HbA1c, We investigated the relationship between glycated hemoglobin and cardiovascular events in older adults with diabetes mellitus using administrative medical databases in Japan. Among approximately 4,000 older patients without a history of dementia, cancer or cardiovascular disease, those with glycated hemoglobin in the highest quartile had a significantly higher risk of hospitalization for cardiovascular disease than those with glycated hemoglobin in the lowest quartile. Age, sex, prescription of antihypertensive drugs and comorbidities were also associated with the risk of cardiovascular events.

Lymphodepleting cytotoxic regimens enhance the antitumor effects of adoptively transferred effector and naive T cells. Although the mechanisms of antitumor immunity augmentation by lymphodepletion have been intensively investigated, the effects of lymphodepletion followed by T cell transfer on immune checkpoints in the tumor microenvironment remain unclear. The current study demonstrated that the expression of immune checkpoint molecules on transferred donor CD4+ and CD8+ T cells was significantly decreased in lymphodepleted tumor-bearing mice. In contrast, lymphodepletion did not reduce immune checkpoint molecule levels on recipient CD4+ and CD8+ T cells. Administration of anti-PD-1 antibodies after lymphodepletion and adoptive transfer of T cells significantly inhibited tumor progression. Further analysis revealed that transfer of both donor CD4+ and CD8+ T cells was responsible for the antitumor effects of a combination therapy consisting of lymphodepletion, T cell transfer and anti-PD-1 treatment. Our findings indicate that a possible mechanism underlying the antitumor effects of lymphodepletion followed by T cell transfer is the prevention of donor T cell exhaustion and dysfunction. PD-1 blockade may reinvigorate exhausted recipient T cells and augment the antitumor effects of lymphodepletion and adoptive T cell transfer.

A self-driven sensor that can detect urine and urine sugar and can be mounted on diapers is desirable to reduce the burden of long-term care. In this study, we created a paper-based glucose biofuel cell that can be mounted on diapers to detect urine sugar. Electrodes for biofuel cells were produced by printing MgO-templated porous carbon on which poly(glycidyl methacrylate) was modified using graft polymerization. A new bioanode was prepared through covalently modifying flavin-adenine-dinucleotide-dependent glucose dehydrogenase and azure A with pendant glycidyl groups of poly(glycidyl methacrylate). We prepared a cathode with covalently bonded bilirubin oxidase. Covalent bonding of enzymes and mediators to both the bioanode and biocathode suppressed elution and improved stability. The biofuel cell could achieve a maximum output density of 0.12 mW cm–2, and by combining it with a wireless transmission device, the concentration of glucose sensed from the transmission frequency was in the range of 0–10 mM. The sensitivity of the sensor was estimated at 0.0030 ± 0.0002 Hz mmol–1 dm3. This device is expected to be a new urine-sugar detection device, composed only of organic materials with a low environmental load and it can be useful for detecting postprandial hyperglycemia.

Aims/Introduction To prevent diabetic complications, strict glucose control and frequent monitoring of blood glucose levels with invasive methods are necessary. We considered the monitoring of tear glucose levels might be a possible method for non‐invasive glucose monitoring. To develop tear glucose monitoring for clinical application, we investigated the precise correlation between the blood and tear glucose concentrations. Materials and Methods A total of 10 participants and 20 participants with diabetes were admitted, and blood and tear samples were collected. Before statistical analysis, we eliminated tear samples contaminated with blood. We observed the daily blood and tear glucose dynamics, and carried out a random intercept model analysis to examine the association between the blood and tear glucose concentrations. Results Tear occult blood tests showed that the tear glucose concentrations and their variation increased in both participants with and without diabetes as contamination of blood increased. In both participants with and without diabetes, fluctuations of the plasma glucose concentrations were observed depending on the timing of collection of the samples, and the dynamics of the tear glucose concentrations paralleled those of the plasma glucose concentrations. The random intercept model analysis showed a significant association between the plasma and tear glucose concentrations in participants with diabetes (P, It is important to eliminate the tear samples contaminated with blood. Tear glucose monitoring might be a reliable and non‐invasive substitute method for monitoring the blood glucose concentrations for diabetes patients, irrespective of glycated hemoglobin levels and the timing of sample collection.

Pegylated liposomal doxorubicin (PLD) is a representative nanomedicine that has improved tumor selectivity and safety profile. However, the therapeutic superiority of PLD over conventional doxorubicin has been reported to be insignificant in clinical medicine. Combination treatment with microbubbles and ultrasound (US) is a promising strategy for enhancing the antitumor effects of chemotherapeutics by improving drug delivery. Recently, several preclinical studies have shown the drug delivery potential of lipid bubbles (LBs), newly developed monolayer microbubbles, in combination with low-intensity US (LIUS). This study aimed to elucidate whether the combined use of LBs and LIUS enhanced the intratumoral accumulation and antitumor effect of PLD in syngeneic mouse tumor models. Contrast-enhanced US imaging using LBs showed a significant decrease in contrast enhancement after LIUS, indicating that LIUS exposure induced the destruction of LBs in the tumor tissue. A quantitative evaluation revealed that the combined use of LBs and LIUS improved the intratumoral accumulation of PLD. Furthermore, tumor growth was inhibited by combined treatment with PLD, LBs, and LIUS. Therefore, the combined use of LBs and LIUS enhanced the antitumor effect of PLD by increasing its accumulation in the tumor tissue. In conclusion, the present study provides important evidence that the combination of LBs and LIUS is an effective method for enhancing the intratumoral delivery and antitumor effect of PLD in vivo.

Dynamic spatiotemporal patterns that arise from out-of-equilibrium biochemical reactions generate forces in living cells. Despite considerable recent efforts, rational design of spatiotemporal patterns in artificial molecular systems remains at an early stage of development. Here, we describe force generation by a propagating wave of supramolecular nanofibers. Inspired by actin dynamics, a reaction network is designed to control the formation and degradation of nanofibers by two chemically orthogonal stimuli. Real-time fluorescent imaging successfully visualizes the propagating wave based on spatiotemporally coupled generation and collapse of nanofibers. Numerical simulation indicates that the concentration gradient of degradation stimulus and the smaller diffusion coefficient of the nanofiber are critical for wave emergence. Moreover, the force (0.005 pN) generated by chemophoresis and/or depletion force of this propagating wave can move nanobeads along the wave direction., The rational design of spatiotemporal patterns in artificial molecular systems remains at an early stage of development. Here, the authors design a reaction network to control the formation and degradation of nanofibers by orthogonal chemical stimuli and describe force generation by a propagating wave of supramolecular nanofibers.

