Ghost Hunter: a Handheld Augmented Reality Game System With Dynamic Environment

Abstract. The progress of handheld devices has encouraged many researchers to make efforts to use handheld devices as an augmented reality platform. However, one of the important problems is a lack of immersion and reality due to small display. To overcome this problem, we introduce dynamic environment which consists of some movable structures and their controller that enable changes of virtual world to affect the real world. It has an effect on expanding the user's view limited by the small display of the handheld device. We have also developed the game content, 'Ghost Hunter', which is suitable for this platform. keywords: handheld augmented reality, dynamic environment, entertainment syste

Ghost Hunter: a Handheld Augmented Reality Game System With Dynamic Environment

Abstract. The progress of handheld devices has encouraged many researchers to make efforts to use handheld devices as an augmented reality platform. However, one of the important problems is a lack of immersion and reality due to small display. To overcome this problem, we introduce dynamic environment which consists of some movable structures and their controller that enable changes of virtual world to affect the real world. It has an effect on expanding the user's view limited by the small display of the handheld device. We have also developed the game content, 'Ghost Hunter', which is suitable for this platform. keywords: handheld augmented reality, dynamic environment, entertainment syste

Microneedle-based minimally-invasive measurement of puncture resistance and fracture toughness of sclera

The sclera provides the structural support of the eye and protects the intraocular contents. Since it covers a large portion of the eye surface and has relatively high permeability for most drugs, the sclera has been used as a major pathway for drug administration. Recently, microneedle (MN) technology has shown the possibility of highly local and minimally-invasive drug delivery to the eye by MN insertion through the sclera or the suprachoroidal space. Although ocular MN needs to be inserted through the sclera, there has been no systematic study to understand the mechanical properties of the sclera, which are important to design ocular MNs. In this study, we investigated a MN-based method to measure the puncture resistance and fracture toughness of the sclera. To reflect the conditions of MN insertion into the sclera, force displacement curves obtained from MN-insertion tests were used to estimate the puncture resistance and fracture toughness of sclera tissue. To understand the effect of the insertion conditions, dependency of the mechanical properties on insertion speeds, pre-strain of the sclera, and MN sizes were analyzed and discussed. Statement of Significance Measurement of mechanical property of soft biological tissue is challenging due to variations between tissue samples or lack of well-defined measurement techniques. Although non-invasive measurement techniques such as nano/micro indentation were employed to locally measure the elastic modulus of soft biological materials, mechanical properties such as puncture resistance or fracture toughness, which requires "invasive" measurement and is important for the application of "microneedles or hypodermic needles", has not been well studied. In this work, we report minimally-invasive measurement of puncture resistance and fracture toughness of sclera using a double MN insertion method. Parametric studies showed that use of MN proved to be advantageous because of minimally-invasive insertion into tissue as well as higher sensitivity to sub-tissue architecture during the measurement. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.N

A Biodegradable Microneedle Cuff for Comparison of Drug Effects through Perivascular Delivery to Balloon-Injured Arteries

Restenosis at a vascular anastomosis site is a major cause of graft failure and is difficult to prevent by conventional treatment. Perivascular drug delivery has advantages as drugs can be diffused to tunica media and subintima while minimizing the direct effect on endothelium. This in vivo study investigated the comparative effectiveness of paclitaxel, sirolimus, and sunitinib using a perivascular biodegradable microneedle cuff. A total of 31 New Zealand white rabbits were used. Rhodamine was used to visualize drug distribution (n = 3). Sirolimus- (n = 7), sunitinib- (n = 7), and paclitaxel-loaded (n = 7) microneedle cuffs were placed at balloon-injured abdominal aortae and compared to drug-free cuffs (n = 7). Basic histological structures were not affected by microneedle devices, and vascular wall thickness of the device-only group was similar to that of normal artery. Quantitative analysis revealed significantly decreased neointima formation in all drug-treated groups (p < 0.001). However, the tunica media layer of the paclitaxel-treated group was significantly thinner than that of other groups and also showed the highest apoptotic ratio (p < 0.001). Proliferating cell nuclear antigen (PCNA)-positive cells were significantly reduced in all drug-treated groups. Sirolimus or sunitinib appeared to be more appropriate for microneedle devices capable of slow drug release because vascular wall thickness was minimally affected

Brazilin Limits Inflammatory Responses through Induction of Prosurvival Autophagy in Rheumatoid Fibroblast-Like Synoviocytes.

