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Bone has multiple functions in animals, such as supporting the body for mobility. The zebrafish skeleton is composed of craniofacial and axial skeletons. It shares a physiological curvature and consists of a similar number of vertebrae as humans. Bone degeneration and malformations have been widely studied in zebrafish as human disease models. High-resolution imaging and different bone properties such as density and volume can be obtained using micro-computed tomography (micro-CT). This study aimed to understand the possible changes in the structure and bone mineral density (BMD) of the vertebrae and craniofacial skeleton with age (4, 12 and 24 months post fertilisation [mpf]) in zebrafish. Our data showed that the BMD in the vertebrae and specific craniofacial skeleton (mandibular arch, ceratohyal and ethmoid plate) of 12 and 24 mpf fish were higher than that of the 4 mpf fish. In addition, we found the age-dependent increase in BMD was not ubiquitously observed in facial bones, and such differences were not correlated with bone type. In summary, such additional information on the craniofacial skeleton could help in understanding bone development throughout the lifespan of zebrafish.

Small cell lung cancer (SCLC) continues to cause poor clinical outcomes due to limited advances in sustained treatments for rapid cancer cell proliferation and progression. The transcriptional factor Forkhead Box M1 (FOXM1) regulates cell proliferation, tumor initiation, and progression in multiple cancer types. However, its biological function and clinical significance in SCLC remain unestablished. Analysis of the Cancer Cell Line Encyclopedia and SCLC datasets in the present study disclosed significant upregulation of FOXM1 mRNA in SCLC cell lines and tissues. Gene set enrichment analysis (GSEA) revealed that FOXM1 is positively correlated with pathways regulating cell proliferation and DNA damage repair, as evident from sensitization of FOXM1-depleted SCLC cells to chemotherapy. Furthermore, Foxm1 knockout inhibited SCLC formation in the Rb1fl/flTrp53fl/flMycLSL/LSL (RPM) mouse model associated with increased levels of neuroendocrine markers, Ascl1 and Cgrp, and decrease in Yap1. Consistently, FOXM1 depletion in NCI-H1688 SCLC cells reduced migration and enhanced apoptosis and sensitivity to cisplatin and etoposide. SCLC with high FOXM1 expression (N = 30, 57.7%) was significantly correlated with advanced clinical stage, extrathoracic metastases, and decrease in overall survival (OS), compared with the low-FOXM1 group (7.90 vs. 12.46 months). Moreover, the high-FOXM1 group showed shorter progression-free survival after standard chemotherapy, compared with the low-FOXM1 group (3.90 vs. 8.69 months). Our collective findings support the utility of FOXM1 as a prognostic biomarker and potential molecular target for SCLC.

We investigated the effects of nickel oxide nanoparticles (NiONPs) on the pulmonary inflammopathology. NiONPs were intratracheally installed into mice, and lung injury and inflammation were evaluated between 1 and 28 days. NiONPs caused significant increases in LDH, total protein, and IL-6 and a decrease in IL-10 in the BALF and increases in 8-OHdG and caspase-3 in lung tissues at 24 h. Airway inflammation was present in a dose-dependent manner from the upper to lower airways at 24 h of exposure as analyzed by SPECT. Lung parenchyma inflammation and small airway inflammation were observed by CT after NiONP exposure. 8-OHdG in lung tissues had increased with formation of fibrosis at 28 days. Focal adhesion was the most important pathways identified at 24 h as determined by protemics, whereas glutathione metabolism was the most important identified at 28 days. Our results demonstrated the pulmonary inflammopathology caused by NiONPs based on image-to-biochemical approaches.

Inhaled zinc oxide nanoparticles (ZnONPs) have high deposition rates in the alveolar region of the lungs; however, the adverse health effects of ZnONPs on the respiratory system are unclear. Herein, pathobiological responses of the respiratory system of mice that received intratracheal administration of ZnONPs were investigated by a combination of molecular and imaging (SPECT and CT) approaches. Also, normal BEAS-2B and adenocarcinoma A549 cells were used to confirm the results in mice. First, female BALB/c mice were administrated a series of doses of 20-nm ZnONPs and were compared to the phosphate-buffered saline control for 24-h and 28-day follow-up observations. Field emission-scanning electron microscopy and an energy-dispersive X-ray microanalysis were first used to characterize ZnONPs. After 24h, instilled ZnONPs had caused significant increases in lactic dehydrogenase (LDH) in bronchoalveolar lavage fluid (BALF) and 8-hydroxy-2'-deoxyguanosine (8-OHdG), caspase-3, and the p63 tumor marker in lung tissues (p

