Targeted Overexpression of Osteoactivin in Cells of Osteoclastic Lineage Promotes Osteoclastic Resorption and Bone Loss in Mice

This study sought to test whether targeted overexpression of osteoactivin (OA) in cells of osteoclastic lineage, using the tartrate-resistant acid phosphase (TRAP) exon 1B/C promoter to drive OA expression, would increase bone resorption and bone loss in vivo. OA transgenic osteoclasts showed ∼2-fold increases in OA mRNA and proteins compared wild-type (WT) osteoclasts. However, the OA expression in transgenic osteoblasts was not different. At 4, 8, and 15.3 week-old, transgenic mice showed significant bone loss determined by pQCT and confirmed by μ-CT. In vitro, transgenic osteoclasts were twice as large, had twice as much TRAP activity, resorbed twice as much bone matrix, and expressed twice as much osteoclastic genes (MMP9, calciton receptor, and ADAM12), as WT osteoclasts. The siRNA-mediated suppression of OA expression in RAW264.7-derived osteoclasts reduced cell size and osteoclastic gene expression. Bone histomorphometry revealed that transgenic mice had more osteoclasts and osteoclast surface. Plasma c-telopeptide (a resorption biomarker) measurements confirmed an increase in bone resorption in transgenic mice in vivo. In contrast, histomorphometric bone formation parameters and plasma levels of bone formation biomarkers (osteocalcin and pro-collagen type I N-terminal peptide) were not different between transgenic mice and WT littermates, indicating the lack of bone formation effects. In conclusion, this study provides compelling in vivo evidence that osteoclast-derived OA is a novel stimulator of osteoclast activity and bone resorption

Through vial impedance spectroscopy (TVIS): A novel approach to process understanding for freeze-drying cycle development

The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link.Through vial impedance spectroscopy (TVIS) provides a new process analytical technology for monitoring a development scale lyophilization process, which exploits the changes in the bulk electrical properties that occur on freezing and subsequent drying of a drug solution. Unlike the majority of uses of impedance spectroscopy, for freeze-drying process development, the electrodes do not contact the product but are attached to the outside of the glass vial which is used to contain the product to provide a non-sample-invasive monitoring technology. Impedance spectra (in frequency range 10 Hz to 1 MHz) are generated throughout the drying cycle by a specially designed impedance spectrometer based on a 1 GΩ trans-impedance amplifier and then displayed in terms of complex capacitance. Typical capacitance spectra have one or two peaks in the imaginary capacitance (i.e., the dielectric loss) and the same number of steps in the real part capacitance (i.e., the dielectric permittivity). This chapter explores the underlying mechanisms that are responsible for these dielectric processes, i.e., the Maxwell-Wagner (space charge) polarization of the glass wall of the vial through the contents of the vial when in the liquid state, and the dielectric relaxation of ice when in the frozen state. In future work, it will be demonstrated how to measure product temperature and drying rates within single vials and multiple (clusters) of vials, from which other critical process parameters, such as heat transfer coefficient and dry layer resistance, may be determined

Ocrelizumab versus Interferon Beta-1a in Relapsing Multiple Sclerosis

Supported by F. Hoffmann–La Roche

Correlations of Interlayer Time with Distortion of Large Ti-6Al-4V Components in Laser Metal Deposition with Wire

Laser metal deposition with wire (LMD-w) is one of the emerging additive manufacturing (AM) technologies for large-scale aerospace components due to high deposition rates and material efficiency. However, it often results in undesired stresses and distortions due to non-uniform expansion and contraction of material during printing. Controlling inter-layer time, preheating, and clamping are the effective methods to mitigate the thermally induced stress and deformation. In this study, the effect of inter-layer cooling time on part distortion is investigated using a finite element method (FEM). The model accounts for actual tool paths, power, and cooling conditions. The results show that the model effectively captures the fluctuation of the Ti-6Al-4V plate during printing. Also, it shows an asymmetric distortion on the plate edges. Ultimately, the sequentially coupled thermal-stress simulation provided a quantitative understanding of the inter-layer cooling time on titanium plate distortion for the large-scale LMD-w process.Mechanical Engineerin

A heterodimeric [RGD-Glu-[64Cu-NO2A]-6-Ahx-RM2] ?vß3/GRPr-targeting antagonist radiotracer for PET imaging of prostate tumors

