Sample records for vivo oxygen-sensing applications

  1. Polymer coating of paramagnetic particulates for in vivo oxygen-sensing applications

    PubMed Central

    Eteshola, Edward; Pandian, Ramasamy P.; Kuppusamy, Periannan

    2009-01-01

    Crystalline lithium phthalocyanine (LiPc) can be used to sense oxygen. To enhance biocompatibility/stability of LiPc, we encapsulated LiPc in Teflon AF (TAF), cellulose acetate (CA), and polyvinyl acetate (PVAc) (TAF, previously used to encapsulate LiPc, was a comparator). We identified water-miscible solvents that don’t dissolve LiPc crystals, but are solvents for the polymers, and encapsulated crystals by solvent evaporation. Oxygen sensitivity of films was characterized in vitro and in vivo. Encapsulation did not change LiPc oximetry properties in vitro at anoxic conditions or varying partial pressures of oxygen (pO2). EPR linewidth of encapsulated particles was linear with pO2, responding to pO2 changes quickly and reproducibly for dynamic measurements. Encapsulated LiPc was unaffected by biological oxidoreductants, stable in vivo for four weeks. Oximetry, stability and biocompatibility properties of LiPc films were comparable, but both CA and PVAc films are cheaper, and easier to fabricate and handle than TAF films, making them superior. PMID:19083100

  2. Solid MRI contrast agents for long-term, quantitative in vivo oxygen sensing

    PubMed Central

    Liu, Vincent H.; Vassiliou, Christophoros C.; Imaad, Syed M.; Cima, Michael J.

    2014-01-01

    Targeted MRI contrast agents have proven useful in research and clinical studies for highlighting specific metabolites and biomarkers [Davies GL, et al. (2013) Chem Commun (Camb) 49(84):9704–9721] but their applicability in serial imaging is limited owing to a changing concentration postinjection. Solid enclosures have previously been used to keep the local concentration of contrast agent constant, but the need to surgically implant these devices limits their use [Daniel K, et al. (2009) Biosens Bioelectron 24(11):3252–3257]. This paper describes a novel class of contrast agent that comprises a responsive material for contrast generation and an injectable polymeric matrix for structural support. Using this principle, we have designed a contrast agent sensitive to oxygen, which is composed of dodecamethylpentasiloxane as the responsive material and polydimethylsiloxane as the matrix material. A rodent inspired-gas model demonstrated that these materials are functionally stable in vivo for at least 1 mo, which represents an order of magnitude improvement over an injection of liquid siloxane [Kodibagkar VD, et al. (2006) Magn Reson Med 55(4):743–748]. We also observed minimal adverse tissue reactions or migration of contrast agents from the initial injection site. This class of contrast agents, thus, represented a new and complementary method to monitor chronic diseases by MRI. PMID:24753603

  3. Optical oxygen sensing systems for drug discovery applications: Respirometric Screening Technology (RST)

    NASA Astrophysics Data System (ADS)

    Papkovsky, Dmitri B.; Hynes, James; Fernandes, Richard

    2005-11-01

    Quenched-fluorescence oxygen sensing allows non-chemical, reversible, real-time monitoring of molecular oxygen and rates of oxygen consumption in biological samples. Using this approach we have developed Respirometric Screening Technology (RST); a platform which facilitates the convenient analysis of cellular oxygen uptake. This in turn allows the investigation of compounds and processes which affect respiratory activity. The RST platform employs soluble phosphorescent oxygen-sensitive probes, which may be assessed in standard microtitter plates on a fluorescence plate reader. New formats of RST assays and time-resolved fluorescence detection instrumentation developed by Luxcel provide improvements in assay sensitivity, miniaturization and overall performance. RST has a diverse range of applications in drug discovery area including high throughput analysis of mitochondrial function; studies of mechanisms of toxicity and apoptosis; cell and animal based screening of compound libraries and environmental samples; and, sterility testing. RST has been successfully validated with a range of practical targets and adopted by several leading pharmaceutical companies.

  4. Solid MRI contrast agents for long-term, quantitative in vivo oxygen sensing

    E-print Network

    Liu, Vincent Hok

    Targeted MRI contrast agents have proven useful in research and clinical studies for highlighting specific metabolites and biomarkers [Davies GL, et al. (2013) Chem Commun (Camb) 49(84):9704–9721] but their applicability ...

  5. Intracellular and in Vivo Oxygen Sensing Using Phosphorescent Ir(III) Complexes with a Modified Acetylacetonato Ligand.

    PubMed

    Yoshihara, Toshitada; Hosaka, Masahiro; Terata, Motoki; Ichikawa, Kazuki; Murayama, Saori; Tanaka, Asami; Mori, Masanobu; Itabashi, Hideyuki; Takeuchi, Toshiyuki; Tobita, Seiji

    2015-03-01

    Small luminescent molecular probes based on the iridium(III) complex BTP, (btp)2Ir(acac) (btp = benzothienylpyridine, acac = acetylacetone) have been developed for sensing intracellular and in vivo O2. These compounds are BTPSA (containing an anionic carboxyl group), BTPNH2 (containing a cationic amino group), and BTPDM1 (containing a cationic dimethylamino group); all substituents are incorporated into the ancillary acetylacetonato ligand of BTP. Introduction of the cationic dimethylamino group resulted in an almost 20-fold increase in cellular uptake efficiency of BTPDM1 by HeLa cells compared with BTP. The phosphorescence intensity of BTPDM1 internalized in living cells provided a visual representation of the O2 gradient produced by placing a coverslip over cultured monolayer cells. The intracellular O2 levels (pO2) inside and outside the edge of the coverslip could be evaluated by measuring the phosphorescence lifetime of BTPDM1. Furthermore, intravenous administration of 25 nmol BTPDM1 to tumor-bearing mice allowed the tumor region to be visualized by BTPDM1 phosphorescence. The lifetime of BTPDM1 phosphorescence from tumor regions was much longer than that from extratumor regions, thereby demonstrating tumor hypoxia (pO2 = 6.1 mmHg for tumor and 50 mmHg for extratumor epidermal tissue). Tissue distribution studies showed that 2 h after injection of BTPDM1 into a mouse, the highest distribution was in liver and kidney, while after 24 h, BTPDM1 was excreted in the feces. These results demonstrate that BTPDM1 can be used as a small molecular probe for measuring intracellular O2 levels in both cultured cells and specific tissues and organs. PMID:25634116

  6. Oxygen sensing in the body

    Microsoft Academic Search

    S. Lahiri; A. Roy; S. M. Baby; T. Hoshi; G. L. Semenza; N. R. Prabhakar

    2006-01-01

    This review is divided into three parts: (a) The primary site of oxygen sensing is the carotid body which instantaneously respond to hypoxia without involving new protein synthesis, and is historically known as the first oxygen sensor and is therefore placed in the first section (Lahiri, Roy, Baby and Hoshi). The carotid body senses oxygen in acute hypoxia, and produces

  7. Injectable polymer for in vivo oxygen sensing

    E-print Network

    Imaad, Syed M. (Syed Muhammad)

    2013-01-01

    This thesis documents the synthesis and characterization of an elastomeric polymer that is oxygen sensitive and can be interrogated using Magnetic Resonance Imaging (MRI) or Magnetic Resonance (MR) technology to report the ...

  8. Influence of Matrices on Oxygen Sensing of Three Sensing Films with Chemically Conjugated Platinum Porphyrin Probes and Preliminary Application for Monitoring of Oxygen Consumption of Escherichia coli (E. coli)

    PubMed Central

    Tian, Yanqing; Shumway, Bradley R.; Gao, Weimin; Youngbull, Cody; Holl, Mark R.; Johnson, Roger H.; Meldrum, Deirdre R.

    2010-01-01

    Oxygen sensing films were synthesized by a chemical conjugation of functional platinum porphyrin probes in silica gel, polystyrene (PS), and poly(2-hydroxyethyl methacrylate) (PHEMA) matrices. Responses of the sensing films to gaseous oxygen and dissolved oxygen were studied and the influence of the matrices on the sensing behaviors was investigated. Silica gel films had the highest fluorescence intensity ratio from deoxygenated to oxygenated environments and the fastest response time to oxygen. PHEMA films had no response to gaseous oxygen, but had greater sensitivity and a faster response time for dissolved oxygen than those of PS films. The influence of matrices on oxygen response, sensitivity and response time was discussed. The influence is most likely attributed to the oxygen diffusion abilities of the matrices. Since the probes were chemically immobilized in the matrices, no leaching of the probes was observed from the sensing films when applied in aqueous environment. One sensing film made from the PHEMA matrix was used to preliminarily monitor the oxygen consumption of Escherichia coli (E. coli) bacteria. E. coli cell density and antibiotics ampicillin concentration dependent oxygen consumption was observed, indicating the potential application of the oxygen sensing film for biological application. PMID:21076638

  9. A miniature inexpensive, oxygen sensing element

    SciTech Connect

    Arenz, R.W.

    1991-10-07

    An exhaustive study was conducted to determine the feasibility of Nernst-type oxygen sensors based on ceramics containing Bi{sub 2}O{sub 3}. The basic sensor design consisted of a ceramic sensing module sealed into a metal tube. The module accommodated an internal heater and thermocouple. Thermal-expansion-matched metals, adhesives, and seals were researched and developed, consistent with sequential firings during sensor assembly. Significant effort was devoted to heater design/testing and to materials' compatibility with Pt electrodes. A systematic approach was taken to develop all sensor components which led to several design modifications. Prototype sensors were constructed and exhaustively tested. It is concluded that development of Nerst-type oxygen sensors based on Bi{sub 2}O{sub 3} will require much further effort and application of specialized technologies. However, during the course of this 3-year program much progress was reported in the literature on amperometric-type oxygen sensors, and a minor effort was devoted here to this type of sensor based on Bi{sub 2}O{sub 3}. These studies were made on Bi{sub 2}O{sub 3}-based ceramic samples in a multilayer-capacitor-type geometry and amperometric-type oxygen sensing was demonstrated at very low temperatures ({approximately} 160{degree}C). A central advantage here is that these types of sensors can be mass-produced very inexpensively ({approximately} 20--50 cents per unit). Research is needed, however, to develop an optimum diffusion-limiting barrier coating. In summary, the original goals of this program were not achieved due to unforeseen problems with Bi{sub 2}O{sub 3}-based Nernst sensors. However, a miniature amperometric sensor base on Bi{sub 2}O{sub 3} was demonstrated in this program, and it is now seen that this latter sensor is far superior to the originally proposed Nernst sensor. 6 refs., 24 figs.

  10. Oxygen Sensing: Getting Pumped by Sterols

    NSDL National Science Digital Library

    Brooke M. Emerling (Northwestern University Medical School; Department of Medicine REV)

    2005-06-21

    Oxygen plays a pivotal role in the maintenance of life for all eukaryotes, with the exception of strict anaerobes. Eukaryotes have developed mechanisms to sense and respond to decreased oxygen levels. How eukaryotes sense oxygen is still not fully understood. What is (or are) the oxygen sensor(s)? This question has vital physiological and pathophysiological implications, because all living aerobic organisms have adaptive mechanisms to maintain oxygen homeostasis. A recent report describes a novel eukaryotic oxygen-sensing mechanism in the fission yeast Schizosaccharomyces pombe, involving the depletion of sterols as a trigger to induce gene expression in response to decreased oxygen levels. It is not yet clear whether this mechanism is involved in the mammalian response to hypoxia, possibly in conjunction with activation of one or both of the hypoxia-inducible factor (HIF-1 or HIF-2) transcription factors.

  11. Evolution and physiology of neural oxygen sensing

    PubMed Central

    Costa, Kauê M.; Accorsi-Mendonça, Daniela; Moraes, Davi J. A.; Machado, Benedito H.