Poly(glycidyl methacrylate) (poly(GMA)) bearing pendant glycidyl groups, grafted on the surface of MgO-templated carbon (MgOC), is useful for forming strong multipoint covalent bonds with amino fun...

To improve severe ketoacidosis with COVID‐19, insulin treatment, invasive mechanical ventilation therapy, and continuous hemodiafiltration with sodium bicarbonate infusion were effective.

Background/aim This study analysed threonine-phosphorylated Smad2/3 (pSmad2/3L-Thr) expression and investigated whether pSmad2/3L-Thr is related to the transition from human colorectal adenoma (CRA) to carcinoma (CRC). Materials and methods Immunofluorescent staining was performed forβ-catenin, p53, CDK4, Ki67, Sox9, aldehyde dehydrogenase (ALDH) 1, and pSmad2/3L-Thr. Results We analysed specimens of diffuse p53-positive CRCs arising from p53-negative CRAs. Percentage of p53, nuclear β-catenin, Ki67, CDK4, and pSmad2/3L-Thr-positive cells at the site of CRCs was significantly higher than that at the site of CRAs. At the site of normal colorectal mucosae, few epithelial cells were stained positively for pSmad2/3L-Thr. At the site of CRCs, pSmad2/3L-Thr-positive cells showed co-localization with p53, nuclear β-catenin, and ALDH1. At any site, pSmad2/3L-Thr-positive cells showed co-localization with CDK4. Conclusion pSmad2/3L-Thr correlates with human CRC carcinogenesis, and pSmad2/3L-Thr-positive cells show human colorectal stem cell-like and cancer stem cell characteristics.

Roundabout guidance receptor 4 (Robo4) is an endothelial-specific membrane protein that suppresses pathological angiogenesis and vascular hyperpermeability by stabilizing endothelial cells. Robo4 suppresses severe systemic inflammation induced by pathogens and endotoxins and inhibits tumor growth and metastasis, therefore serving as a potential therapeutic target. Although the regulation of Robo4 expression through transcription factors and epigenetic mechanisms has been studied, the role of histone deacetylases (HDACs) has not been explored. In the present study, we investigated the involvement of HDACs in the regulation of Robo4 expression. An HDAC inhibitor, MS-275, which inhibits HDAC1, HDAC2, and HDAC3, was found to suppress Robo4 expression in endothelial cells. Small interfering RNA (siRNA)-mediated knockdown of HDAC3, but not of HDAC1 and 2, also decreased its expression level. MS-275 downregulated the expression of the transcription factor complex GABP, in addition to suppressing Robo4 promoter activity. GABP expression was also downregulated by the siRNA against HDAC3. MS-275 decreased the transendothelial electrical resistance of a monolayer of mouse endothelial cells and increased the rate of leakage of Evans blue dye in the mouse lungs. In addition, MS-275 accelerated cell migration through the endothelial cell monolayer and augmented cell extravasation in the mouse lungs. Taken together, we demonstrated that MS-275 suppresses Robo4 expression by inhibiting HDAC3 in endothelial cells and enhances endothelial and vascular permeability. Thus, we demonstrated a novel mechanism regulating Robo4 expression and vascular permeability, which is anticipated to contribute to future therapies for infectious and inflammatory diseases.

Glutathione (GSH) is an important antioxidant that plays a critical role in neuroprotection. GSH depletion in neurons induces oxidative stress and thereby promotes neuronal damage, which in turn is regarded as a hallmark of the early stage of neurodegenerative diseases. The neuronal GSH level is mainly regulated by cysteine transporter EAAC1 and its inhibitor, GTRAP3-18. In this study, we found that the GTRAP3-18 level was increased by up-regulation of the microRNA miR-96-5p, which was found to decrease EAAC1 levels in our previous study. Since the 3’-UTR region of GTRAP3-18 lacks the consensus sequence for miR-96-5p, an unidentified protein should be responsible for the intermediate regulation of GTRAP3-18 expression by miR-96-5p. Here, we discovered that RNA-binding protein NOVA1 functions as an intermediate protein for GTRAP3-18 expression via miR-96-5p. Moreover, we show that intra-arterial injection of a miR-96-5p-inhibiting nucleic acid to living mice by a drug delivery system using microbubbles and ultrasound decreased the level of GTRAP3-18 via NOVA1 and increased the levels of EAAC1 and GSH in the dentate gyrus of the hippocampus. These findings suggest that the delivery of a miR-96-5p inhibitor to the brain would efficiently increase the neuroprotective activity by increasing GSH levels via EAAC1, GTRAP3-18 and NOVA1., Chisato Kinoshita et al. use a microbubble and ultrasound delivery system to inhibit miR-96-5p in the mouse brain. They demonstrate that miR96-5p regulates glutathione levels through the NOVA1 RNA binding protein, suggesting that this microRNA could be a target for altering glutathione levels in neurodegenerative disease.

Bronchopleural fistula is one of the most serious postoperative complications caused by the incomplete healing of a bronchial stump. Fibroblasts play an important role in wound healing by facilitating connective tissue formation and inducing angiogenesis. We developed a method for production of multilayered fibroblast sheets that secreted some growth factors and promoted wound healing. The present study aimed to assess the treatment effect of multilayered fibroblast sheets on bronchial stump healing. In this rat model, left pneumonectomy was performed, and multilayered fibroblast sheets derived from autologous oral mucosal tissues were transplanted to the bronchial stump. The changes in the bronchial stump were examined macroscopically, histologically, and mechanically. The fibroblast sheets promoted the formation of thick connective tissues around the bronchial stump. The formed connective tissues were accompanied by new blood vessels, and fibrosis was observed over time. Then, 7 days after the transplantation of the fibroblast sheets, the bronchial wall became significantly thicker, and the area of the blood vessels for the bronchial wall tissues was significantly larger in the experimental group than in the control group. In addition, the burst pressure in the bronchial stump was significantly higher in the experimental group than in the control group. Bronchial stumps were reinforced by the transplantation of multilayered fibroblast sheets derived from autologous oral mucosal tissues.