PubMed ID: 24480266Introduction: In the present study, we describe a 64Cu-radiolabeled heterodimeric peptide conjugate for dual ?vß3/GRPr (?vß3 integrin/gastrin releasing peptide receptor) targeting of the form [RGD-Glu-[64Cu-NO2A]-6-Ahx-RM2] (RGD: the amino acid sequence [Arg-Gly-Asp], a nonregulatory peptide used for ?vß3 integrin receptor targeting; Glu: glutamic acid; NO2A: 1,4,7-triazacyclononane-1,4-diacetic acid; 6-Ahx: 6-amino hexanoic acid; and RM2: (D-Phe-Gln-Trp-Ala-Val-Gly-His-Sta-Leu-NH2), an antagonist analogue of bombesin (BBN) peptide used for GRPr targeting). Methods: RGD-Glu-6Ahx-RM2] was conjugated to a NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) complexing agent to produce [RGD-Glu-[NO2A]-6-Ahx-RM2], which was purified by reversed-phase high-performance liquid chromatography (RP-HPLC) and characterized by electrospray ionization-mass spectrometry (ESI-MS). Radiolabeling of the conjugate with 64Cu produced [RGD-Glu-[64Cu-NO2A]-6-Ahx-RM2 in high radiochemical yield (?95%). In vivo behavior of the radiolabeled peptide conjugate was investigated in normal CF-1 mice and in the PC-3 human prostate cancer experimental model. Results: A competitive displacement receptor binding assay in human prostate PC-3 cells using 125I-[Tyr4]BBN as the radioligand showed high binding affinity of [RGD-Glu-[natCu-NO2A]-6-Ahx-RM2] conjugate for the GRPr (3.09±0.34 nM). A similar assay in human, glioblastoma U87-MG cells using 125I-Echistatin as the radioligand indicated a moderate receptor-binding affinity for the ?vß3 integrin (518±37.5 nM). In vivo studies of [RGD-Glu-[64Cu-NO2A]-6-Ahx-RM2] showed high accumulation (4.86±1.01 %ID/g, 1h post-intravenous injection (p.i.)) and prolonged retention (4.26±1.23 %ID/g, 24h p.i.) of tracer in PC-3 tumor-bearing mice. Micro-positron emission tomography (microPET) molecular imaging studies produced high-quality, high contrast images in PC-3 tumor-bearing mice at 4h p.i. Conclusions: The favorable pharmacokinetics and enhanced tumor uptake of 64Cu-NOTA-RGD-Glu-6Ahx-RM2 warrant further investigations for dual integrin and GRPr-positive tumor imaging and possible radiotherapy. © 2014 Elsevier Inc.This material was the result of work supported with resources and the use of facilities at the Truman VA, Columbia, MO, 65201 and the University of Missouri School of Medicine, Columbia, MO 65211, USA. Dr. Kubra Durkan acknowledges financial support from the Scientific and Technological Research Council of Turkey (TUBITAK) Science Fellowships and Grant Programmes Department (BIDEB). This work was also funded in part by The United States Department of Veterans’ Affairs, VA Merit Bridge Funding Award Mechanism. -

Self-Assembled Flexible and Integratable 3D Microtubular Asymmetric Supercapacitors

The rapid development of microelectronics has equally rapidly increased the demand for miniaturized energy storage devices. On-chip microsupercapacitors (MSCs), as promising power candidates, possess great potential to complement or replace electrolytic capacitors and microbatteries in various applications. However, the areal capacities and energy densities of the planar MSCs are commonly limited by the low voltage window, the thin layer of the electrode materials and complex fabrication processes. Here, a new-type three-dimensional (3D) tubular asymmetric MSC with small footprint area, high potential window, ultrahigh areal energy density, and long-term cycling stability is fabricated with shapeable materials and photolithographic technologies, which are compatible with modern microelectronic fabrication procedures widely used in industry. Benefiting from the novel architecture, the 3D asymmetric MSC displays an ultrahigh areal capacitance of 88.6 mF cm−2 and areal energy density of 28.69 mW h cm−2, superior to most reported interdigitated MSCs. Furthermore, the 3D tubular MSCs demonstrate remarkable cycling stability and the capacitance retention is up to 91.8% over 12 000 cycles. It is believed that the efficient fabrication methodology can be used to construct various integratable microscale tubular energy storage devices with small footprint area and high performance for miniaturized electronics

Self‐Assembled Flexible and Integratable 3D Microtubular Asymmetric Supercapacitors

The rapid development of microelectronics has equally rapidly increased the demand for miniaturized energy storage devices. On-chip microsupercapacitors (MSCs), as promising power candidates, possess great potential to complement or replace electrolytic capacitors and microbatteries in various applications. However, the areal capacities and energy densities of the planar MSCs are commonly limited by the low voltage window, the thin layer of the electrode materials and complex fabrication processes. Here, a new-type three-dimensional (3D) tubular asymmetric MSC with small footprint area, high potential window, ultrahigh areal energy density, and long-term cycling stability is fabricated with shapeable materials and photolithographic technologies, which are compatible with modern microelectronic fabrication procedures widely used in industry. Benefiting from the novel architecture, the 3D asymmetric MSC displays an ultrahigh areal capacitance of 88.6 mF cm−2 and areal energy density of 28.69 mW h cm−2, superior to most reported interdigitated MSCs. Furthermore, the 3D tubular MSCs demonstrate remarkable cycling stability and the capacitance retention is up to 91.8% over 12 000 cycles. It is believed that the efficient fabrication methodology can be used to construct various integratable microscale tubular energy storage devices with small footprint area and high performance for miniaturized electronics