    2014-01-01

    Major evolutionary trends in animal physiology have been heavily influenced by atmospheric O2 levels. Amongst other important factors, the increase in atmospheric O2 which occurred in the Pre-Cambrian and the development of aerobic respiration beckoned the evolution of animal organ systems that were dedicated to the absorption and transportation of O2, e.g., the respiratory and cardiovascular systems of vertebrates. Global variations of O2 levels in post-Cambrian periods have also been correlated with evolutionary changes in animal physiology, especially cardiorespiratory function. Oxygen transportation systems are, in our view, ultimately controlled by the brain related mechanisms, which senses changes in O2 availability and regulates autonomic and respiratory responses that ensure the survival of the organism in the face of hypoxic challenges. In vertebrates, the major sensorial system for oxygen sensing and responding to hypoxia is the peripheral chemoreflex neuronal pathways, which includes the oxygen chemosensitive glomus cells and several brainstem regions involved in the autonomic regulation of the cardiovascular system and respiratory control. In this review we discuss the concept that regulating O2 homeostasis was one of the primordial roles of the nervous system. We also review the physiology of the peripheral chemoreflex, focusing on the integrative repercussions of chemoreflex activation and the evolutionary importance of this system, which is essential for the survival of complex organisms such as vertebrates. The contribution of hypoxia and peripheral chemoreflex for the development of diseases associated to the cardiovascular and respiratory systems is also discussed in an evolutionary context. PMID:25161625

  12. Fabrication of Eu(III) complex doped nanofibrous membranes and their oxygen-sensing properties.

    PubMed

    Songzhu, Lin; Xiangting, Dong; Jinxian, Wang; Guixia, Liu; Wenshen, Yu; Ruokun, Jia

    2010-11-01

    In this paper, we report the synthesis, characterization, and photophysical properties of Eu(TTA)?ECIP, where TTA=2-thenoyltrifluoroacetonate, and ECIP=1-ethyl-2-(N-ethyl-carbazole-yl-4-)imidazo[4,5-f]1,10-phenanthroline. Its elementary application for oxygen-sensing application is also investigated by doping it into a polymer matrix of polystyrene (PS). Experimental data suggest that the 2.5 wt% doped Eu(TTA)?ECIP/PS nanofibrous membrane exhibits a high sensitivity of 3.4 towards oxygen with a good linear relationship of R² = 0.9962. In addition, the 2.5 wt% doped Eu(TTA)?ECIP/PS nanofibrous membrane owns a quick response of 8s towards oxygen, along with its excellent atmosphere insensitivity and photobleaching resistance. All these results suggest that both Eu(TTA)?ECIP and Eu(TTA)?CIP/PS system are promising candidates for oxygen-sensing optical sensors. PMID:20843734

  13. Dissolved oxygen sensing using organometallic dyes deposited within a microfluidic environment

    Microsoft Academic Search

    Q. L. Chen; H. P. Ho; L. Jin; B. W.-K. Chu; M. J. Li; V. W.-W. Yam

    2008-01-01

    This work primarily aims to integrate dissolved oxygen sensing capability with a microfluidic platform containing arrays of micro bio-reactors or bio-activity indicators. The measurement of oxygen concentration is of significance for a variety of bio-related applications such as cell culture and gene expression. Optical oxygen sensors based on luminescence quenching are gaining much interest in light of their low power

  14. Quality assessment of packaged foods by optical oxygen sensing

    NASA Astrophysics Data System (ADS)

    Papkovsky, Dmitri B.; O'Mahony, Fiach C.; Kerry, Joe P.; Ogurtsov, Vladimir I.

    2005-11-01

    A phase-fluorometric oxygen sensor system has been developed, which allows non-destructive measurement of residual oxygen levels in sealed containers such as packaged foods. It operates with disposable solid-state sensors incorporated in each pack, and a portable detector which interrogates with the sensors through a (semi)transparent packaging material. The system has been optimized for packaging applications and validated in small and medium scale trials with different types of food, including MAP hams, cheese, convenience foods, smoked fish, bakery. It has demonstrated high efficiency in monitoring package integrity, oxygen profiles in packs, performance of packaging process and many other research and quality control tasks, allowing control of 100% of packs. The low-cost batch-calibrated sensors have demonstrated reliability, safety, stability including direct contact with food, high efficiency in the low oxygen range. Another system, which also employs the fluorescence-based oxygen sensing approach, provides rapid assessment of microbial contamination (total viable counts) in complex samples such as food homogenates, industrial waste, environmental samples, etc. It uses soluble oxygen-sensitive probes, standard microtitter plates and fluorescence measurements on conventional plate reader to monitor growth of aerobic bacteria in small test samples (e.g. food homogenates) via their oxygen respiration. The assay provides high sample through put, miniaturization, speed, and can serve as alternative to the established methods such as agar plate colony counts and turbidimetry.

  15. Dissolved oxygen sensing using organometallic dyes deposited within a microfluidic environment

    NASA Astrophysics Data System (ADS)

    Chen, Q. L.; Ho, H. P.; Jin, L.; Chu, B. W.-K.; Li, M. J.; Yam, V. W.-W.

    2008-02-01

    This work primarily aims to integrate dissolved oxygen sensing capability with a microfluidic platform containing arrays of micro bio-reactors or bio-activity indicators. The measurement of oxygen concentration is of significance for a variety of bio-related applications such as cell culture and gene expression. Optical oxygen sensors based on luminescence quenching are gaining much interest in light of their low power consumption, quick response and high analyte sensitivity in comparison to similar oxygen sensing devices. In our microfluidic oxygen sensor device, a thin layer of oxygen-sensitive luminescent organometallic dye is covalently bonded to a glass slide. Micro flow channels are formed on the glass slide using patterned PDMS (Polydimethylsiloxane). Dissolved oxygen sensing is then performed by directing an optical excitation probe beam to the area of interest within the microfluidic channel. The covalent bonding approach for sensor layer formation offers many distinct advantages over the physical entrapment method including minimizing dye leaching, ensuring good stability and fabrication simplicity. Experimental results confirm the feasibility of the device.

  16. Fabrication of Eu(III) complex doped nanofibrous membranes and their oxygen-sensing properties

    Microsoft Academic Search

    Lin Songzhu; Dong Xiangting; Wang Jinxian; Liu Guixia; Yu Wenshen; Jia Ruokun

    2010-01-01

    In this paper, we report the synthesis, characterization, and photophysical properties of Eu(TTA)3ECIP, where TTA=2-thenoyltrifluoroacetonate, and ECIP=1-ethyl-2-(N-ethyl-carbazole-yl-4-)imidazo[4,5-f]1,10-phenanthroline. Its elementary application for oxygen-sensing application is also investigated by doping it into a polymer matrix of polystyrene (PS). Experimental data suggest that the 2.5wt% doped Eu(TTA)3ECIP\\/PS nanofibrous membrane exhibits a high sensitivity of 3.4 towards oxygen with a good linear relationship of

  17. Superhydrophobic porous surfaces: dissolved oxygen sensing.

    PubMed

    Gao, Yu; Chen, Tao; Yamamoto, Shunsuke; Miyashita, Tokuji; Mitsuishi, Masaya

    2015-02-18

    Porous polymer films are necessary for dissolved gas sensor applications that combine high sensitivity with selectivity. This report describes a greatly enhanced dissolved oxygen sensor system consisting of amphiphilic acrylamide-based polymers: poly(N-(1H, 1H-pentadecafluorooctyl)-methacrylamide) (pC7F15MAA) and poly(N-dodecylacrylamide-co-5- [4-(2-methacryloyloxyethoxy-carbonyl)phenyl]-10,15,20-triphenylporphinato platinum(II)) (p(DDA/PtTPP)). The nanoparticle formation capability ensures both superhydrophobicity with a water contact angle greater than 160° and gas permeability so that molecular oxygen enters the film from water. The film was prepared by casting a mixed solution of pC7F15MAA and p(DDA/PtTPP) with AK-225 and acetic acid onto a solid substrate. The film has a porous structure comprising nanoparticle assemblies with diameters of several hundred nanometers. The film shows exceptional performance as the oxygen sensitivity reaches 126: the intensity ratio at two oxygen concentrations (I0/I40) respectively corresponding to dissolved oxygen concentration 0 and 40 (mg L(-1)). Understanding and controlling porous nanostructures are expected to provide opportunities for making selective penetration/separation of molecules occurring at the superhydrophobic surface. PMID:25659178

  18. Genetic Causes of Erythrocytosis and the Oxygen Sensing Pathway

    PubMed Central

    Lee, Frank S.

    2012-01-01

    Idiopathic erythrocytosis is an uncommon disease, and is defined by an increase in red blood cell mass. The differential diagnosis of erythrocytosis is extensive, and can be divided into primary and secondary forms. Primary erythrocytoses are due to intrinsic defects in erythroid precursor cells and are characterized by low erythropoietin levels. Secondary erythrocytoses are extrinsic to erythroid progenitors and are characterized by either high or inappropriately normal erythropoietin levels. A distinct subset of secondary erythrocytoses are due to genetic mutations in key proteins of the oxygen sensing pathway. These proteins constitute the core molecular machinery of oxygen-sensing with respect to red blood cell control. Apart from assigning physiologic roles for these proteins, studies of these rare mutations have (i) revealed the exquisite sensitivity of this pathway to genetic perturbations, (ii) highlighted important functional regions of the proteins, and (iii) provided a basis for potentially targeting this pathway for therapeutic benefit. PMID:18538455

  19. Ceramide Mediates Acute Oxygen Sensing in Vascular Tissues

    PubMed Central

    Moreno, Laura; Moral-Sanz, Javier; Morales-Cano, Daniel; Barreira, Bianca; Moreno, Enrique; Ferrarini, Alessia; Pandolfi, Rachele; Ruperez, Francisco J.; Cortijo, Julio; Sanchez-Luna, Manuel; Villamor, Eduardo; Perez-Vizcaino, Francisco

    2014-01-01

    Abstract Aims: A variety of vessels, such as resistance pulmonary arteries (PA) and fetoplacental arteries and the ductus arteriosus (DA) are specialized in sensing and responding to changes in oxygen tension. Despite opposite stimuli, normoxic DA contraction and hypoxic fetoplacental and PA vasoconstriction share some mechanistic features. Activation of neutral sphingomyelinase (nSMase) and subsequent ceramide production has been involved in hypoxic pulmonary vasoconstriction (HPV). Herein we aimed to study the possible role of nSMase-derived ceramide as a common factor in the acute oxygen-sensing function of specialized vascular tissues. Results: The nSMase inhibitor GW4869 and an anticeramide antibody reduced the hypoxic vasoconstriction in chicken PA and chorioallantoic arteries (CA) and the normoxic contraction of chicken DA. Incubation with interference RNA targeted to SMPD3 also inhibited HPV. Moreover, ceramide and reactive oxygen species production were increased by hypoxia in PA and by normoxia in DA. Either bacterial sphingomyelinase or ceramide mimicked the contractile responses of hypoxia in PA and CA and those of normoxia in the DA. Furthermore, ceramide inhibited voltage-gated potassium currents present in smooth muscle cells from PA and DA. Finally, the role of nSMase in acute oxygen sensing was also observed in human PA and DA. Innovation: These data provide evidence for the proposal that nSMase-derived ceramide is a critical player in acute oxygen-sensing in specialized vascular tissues. Conclusion: Our results indicate that an increase in ceramide generation is involved in the vasoconstrictor responses induced by two opposite stimuli, such as hypoxia (in PA and CA) and normoxia (in DA). Antioxid. Redox Signal. 20, 1–14. PMID:23725018

  20. Two–Photon Oxygen Sensing with Quantum Dot–Porphyrin Conjugates

    PubMed Central

    Lemon, Christopher M.; Karnas, Elizabeth; Bawendi, Moungi G.; Nocera, Daniel G.

    2013-01-01

    Supramolecular assemblies of a quantum dot (QD) associated to palladium(II) porphyrins have been developed to detect oxygen (pO2) in organic solvents. Palladium porphyrins are sensitive in the 0–160 torr range, making them ideal phosphors for in vivo biological oxygen quantification. Porphyrins with meso pyridyl substituents bind to the surface of the QD to produce self–assembled nanosensors. Appreciable overlap between QD emission and porphyrin absorption features results in efficient Förster resonance energy transfer (FRET) for signal transduction in these sensors. The QD serves as a photon antenna, enhancing porphyrin emission under both one– and two–photon excitation, demonstrating that QD–palladium porphyrin conjugates may be used for oxygen sensing over physiological oxygen ranges. PMID:23978247

  1. Biomedical Applications of Sodium MRI In Vivo

    PubMed Central

    Madelin, Guillaume; Regatte, Ravinder R.