Nanoparticle-based imaging contrast agents have drawn tremendous attention especially in multi-modality imaging. In this study, we developed mesoporous silica nanoparticles (MSNs) for use as dual-modality contrast agents for computed tomography (CT) and near-infrared (NIR) optical imaging (OI). A microwave synthesis for preparing naked platinum nanoparticles (nPtNPs) on MSNs (MSNs-Pt) was developed and characterized with physicochemical analysis and imaging systems. The high density of nPtNPs on the surface of the MSNs could greatly enhance the CT contrast. Inductively coupled plasma mass spectrometry (ICP-MS) revealed the MSNs-Pt compositions to be ~14% Pt by weight and TEM revealed an average particle diameter of ~50 nm and covered with ~3 nm diameter nPtNPs. To enhance the OI contrast, the NIR fluorescent dye Dy800 was conjugated to the MSNs-Pt nanochannels. The fluorescence spectra of MSNs-Pt-Dy800 were very similar to unconjugated Dy800. The CT imaging demonstrated that even modest degrees of Pt labeling could result in substantial X-ray attenuation. In vivo imaging of breast tumor-bearing mice treated with PEGylated MSNs-Pt-Dy800 (PEG-MSNs-Pt-Dy800) showed significantly improved contrasts in both fluorescence and CT imaging and the signal intensity within the tumor retained for 24 h post-injection.

Finding an effective therapeutic regimen is an urgent demand for various neurodegenerative disorders including Huntington's disease (HD). For the difficulties in observing the dynamic aggregation and oligomerization process of mutant Huntingtin (mHtt) in vivo, the evaluation of potential drugs at the molecular protein level is usually restricted. By combing lifetime‐based fluorescence microscopies and biophysical tools, it is showcased that a designed amphiphilic peptide, which targets the mHtt at an early stage, can perturb the oligomer assembly process nanoscopically, suppress the amyloid property of mHtt, conformationally transform the oligomers and/or aggregates of mHtt, and ameliorate mHtt‐induced neurological damage and aggregation in cell and HD mouse models. It is also found that this amphiphilic peptide is able to transport to the brain and rescue the memory deficit through intranasal administration, indicating its targeting specificity in vivo. In summary, a biophotonic platform is provided to investigate the oligomerization/aggregation process in detail that offers insight into the design and effect of a targeted therapeutic agent for Huntington's disease., An amphiphilic peptide (8R10Q) that assembles into nanovesicles and targets mutant Huntingtin (mHtt) protein is designed. By applying various advanced nanoscopic tools, it is shown that 8R10Q perturbs the mHtt oligomer assembly process, conformationally transforms mHtt inclusions into puncta, and reduces mHtt toxicity. Moreover, 8R10Q can be transported into the brain to ameliorate mHtt‐induced neurological damage and rescue memory deficit through intranasal administration.