Ephedra herb (Mao) exerts potent anti-allergic effects. This study aimed to examine the underlying mechanisms of Mao on allergic inflammation using in vitro cultured mast cells (MCs) and an in vivo model of MC-dependent anaphylaxis. Bone marrow-derived MCs (BMMCs) were presensitized with anti-2,4-dinitrophenol (DNP) immunoglobulin E (IgE) and challenged with antigens (Ag; DNP-human serum albumin). Degranulation responses and cell surface high-affinity receptor for IgE (FceRI) expression were assessed with/without Mao treatment. Passive systemic anaphylaxis (PSA)-treated mice were administered Mao and the pathophysiological responses were evaluated. Mao inhibited Ag-induced BMMC degranulation, but not polyclonal activation with phorbol 12-myristate 13-acetate (PMA) and ionomycin, indicating that Mao inhibits IgE-dependent activation of BMMCs. Mao-treated BMMCs exhibited significant reductions in expression of surface IgE and its receptor FceRI. Analysis of subcellular localization revealed that Mao induces FceRI internalization in BMMCs without degranulation. In the PSA mouse model, Mao administration prevented antigen-induced hypothermia. Mao administration significantly reduced cell surface expression of IgE-bound FceRI on peritoneal MCs. Mao induced FceRI internalization in MCs, thereby inhibiting Ag-induced IgE-dependent degranulation. The inhibitory effects of Mao on MC degranulation may offer a novel therapeutic approach for allergic diseases.

We have previously reported methods for the active control of microbubbles, and the use of cells surrounded by microbubbles to deliver drugs in human blood vessels using acoustic radiation force. However, the effect and degree of damage to blood vessel walls has not previously been studied with respect to the application of continuous and burst ultrasound emissions. We examined the destruction of lipid bubbles, which were injected into the circulation path, to measure the fragility under various conditions of ultrasound exposure. Next, the cell viability of vascular endothelial cells in the presence of bubbles was verified, where the various conditions of ultrasound were irradiated to suspension composed with the cells and the bubbles. We confirmed that cell viability decreased in the presence of bubbles in proportion to a sound pressure and a duty ratio. Furthermore, we found a significant relationship between the cell viability and the destruction coefficient of bubbles.

Mechanisms of BBB opening were investigated using a capillary model created by a three-dimensional cell culture technique. The model was made using human umbilical vein endothelial cells (HUVECs) suspended in collagen gel. Incubation with endothelial growth factor for one week resulted in the creation of capillary lumens, and a suspension of microbubbles supplemented with FITC-dextran, SYTOX Blue, and Calcein-AM was introduced inside the lumens. Bubble-cell interaction under exposure to short pulsed ultrasound of 1.0 MHz in center frequency and 0.4 MPa in peak negative pressure was captured using a high-speed camera, and lumen and endothelial cell damage was visualized using fluorescence microscopy. High-speed observation showed that non-uniform contraction of a bubble caused significant local deformation of the lumen beside the bubble, and strong extravasation from the lumen (FITC-dextran) and significant membrane damage of the endothelial cells (SYTOX Blue) were confirmed at this location. Lethal damage was also observed frequently (Calcein). On the other hand, cells on the opposite side of the lumen received stretching force during bubble expansion, and smaller extravasation and cell damage maintained the integrity of endothelial cells. These suggested the presence of different mechanisms of BBB opening.

Diagnostic ultrasound is non-invasive and provides real-time imaging. Microbubbles (MBs) are ultrasound contrast agents used to observe small blood flow, such as tumor tissue. However, MBs have short blood flow imaging time. This study developed lipid-based microbubbles (LMBs) with longer blood flow imaging time by focusing on their shell composition. Liposome research reported that addition 1,2-distearoyl-sn-glycero-3-phosphoglycerol (DSPG) to the lipid composition enhances liposome membrane stability. Therefore, we introduced DSPG at different ratios into the LMBs lipid shell. Results showed that the lipid shell composition of MBs affects stability in vivo. 60% DSPG-containing LMBs (DSPG60-LMBs) have sustained blood flow imaging time compared with LMBs, which have other DSPG ratios, Sonazoid® and SonoVue®. DSPG60-LMBs also showed less uptake into the liver compared with Sonazoid®. Therefore, DSPG60-LMBs can have long blood flow imaging time and can be effective diagnostic agents in ultrasound imaging.

Here, we present the design, synthesis, and SAR of dual orexin 1 and 2 receptor antagonists, which were optimized by balancing the antagonistic activity for orexin receptors and lipophilicity. Based on the prototype compound 1, ring construction and the insertion of an additional heteroatom into the resulting ring led to the discovery of orexin 1 and 2 receptor antagonists, which were 3-benzoyl-1,3-oxazinane derivatives. Within these derivatives, (−)-3h enabled a high dual orexin receptor antagonistic activity and a low lipophilicity. Compound (−)-3h exhibited potent sleep-promoting effects at a po dose of 1 mg/kg in a rat polysomnogram study, and optimal PK properties with a rapid Tmax and short half-lives in rats and dogs were observed, indicating a predicted human half-life of 0.9–2.0 h. Thus, (−)-3h (ORN0829; investigation code name, TS-142) was selected as a viable candidate and is currently in clinical development for the treatment of insomnia.

Objectives/hypothesis To elucidate the aging physiology of the vocal folds, we examined the characters of aged vocal fold fibroblasts (VFFs) in various conditions. Study design In vitro study. Methods VFFs from young (12-week-old) and aged (19-month-old) Sprague-Dawley rats were compared. Proliferative capacity, ratio of myofibroblast to fibroblast, myofibroblast function, and extracellular matrix production were examined in the following conditions: naive, basic fibroblast growth factor (bFGF) supplemented, and hepatocyte growth factor (HGF) supplemented. Results Aged VFFs demonstrated reduced proliferation by cell counting, though the ratio of Ki-67-positive cells showed no difference. Aged VFFs exhibited an increased expression of α-smooth muscle actin (α-SMA); however, they demonstrated no enhanced contractile ability in a gel contraction assay. Type I collagen protein was increased age dependently, accompanied with decreased Mmp1 and unchanged Col1a1 transcription. Type I collagen protein and α-SMA represented quite similar reduction patterns to bFGF or HGF administration. Conclusions The following possible characteristics of aged VFFs were implied: long duration of mitosis, increased myofibroblast population size with certain dysfunctions, reduced type I collagen turnover, and correlation between α-SMA expression and type I collagen metabolism. Further investigations of these features will help to clarify presbyphonia's pathology and establish treatment strategies. Level of evidence NA Laryngoscope, 129:E94-E101, 2019.