    2013-01-01

    In this article, we present an up-to-date overview of the potential biomedical applications of sodium MRI in vivo. Sodium MRI is a subject of increasing interest in translational imaging research as it can give some direct and quantitative biochemical information on the tissue viability, cell integrity and function, and therefore not only help the diagnosis but also the prognosis of diseases and treatment outcomes. It has already been applied in vivo in most of human tissues, such as brain for stroke or tumor detection and therapeutic response, in breast cancer, in articular cartilage, in muscle and in kidney, and it was shown in some studies that it could provide very useful new information not available through standard proton MRI. However, this technique is still very challenging due to the low detectable sodium signal in biological tissue with MRI and hardware/software limitations of the clinical scanners. The article is divided in three parts: (1) the role of sodium in biological tissues, (2) a short review on sodium magnetic resonance, and (3) a review of some studies on sodium MRI on different organs/diseases to date. PMID:23722972

  2. Dissolved Oxygen Sensing in a Flow Stream using Molybdenum Chloride Optical Indicators

    E-print Network

    Ghosh, Ruby N.

    Dissolved Oxygen Sensing in a Flow Stream using Molybdenum Chloride Optical Indicators Reza Loloee1@msu.edu Abstract--Dissolved oxygen concentration is considered the most important water quality variable in fish culture. Reliable and continuous (24/7) oxygen monitoring of dissolved oxygen (DO) in the 1 ­ 11 mg

  3. Sensors and Actuators B 113 (2006) 162168 Dependence of potentiometric oxygen sensing

    E-print Network

    Dutta, Prabir K.

    2006-01-01

    in mixed potential NOx and CO sensors [5]. The output voltage of these sensors is determined by the mixedSensors and Actuators B 113 (2006) 162­168 Dependence of potentiometric oxygen sensing for Industrial Sensors and Measurements (CISM), Department of Materials Science and Engineering, 2041 College

  4. Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads.

    PubMed

    Wang, Lin; Acosta, Miguel A; Leach, Jennie B; Carrier, Rebecca L

    2013-04-21

    Capability of measuring and monitoring local oxygen concentration at the single cell level (tens of microns scale) is often desirable but difficult to achieve in cell culture. In this study, biocompatible oxygen sensing beads were prepared and tested for their potential for real-time monitoring and mapping of local oxygen concentration in 3D micro-patterned cell culture systems. Each oxygen sensing bead is composed of a silica core loaded with both an oxygen sensitive Ru(Ph2phen3)Cl2 dye and oxygen insensitive Nile blue reference dye, and a poly-dimethylsiloxane (PDMS) shell rendering biocompatibility. Human intestinal epithelial Caco-2 cells were cultivated on a series of PDMS and type I collagen based substrates patterned with micro-well arrays for 3 or 7 days, and then brought into contact with oxygen sensing beads. Using an image analysis algorithm to convert florescence intensity of beads to partial oxygen pressure in the culture system, tens of microns-size oxygen sensing beads enabled the spatial measurement of local oxygen concentration in the microfabricated system. Results generally indicated lower oxygen level inside wells than on top of wells, and local oxygen level dependence on structural features of cell culture surfaces. Interestingly, chemical composition of cell culture substrates also appeared to affect oxygen level, with type-I collagen based cell culture systems having lower oxygen concentration compared to PDMS based cell culture systems. In general, results suggest that oxygen sensing beads can be utilized to achieve real-time and local monitoring of micro-environment oxygen level in 3D microfabricated cell culture systems. PMID:23443975

  5. Spatially monitoring oxygen level in 3D microfabricated cell culture systems using optical oxygen sensing beads

    PubMed Central

    Wang, Lin; Acosta, Miguel A.; Leach, Jennie B.; Carrier, Rebecca L.

    2013-01-01

    Capability of measuring and monitoring local oxygen concentration at the single cell level (tens of microns scale) is often desirable but difficult to achieve in cell culture. In this study, biocompatible oxygen sensing beads were prepared and tested for their potential for real-time monitoring and mapping of local oxygen concentration in 3D micro-patterned cell culture systems. Each oxygen sensing bead is composed of a silica core loaded with both an oxygen sensitive Ru(Ph2phen3)Cl2 dye and oxygen insensitive Nile blue reference dye, and a poly-dimethylsiloxane (PDMS) shell rendering biocompatibility. Human intestinal epithelial Caco-2 cells were cultivated on a series of PDMS and type I collagen based substrates patterned with micro-well arrays for 3 or 7 days, and then brought into contact with oxygen sensing beads. Using an image analysis algorithm to convert florescence intensity of beads to partial oxygen pressure in the culture system, tens of microns-size oxygen sensing beads enabled the spatial measurement of local oxygen concentration in the microfabricated system. Results generally indicated lower oxygen level inside wells than on top of wells, and local oxygen level dependence on structural features of cell culture surfaces. Interestingly, chemical composition of cell culture substrates also appeared to affect oxygen level, with type-I collagen based cell culture systems having lower oxygen concentration compared to PDMS based cell culture systems. In general, results suggest that oxygen sensing beads can be utilized to achieve real-time and local monitoring of micro-environment oxygen level in 3D microfabricated cell culture systems. PMID:23443975

  6. Oxygen Sensing for Industrial Safety — Evolution and New Approaches

    PubMed Central

    Willett, Martin

    2014-01-01

    The requirement for the detection of oxygen in industrial safety applications has historically been met by electrochemical technologies based on the consumption of metal anodes. Products using this approach have been technically and commercially successful for more than three decades. However, a combination of new requirements is driving the development of alternative approaches offering fresh opportunities and challenges. This paper reviews some key aspects in the evolution of consumable anode products and highlights recent developments in alternative technologies aimed at meeting current and anticipated future needs in this important application. PMID:24681673

  7. Dissolved oxygen sensing based on fluorescence quenching of ceria nanoparticles

    NASA Astrophysics Data System (ADS)

    Shehata, Nader; Meehan, Kathleen; Leber, Donald

    2012-10-01

    The development of oxygen sensors has positively impacted the fields of medical science, bioengineering, environmental monitoring, solar cells, industrial process control, and a number of military applications. Fluorescent quenching sensors have an inherent high sensitivity, chemical selectivity, and stability when compared to other types of sensors. While cerium oxide thin films have been used to monitor oxygen in the gas phase, the potential of cerium oxide (ceria) nanoparticles as the active material in sensor for oxygen gas has only recently been investigated. Ceria nanoparticles are one of the most unique nanomaterials that are being studied today due to the diffusion and reactivity of its oxygen vacancies, which contributes to its high oxygen storage capability. The reactivity of the oxygen vacancies, which is also related to conversion of cerium ion from the Ce+4 to Ce+3 state, affects the fluorescence properties of the ceria nanoparticles. Our research demonstrates that the ceria nanoparticles (~7 nm in diameter) have application as a fluorescence quenching sensor to measure dissolved oxygen in water. We have found a strong inverse correlation between the amplitude of the fluorescence emission (?excitation = 430 nm and ?peak = 520 nm) and the dissolved oxygen concentration between 5 - 13 mg/L. The Stern-Volmer constant, which is an indication of the sensitivity of gas sensing is 184 M-1 for the ceria nanoparticles. The results show that ceria nanoparticles can be used in an improved, robust fluorescence sensor for dissolved oxygen in a liquid medium.

  8. Ancient Atmospheres and the Evolution of Oxygen Sensing Via the Hypoxia-Inducible Factor in Metazoans

    NSDL National Science Digital Library

    Cormac Taylor (UCD Conway Institute)

    2010-10-01

    Metazoan diversification occurred during a time when atmospheric oxygen levels fluctuated between 15 and 30%. The hypoxia-inducible factor (HIF) is a primary regulator of the adaptive transcriptional response to hypoxia. Although the HIF pathway is highly conserved, its complexity increased during periods when atmospheric oxygen concentrations were increasing. Thus atmospheric oxygen levels may have provided a selection force on the development of cellular oxygen-sensing pathways.

  9. Dual emission probe for luminescence oxygen sensing: a critical comparison between intensity, lifetime and ratiometric measurements

    Microsoft Academic Search

    Hannes Hochreiner; Israel Sánchez-Barragán; José M. Costa-Fernández; Alfredo Sanz-Medel

    2005-01-01

    The characterization of a dual emission sensing luminescence material for water-dissolved oxygen sensing is presented in this paper. The oxygen-sensitive material is based on a dual-emitting luminescent molecule immobilized onto an adequate solid support. The metal chelate formed between the 8-hydroxy-7-iodo-5-quinolinesulphonic acid (Ferron) and aluminium (Al-Ferron) was the selected oxygen-sensitive dual-emitting luminescent complex, while the anionic exchanger Dowex 1X2-200 resin

  10. Activation of the oxygen-sensing signal cascade prevents mitochondrial injury after mouse liver ischemia-reperfusion

    PubMed Central

    Zhong, Zhi; Ramshesh, Venkat K.; Rehman, Hasibur; Currin, Robert T.; Sridharan, Vijayalakshmi; Theruvath, Tom P.; Kim, Insil; Wright, Gary L.; Lemasters, John J.

    2008-01-01

    The mitochondrial permeability transition (MPT) plays an important role in hepatocyte death caused by ischemia-reperfusion (IR). This study investigated whether activation of the cellular oxygen-sensing signal cascade by prolyl hydroxylase inhibitors (PHI) protects against the MPT after hepatic IR. Ethyl 3,4-dihyroxybenzoate (EDHB, 100 mg/kg ip), a PHI, increased mouse hepatic hypoxia-inducible factor-1? and heme oxygenase-1 (HO-1). EDHB-treated and untreated mice were subjected to 1 h of warm ischemia to ?70% of the liver followed by reperfusion. Mitochondrial polarization, cell death, and the MPT were assessed by intravital confocal/multiphoton microscopy of rhodamine 123, propidium iodide, and calcein. EDHB largely blunted alanine aminotransferase (ALT) release and necrosis after reperfusion. In vehicle-treated mice at 2 h after reperfusion, viable cells with depolarized mitochondria were 72%, and dead cells were 2%, indicating that depolarization preceded necrosis. Mitochondrial voids excluding calcein disappeared, indicating MPT onset in vivo. NIM811, a specific inhibitor of the MPT, blocked mitochondrial depolarization after IR, further confirming that mitochondrial depolarization was due to MPT onset. EDHB decreased mitochondrial depolarization to 16% and prevented the MPT. Tin protoporphyrin (10 ?mol/kg sc), an HO-1 inhibitor, partially abrogated protection by EDHB against ALT release, necrosis, and mitochondrial depolarization. In conclusion, IR causes the MPT and mitochondrial dysfunction, leading to hepatocellular death. PHI prevents MPT onset and liver damage through an effect mediated partially by HO-1. PMID:18772364

  11. Silicon-on-glass pore network micromodels with oxygen-sensing fluorophore films for chemical imaging and defined spatial structure

    SciTech Connect

    Grate, Jay W.; Kelly, Ryan T.; Suter, Jonathan D.; Anheier, Norman C.

    2012-11-21

    Pore network microfluidic models were fabricated by a silicon-on-glass technique that provides the precision advantage of dry etched silicon while creating a structure that is transparent across all microfluidic channels and pores, and can be imaged from either side. A silicon layer is bonded to an underlying borosilicate glass substrate and thinned to the desired height of the microfluidic channels and pores. The silicon is then patterned and through-etched by deep reactive ion etching (DRIE), with the underlying glass serving as an etch stop. After bonding on a transparent glass cover plate, one obtains a micromodel in oxygen impermeable materials with water wet surfaces where the microfluidic channels are transparent and structural elements such as the pillars creating the pore network are opaque. The micromodel can be imaged from either side. The advantageous features of this approach in a chemical imaging application are demonstrated by incorporating a Pt porphyrin fluorophore in a PDMS film serving as the oxygen sensing layer and a bonding surface, or in a polystyrene film coated with a PDMS layer for bonding. The sensing of a dissolved oxygen gradient was demonstrated using fluorescence lifetime imaging, and it is shown that different matrix polymers lead to optimal use in different ranges dissolved oxygen concentration. Imaging with the opaque pillars in between the observation direction and the continuous fluorophore film yields images that retain spatial information in the sensor image.