Brazilin is an active compound of Caesalpinia sappan L. (Leguminosae), which possesses pro-apoptotic and anti-inflammation potentials depending on the specific cell type. However, it is largely unknown whether autophagy is implicated in the mechanism underlying its chemotherapeutic and anti-inflammatory effects in rheumatoid arthritis (RA). Here, we show that treatment of RA fibroblast-like synoviocytes (FLS) with brazilin results in enhanced level of autophagic flux, evidenced by accumulation of autophagosome and increased level of lipidated LC3 (LC3-II), which is mainly mediated by enhanced production of reactive oxygen species (ROS). Interestingly, long-term exposure of brazilin was able to restore cell survival against the cytotoxity, exclusively in RA FLS, but not in normal fibroblast. Importantly, such a restoration from brazilin-induced cytotoxity in RA FLS was completely abrogated after co-treatment with autophagy inhibitors including NH4Cl or chloroquine. Furthermore, we found that the pretreatment of RA FLS with brazilin reduced LPS- or TNF-induced NF-κB activation and the secretion of inflammatory cytokines in parallel with the enhanced autophagic flux. Such anti-NF-κB potentials of brazilin were drastically masked in RA FLS when autophagy was suppressed. These results suggest that brazilin is capable of activating autophagy exclusively in RA FLS, and such inducible autophagy promotes cell survival and limits inflammatory response

Self-Plugging Microneedle (SPM) for Intravitreal Drug Delivery

© 2022 Wiley-VCH GmbHIntravitreal injection (IVI) is a common technology which is used to treat ophthalmic diseases inside eyeballs by delivering various drugs into the vitreous cavity using hypodermic needles. However, in some cases, there are possible side effects such as ocular tissue damage due to repeated injection or eyeball infection through the hole created during the needle retraction process. The best scenario of IVI is a one-time injection of drugs without needle retraction, keeping the system of the eyeball closed. Microneedles (MNs) have been applied to ocular tissues over 10 years, and no serious side effects on ocular tissue due to MN injection have been reported. Therefore, a self-plugging MN (SPM) is developed to perform intraocular drug delivery and to seal the scleral puncture simultaneously. The SPMs are fabricated by a thermal drawing process and then coated with a polymeric carrier of drugs and a hydrogel-based scleral plugging component. Each coated functional layer is characterized and demonstrated by in vitro and ex vivo experiments. Finally, in vivo tests using a porcine model confirms prompt sealing of SPM and sustained intraocular drug delivery.N

Autophagy induced by brazilin limits NF-κB activation and inflammatory response in RA FLS.

<p>(A, B) After pretreatment with brazilin for 48 h, RA FLS were treated with 1 μg/ml of LPS (A) or 15 ng/ml of TNF (B) as for various times as indicated. Whole cell lysates were separated by SDS-PAGE and the immunoblotting was performed using indicated antibodies. (C) RA FLS were pretreated with brazilin for 48 h in the absence or presence of NAC (10 mM), and then treated with TNF as indicated times. Whole cell lysates were separated by SDS-PAGE and the immunoblotting was performed, as in (A). (D, E). RA FLS were pretreated with brazilin for 48 h in the absence or presence of NAC (10 mM). Supernatants were harvested at the indicated times after the commencement of LPS stimulation, and the levels of IL-6 and IL-8 were measured by ELISA. (F) RA FLS transfected with either a lentivirus expressing control shRNA (shCont) or shRNA specific for Atg5 (shAtg5) were cultured for 48 h. Consequently, cells were infected with recombinant adeno-virus expressing NF-κB-Luc at 200 PFU/cells for 2h, and then replaced with DMEM containing 10% FBS. After 24 h of infection. cells were treated for 6 h with LPS (1 μg/ml), and luciferase assay was performed as described in Materials and Methods. Knock down efficiency of Atg5 in RA FLS were assessed by immunoblotting (inset). (G) After RA FLS were transduced with shCont or shAtg5, the cells were further treated with LPS (1 μg/ml) for 24 h. The levels of IL-6 and IL-8 in the supernatant were measured by ELISA. Each columns shows mean ± S.E. from the three independent experiments. *<i>P</i><0.05, compared with shCont-transfected cells.</p