Rumen responses to cashew nut shell liquid (CNSL) were evaluated in a feeding study. Four wethers were fed a hay and concentrate diet for 4 weeks (pre-CNSL period), and then fed the same diet supplemented with low and high levels of CNSL for 2 weeks each (L-CNSL and H-CNSL periods respectively). The diet was then reverted to the unsupplemented control diet for another 2 weeks (post-CNSL period). Rumen parameters were monitored in each feeding period. CNSL, regardless of the two levels tested, did not show any adverse effects on total short chain fatty acid concentration and dry matter digestibility in the rumen. Propionate proportion increased in the H-CNSL period, while methane production potential, acetate and butyrate proportions, viscosity, foam formation and its stability, and ammonia concentration decreased. Values of these parameters returned to those in the unsupplemented control period after cessation of CNSL supplementation. Clone library analysis of 16S rRNA genes revealed the following shifts in the H-CNSL period. For bacteria, Firmicutes was frequently detected, while Bacteroidetes and Spirochetes were not. For archaea, Methanobrevibacter wolinii was predominant. These results indicate that CNSL could be a methane inhibitor and propionate enhancer by altering the rumen microbial community.

Choline is used to synthesize phospholipids and a lack of choline induces a number of liver-related diseases, including non-alcoholic steatohepatitis. The current study characterized the choline uptake system, at molecular and functional levels, in the immortalized human hepatic cell line, Fa2N-4, to identify the specific choline transporter involved in choline uptake. The present study also assesed whether choline deficiency or the inhibited choline uptake affected cell viability and apoptosis. Reverse transcription-quantitative polymerase chain reaction (PCR) revealed choline transporter-like protein 1 (CTL1) and CTL2 mRNA and protein expression in Fa2N-4 cells. [Methyl-3H]choline studies revealed choline uptake was saturable and mediated by a single transport system that functioned in a Na+-independent but pH-dependent manner, which was similar to CTL1. Hemicholinium-3 (HC-3), which is a choline uptake inhibitor, and choline deficiency inhibited cell viability, increased caspase-3 and −7 activities, and increased fluorescein isothiocyanate-Annexin V immunofluorescent staining indicated apoptosis. Immunofluorescent staining also revealed CTL1 and CTL2 localized in plasma and mitochondrial membranes, respectively. [Methyl-3H]choline uptake was enhanced by a protein kinase C (PKC) activator, phorbol-12-myristate 13-acetate (PMA). Immunofluorescence staining and western blot analysis demonstrated increased CTL1 expression on the cell membrane following PMA treatment. The results of current study indicated that extracellular choline is primarily transported via CTL1, relying on a direct H+ gradient that functions as a driving force in Fa2N-4 cells. Furthermore, it was hypothesized that CTL1 and the choline uptake system are strongly associated with cell survival, and that the choline uptake system is modulated by PKC signaling via increased CTL1 expression on the cell surface. These findings provide further insights into the pathogenesis of liver disease involving choline metabolism.

The epidermis, the outermost layer of the skin, retains moisture and functions as a physical barrier against the external environment. Epidermal cells are continuously replaced by turnover, and thus to understand in detail the dynamic cellular events in the epidermis, techniques to observe live tissues in 3D are required. Here, we established a live 3D imaging technique for epidermis models. We first obtained immortalized human epidermal cell lines which have a normal differentiation capacity and fluorescence-labelled cytoplasm or nuclei. The reconstituted 3D epidermis was prepared with these lines. Using this culture system, we were able to observe the structure of the reconstituted epidermis live in 3D, which was similar to an in vivo epidermis, and evaluate the effect of a skin irritant. This technique may be useful for dermatological science and drug development.

To realize highly efficient solid-state dye-sensitized solar cells (SDSCs), the absorption range of the dye should be extended to the near-IR range to increase short-circuit current density (Jsc); a high Jsc in turn requires a highly conductive p-type semiconductor. A newly developed dye (DIPDAB2) with a porphyrin dimer structure provided higher absorption coefficients than the conventional dye with a similar framework (DTBC) in the long wavelength range of 700–800 nm, leading to higher incident photon-to-current conversion efficiencies. The dip in the absorption spectrum of DIPDAB2 located at 500–700 nm between the Soret band and Q band was filled by combining with two kinds of organic dyes (D131 and D358). The multi-dye consisting of the three dyes realized a high Jsc over 20 mA cm−2. The use of copper iodide that has a higher conductivity than p-type organic semiconductors and copper complexes secured a high filling factor. Introduction of Li ions into the TiO2 photoelectrodes improved the open-circuit voltage (Voc) along with a slight increase in Jsc. Light soaking also contributed to a higher Voc. The conversion efficiency of the present SDSC was as high as 10%.

The discovery of 1-({6-[(2-methoxy-4-propylbenzyl)oxy]-1-methyl-3,4-dihydronaphthalen-2-yl}methyl)azetidine-3-carboxylic acid 13n (ceralifimod, ONO-4641), a sphingosine-1-phosphate (S1P) receptor agonist selective for S1P1 and S1P5, is described. While it has been revealed that the modulation of the S1P1 receptor is an effective way to treat autoimmune diseases such as relapsing–remitting multiple sclerosis (RRMS), it was also reported that activation of the S1P3 receptor is implicated in some undesirable effects. We carried out a structure–activity relationship (SAR) study of hit compound 6 with an amino acid moiety in the hydrophilic head region. Following identification of a lead compound with a dihydronaphthalene central core by inducing conformational constraint, optimization of the lipophilic tail region led to the discovery of 13n as a clinical candidate that exhibited >30 000-fold selectivity for S1P1 over S1P3 and was potent in a peripheral lymphocyte lowering (PLL) test in mice (ED50 = 0.029 mg/...

It is now well known that immunoglobulin E (IgE) and mast cells (MCs) are important participants in allergic diseases. MCs contain electron-dense secretory granules which are filled with inflammatory mediators. The interaction of an allergen (antigen) with an antigen-specific IgE-bound high-affinity receptor for IgE (FcεRI) is an essential step in MC activation as well as subsequent downstream signaling events. What we know is that IgE and FcεRI activate a complex regulatory network (i.e., signaling molecules and messengers) that governs both the type of MC activation and the symptoms of allergic disease. This review focuses on recent discoveries that shed new light on FcεRI signaling networks, holding promise for the development of new therapeutic solutions in the treatment of allergic diseases.