  12. Silicon-on-glass pore network micromodels with oxygen-sensing fluorophore films for chemical imaging and defined spatial structure.

    PubMed

    Grate, Jay W; Kelly, Ryan T; Suter, Jonathan; Anheier, Norm C

    2012-11-21

    Pore network microfluidic models were fabricated by a silicon-on-glass technique that provides the precision advantage of dry etched silicon while creating a structure that is transparent across all microfluidic channels and pores, and can be imaged from either side. A silicon layer is bonded to an underlying borosilicate glass substrate and thinned to the desired height of the microfluidic channels and pores. The silicon is then patterned and through-etched by deep reactive ion etching (DRIE), with the underlying glass serving as an etch stop. After bonding on a transparent glass cover plate, one obtains a micromodel in oxygen impermeable materials with water-wet surfaces where the microfluidic channels are transparent and structural elements such as the pillars creating the pore network are opaque. The advantageous features of this approach in a chemical imaging application are demonstrated by incorporating a Pt porphyrin fluorophore in a PDMS film serving as the oxygen-sensing layer and a bonding surface, or in a polystyrene film coated with a PDMS layer for bonding. The sensing of a dissolved oxygen gradient was demonstrated using fluorescence lifetime imaging, and it is shown that different matrix polymers lead to optimal use in different ranges of oxygen concentration. Imaging with the opaque pillars in between the observation direction and the continuous fluorophore film yields images that retain defined spatial structure in the sensor image. PMID:22995983

  13. Handheld multispectral fluorescence lifetime imaging system for in vivo applications

    PubMed Central

    Cheng, Shuna; Cuenca, Rodrigo M.; Liu, Boang; Malik, Bilal H.; Jabbour, Joey M.; Maitland, Kristen C.; Wright, John; Cheng, Yi-Shing Lisa; Jo, Javier A.

    2014-01-01

    There is an increasing interest in the application of fluorescence lifetime imaging (FLIM) for medical diagnosis. Central to the clinical translation of FLIM technology is the development of compact and high-speed clinically compatible systems. We present a handheld probe design consisting of a small maneuverable box fitted with a rigid endoscope, capable of continuous lifetime imaging at multiple emission bands simultaneously. The system was characterized using standard fluorescent dyes. The performance was then further demonstrated by imaging a hamster cheek pouch in vivo, and oral mucosa tissue both ex vivo and in vivo, all using safe and permissible exposure levels. Such a design can greatly facilitate the evaluation of FLIM for oral cancer imaging in vivo. PMID:24688824

  14. Theory and in Vivo Application of Electroporative Gene Delivery

    Microsoft Academic Search

    Stella Somiari; Jill Glasspool-Malone; Joseph J. Drabick; Richard A. Gilbert; Richard Heller; Mark J. Jaroszeski; Robert W. Malone

    2000-01-01

    Efficient and safe methods for delivering exogenous genetic material into tissues must be developed before the clinical potential of gene therapy will be realized. Recently, in vivo electroporation has emerged as a leading technology for developing nonviral gene therapies and nucleic acid vaccines (NAV). Electroporation (EP) involves the application of pulsed electric fields to cells to enhance cell permeability, resulting

  15. From spin-labeled proteins to in vivo EPR applications

    Microsoft Academic Search

    Lawrence J. Berliner

    2010-01-01

    This is a historical overview of the advent of applications of spin labeling to biological systems and the subsequent developments\\u000a from the perspective of a scientist who was working as a Ph.D. student when the technique was conceived and was fortunate\\u000a enough to participate in its development. In addition, the historical development of in vivo applications of EPR on animals

  16. Application of in vivo laser scanning microscope in dermatology

    NASA Astrophysics Data System (ADS)

    Lademann, Juergen; Richter, H.; Otberg, N.; Lawrenz, F.; Blume-Peytavi, U.; Sterry, W.

    2003-10-01

    The state of the art of in-vivo and in-vitro penetration measurements of topically applied substances is described. Only optical techniques represent online measuring methods based on the absorption or scattering properties of the topically applied substances. Laser scanning microscopy (LSM) has become a promising method for investigations in dermatology and skin physiology, after it was possible to analyze the skin surface on any body side in-vivo. In the present paper the application of a dermatological laser scanning microscope for penetration and distribution measurements of topically applied substances is described. The intercellular and follicular penetration pathways were studied.

  17. Comparative study of optical fluorescent nanosensors (PEBBLEs) and fiber optic microsensors for oxygen sensing

    NASA Astrophysics Data System (ADS)

    Chen-Esterlit, Zoe; Peteu, Serban F.; Clark, Heather A.; McDonald, William; Kopelman, Raoul

    1999-05-01

    In this paper we report the use of phase sensitive fluorometry to obtain preliminary results from opto-chemical fluorescent oxygen nanosensors. PEBBLE (Probe Encapsulated By Biologically Localized Embedding) sensors were fabricated by immobilizing tris(4,7-diphenyl-1,10-phenanthroline)Ru(II) chloride and tris(1,10-phenanthroline)Ru(II) chloride within a polyacrylamide matrix. PEBBLEs have diameters of 20 - 200 nm and exhibit excellent performance for dissolved oxygen detection. Their performance is compared with micrometer-sized (10 - 20 micrometer) optical fiber sensors and free dye in solution. Oxygen sensing ability of PEBBLEs was tested in the presence of other quenchers and compared with free dyes in solution. While PEBBLEs have been developed for minimally invasive intracellular chemical analysis they show additional advantages, such as increased dynamic range, compared to microsensors, and an absence of interference (quenching) by heavy ions in contrast to free dye solutions.

  18. Biodegradable luminescent porous silicon nanoparticles for in vivo applications

    PubMed Central

    Park, Ji-Ho; Gu, Luo; von Maltzahn, Geoffrey; Ruoslahti, Erkki; Bhatia, Sangeeta N.; Sailor, Michael J.

    2011-01-01

    Nanomaterials that can circulate in the body hold great potential to diagnose and treat disease1–4. For such applications, it is important that the nanomaterials be harmlessly eliminated from the body in a reasonable period of time after they carry out their diagnostic or therapeutic function. Despite efforts to improve their targeting efficiency, significant quantities of systemically administered nanomaterials are cleared by the mononuclear phagocytic system before finding their targets, increasing the likelihood of unintended acute or chronic toxicity. However, there has been little effort to engineer the self-destruction of errant nanoparticles into non-toxic, systemically eliminated products. Here, we present luminescent porous silicon nanoparticles (LPSiNPs) that can carry a drug payload and of which the intrinsic near-infrared photoluminescence enables monitoring of both accumulation and degradation in vivo. Furthermore, in contrast to most optically active nanomaterials (carbon nanotubes, gold nanoparticles and quantum dots), LPSiNPs self-destruct in a mouse model into renally cleared components in a relatively short period of time with no evidence of toxicity. As a preliminary in vivo application, we demonstrate tumour imaging using dextran-coated LPSiNPs (D-LPSiNPs). These results demonstrate a new type of multifunctional nanostructure with a low-toxicity degradation pathway for in vivo applications. PMID:19234444

  19. Clinical applications of in vivo fluorescence confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Oh, Chilhwan; Park, Sangyong; Kim, Junhyung; Ha, Seunghan; Park, Gyuman; Lee, Gunwoo; Lee, Onseok; Chun, Byungseon; Gweon, Daegab

    2008-02-01

    Living skin for basic and clinical research can be evaluated by Confocal Laser Scanning Microscope (CLSM) non-invasively. CLSM imaging system can achieve skin image its native state either "in vivo" or "fresh biopsy (ex vivo)" without fixation, sectioning and staining that is necessary for routine histology. This study examines the potential fluorescent CLSM with a various exogenous fluorescent contrast agent, to provide with more resolution images in skin. In addition, in vivo fluorescent CLSM researchers will be extended a range of potential clinical application. The prototype of our CLSM system has been developed by Prof. Gweon's group. The operating parameters are composed of some units, such as illuminated wavelength 488 nm, argon illumination power up to 20mW on the skin, objective lens, 0.9NA oil immersion, axial resolution 1.0?m, field of view 200?m x 100?m (lateral resolution , 0.3?m). In human volunteer, fluorescein sodium was administrated topically and intradermally. Animal studies were done in GFP transgenic mouse, IRC mouse and pig skin. For imaging of animal skin, fluorescein sodium, acridine orange, and curcumine were used for fluorescein contrast agent. We also used the GFP transgenic mouse for fluorescein CLSM imaging. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. Acridin Orange can be highlight nuclei in viable keratinocyte. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, hair and eccrine gland. In intact skin, absorption of fluorescein sodium by individual corneocyte and hair. Intradermal administrated the fluorescein sodium, distinct outline of keratinocyte cell border could be seen. In papillary dermis, fluorescein distribution is more homogeneous. Curcumin is a yellow food dye that has similar fluorescent properties to fluorescein sodium. In vivo CLSM of transgenic GFP mouse enable on in vivo, high resolution view of GFP expressing skin tissue. GFP signals are brightest in corneocyte, kertinocyte, skin appendage and blood vessels. In conclusion, this study demonstrates the usefulness of CLSM as technique for imaging skin in vivo. In addition, CLSM is non-invasive, the same tissue site may be imaged over a period of time to monitor the various change such as wound healing, severity of skin diseases and effect of therapeutic management.

  20. Applications of nuclear technologies for in-vivo elemental analysis

    SciTech Connect

    Cohn, S.H.; Ellis, K.J.; Vartsky, D.; Wielopolski, L.

    1982-01-01

    Measurement facilities developed, to date, include a unique whole-body-counter, (WBC); a total-body neutron-activation facility (TBNAA); and a partial-body activation facility (PBNAA). A variation of the prompt-gamma neutron-activation technique for measuring total-body nitrogen was developed to study body composition of cancer patients and the effect of nutritional regimens on the composition. These new techniques provide data in numerous clinical studies not previously amenable to investigation. The development and perfection of these techniques provide unique applications of radiation and radioisotopes to the early diagnosis of certain diseases and the evaluation of therapeutic programs. The PBNAA technique has been developed and calibrated for in-vivo measurement of metals. Development has gone forward on prompt-gamma neutron activation for the measurement of cadmium, x-ray fluorescence (XRF) for measurement of iron. Other techniques are being investigated for in-vivo measurement of metals such as silicon and beryllium.

  1. Engineering the oxygen sensing regulation results in an enhanced recombinant human hemoglobin production by Saccharomyces cerevisiae.

    PubMed

    Martínez, José L; Liu, Lifang; Petranovic, Dina; Nielsen, Jens

    2015-01-01

    Efficient production of appropriate oxygen carriers for transfusions (blood substitutes or artificial blood) has been pursued for many decades, and to date several strategies have been used, from synthetic polymers to cell-free hemoglobin carriers. The recent advances in the field of metabolic engineering also allowed the generation of different genetically modified organisms for the production of recombinant human hemoglobin. Several studies have showed very promising results using the bacterium Escherichia coli as a production platform, reporting hemoglobin titers above 5% of the total cell protein content. However, there are still certain limitations regarding the protein stability and functionality of the recombinant hemoglobin produced in bacterial systems. In order to overcome these limitations, yeast systems have been proposed as the eukaryal alternative. We recently reported the generation of a set of plasmids to produce functional human hemoglobin in Saccharomyces cerevisiae, with final titers of active hemoglobin exceeding 4% of the total cell protein. In this study, we propose a strategy for further engineering S. cerevisiae by altering the oxygen sensing pathway by deleting the transcription factor HAP1, which resulted in an increase of the final recombinant active hemoglobin titer exceeding 7% of the total cellular protein. PMID:25082441

  2. Cellular oxygen sensing: Crystal structure of hypoxia-inducible factor prolyl hydroxylase (PHD2)

    PubMed Central

    McDonough, Michael A.; Li, Vivian; Flashman, Emily; Chowdhury, Rasheduzzaman; Mohr, Christopher; Liénard, Benoît M. R.; Zondlo, James; Oldham, Neil J.; Clifton, Ian J.; Lewis, Jeffrey; McNeill, Luke A.; Kurzeja, Robert J. M.; Hewitson, Kirsty S.; Yang, Evelyn; Jordan, Steven; Syed, Rashid S.; Schofield, Christopher J.