// Arivajiagane Arundhathi 1, * , Wen-Han Chuang 1, * , Jen-Kun Chen 2, * , Shin-E Wang 3, * , Yi-Ming Shyr 3 , Jiun-Yu Chen 2 , Wei-Neng Liao 2 , Hsin-Wei Chen 1 , Yi-Min Teng 1 , Chiao-Chih Pai 1 , Chih-Hong Wang 1 1 Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan 2 Institute of Biomedical Engineering & Nanomedicine, National Health Research Institutes, Miaoli, Taiwan 3 General Surgery, Taipei Veterans General Hospital, and National Yang Ming University, Taipei, Taiwan * These authors have contributed equally to this work Correspondence to: Chih-Hong Wang, email: chihhong@nctu.edu.tw Keywords: prorenin receptor, pancreatic cancer, PanIN lesions, Kras, SPECT/CT Received: August 21, 2015 Accepted: July 05, 2016 Published: July 13, 2016 ABSTRACT Recent studies have implicated the prorenin receptor (PRR) is associated with pancreatic tumorigenesis. We therefore investigated the role of PRR in pancreatic tumorigenesis and assessed whether PRR can serve as a target for imaging diagnosis at early stages of PDAC. Here we show that aberrant expression of PRR in premalignant PanIN lesions, and human PDAC samples, and PDAC cell lines, particularly in Panc-1 cells. Interestingly, PRR expression was positively associated with PDAC progression. Moreover, overexpression of human PRR resulted in increased cell proliferation and decreased apoptosis, while knockdown of human PRR caused decreased cell proliferation and enhanced apoptosis in pancreatic cancer cells. We also observed that overexpression of human PRR enhanced MAPK and PI3K/Akt signaling pathways in PDAC cells, while knockdown of human PRR suppressed both of pathways. The confocal imaging analysis showed that human PRR was highly expressed in Panc-1, ASPC, and Miapaca cells, whereas BXPC-3, and HPAC cells had a significantly lower fluorescent signals. Consistently, the single-photon emission computed tomography (SPET/CT) showed that the uptake of anti-PRR labelled with 125 I was higher in Panc-1 and ASPC tumors-bearing mice after 96 hours injection. Importantly, tumors in pancreas of Pdx1-cre; LSL-Kras G12D mice had a significant increased PRR expression and accumulation of radioactivity at 96 h after injection. These data suggest that 125 I-anti-PRR can detect the orthotopic tumors in Pdx1-cre; LSL-Kras G12D mice. Therefore, anti-PRR labelled with 125 I is a promising radiotracer for imaging diagnosis at early stages of pancreatic cancer.

This study aims to establish a (198)Au-radiotracer technique for in vivo tracing, rapid quantification, and ex vivo visualization of PEGylated gold nanoparticles (GNPs) in animals, organs and tissue dissections. The advantages of GNPs lie in its superior optical property, biocompatibility and versatile conjugation chemistry, which are promising to develop diagnostic probes and drug delivery systems. (198)Au is used as a radiotracer because it simultaneously emits beta and gamma radiations with proper energy and half-life; therefore, (198)Au can be used for bioanalytical purposes. The (198)Au-tagged radioactive gold nanoparticles ((198)Au-GNPs) were prepared simply by irradiating the GNPs in a nuclear reactor through the (197)Au(n,γ)(198)Au reaction and subsequently the (198)Au-GNPs were subjected to surface modification with polyethylene glycol to form PEGylated (198)Au-GNPs. The (198)Au-GNPs retained physicochemical properties that were the same as those of GNP before neutron irradiation. Pharmacokinetic and biodisposition studies were performed by intravenously injecting three types of (198)Au-GNPs with or without PEGylation into mice; the γ radiation in blood specimens and dissected organs was then measured. The (198)Au-radiotracer technique enables rapid quantification freed from tedious sample preparation and shows more than 95% recovery of injected GNPs. Clinical gamma scintigraphy was proved feasible to explore spatial- and temporal-resolved biodisposition of (198)Au-GNPs in living animals. Moreover, autoradiography, which recorded beta particles from (198)Au, enabled visualizing the heterogeneous biodisposition of (198)Au-GNPs in different microenvironments and tissues. In this study, the (198)Au-radiotracer technique facilitated creating a trimodality analytical platform for tracing, quantifying and imaging GNPs in animals.

Periodontal diseases are chronic inflammatory diseases characterized by the destruction of the tooth-supporting structures. They are the most prevalent form of bone pathology in humans and act as a modifying factor of the systemic health of patients. Accumulating evidence has provided insight into mechanisms of periodontal inflammation revealing that oral pathogens induce inflammatory cascades, including a variety of cytokines produced by different cell types, which promotes host-mediated tissue destruction. Cytokine networks established in diseased periodontal tissues are extremely complex, and substances regulating immuno-inflammatory reactions and signaling pathways, in addition to traditional periodontal treatment, could potentially be targeted as an approach for prevention and treatment of periodontal diseases. Diacerein, a purified anthraquinone derivative, was derived originally from plants with profound anti-inflammatory and analgesic activities. Its wide range of biological activities have been applied and discussed for several decades; however, studies of diacerein have mainly concentrated on effects on joint-derived tissues/cells, which suggest a beneficial role in osteoarthritis treatment. Diacerein reduces association of the IL-1 receptor to form heterodimer complexes, repressing IL-1 and its related downstream events and impairing active IL-1 release due to the inhibition of the IL-1-converting enzyme (ICE). To date, there are no reports describing the therapeutic effect of diacerein for treatment of periodontitis. Given the involvement of inflammation and occurrence of tissue destruction in periodontal disease, we propose that diacerein might be a promising biological drug for periodontal disease due to its therapeutic advantages. In addition, we hypothesize that the underlying mechanisms might involve the capacity of diacerein to selectively inhibit signal transduction to affect the cytokine profiles and, consequently, produce the outcome of ameliorating disease breakdown.