Type-A CpG oligodeoxynucleotides (ODNs), which have a natural phosphodiester backbone, is one of the highest IFN-α inducer from plasmacytoid dendritic cells (pDC) via Toll-like receptor 9 (TLR9)-dependent signaling. However, the in vivo application of Type-A CpG has been limited because the rapid degradation in vivo results in relatively weak biological effect compared to other Type-B, -C, and -P CpG ODNs, which have nuclease-resistant phosphorothioate backbones. To overcome this limitation, we developed lipid nanoparticles formulation containing a Type-A CpG ODN, D35 (D35LNP). When tested in a mouse tumor model, intratumoral and intravenous D35LNP administration significantly suppressed tumor growth in a CD8 T cell-dependent manner, whereas original D35 showed no efficacy. Tumor suppression was associated with Th1-related gene induction and activation of CD8 T cells in the tumor. The combination of D35LNP and an anti-PD-1 antibody increased the therapeutic efficacy. Importantly, the therapeutic schedule and dose of intravenous D35LNP did not induce apparent liver toxicity. These results suggested that D35LNP is a safe and effective immunostimulatory drug formulation for cancer immunotherapy.

Ultrahigh-Q crystalline microresonators have various potential applications including as microresonator frequency combs [1] and low-noise radio frequency oscillators [2]. Since crystalline materials are transparent in the visible to mid-infrared wavelength region, magnesium fluoride (MgF 2 ) and calcium fluoride (CaF 2 ) microresonators have been expected to achieve broad bandwidth Kerr comb generation. The demand for precise geometrical dispersion control to compensate for material dispersion has recently been increasing because material dispersion in the visible or mid-infrared region is not feasible for obtaining a Kerr comb [3,4]. However, the dispersion engineering of crystalline microresonators remains a major challenge since hand polishing is generally needed after diamond turning to obtain a high Q [5]. This restricts the possibility of tailoring the resonator dispersion because it degrades the controllability of the cross-sectional shape.

The combination of Ultrasound (US) and US contrast agent (microbubbles, MBs), which is gas stabilized by a shell such as phospholipids or proteins, has potential as a useful innovative diagnostic and therapeutic tool. Previous studies have evaluated how particle size or shell components of MBs affect their physical characteristics, imaging ability, and drug delivery efficacy. We reported that MBs composed of neutral, anionic phospholipids, and polyethylene glycol-conjugated phospholipids at appropriate ratios were highly stable for US imaging. However, the effects of encapsulated gas on stability and drug delivery efficacy have not been characterized. Therefore, we developed several gas-loaded MBs with identical shell compositions and assessed their stability by US imaging (LOGIQ E9 with ML6-15 probe, MI 0.20). In addition, we assessed the effects of gas encapsulated in MBs on brain-targeted drug delivery, because the brain requires an efficient drug delivery system. Perfluoropropane and perfluorobutane-loaded MBs (MB-C3F8 and MB-C4F10) showed sustained US imaging in vitro and in vivo compared with sulfur hexafluoride-loaded MBs (MB-SF6). In addition, treatment of MB-C3F8 and MB-C4F10 with non-focused US efficiently delivered Evans blue, which was used as a model drug, to the brain to a greater extent than MB-SF6. In these treatments, notable damage to brain was not observed, which was assessed by HE staining and denatured neuron staining. Our results suggested that perfluoropropane and perfluorobutane could be useful for the production of MBs with high stability to allow for US imaging and drug delivery.

The photovoltaic effect in traditional p–n junctions—where a p-type material (with an excess of holes) abuts an n-type material (with an excess of electrons)—involves the light-induced creation of electron–hole pairs and their subsequent separation, generating a current. This photovoltaic effect is particularly important for environmentally benign energy harvesting, and its efficiency has been increased dramatically, almost reaching the theoretical limit1. Further progress is anticipated by making use of the bulk photovoltaic effect (BPVE)2, which does not require a junction and occurs only in crystals with broken inversion symmetry3. However, the practical implementation of the BPVE is hampered by its low efficiency in existing materials4–10. Semiconductors with reduced dimensionality2 or a smaller bandgap4,5 have been suggested to be more efficient. Transition-metal dichalcogenides (TMDs) are exemplary small-bandgap, two-dimensional semiconductors11,12 in which various effects have been observed by breaking the inversion symmetry inherent in their bulk crystals13–15, but the BPVE has not been investigated. Here we report the discovery of the BPVE in devices based on tungsten disulfide, a member of the TMD family. We find that systematically reducing the crystal symmetry beyond mere broken inversion symmetry—moving from a two-dimensional monolayer to a nanotube with polar properties—greatly enhances the BPVE. The photocurrent density thus generated is orders of magnitude larger than that of other BPVE materials. Our findings highlight not only the potential of TMD-based nanomaterials, but also more generally the importance of crystal symmetry reduction in enhancing the efficiency of converting solar to electric power. A bulk photovoltaic effect is observed in devices based on tungsten disulfide, and is enhanced if the devices take the form of polar nanotubes, showing the importance of reducing crystal symmetry to a polar structure in achieving higher efficiencies.