    2006-01-01

    Cellular and physiological responses to changes in dioxygen levels in metazoans are mediated via the posttranslational oxidation of hypoxia-inducible transcription factor (HIF). Hydroxylation of conserved prolyl residues in the HIF-? subunit, catalyzed by HIF prolyl-hydroxylases (PHDs), signals for its proteasomal degradation. The requirement of the PHDs for dioxygen links changes in dioxygen levels with the transcriptional regulation of the gene array that enables the cellular response to chronic hypoxia; the PHDs thus act as an oxygen-sensing component of the HIF system, and their inhibition mimics the hypoxic response. We describe crystal structures of the catalytic domain of human PHD2, an important prolyl-4-hydroxylase in the human hypoxic response in normal cells, in complex with Fe(II) and an inhibitor to 1.7 Å resolution. PHD2 crystallizes as a homotrimer and contains a double-stranded ?-helix core fold common to the Fe(II) and 2-oxoglutarate-dependant dioxygenase family, the residues of which are well conserved in the three human PHD enzymes (PHD 1–3). The structure provides insights into the hypoxic response, helps to rationalize a clinically observed mutation leading to familial erythrocytosis, and will aid in the design of PHD selective inhibitors for the treatment of anemia and ischemic disease. PMID:16782814

  3. A green-emitting Cu complex for oxygen-sensing purpose: Synthesis, characterization and photophysical features.

    PubMed

    Hui, Han; Wei, Li; Zhentao, Liu; Xiangen, Han

    2015-05-01

    In the present work, a green-emitting Cu(I) complex [Cu(BT-Et)(POP)]BF4 was synthesized and fully characterized, where BT-Et=4-(1-ethyl-1H-benzo[d]imidazol-2-yl)thiazole, POP=bis(2-(diphenylphosphanyl)phenyl) ether, respectively. An ethyl group was connected onto the diamine ligand to breach ?-? attraction within solid [Cu(BT-Et)(POP)]BF4, favoring O2 molecule attack and sensitivity improvement. Its molecular identity was confirmed by single crystal analysis and theoretical calculation. [Cu(BT-Et)(POP)]BF4 emitted long-lived green emission peaking at 521nm upon photoexcitation which was vulnerable towards O2 molecule, making itself a potential oxygen sensing material. [Cu(BT-Et)(POP)]BF4 was then doped into a silica supporting matrix MCM-41. The resulting composite samples showed sensing behavior towards O2 molecule, with short response time of 10s and sensitivity of 5.56. PMID:25706596

  4. Diversity of magneto-aerotactic behaviors and oxygen sensing mechanisms in cultured magnetotactic bacteria.

    PubMed

    Lefèvre, Christopher T; Bennet, Mathieu; Landau, Livnat; Vach, Peter; Pignol, David; Bazylinski, Dennis A; Frankel, Richard B; Klumpp, Stefan; Faivre, Damien

    2014-07-15

    Microorganisms living in gradient environments affect large-scale processes, including the cycling of elements such as carbon, nitrogen or sulfur, the rates and fate of primary production, and the generation of climatically active gases. Aerotaxis is a common adaptation in organisms living in the oxygen gradients of stratified environments. Magnetotactic bacteria are such gradient-inhabiting organisms that have a specific type of aerotaxis that allows them to compete at the oxic-anoxic interface. They biomineralize magnetosomes, intracellular membrane-coated magnetic nanoparticles, that comprise a permanent magnetic dipole that causes the cells to align along magnetic field lines. The magnetic alignment enables them to efficiently migrate toward an optimal oxygen concentration in microaerobic niches. This phenomenon is known as magneto-aerotaxis. Magneto-aerotaxis has only been characterized in a limited number of available cultured strains. In this work, we characterize the magneto-aerotactic behavior of 12 magnetotactic bacteria with various morphologies, phylogenies, physiologies, and flagellar apparatus. We report six different magneto-aerotactic behaviors that can be described as a combination of three distinct mechanisms, including the reported (di-)polar, axial, and a previously undescribed mechanism we named unipolar. We implement a model suggesting that the three magneto-aerotactic mechanisms are related to distinct oxygen sensing mechanisms that regulate the direction of cells' motility in an oxygen gradient. PMID:25028894

  5. Reversed oxygen sensing using colloidal quantum wells towards highly emissive photoresponsive varnishes

    PubMed Central

    Lorenzon, Monica; Christodoulou, Sotirios; Vaccaro, Gianfranco; Pedrini, Jacopo; Meinardi, Francesco; Moreels, Iwan; Brovelli, Sergio

    2015-01-01

    Colloidal quantum wells combine the advantages of size-tunable electronic properties with vast reactive surfaces that could allow one to realize highly emissive luminescent-sensing varnishes capable of detecting chemical agents through their reversible emission response, with great potential impact on life sciences, environmental monitoring, defence and aerospace engineering. Here we combine spectroelectrochemical measurements and spectroscopic studies in a controlled atmosphere to demonstrate the ‘reversed oxygen-sensing’ capability of CdSe colloidal quantum wells, that is, the exposure to oxygen reversibly increases their luminescence efficiency. Spectroelectrochemical experiments allow us to directly relate the sensing response to the occupancy of surface states. Magneto-optical measurements demonstrate that, under vacuum, heterostructured CdSe/CdS colloidal quantum wells stabilize in their negative trion state. The high starting emission efficiency provides a possible means to enhance the oxygen sensitivity by partially de-passivating the particle surfaces, thereby enhancing the density of unsaturated sites with a minimal cost in term of luminescence losses.

  6. Cellular Oxygen Sensing: Crystal Structure of Hypoxia-Inducible Factor Prolyl Hydroxylase (PHD2)

    SciTech Connect

    McDonough,M.; Li, V.; Flashman, E.; Chowdhury, R.; Mohr, C.; Lienard, B.; Zondlo, J.; Oldham, N.; Clifton, I.; et al.

    2006-01-01

    Cellular and physiological responses to changes in dioxygen levels in metazoans are mediated via the posttranslational oxidation of hypoxia-inducible transcription factor (HIF). Hydroxylation of conserved prolyl residues in the HIF-{alpha} subunit, catalyzed by HIF prolyl-hydroxylases (PHDs), signals for its proteasomal degradation. The requirement of the PHDs for dioxygen links changes in dioxygen levels with the transcriptional regulation of the gene array that enables the cellular response to chronic hypoxia; the PHDs thus act as an oxygen-sensing component of the HIF system, and their inhibition mimics the hypoxic response. We describe crystal structures of the catalytic domain of human PHD2, an important prolyl-4-hydroxylase in the human hypoxic response in normal cells, in complex with Fe(II) and an inhibitor to 1.7 Angstroms resolution. PHD2 crystallizes as a homotrimer and contains a double-stranded {beta}-helix core fold common to the Fe(II) and 2-oxoglutarate-dependant dioxygenase family, the residues of which are well conserved in the three human PHD enzymes (PHD 1-3). The structure provides insights into the hypoxic response, helps to rationalize a clinically observed mutation leading to familial erythrocytosis, and will aid in the design of PHD selective inhibitors for the treatment of anemia and ischemic disease.

  7. Thickness Dependency of Thin Film Samaria Doped Ceria for Oxygen Sensing

    SciTech Connect

    Sanghavi, Rahul P.; Nandasiri, Manjula I.; Kuchibhatla, Satyanarayana V N T; Jiang, Weilin; Varga, Tamas; Nachimuthu, Ponnusamy; Engelhard, Mark H.; Shutthanandan, V.; Thevuthasan, Suntharampillai; Kayani, Asghar N.; Prasad, Shalini

    2011-01-01

    High temperature oxygen sensors are widely used for exhaust gas monitoring in automobiles. This particular study explores the use of thin film single crystalline samaria doped ceria as the oxygen sensing material. Desired signal to noise ratio can be achieved in a material system with high conductivity. From previous studies it is established that 6 atomic percent samarium doping is the optimum concentration for thin film samaria doped ceria to achieve high ionic conductivity. In this study, the conductivity of the 6 atomic percent samaria doped ceria thin film is measured as a function of the sensing film thickness. Hysteresis and dynamic response of this sensing platform is tested for a range of oxygen pressures from 0.001 Torr to 100 Torr for temperatures above 673 K. An attempt has been made to understand the physics behind the thickness dependent conductivity behavior of this sensing platform by developing a hypothetical operating model and through COMSOL simulations. This study can be used to identify the parameters required to construct a fast, reliable and compact high temperature oxygen sensor.

  8. Microwave applicator for hyperthermia treatment on in vivo melanoma model.

    PubMed

    Togni, Paolo; Vrba, Jan; Vannucci, Luca

    2010-03-01

    In this article, we evaluated a planar microwave applicator for in vivo superficial hyperthermia treatments on small tumors in the mouse mimicking treatments for human neoplasms. The design of the applicator, was challenged by the small dimensions of the tumors and unwanted diffusion of heating in the tumor-bearing animals. The required solution was to limit the penetration of microwaves in the depth of the tissue maintaining the full efficacy of hyperthermia. The study was firstly performed by computer simulations of SAR distribution inside a flat homogeneous phantom, considering various thicknesses of the integrated water bolus. Simulations, validated by the measurements, were also used to evaluate the impedance matching. Further tests were performed on homogeneous agar phantom to simulate the temperature distribution in the biological tissue and to preliminary assess the possible modality and schedule of microwave hyperthermia delivery. The in vivo experiments showed the evidence of direct microwave-induced heating and damage of the melanoma tissue in a range of penetration coherent both with computer simulations and phantom studies. The described approach appears perspective for designing limited-microwave-delivery applicators tailored for treatments of human superficial tumors and pre-tumoral lesions. PMID:20033789

  9. In vivo Coherent Raman Imaging for Neuroscience Applications

    NASA Astrophysics Data System (ADS)

    Cote, Daniel

    2010-08-01

    The use of coherent Raman imaging is described for applications in neuroscience. Myelin imaging of the spinal cord can be performed with Raman imaging through the use of the vibration in carbon-hydrogen bonds, dominant in lipids. First, we demonstrate in vivo histomorphometry in live animal for characterization of myelin-related nervous system pathologies. This is used to characterize spinal cord health during multiple sclerosis. Second, Raman spectroscopy of tissue is discussed. We discuss the challenges that live animal imaging brings, together with important aspects of coherent Raman imaging in tissue.

  10. Fluorescence-lifetime-based sensors: oxygen sensing and other biomedical applications

    NASA Astrophysics Data System (ADS)

    Randers-Eichhorn, Lisa; Bartlett, Roscoe A.; Sipior, Jeffrey; Frey, Douglas D.; Carter, Gary M.; Lakowicz, Joseph R.; Rao, Govind

    1996-05-01

    Murine hybridomas were cultivated in tissue culture flasks. Dissolved oxygen tensions in the gas and liquid phases during cell growth were measured non-invasively by an optical oxygen sensor. Readings were made with caps both cracked open and completely closed. During cell growth, gas phase oxygen concentrations remained near atmospheric levels, while the oxygen tension at the bottom of the flasks eventually reached zero. These results suggest that the widespread practice of cracking open tissue culture flask caps during cell growth with a view to supplying adequate oxygen to cells is ineffective and unnecessary. The mass transfer characteristics of the tissue culture flask indicate the dominant resistance to oxygen mass transfer to the cells was the liquid media. The mass transfer rates through the liquid layer under standard laboratory conditions were found to be greater than those predicted by diffusion alone, suggesting microscale mixing. Volumetric and specific oxygen consumption rates were calculated from the sensor data, and were comparable to published values. A recently developed single fiber optic oxygen sensor is described. This new sensor will provide oxygen concentrations at various levels in the tissue culture flasks, allowing more accurate modeling of oxygen diffusion.