In this study, novel aminothiazole-paeonol derivatives were synthesized and characterized using ¹H-NMR, (13)C-NMR, IR, mass spectroscopy, and high performance liquid chromatography. All the new synthesized compounds were evaluated according to their anticancer effect on seven cancer cell lines. The experimental results indicated that these compounds possess high anticancer potential regarding human gastric adenocarcinoma (AGS cells) and human colorectal adenocarcinoma (HT-29 cells). Among these compounds, N-[4-(2-hydroxy-4-methoxyphenyl)thiazol-2-yl]-4-methoxybenzenesulfonamide (13c) had the most potent inhibitory activity, with IC50 values of 4.0 µM to AGS, 4.4 µM to HT-29 cells and 5.8 µM to HeLa cells. The 4-fluoro-N-[4-(2-hydroxy-4-methoxyphenyl)thiazol-2-yl]benzenesulfonamide (13d) was the second potent compound, showing IC50 values of 7.2, 11.2 and 13.8 µM to AGS , HT-29 and HeLa cells, respectively. These compounds are superior to 5-fluorouracil (5-FU) for relatively higher potency against AGS and HT-29 human cancer cell lines along with lower cytotoxicity to fibroblasts. Novel aminothiazole-paeonol derivatives in this work might be a series of promising lead compounds to develop anticancer agents for treating gastrointestinal adenocarcinoma.

We designed a novel cis-platin (CP) delivery system by modification of mesoporous silica nanoparticle (MSN) surfaces with a carboxylate group through a hydrazone bond. The further immobilization of CP can be achieved through the coordination of the carboxylate-modified MSN surfaces with the hydroxo-substituted CP. This new formulation can efficiently increase efficiency of both the cellular uptake and the drug release under endosomal or lysosomal pHs; therefore, the anti-proliferative effect of this new formulation on the colon cancer cell line (HT-29) was twenty times more than the free CP molecules. In addition, the encapsulation of CP complexes in the confined spaces of MSNs can decrease non-specific release from enzymatic hydrolysis because most hydrolytic enzymes have diameters considerably greater than the pore size of MSNs. The DNA fragmentation and caspase-3 activity assay showed that the apoptosis was induced by DNA damages and then an increase in caspase-3 activity. Thus, the TA-MSN-carboxylate-CP samples were induced cell apoptosis through the caspase-3 dependent pathway. Moreover, the hemolysis assay also indicated that the exposure of the carboxylate-modified MSNs in red blood cells (RBCs) did not observe the release of red hemoglobin from the cell lysis, and the further exposure of the TA-MSN-carboxylate-CP complexes to RBCs also did not observe notably the lysis of RBCs under the effectively therapeutic dosage. Therefore, our design of MSN with controllable release of CP has highly therapeutic effects and is highly biocompatible; however, a low cytotoxicity and site effect were observed.