It is known that Phosphatidyl choline-Phosphatidyl glycerol mixtures can be used for liposome formulations, making them less leaky than liposomes with only one lipid. We hypothesized that this might also be the case for bubbles, which can be used as ultrasound (US) contrast agents. Therefore, we have compared a series of mixed distearoyl phosphatidylcholine-distearoyl phosphatidylglycerol (DSPC-DPSG) bubbles and with bubbles containing either DSPC or DSPG (and distearoyl ethanolamine-polyethyleneglycol 2000, DSPE-PEG2k). Here, we describe the development, examination of stability

This paper proposes natural drug candidate compounds for the treatment of coronavirus disease 2019 (COVID-19). We investigated the binding properties between the compounds in the Moringa oleifera plant and the main protease (Mpro) of severe acute respiratory syndrome coronavirus 2 using molecular docking and ab initio fragment molecular orbital calculations. Among the 12 compounds, niaziminin was found to bind the strongest to Mpro. We furthermore proposed novel compounds based on niaziminin and investigated their binding properties to Mpro. The results reveal that the introduction of a hydroxyl group into niaziminin enhances its binding affinity to Mpro. These niaziminin derivatives can be promising candidate drugs for the treatment of COVID-19., Graphical abstract The interacting structure between our proposed compound 9d and selected important residues of SARS-CoV-2 main protease. Red and blue lines indicate hydrogen bonding and electrostatic interactions, respectively.Unlabelled Image

Hardness and slip systems by an indentation method were investigated on different habit planes of orthorhombic hen egg-white lysozyme (O-HEWL) crystals containing water. A dependence of the hardness on the water-evaporation time exhibits three stages as incubation, transition and saturated ones, as tetragonal (T)-HEWL crystals reported previously. The hardness values of (1 1 0), (0 1 0) and (0 1 1) habit planes of O-HEWL in the incubation stage or wet condition exhibits 6, 8 and 10 MPa, respectively. The hardness depends on indented planes but it is independent of the air-humidity and crystal volumes. These values correspond to the intrinsic hardness for O-HEWL crystals containing water. In the incubation stage, the slip traces are clearly observed around the indentation mark and the corresponding six kinds of slip systems are identified to be {0 1 1} , {1 1 0} , {0 1 1} , {1 1 0} , {1 0 0} and {0 1 0} .

Objectives/Hypothesis Vocal fold scarring, which causes severe hoarseness, is intractable. The optimal treatment for vocal fold scarring has not been established; therefore, prevention of scarring is important. The aim of this study was to clarify the effectiveness of basic fibroblast growth factor (bFGF) for prevention of postsurgical vocal fold scarring. Study Design Prospective animal experiments with controls. Method The vocal folds of Sprague-Dawley rats were injured unilaterally or bilaterally after local application of a 10 μL solution of bFGF. Larynges ware harvested for histological and immunohistochemical examination 2 months postoperation and for quantitative real-time polymerase chain reaction (qRT-PCR) analysis 1 week postoperation. Results Histological examination showed significantly increased hyaluronic acid and decreased deposition of dense collagen in the bFGF-treated group at 100 ng/10 μL compared with the sham-treated group. Immunohistochemical examination showed significantly decreased collagen type III deposition in the bFGF-treated group at 100 ng/10 μL compared with the sham-treated group. qRT-PCR revealed that hyaluronan synthase 2 (Has2), Has3, and hepatocyte growth factor were upregulated in bFGF-treated groups compared with sham-treated group. Conclusion The current results suggest that local application of bFGF at the time of injury has the potential to prevent vocal fold scarring. Preventive injection of bFGF could be applied at the time of phonomicrosurgery to avoid postoperative scar formation. Level of Evidence N/A. Laryngoscope, 2016 127:E67–E74, 2017

We previously developed novel liposomal nanobubbles (Bubble liposomes [BL]) that oscillate and collapse in an ultrasound field, generating heat and shock waves. We aimed to investigate the feasibility of cancer therapy using the combination of BL and ultrasound. In addition, we investigated the anti-tumor mechanism of this cancer therapy. Colon-26 cells were inoculated into the flank of BALB/c mice to induce tumors. After 8 days, BL or saline was intratumorally injected, followed by transdermal ultrasound exposure of tumor tissue (1 MHz, 0–4 W/cm[2], 2 min). The anti-tumor effects were evaluated by histology (necrosis) and tumor growth. In vivo cell depletion assays were performed to identify the immune cells responsible for anti-tumor effects. Tumor temperatures were significantly higher when treated with BL + ultrasound than ultrasound alone. Intratumoral BL caused extensive tissue necrosis at 3–4 W/cm[2] of ultrasound exposure. In addition, BL + ultrasound significantly suppressed tumor growth at 2–4 W/cm[2]. In vivo depletion of CD8[+] T cells (not NK or CD4[+] T cells) completely blocked the effect of BL + ultrasound on tumor growth. These data suggest that CD8[+] T cells play a critical role in tumor growth suppression. Finally, we concluded that BL + ultrasound, which can prime the anti-tumor cellular immune system, may be an effective hyperthermia strategy for cancer treatment.

Aim. Callus is a risk factor, leading to severe diabetic foot ulcer; thus, prevention of callus formation is important. However, normal stress (pressure) and shear stress associated with callus have not been clarified. Additionally, as new valuables, a shear stress-normal stress (pressure) ratio (SPR) was examined. The purpose was to clarify the external force associated with callus formation in patients with diabetic neuropathy.Methods. The external force of the 1st, 2nd, and 5th metatarsal head (MTH) as callus predilection regions was measured. The SPR was calculated by dividing shear stress by normal stress (pressure), concretely, peak values (SPR-p) and time integral values (SPR-i). The optimal cut-off point was determined.Results. Callus formation region of the 1st and 2nd MTH had high SPR-i rather than noncallus formation region. The cut-off value of the 1st MTH was 0.60 and the 2nd MTH was 0.50. For the 5th MTH, variables pertaining to the external forces could not be determined to be indicators of callus formation because of low accuracy.Conclusions. The callus formation cut-off values of the 1st and 2nd MTH were clarified. In the future, it will be necessary to confirm the effect of using appropriate footwear and gait training on lowering SPR-i.