  11. Sol-gel-Derived highly sensitive optical oxygen sensing materials using Ru(II) complex via covalent grafting strategy.

    PubMed

    Zhang, Haoran; Lei, Bingfu; Liu, Yingliang; Liu, Xiaotang; Zheng, Mingtao; Dong, Hanwu; Xiao, Yong; Zhang, Jianying

    2014-06-01

    The preparation and oxygen sensing properties of Ru(ll) covalently-grafted and physically-incorporated silica based hybrid materials by sol-gel technique are described in this article. The Ru(II) complexes are successfully grafted onto the backbone of the silica via the condensation reaction of the tetraethoxysilane and the functionalized Ru(II) complex 2-[4'-{3-(Triethoxysilyl)propyl}phenyl]imidazo [4,5-f]-1,10-phenanthroline that contains the hydrolysable tri-alkoxylsilyl group. The luminescence quenching of Ru(II) complex by oxygen within the silica matrix is efficient. The oxygen quenching sensitivity of the covalently-grafted sample is higher than that of the physically-incorporated one due to the strong Si-CH2 bond that is useful to prolong the excited state lifetimes and enhance the photobleaching of the luminophore. The downward oxygen sensing Stern-Volmer plots can be well fitted using the Demas two-site model and the Lehrer model due to the heterogeneous distribution of the Ru(ll) complex within the sol-gel derived silica. PMID:24738438

  12. In vitro - In vivo Correlation: From Theory to Applications

    Microsoft Academic Search

    Jaber Emami

    A key goal in pharmaceutical development of dosage forms is a good understanding of the in vitro and in vivo performance of the dosage forms. One of the challenges of biopharmaceutics research is correlating in vitro drug release information of various drug formulations to the in vivo drug profiles (IVIVC). Thus the need for a tool to reliably correlate in

  13. An optical biopsy system with miniaturized Raman and spectral imaging probes; in vivo animal and ex vivo clinical application studies

    NASA Astrophysics Data System (ADS)

    Sato, Hidetoshi; Suzuki, Toshiaki; Andriana, Bibin B.; Morita, Shin'ichi; Maruyama, Atsushi; Shinzawa, Hideyuki; Komachi, Yuichi; Kanai, Gen'ichi; Ura, Nobuo; Masutani, Koji; Matsuura, Yuji; Toi, Masakazu; Shimosegawa, Toru; Ozaki, Yukihiro

    2009-02-01

    An optical biopsy system which equips miniaturized Raman probes, a miniaturized endoscope and a fluorescent image probe has been developed for in vivo studies of live experimental animals. The present report describes basic optical properties of the system and its application studies for in vivo cancer model animals and ex vivo human cancer tissues. It was developed two types of miniaturized Raman probes, micro Raman probe (MRP) made of optical fibers and ball lens hollow optical fiber Raman probe (BHRP) made of single hollow optical fiber (HOF) with a ball lens. The former has rather large working distance (WD), up to one millimeter. The latter has small WD (~300?m) which depends on the focal length of the ball lens. Use of multiple probes with different WD allows one to obtain detailed information of subsurface tissues in the totally noninvasive manner. The probe is enough narrow to be inserted into a biopsy needle (~19G), for observations of the lesion at deeper inside bodies. The miniaturized endoscope has been applied to observe progression of a stomach cancer in the same rat lesion. It was succeeded to visualize structure of non-stained cancer tissue in live model animals by the fluorescent image technique. The system was also applied to ex vivo studies of human breast and stomach cancers.

  14. In vivo and ex vivo applications of gold nanoparticles for biomedical SERS imagingi

    PubMed Central

    Yigit, Mehmet V; Medarova, Zdravka

    2012-01-01

    Surface enhanced Raman scattering (SERS) is a signal-increasing phenomenon that occurs whenever Raman scattering on a metal surface is enhanced many orders of magnitude. Recently SERS has received considerable attention due to its ultrasensitive multiplex imaging capability with strong photostability. It provides rich molecular information on any Raman molecule adsorbed to rough metal surfaces. The signal enhancement is so remarkable that identification of a single molecule is possible. SERS has become a genuine molecular imaging technique. Gold nanoparticles, encoded with Raman reporters, provide a SERS signal and have been used as imaging probes, often referred to as SERS nanoparticles. They have been used for molecular imaging in vivo, ex vivo and in vitro. Detection of picomolar concentrations of target molecules has been achieved by functionalizing the nanoparticles with target recognition ligands. This review focuses on recent achievements in utilizing SERS nanoparticles for in vivo molecular imaging. In the near future, SERS technology may allow detection of disease markers at the single cell level. PMID:23133814

  15. Ex vivo culture of the intestinal epithelium: strategies and applications.

    PubMed

    Leushacke, Marc; Barker, Nick

    2014-08-01

    Limited pools of resident adult stem cells are critical effectors of epithelial renewal in the intestine throughout life. Recently, significant progress has been made regarding the isolation and in vitro propagation of fetal and adult intestinal stem cells in mammals. It is now possible to generate ever-expanding, three-dimensional epithelial structures in culture that closely parallel the in vivo epithelium of the intestine. Growing such organotypic epithelium ex vivo facilitates a detailed description of endogenous niche factors or stem-cell characteristics, as they can be monitored in real time. Accordingly, this technology has already greatly contributed to our understanding of intestinal adult stem-cell renewal and differentiation. Transplanted organoids have also been proven to readily integrate into, and effect the long-term repair of, mouse colonic epithelia in vivo, establishing the organoid culture as a promising tool for adult stem cell/gene therapy. In another exciting development, novel genome-editing techniques have been successfully employed to functionally repair disease loci in cultured intestinal stem cells from human patients with a hereditary defect. It is anticipated that this technology will be instrumental in exploiting the regenerative medicine potential of human intestinal stem cells for treating human disorders in the intestinal tract and for creating near-physiological ex vivo models of human gastrointestinal disease. PMID:24841573

  16. Application of in vivo laser scanning microscope in dermatology

    Microsoft Academic Search

    Juergen Lademann; H. Richter; N. Otberg; F. Lawrenz; U. Blume-Peytavi; W. Sterry

    2003-01-01

    The state of the art of in-vivo and in-vitro penetration measurements of topically applied substances is described. Only optical techniques represent online measuring methods based on the absorption or scattering properties of the topically applied substances. Laser scanning microscopy (LSM) has become a promising method for investigations in dermatology and skin physiology, after it was possible to analyze the skin

  17. A single-unit carbon dioxide-oxygen sensing microelectrode system.

    PubMed

    Kempen, L H; Kreuzer, F

    1975-04-01

    A membrane-covered CO2 microelectrode system is described; it consists of a platinum electrode surronded by a ring-shaped Ag-AgCl electrode, both in contact with an electrolyte layer composed 10 minus 3 M/l quinhydrone in 0.1 N KCl. The resulting oxidation-reduction potential is shown to vary linearly with the logarithm of the PCO2 of the medium. Response time for 95% deflection to a step change in PCO2 of 65 mm Hg is about 1 min i.e. considerably less than in the macroelectrode described previously, but stability is decreased at the same time. Since oxygen in concentrations up to about 21% (air) did not influence the CO2 response and the presence of both CO2 (10.33%) and quinhydrone (10 minus 3 M/l) did not alter the oxygen polarogran, this electorde may be used independently as an oxygen electrode as well. Stability and response time for oxygen were similar to those of the Clark electrode. Possibilities and limitations for in vivo estimation of PCO2 and PO2 are discussed. PMID:1144948

  18. In vivo imaging of free radicals: Applications from mouse to man

    Microsoft Academic Search

    Guanglong He; Alexandre Samouilov; Periannan Kuppusamy; Jay L. Zweier

    2002-01-01

    Free radicals and other paramagnetic species, play an important role in cellular injury and pathophysiology. EPR spectroscopy and imaging has emerged as an important tool for non-invasive in vivo measurement and spatial mapping of free radicals in biological tissues. Extensive applications have been performed in small animals such as mice and recently applications in humans have been performed. Spatial EPR

  19. Transesophageal ultrasound applicator for sector-based thermal ablation: First in vivo experiments

    E-print Network

    Paris-Sud XI, Université de

    Transesophageal ultrasound applicator for sector-based thermal ablation: First in vivo experiments@lyon.inserm.fr Running title: Transesophageal plane transducer 1 #12;Transesophageal ultrasound applicator for sector that High Intensity Ultrasound (HIU) can induce rapid, complete and well-defined coagulation necrosis

  20. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III

    2004-10-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. High temperature measurements of the emission of clusters in sol gel films show that the luminescence intensity from the films follow a 1/T relationship from room temperature to 150 C, and then declines at a slower rate at higher temperatures. The large number of photons available at 230 C is consistent with simple low cost optics for fiber optic probes based on the emission from clusters in sol gel films.

  1. In Vivo Application of Optogenetics for Neural Circuit Analysis

    PubMed Central

    2012-01-01

    Optogenetics combines optical and genetic methods to rapidly and reversibly control neural activities or other cellular functions. Using genetic methods, specific cells or anatomical pathways can be sensitized to light through exogenous expression of microbial light activated opsin proteins. Using optical methods, opsin expressing cells can be rapidly and reversibly controlled by pulses of light of specific wavelength. With the high spatial temporal precision, optogenetic tools have enabled new ways to probe the causal role of specific cells in neural computation and behavior. Here, we overview the current state of the technology, and provide a brief introduction to the practical considerations in applying optogenetics in vivo to analyze neural circuit functions. PMID:22896801

  2. Somatic pairing of chromosome 19 in renal oncocytoma is associated with deregulated EGLN2-mediated [corrected] oxygen-sensing response.

    PubMed

    Koeman, Julie M; Russell, Ryan C; Tan, Min-Han; Petillo, David; Westphal, Michael; Koelzer, Katherine; Metcalf, Julie L; Zhang, Zhongfa; Matsuda, Daisuke; Dykema, Karl J; Houseman, Heather L; Kort, Eric J; Furge, Laura L; Kahnoski, Richard J; Richard, Stéphane; Vieillefond, Annick; Swiatek, Pamela J; Teh, Bin Tean; Ohh, Michael; Furge, Kyle A

    2008-01-01

    Chromosomal abnormalities, such as structural and numerical abnormalities, are a common occurrence in cancer. The close association of homologous chromosomes during interphase, a phenomenon termed somatic chromosome pairing, has been observed in cancerous cells, but the functional consequences of somatic pairing have not been established. Gene expression profiling studies revealed that somatic pairing of chromosome 19 is a recurrent chromosomal abnormality in renal oncocytoma, a neoplasia of the adult kidney. Somatic pairing was associated with significant disruption of gene expression within the paired regions and resulted in the deregulation of the prolyl-hydroxylase EGLN2 [corrected] a key protein that regulates the oxygen-dependent degradation of hypoxia-inducible factor (HIF). Overexpression of EGLN2 [corrected] in renal oncocytoma increased ubiquitin-mediated destruction of HIF and concomitantly suppressed the expression of several HIF-target genes, including the pro-death BNIP3L gene. The transcriptional changes that are associated with somatic pairing of chromosome 19 mimic the transcriptional changes that occur following DNA amplification. Therefore, in addition to numerical and structural chromosomal abnormalities, alterations in chromosomal spatial dynamics should be considered as genomic events that are associated with tumorigenesis. The identification of EGLN2 as a significantly deregulated gene that maps within the paired chromosome region directly implicates defects in the oxygen-sensing network to the biology of renal oncocytoma. PMID:18773095

  3. Somatic Pairing of Chromosome 19 in Renal Oncocytoma Is Associated with Deregulated ELGN2-Mediated Oxygen-Sensing Response

    PubMed Central

    Petillo, David; Westphal, Michael; Koelzer, Katherine; Metcalf, Julie L.; Zhang, Zhongfa; Matsuda, Daisuke; Dykema, Karl J.; Houseman, Heather L.; Kort, Eric J.; Furge, Laura L.; Kahnoski, Richard J.; Richard, Stéphane; Vieillefond, Annick; Swiatek, Pamela J.; Teh, Bin Tean; Ohh, Michael; Furge, Kyle A.