We incorporated a near-infrared (NIR) fluorescent dye, indocyanine green (ICG), in amine-modified layered double hydroxide nanoparticles (LDHs) by electrostatic attractions to render LDHs–ICG as an efficient NIR contrast agent for in vivo optical imaging. The further coating of chitosan on the external surfaces of LDHs–ICG was achieved through the cross-linking of amine-modified LDHs and different amounts of chitosan by using glutaraldehyde (GA) as a cross-linked agent. The hybridization of this organic–inorganic nanocomposite produced an efficient NIR optical contrast agent because the adsorbed ICG molecules were stabilized in the layered structures of LDHs to prevent them from leaching and/or metabolizing in physiological conditions. The cell viability and hemolysis assay also showed low cytotoxicity and low release of hemoglobin from the cell lysis of red blood cells (RBCs). The in vivo biodistribution results demonstrated that the coating of LDHs with different amounts of chitosan can develop organ-specific drug delivery systems, which can efficiently regulate the nanoparticle accumulation in various organs with un-coated LDHs–ICG targeting the liver and spleen, mono-chitosan-coated LDHs–ICG targeting the lungs, double chitosan-coated LDHs–ICG targeting the lungs and liver, and trimethylammonium (TA) modified double chitosan-coated LDHs–ICG samples targeting the liver. In addition to the high potential for the employment of chitosan-coated LDHs–ICG samples for developing contrast agents for in vivo imaging, the LDH nanoparticles can deliver therapeutic drugs to desired target organs by controlling the coating amounts of chitosan. Therefore, this approach can improve efficiency for traditional cancer diagnosis and cancer chemotherapy.

We report a hydrothermally stable and highly reactive cytochrome c (cyt c) immobilized in the nanochannels of mesoporous silica (SBA-15) through a metal affinity interaction. Due to the strong affinity of mercury–sulfuric bonds, we modified the SBA-15 surfaces with 4-aminophenylmercuric acetate (APMA) groups. As a result, an enzyme molecule (yeast cyt c) with a cysteine residue (cys-102) demonstrated strong adsorption, which provided high cyt c loading amounts, highly catalytic activity, and high stability against hydrothermal processes and organic solvents. To compare the immobilization of cysteine-containing cyt c through metal affinity interactions and a traditional covalent bond (a disulfide bond), we modified the SBA-15 surfaces with 3-mercaptopropyl-trimethoxysilane (MPTS) for further production of a disulfide bond with the cysteine residue of cyt c. The cysteine residue of the cyt c can covalently link to thiol-modified SBA-15 through the formation of a disulfide bond. In addition, a non-specific coordination from the thiol groups of SBA-15 to the heme Fe(III) of cyt c may destroy the catalytic center and cause the leaching of Fe(III) ions. Our previous studies of the molecular model have shown that the immobilization of cyt c through the cysteine residue can provide a correct orientation of the catalytic center, where the active site can easily approach the substrate molecules. Therefore, we have developed rapid and highly efficient approaches to immobilize a cysteine-containing enzyme through APMA ligands, which can both protect the protein folding and control the orientation to optimize the stability and catalytic activity.

We designed a novel oral colon-specific drug delivery system (OCDDS) using a modification of mesoporous silica nanoparticle (MSN) surfaces with pH-sensitive trimethylammonium (TA) groups through a pH-sensitive hydrazone bond. The pH-sensitive TA groups can efficiently increase the loading amounts of anionic drugs by a strong electrostatic attraction. After oral administration, the acidic pH of gastric juice can fully hydrolyze the TA–hydrazone bonds and further eliminate the positive charges of TA groups from MSN surfaces. When the hydrolyzed complexes were further delivered to the colon's pH of 7–8, a rapid and complete release of adsorbed drugs was observed. From the studies of spectroscopic characterizations, we demonstrated that the combination of pH-sensitive hydrazone–TA groups and nano-sized particles of the MSN carriers took the advantages of increasing the accessible surface areas of drug molecules and varying the charges of MSN surfaces, which can increase dissolution and release rate of hydrophobic drug molecules. In addition, a cell viability assay also indicated that no cytotoxicity of MSN–hydrazone–TA complexes was observed even with treatment in an extremely high nanoparticle concentration. Consequently, our new formulation is highly biocompatible for the OCDDS, and we can completely solve the low stability, low solubility, and low drug bioavailability in the free form of drug molecules for the design of OCDDS.

A double-surfactant-layer-assisted polymerization method was designed to prepare well-controlled core/shell metal oxide/polyaniline (c/s-MO/PANi) nanocomposites. Monodispersed and uniform core/shell nanocomposites including c/s-CuO/PANi, −Fe2O3/PANi, −In2O3/PANi and −Fe2O3/SiO2/PANi were successfully prepared using this polymerization method. By the removal of the cores for the resulting c/s-MO/PANi nanocomposites, hollow PANi capsules with different shapes and sizes were obtained. Our method provides a facile and effective way to design novel core/shell nanostructures with diverse functionality and high colloidal stability.