ROS production plays an important role in autophagy activation and early event of cell death in response to brazilin.

<p>(A) RA FLS were treated with 25 μg/ml of brazilin for various times, as indicated. CM-H₂-DCF-DA (1 μM) was added 30 min before end of treatment. ROS were measured with a flow cytometer (left panel) as described in materials and methods. Data were processed and quantified with the FlowJo software (right panel). (B) RA FLS were treated with brazilin (25 μg/ml) in the absence or presence of NAC (10 mM), Mito-TEMPO (100 μM) and DPI (10 μM) for 9 h. Generation of ROS were measured, as in (A) (C) RA FLS were pretreated with NAC (10 mM), Mito-TEMPO (100 μM) and DPI (10 μM), and then followed by brazilin (25 μg/ml) for 24 h. Whole cell lysates were separated by SDS-PAGE and the immunoblotting was performed with anti-LC3 and actin antibodies. (D, E) RA FLS were treated with brazilin (25 μg/ml) in the absence or presence of NAC (10 mM) or Mito-TEMPO (100 μM) for indicated times. Cell were visualized with a normal inverted microscope (D) and the cell viability was determined (E), as describe in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136122#pone.0136122.g001" target="_blank">Fig 1A</a>. (F) HDF were treated with brazilin (25 μg/ml) for various times, as indicated and the levels of ROS were determined, as in (A).</p

Failure to restoration from brazilin-induced cytotoxicity by blockade of autophagy in RA FLS.

<p>(A) RA FLS were treated with brazilin (25 μg/ml), Fas ligand (FasL, 1 μg/ml) plus cycloheximide (CHX, 10 μg/ml), MNNG (0.5 mM) and <i>t</i>-BHP (100 μM) in the absence or presence of pancaspase inhibitor (z-VAD-FMK, 20 μM) or necrosis inhibitor (IM54, 10 μM). The cells were collected and cell viability was then determined by trypan blue exclusion assay. The results are presented as the mean ± S.E. from the three independent experiments. (B) HDF were treated with brazilin (25 μg/ml) in the absence or presence of z-VAD-FMK (20 μM) or IM54 (10 μM) as indicated times, and cell viability was determined, as in (A). *<i>P</i><0.05, compared with brazilin only treated group. (C, D) RA FLS were pretreated with autophagy inhibitor, NH₄Cl (5 mM) and chloroquine (CQ, 5 mM) for 30 min, and then followed by 25 μg/ml of brazilin as indicated times. (E) HDF were pretreated with rapamycin (100 nM) for 30 min, and then followed by 25 μg/ml of brazilin as indicated times. The cell viability was determined, as describe in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0136122#pone.0136122.g001" target="_blank">Fig 1A</a>.</p

Restoration of cellular survival against from brazilin-induced cytotoxicity in RA FLS, but not in normal fibroblasts.

<p>FLS from RA patients (A) and several normal fibroblasts (dermal fibroblast from human foreskin tissues, NIH3T3, COS-7 and MEF) (B) were treated with indicated concentrations of brazilin. The cells were collected and cell viability was then determined by trypan blue exclusion assay. The results are presented as the mean ± S.E. from the three independent experiments.</p