    2008-01-01

    Chromosomal abnormalities, such as structural and numerical abnormalities, are a common occurrence in cancer. The close association of homologous chromosomes during interphase, a phenomenon termed somatic chromosome pairing, has been observed in cancerous cells, but the functional consequences of somatic pairing have not been established. Gene expression profiling studies revealed that somatic pairing of chromosome 19 is a recurrent chromosomal abnormality in renal oncocytoma, a neoplasia of the adult kidney. Somatic pairing was associated with significant disruption of gene expression within the paired regions and resulted in the deregulation of the prolyl-hydroxylase ELGN2, a key protein that regulates the oxygen-dependent degradation of hypoxia-inducible factor (HIF). Overexpression of ELGN2 in renal oncocytoma increased ubiquitin-mediated destruction of HIF and concomitantly suppressed the expression of several HIF-target genes, including the pro-death BNIP3L gene. The transcriptional changes that are associated with somatic pairing of chromosome 19 mimic the transcriptional changes that occur following DNA amplification. Therefore, in addition to numerical and structural chromosomal abnormalities, alterations in chromosomal spatial dynamics should be considered as genomic events that are associated with tumorigenesis. The identification of EGLN2 as a significantly deregulated gene that maps within the paired chromosome region directly implicates defects in the oxygen-sensing network to the biology of renal oncocytoma. PMID:18773095

  4. Linear oxygen-sensing response from a rhenium complex induced by heavy atom: synthesis, characterization, photophysical study and sensing performance.

    PubMed

    Wan, Pu; Zhao, Lun; Wang, Lisha; Xu, Guangyang

    2013-08-01

    In this paper, we synthesized a Br-containing ligand of 2-(4-bromophenyl)-5-(pyridin-2-yl)-1,3,4-oxadiazole and its corresponding Re(I) complex. Their synthesis, characterization, single crystal structure, electronic transitions and photophysical property were presented and discussed in detail. This Re(I) complex was found to be a yellow emitter with slim ???* radiative decay contribution, and its emission was also found to be sensitive towards O2. By doping this Re(I) complex into a polymer matrix, the oxygen-sensing performance of the resulted composite nanofibers was also investigated. Owing to the porous structure of the supporting matrix, the optimal sample gave the highest sensitivity of 3.91 with short response time of only 9 s. In addition, the linearity of the Stern-Volmer plots was greatly improved due to the highly pure emissive center triggered by heavy-atom turbulence effect from Br atom, as indicted by theoretical calculation result. PMID:23673241

  5. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D. J. Osborn; Po Zhang

    2006-09-30

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. Our approach towards immobilizing the potassium salt of the molybdenum cluster, K{sub 2}Mo{sub 6}Cl{sub 14}, at the far end of an optical fiber is to embed the cluster in a thermally cured sol-gel matrix particle. Due to the improved mechanical properties of this approach high temperature sensor measurements were performed up to 100 C. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  6. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn; Po Zhang

    2006-06-30

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. Our approach towards immobilizing the potassium salt of the molybdenum cluster, K{sub 2}Mo{sub 6}Cl{sub 14}, at the far end of an optical fiber is to embed the cluster in a thermally cured sol-gel matrix particle. This particle-in-binder approach affords fibers with greatly improved mechanical properties, as compared to previous approaches. The sensor was characterized in 2-21% gas phase oxygen at 40, 70 and 100 C. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  7. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D. J. Osborn; Po Zhang

    2006-09-30

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications has been developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. We report on a fiber optic technique for detection of gas phase oxygen up to 100 C based on the {sup 3}O{sub 2} quenching of the luminescence from molybdenum chloride clusters, K{sub 2}Mo{sub 6}Cl{sub 14}. The inorganic sensing film is a composite of sol-gel particles embedded in a thin, oxygen permeable sol-gel binder. The particles are comprised of thermally stable, luminescent K{sub 2}Mo{sub 6}Cl{sub 14} clusters dispersed in a fully equilibrated sol-gel matrix. From 40 to 100 C, the fiber sensor switches {approx}6x in intensity in response to alternating pulses of <0.001% O2 and 21% O{sub 2} between two well defined levels with a response time of 10 s. The sensor signal is a few nW for an input pump power of 250 {micro}W. The normalized sensor signal is linear with molar oxygen concentration and fits the theoretical Stern-Volmer relationship. Although the sensitivity decreases with temperature, sensitivity at 100 C is 160 [O{sub 2}]{sup -1}. These parameters are well suited for in-situ, real-time monitoring of oxygen for industrial process control applications.

  8. A Telemedicine Application to Schedule Temperature in an In Vivo Sensor Network for Cancer Treatment

    PubMed Central

    Kamal, Rossi; Lee, Seok-Geun

    2012-01-01

    Abstract Wireless communication has played a significant role in modern healthcare systems. However, the death toll from chronic diseases, such as cancer, continues to increase. Hyperthermia combined with radiotherapy and/or chemotherapy is a promising strategy for cancer treatment, and temperature control is critical for the success of this intervention. In vivo sensors are an emerging technology in healthcare. Thermal awareness has also received attention in in vivo sensor research. In this context, we have been motivated to use in vivo sensors to regulate the temperature changes in cancer cells during combined treatment. Limitations in existing in vivo thermal-aware routing algorithms motivated us to use the in vivo “lightweight rendezvous routing” approach. However, smartphone-driven telemedicine applications are proliferating to provide remote healthcare and collaborative consultation, required in combined therapies. In this context, we have proposed a telemedicine application where a smartphone not only regulates temperature scheduling in in vivo sensors, but also communicates with local or remote clinicians to maintain collaborative efforts for combined therapies against cancer. PMID:23234425

  9. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III

    2003-07-01

    Mo{sub 6}Cl{sub 12}, a cluster compound whose luminescence depends on the ambient concentration of oxygen, is the basis for a real-time oxygen sensor for combustion applications. Previously, the properties of Mo{sub 6}Cl{sub 12} have largely been studied at room temperature; these studies have now been extended to 200 C. Optical microscopy shows that Mo{sub 6}Cl{sub 12} undergoes a steady change in color as it is heated from room temperature to 200 C, changing from canary yellow to crimson and then back to canary yellow. Concurrent thermal gravimetric analyses show a small weight loss for Mo{sub 6}Cl{sub 12} that is consistent with loss of water or HCl from the clusters. These changes are reversible. Absorption and fluorescence emission spectroscopy of Mo{sub 6}Cl{sub 12} heated to 200 C for two hours shows no change in the photophysical parameters compared to the control sample that was not heat cycled.

  10. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III; Po Zhang

    2005-04-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. One of the critical materials issues is to demonstrate that the luminescent cluster immobilized in the sol-gel porous support can withstand high temperature. At the same time the sol-gel matrix must have a high permeability to oxygen. Using a potassium salt of the molybdenum clusters, K{sub 2}Mo{sub 6}Cl{sub 14}, we have established the conditions necessary for deposition of optical quality sol-gel films. From spectroscopic measurements of the film we have shown that the cluster luminescence is stable following heat cycling of 54 hours at 200 C. Quenching of a factor of 1.5X between pure nitrogen and 21% oxygen was observed from in-situ measurements of films heated directly at 200 C. An automated system for characterizing fiber optic oxygen sensors up to 220 C with a temporal resolution better than 10 s is under construction. We estimate a signal of 6 x 10{sup 8} photons/s after complete quenching in 21% oxygen. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  11. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III; Po Zhang

    2005-01-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. One of the critical materials issues is to demonstrate that the luminescent cluster immobilized in the sol-gel porous support can withstand high temperature. At the same time the sol-gel matrix must have a high permeability to oxygen. Using a potassium salt of the molybdenum clusters, K{sub 2}Mo{sub 6}Cl{sub 14}, we have established the conditions necessary for deposition of optical quality sol-gel films. From spectroscopic measurements of the film we have shown that the cluster luminescence is stable following heat cycling of 1 hour at 250 C. Quenching of a factor of 4X between pure nitrogen and 21% oxygen was observed for films cured directly at 200 C. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  12. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III; Po Zhang

    2005-07-01

    A reflection mode fiber optic oxygen sensor is being developed that can operate at high temperatures for power plant applications. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. Two critical materials issues are the cluster's ability to withstand high temperatures when immobilized in a porous the sol-gel support, and whether after heating to high temperatures, the sol-gel matrix maintains a high and constant permeability to oxygen to support rapid quenching of luminescence. We used a composite materials approach to prepare stable sensing layers on optical fibers. We dispersed 60 w/w% of a pre-cured sol-gel composite containing the potassium salt of molybdenum clusters (K{sub 2}Mo{sub 6}Cl{sub 14}) into a sol-gel binder solution, and established the conditions necessary for deposition of sol-gel films on optical fibers and planar substrates. The fiber sensor has an output signal of 5 nW when pumped with an inexpensive commercial 365 nm ultraviolet light emitting diode (LED). Quenching of the sensor signal by oxygen was observed up to a gas temperature of 175 C with no degradation of the oxygen permeability of the composite after high temperature cycling. On planar substrates the cluster containing composite responds within <1 second to a gas exchange from nitrogen to oxygen, indicating the feasibility of real-time oxygen detection.

  13. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III

    2004-04-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. The luminescence of Mo{sub 6}Cl{sub 12} immobilized in a sol-gel matrix was measured as a function of heater temperature up to 200 C, in an inert environment. While the luminescence decreased with temperature, the integrated intensity at 200 C should be sufficient to enable detection of the luminescence in a fiber geometry. Previously we found that aging Mo{sub 6}Cl{sub 12} at temperatures above 250 C converts the canary yellow Mo{sub 6}Cl{sub 12} to a non-luminescent gray solid. Optical and thermal aging experiments show that the alkali metal salts of Mo{sub 6}Cl{sub 12} have higher thermal stabilities and remain luminescent after aging at 280 C.

  14. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III; Po Zhang

    2006-05-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. Previously we described a particle-in-binder approach to immobilizing the potassium salt of the molybdenum cluster, K{sub 2}Mo{sub 6}Cl{sub 14}, at the tips of optical fibers. Compared to previous methods, the particle-in-binder approach affords fibers with greatly improved mechanical properties. The response of the sensor to oxygen at 40, 70 and 100 C was measured in 2-21% gas phase oxygen. The normalized sensor signal is linear with molar oxygen concentration and fits the theoretical Stern-Volmer relationship. Although the sensitivity decreases with temperature, at 100 C the sensitivity is 160 [O{sub 2}]{sup -1}. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  15. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III; Po Zhang

    2006-01-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. Previously we described a particle-in-binder approach to immobilizing the potassium salt of a molybdenum cluster, K{sub 2}Mo{sub 6}Cl{sub 14}, at the tips of optical fibers. Compared to previous methods, the particle-in-binder approach affords fibers with greatly improved mechanical properties. We have extensively characterized two fiber sensors at high temperature. We obtain quenching ratios between pure nitrogen and 21% oxygen as high as 3.9 x at 70 C. For the first sensor at 60 C we obtained a {+-} 1% variation in the quenching ratio over 6 cycles of measurement, and monitored the device performance over 23 days. We were able to operate the second sensor continuously for 14 hours at 70 C, and the sensor quenching ratio was stable to 5% over that time period. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  16. Clinical applications of in vivo neutron-activation analysis

    SciTech Connect

    Cohn, S.H.

    1982-01-01

    In vivo neutron activation has opened a new era of both clinical diagnosis and therapy evaluation, and investigation into and modelling of body composition. The techniques are new, but it is already clear that considerable strides can be made in increasing accuracy and precision, increasing the number of elements susceptible to measurement, enhancing uniformity, and reducing the dose required for the measurement. The work presently underway will yield significant data on a variety of environmental contaminants such as Cd. Compositional studies are determining the level of vital constituents such as nitrogen and potassium in both normal subjects and in patients with a variety of metabolic disorders. Therapeutic programs can be assessed while in progress.

  17. In-vivo neutron activation analysis: principles and clinical applications

    SciTech Connect

    Cohn, S.H.