There are a lot of recent researches on high power enzymatic glucose/O2 biofuel cells (BFCs). We have reported that the use of MgO-templated porous carbon (MgOC) at the electrodes of biofuel cells dramatically improves the output power [1-4]. Using MgOC can improve the effective electrode surface area, leading to increased current density and output density. Supporting enzymes on these types of porous materials is expected to lead to increased output density. In the past researches [1-4], the enzymes physically adsorbed inside the porous carbon gradually leach from the carbon surface after long-term operation or during the replacement of the fuel solution. In this study, to prevent enzyme leakage, the enzyme or mediator were immobilized on the MgOC surface via multipoint covalent bonding; the grafting of polymers bearing pendant glycidyl groups was achieved by the polymerization of glycidyl methacrylate (GMA), initiated by electron beam irradiation on the carbon surface. Poly(glycidyl methacrylate) (poly(GMA)) bearing pendant glycidyl groups, grafted on the surface of MgO-templated carbon, is useful for forming strong multipoint covalent bonds with amino functional groups on the surface of flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH), Azure A and aminoferrocene. For example, the immobilized FAD-GDH can generate glucose-oxidation catalytic current using 1,2-naphthoquinone (12NQ) as the redox mediator, which is also captured on the electrode surface. The catalytic current is more stable than that obtained using the FAD-GDH-MgOC electrode without poly(GMA) because the leaching of FAD-GDH and 12NQ is suppressed by the enzyme capping effect. The BFC performance using the anode measured in 1.0 M phosphate buffer (PBS) containing 100 mM glucose exhibited the open circuit voltage of 0.8 V, maximum power density of 1.4 mW/cm2, and maximum current density of 4.25 mA/cm2. In the present paper, we totally discuss three types glucose/oxygen biofuel cells based on the poly(glycidyl methacrylate)-grafted MgO-templated porous carbon. References [1] S. Tsujimura et al., Electrochemistry 83 (2015) 372. [2] I. Shitanda et al., Electrochemistry, 83 (2015) 329. [3] I. Shitanda et al., Journal of Electrochemical Society, 166 (2019) B1063. [4] I. Shitanda et al., Journal of Power Sources, 436 (2019) 226844. Acknowledgement This work was partially supported by JST-ASTEP Grant Number JPMJTS1513, JSPS Grant Number 17H02162 and Private University Research Branding Project (2017-2019) from Ministry of Education, Culture, Sports, Science and Technology, and Tokyo University of Science Grant for President's Research Promotion．

Although chemical-induced animal models of colorectal cancer (CRC) suggest a lot about the disease, more efforts are required to establish metastasis models. Azoxymethane (AOM) and dextran sodium sulfate (DSS)-treated (AOM/DSS) Crl:CD-1 mice were sacrificed after 10 or 20 weeks in our previous study, and most colon tumors exhibited intramucosal adenocarcinomas. Our observations were extended until 30 weeks to study a colitis-associated advanced CRC mouse model, and explore whether linker threonine-phosphorylated Smad2/3 (pSmad2/3L-Thr) immunostaining-positive cells were involved in the progressive course of colitis-associated CRC as cancer stem cells. AOM/DSS mice were sacrificed at 10, 20 and 30 weeks after AOM administration. Following the histopathological analysis, immunohistochemical staining was performed for the following markers: CD34, podoplanin, β-catenin, E-cadherin, Ki67, Bmi1 and pSmad2/3L-Thr. Compared with AOM/DSS mice at 10 and 20 weeks, submucosal tumor infiltration and tumor invasion into vessels were markedly increased at 30 weeks. In the parts of colon tumors from AOM/DSS mice, particularly in mice at 30 weeks, the positive signal of E-cadherin was clearly reduced in the cell membranes. The percentage of Ki67-positive tumor cells in mucosal areas of AOM/DSS mice was higher than that in the sites of submucosal infiltration. In mucosal areas of colon tumors, pSmad2/3L-Thr-positive cells were scattered among tumor cells. At sites of submucosal infiltration and vessel invasion of these tumors, pSmad2/3L-Thr-positive cells were also observed among tumor cells. In colon tumors from AOM/DSS mice at 30 weeks, the percentage of pSmad2/3L-Thr-positive cells among the nuclear β-catenin-positive tumor cells was higher than that among the cytoplasmic β-catenin-positive tumor cells. For both non-neoplastic and neoplastic epithelial cells, pSmad2/3L-Thr-positive cells exhibited immunohistochemical co-localization with Bmi1. The present study developed an advanced CRC mouse model that exhibited tumor infiltration into the submucosa and invasion into vessels. The present study re-confirmed the theory that pSmad2/3L-Thr-positive cells may be cancer stem cells.

The Cancer Genome Atlas project described a robust gene expression-based molecular classification of glioblastoma with the functional and biological significance of the subclasses yet to be determined. Here, we show that a comprehensive analysis of a panel of glioma initiating cell (GIC) lines can identify a group of stem cells with high OLIGg2 expression as in Proneural-like GBM subtype. In vitro differentiation studies showed that proneural GIC lines possess the potential to differentiate into astrocytic, neuronal, and oligodendrocytic lineages, whereas mesenchymal GICs exhibited limited potential for neural lineage differentiation following retinoic acid induction. A considerable decline of OLIG2 in proneural GIC lines was observed following retinoic acid treatment. We also showed that OLIG2 is a functional marker associated with cell proliferation in Olig2-high GIC lines. In addition, OLIG2 inhibition disrupted cell-cycle control mechanism by decreasing CDK2 and CDK4 and elevating apoptosis-related molecules. Mechanistic investigations revealed molecular interactions between CDK2/CDK4 and OLIG2. Inhibition of CDK2/CDK4 activity disrupted OLIG2-CDK2/CDK4 interactions and attenuated OLIG2 protein stability. Further investigation on these mechanisms may lead to novel targeted therapy on GBMs with high OLIG2 expression.

Background/Aims: We aimed to unravel how symptoms related to gastrointestinal (GI) dysfunction impair quality of life (QoL) in patients with diabetes mellitus (DM). Methods: We enrolled 134 outpatients with type 2 DM and 16 non-DM. In DM patients, mean age was 64.7 years, mean body mass index (BMI) 24.7 kg/m2, mean glycated hemoglobin (HbA1c) 7.1%, and mean DM duration 13.7 years. In non-DM, mean age was 66.6 years and mean BMI was 25.2 kg/m2. GI symptom-related QoL was determined using the Izumo scale, based on five factors, i.e., heartburn, gastralgia, postprandial fullness, constipation and diarrhea. The sum of scores obtained for the three questions in each domain was calculated, and subjects with a score of 5 or higher were considered to be symptomatic with impaired QoL. JMP Clinical version 5.0 was used for all statistical analyses. Results: In DM patients, constipation was much greater than non-DM, diarrhea was somewhat higher, other symptoms did not increase significantly. In DM, lower abdominal symptoms were found to be more frequent than those affecting the upper abdomen. Diabetic duration and medications showed associations with GI symptoms. We identified differences in peak prevalences of the five symptoms. Gastralgia and total GI symptoms peaked at a diabetes duration of 15-19 years. Heartburn and postprandial fullness tended to increase with disease duration. Constipation and diarrhea showed bimodal peaks, with the first early and the second late in the disease course. Finally, GI symptoms showed clustering that reflected the region of the GI tract affected, i.e., constipation and diarrhea had similar frequencies. Conclusions: Our study highlights the importance of questioning patients about QoL impairment due to abdominal symptoms, especially in the early and the late periods of diabetes. The heterogeneous nature of the underlying GI symptoms, including medication usage, should be taken into consideration when managing patients with T2DM. Disclosure M. Fujishiro: Research Support; Self; Johnson & Johnson Services, Inc. A. Kushiyama: Research Support; Self; Teijin Pharma Limited, Sanwa Kagaku Kenkyusho Co., Ltd., Kowa Pharmaceutical Co., Ltd, StaGen. Consultant; Self; RIZAP. H. Yamazaki: None. T. Yamamotoya: None. T. Kikuchi: None. H. Sakoda: None. R. Suzuki: Research Support; Self; Daiichi Sankyo Company, Limited, Kyowa Hakko Kirin Co., Ltd.. Other Relationship; Self; MSD K.K., Novo Nordisk Pharma Ltd., Daiichi Sankyo Company, Limited, Eli Lilly Japan K.K. T. Kadowaki: Consultant; Self; Novo Nordisk A/S, AstraZeneca, Merck Sharp & Dohme Corp.. Research Support; Self; Kissei Pharmaceutical Co., Ltd., Taisho Pharmaceutical Co., Ltd., Sanofi, Kyowa Hakko Kirin Co., Ltd., Novo Nordisk A/S, Astellas Pharma, Daiichi Sankyo Company, Limited, Takeda, Mitsubishi Tanabe Pharma Corporation, Ono Pharmaceutical Co., Ltd., Merck Sharp & Dohme Corp., Nippon Boehringer Ingelheim Co. Ltd.. Speaker's Bureau; Self; Astellas Pharma, AstraZeneca, Merck Sharp & Dohme Corp., Ono Pharmaceutical Co., Ltd., Takeda, Eli Lilly and Company, Nippon Boehringer Ingelheim Co. Ltd., Novo Nordisk A/S.

Objectives/Hypothesis Vocal fold scar remains a therapeutic challenge. Mesenchymal stromal cells (MSCs) are promising tools for regenerative medicine. Nevertheless, few in vivo studies have directly compared various sources of MSCs. The aim of this study was to investigate the therapeutic potential of adipose-derived stromal cells (ASCs) in comparison with bone marrow–derived stromal cells (BMSCs) for vocal fold regeneration. Study Design Prospective animal experiments with controls. Methods Two months after stripping of the lamina propria, 18 beagles were divided into four implantation groups: atelocollagen alone (collagen group), atelocollagen with BMSCs (BMSC-collagen), atelocollagen with ASCs (ASC-collagen), or a sham-treated group. One or 6 months after implantation, vibratory and histological examinations were performed. Results Mucosal vibration was significantly improved in both of the MSC-implanted groups compared with the sham-treated group, whereas only the ASC-collagen group showed a significantly smaller glottal gap than the collagen group. Moreover, in the ASC-collagen group, a significant reduction of collagen density was observed compared to the sham-treated group, and there was a trend for better restoration of hyaluronic acid (HA). Implanted MSCs were detected 1 month postimplantation; however, none survived 6 months postimplantation. Conclusions Although implantation of an atelocollagen sponge and ASCs into vocal fold scars induced vibratory recovery comparable to that of BMSCs, ASCs might have more potential in terms of restoration of HA and suppression of excessive collagen deposition. Level of Evidence NA Laryngoscope, 2015

We have successfully developed two types of cellulose triacetate (TAC) polarizer protection films that meet the LCDs trends toward thinner and larger panels. The novel 25µm TAC film, which has high-hardness and high-durability, solves the problems caused by the reduction in the thickness of polarizers and is suitable for small-sized LCDs. The novel Z-TAC film, which has high-dimensional-stability, reduces warpage of large sized IPS-TVs.

Nano-/microbubbles are expected not only to function as ultrasound contrast agents but also as ultrasound-triggered enhancers in gene and drug delivery. Notably, nanobubbles have the ability to pass through tumor vasculature and achieve passive tumor targeting. Thus, nanobubbles would be an attractive tool for use as ultrasound-mediated cancer theranostics. However, the amounts of gas carried by nanobubbles are generally lower than those carried by microbubbles because nanobubbles have inherently smaller volumes. In order to reduce the injection volume and to increase echogenicity, it is important to develop nanobubbles with higher gas content. In this study, we prepared 5 kinds of fluoro-lipids and used these reagents as surfactants to generate “Bubble liposomes”, that is, liposomes that encapsulate nanobubbles such that the lipids serve as stabilizers between the fluorous gas and water phases. Bubble liposome containing 1-stearoyl-2-(18,18-difluoro)stearoyl-sn-glycero-3-phosphocholine carried 2-fold higher amounts of C3F8 compared to unmodified Bubble liposome. The modified Bubble liposome also exhibited increased echogenicity by ultrasonography. These results demonstrated that the inclusion of fluoro-lipid is a promising tool for generating nanobubbles with increased efficiency of fluorous gas carrier.

In this study, a novel phospholipid-based microbubble formulation containing doxorubicin and perfluoropropane gas (DLMB) was developed. The DLMBs were prepared by mechanical agitation of a phospholipid dispersion in the presence of perfluoropropane (PFP) gas. An anionic phospholipid, distearoyl phosphatidylglycerol (DSPG) was selected to load doxorubicin in the microbubbles by means of electrostatic interaction. The particle size, zeta potential, echogenicity and stability of the DLMBs were measured. Drug loading was ?92%. The potential of the DLMBs for use as a theranostic modality was evaluated in tumor bearing mice. Gas chromatography analysis of PFP showed significant enhancement of PFP retention when doxorubicin was used at concentrations of 10-82% equivalent to DSPG. The inhibitory effects on the proliferation of B16BL6 melanoma murine cells in vitro were enhanced using a combination of ultrasound (US) irradiation and DLMBs. Moreover, in vivo DLMBs in combination with (US) irradiation significantly inhibited the growth of B16BL6 melanoma tumor in mice. Additionally, US echo imaging showed high contrast enhancement of the DLMBs in the tumor vasculature. These results suggest that DLMBs could serve as US triggered carriers of doxorubicin as well as tumor imaging agents in cancer therapy., Acta Biomaterialia, 19, pp.112-118; 2015