    1982-01-01

    In vivo neutron activation has opened a new era of both clinical diagnosis and therapy evaluation, and investigation into and modelling of body composition. The techniques are new, but it is already clear that considerable strides can be made in increasing accuracy and precision, increasing the number of elements susceptible to measurement, enhancing uniformity, and reducing the dose required for the measurement. The work presently underway will yield significant data on a variety of environmental contaminants such as Cd. Compositional studies are determining the level of vital constituents such as nitrogen and potassium in both normal subjects and in patients with a variety of metabolic disorders. Therapeutic programs can be assessed while in progress. It seems likely that by the end of this century there will have been significant progress with this research tool, and exciting insights obtained into the nature and dynamics of human body composition.

  18. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III; Po Zhang

    2005-10-01

    A reflection mode fiber optic oxygen sensor that can operate at high temperatures for power plant applications is being developed. The sensor is based on the {sup 3}O{sub 2} quenching of the red emission from hexanuclear molybdenum chloride clusters. Previously we immobilized the potassium salt of a molybdenum cluster, K{sub 2}M{sub 6}Cl{sub 14}, in a sol-gel matrix and showed that the luminescence is stable after 54 hours at 200 C, but the quenching ratios were low and the films delaminated after thermal cycling due to densification of the matrix. Three new approaches to solve decreased quenching over time and delamination of films off fiber tips were investigated. In the first approach K{sub 2}Mo{sub 6}Cl{sub 14} embedded in cured sol-gel particles were incorporated into a TEOS based sol-gel. These gave enhanced quenching (6x), but delaminated. Our second approach was to use a commercial cyanoacrylate glue to immobilize the particles onto the tip of an optical fiber. This gave better adhesion and good quenching initially, but eventually the glue degraded upon heating. Our third approach was to use a 55% OtMOS/ TEOS sol-gel binder. Films based on this new sol-gel binder show high quenching ({approx}6x) and superior mechanical stability even after thermal cycling. Sensor measurements on an optical fiber containing K{sub 2}Mo{sub 6}Cl{sub 14} embedded in cured sol-gel particles were obtained from 100 to 25 C. The signal intensity in nitrogen was stable at 2.8 {+-} 0.2 nW, and the quenching ratio (ratio of signal in N{sub 2} vs. 21 % O{sub 2}) varied from 4.4 to 6.9X. These are promising results for a high temperature fiber optical oxygen sensor based on molybdenum chloride clusters.

  19. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    NASA Astrophysics Data System (ADS)

    Darvin, M. E.; Richter, H.; Zhu, Y. J.; Meinke, M. C.; Knorr, F.; Gonchukov, S. A.; Koenig, K.; Lademann, J.

    2014-07-01

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted.

  20. Deep tissue fluorescence imaging and in vivo biological applications

    NASA Astrophysics Data System (ADS)

    Crosignani, Viera; Dvornikov, Alexander; Aguilar, Jose S.; Stringari, Chiara; Edwards, Robert; Mantulin, William W.; Gratton, Enrico

    2012-11-01

    We describe a novel technical approach with enhanced fluorescence detection capabilities in two-photon microscopy that achieves deep tissue imaging, while maintaining micron resolution. Compared to conventional two-photon microscopy, greater imaging depth is achieved by more efficient harvesting of fluorescence photons propagating in multiple-scattering media. The system maintains the conventional two-photon microscopy scheme for excitation. However, for fluorescence collection the detection system harvests fluorescence photons directly from a wide area of the turbid sample. The detection scheme relies on a wide area detector, minimal optical components and an emission path bathed in a refractive-index-matching fluid that minimizes emission photon losses. This detection scheme proved to be very efficient, allowing us to obtain high resolution images at depths up to 3 mm. This technique was applied to in vivo imaging of the murine small intestine (SI) and colon. The challenge is to image normal and diseased tissue in the whole live animal, while maintaining high resolution imaging at millimeter depth. In Lgr5-GFP mice, we have been successful in imaging Lgr5-eGFP positive stem cells, present in SI and colon crypt bases.

  1. In vitro and in vivo Study of 5Methoxypsoralen Skin Concentration after Topical Application

    Microsoft Academic Search

    G. Colombo; A. Zucchi; F. Allegra; P. Colombo; F. Zani; P. Santi

    2003-01-01

    The aim of this work was to study the skin distribution of 5-methoxypsoralen (5-MOP) after application of topical gels, in vitro and in vivo, in both healthy and psoriatic skin sites of 6 psoriatic patients. Drug skin distribution was determined using the thin slicing technique and subsequent HPLC analysis. In the presence of dermatological disease, i.e. psoriasis, the permeability of

  2. Tracking of stem cells in vivo for cardiovascular applications

    PubMed Central

    2014-01-01

    In the past ten years, the concept of injecting stem and progenitor cells to assist with rebuilding damaged blood vessels and myocardial tissue after injury in the heart and peripheral vasculature has moved from bench to bedside. Non-invasive imaging can not only provide a means to assess cardiac repair and, thereby, cellular therapy efficacy but also a means to confirm cell delivery and engraftment after administration. In this first of a two-part review, we will review the different types of cellular labeling techniques and the application of these techniques in cardiovascular magnetic resonance and ultrasound. In addition, we provide a synopsis of the cardiac cellular clinical trials that have been performed to-date. PMID:24406054

  3. Application of in vivo measurements for the management of cyanobacteria breakthrough into drinking water treatment plants.

    PubMed

    Zamyadi, Arash; Dorner, Sarah; Ndong, Mouhamed; Ellis, Donald; Bolduc, Anouka; Bastien, Christian; Prévost, Michèle

    2014-02-01

    The increasing presence of potentially toxic cyanobacterial blooms in drinking water sources and within drinking water treatment plants (DWTPs) has been reported worldwide. The objectives of this study are to validate the application of in vivo probes for the detection and management of cyanobacteria breakthrough inside DWTPs, and to verify the possibility of treatment adjustment based on intensive real-time monitoring. In vivo phycocyanin YSI probes were used to monitor the fate of cyanobacteria in raw water, clarified water, filtered water, and chlorinated water in a full scale DWTP. Simultaneous samples were also taken for microscopic enumeration. The in vivo probe was successfully used to detect the incoming densities of high cyanobacterial cell number into the clarification process and their breakthrough into the filtered water. In vivo probes were used to trace the increase in floating cells over the clarifier, a robust sign of malfunction of the coagulation-sedimentation process. Pre-emptive treatment adjustments, based on in vivo probe monitoring, resulted in successful removal of cyanobacterial cells. The field results on validation of the probes with cyanobacterial bloom samples showed that the probe responses are highly linear and can be used to trigger alerts to take action. PMID:24429778

  4. Applications of the direct photon absorption technique for measuring bone mineral content in vivo. Determination of body composition in vivo

    NASA Technical Reports Server (NTRS)

    Cameron, J. R.

    1972-01-01

    The bone mineral content, BMC, determined by monoenergetic photon absorption technique, of 29 different locations on the long bones and vertebral columns of 24 skeletons was measured. Compressive tests were made on bone from these locations in which the maximum load and maximum stress were measured. Also the ultimate strain, modulus of elasticity and energy absorbed to failure were determined for compact bone from the femoral diaphysis and cancellous bone from the eighth through eleventh thoracic vertebrae. Correlations and predictive relationships between these parameters were examined to investigate the applicability of using the BMC at sites normally measured in vivo, i.e. radius and ulna in estimating the BMC and/or strength of the spine or femoral neck. It was found that the BMC at sites on the same bone were highly correlated r = 0.95 or better; the BMC at sites on different bones were also highly interrelated, r = 0.85. The BMC at various sites on the long bones could be estimated to between 10 and 15 per cent from the BMC of sites on the radius or ulna.

  5. Time-Resolved Microdialysis for In Vivo Neurochemical Measurements and Other Applications

    NASA Astrophysics Data System (ADS)

    Schultz, Kristin N.; Kennedy, Robert T.

    2008-07-01

    Monitoring changes in chemical concentrations over time in complex environments is typically performed using sensors and spectroscopic techniques. Another approach is to couple sampling methods, such as microdialysis, with chromatographic, electrophoretic, or enzymatic assays. Recent advances of such coupling have enabled improvements in temporal resolution, multianalyte capability, and automation. In a sampling and analysis method, the temporal resolution is set by the mass sensitivity of the analytical method, analysis time, and zone dispersion during sampling. Coupling methods with high speed and mass sensitivity to microdialysis sampling help to reduce some of these contributions to yield methods with temporal resolution of seconds. These advances have been primarily used in monitoring neurotransmitters in vivo. This review covers the problems associated with chemical monitoring in the brain, recent advances in using microdialysis for time-resolved in vivo measurements, sample applications, and other potential applications of the technology such as determining reaction kinetics and process monitoring.

  6. In vitro topical application and in vivo pharmacodynamic evaluation of nonivamide hydrogels using Wistar rat as an animal model

    Microsoft Academic Search

    Jia-You Fang; Yann-Lii Leu; Ying-Yue Wang; Yi-Hung Tsai

    2002-01-01

    Nonivamide, a so-called synthetic capsaicin, is a substitute for capsaicin which has a similar chemical structure and pharmacological activities as those of capsaicin. The purposes of this study were to explore the in vivo pharmacodynamic responses of nonivamide in hydrogels using Wistar rat as an animal model and to correlate the in vivo results with in vitro topical application. The

  7. A Biocompatible in Vivo Ligation Reaction and Its Application for Noninvasive Bioluminescent Imaging of Protease Activity in Living

    E-print Network

    Bogyo, Matthew

    reaction has important implications for both in vivo imaging and biocompatible labeling strategies. First studies of biochemical processes on the level of the whole organism. Since multiple animal modelsA Biocompatible in Vivo Ligation Reaction and Its Application for Noninvasive Bioluminescent

  8. In vivo toxicity of enoxaparin encapsulated in mucoadhesive nanoparticles: Topical application in a wound healing model

    NASA Astrophysics Data System (ADS)

    Huber, S. C.; Marcato, P. D.; Barbosa, R. M.; Duran, N.; Annichino-Bizzacchi, J. M.

    2013-04-01

    Wound healing comprises four distinct phases and involves many cell events and biologic markers. The use of nanoparticles for topical application has gaining attention due to its deeper penetration in the skin and the retention capacity of the drug in the site of application. In this study the effect and toxicity of mucoadhesive polymeric nanoparticles loaded with enoxaparin was evaluated in in vivo model of skin ulcer. Our results showed an interesting formulation based on mucoadhesive nanoparticles with enoxaparin that improved wound healing without cytotoxicity in vitro in all endpoint evaluated. Then, this semi-solid formulation is a promising option for skin ulcer treatment.

  9. The application of dermal papillary rings in dermatology by in vivo confocal laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Xiang, W. Z.; Xu, A. E.; Xu, J.; Bi, Z. G.; Shang, Y. B.; Ren, Q. S.

    2010-08-01

    Confocal laser scanning microscopy (CLSM) allows noninvasive visualization of human skin in vivo, without needing to fix or section the tissue. Melanocytes and pigmented keratinocytes at the level of the basal layer form bright dermal papillary rings which are readily amenable to identify in confocal images. Our purpose was to explore the role of dermal papillary rings in assessment of lesion location, the diagnosis, differential diagnosis of lesions and assessment of therapeutic efficacy by in vivo CLSM. Seventy-one patients were imaged with the VivaScope 1500 reflectance confocal microscope provided by Lucid, Inc. The results indicate that dermal papillary rings can assess the location of lesion; the application of dermal papillary rings can provide diagnostic support and differential diagnosis for vitiligo, nevus depigmentosus, tinea versicolor, halo nevus, common nevi, and assess the therapeutic efficacy of NBUVB phototherapy plus topical 0.1 percent tacrolimus ointment for vitiligo. In conclusion, our findings indicate that the dermal papillary rings play an important role in the assessment the location of lesion, diagnosis, differential diagnosis of lesions and assessment of therapeutic efficacy by in vivo CLSM. CLSM may be a promising tool for noninvasive examination in dermatology. However, larger studies are needed to expand the application of dermal papillary rings in dermatology.

  10. In vivo studies of polyacrylate nanoparticle emulsions for topical and systemic applications.

    PubMed

    Greenhalgh, Kerriann; Turos, Edward

    2009-03-01

    We have recently reported on a new nanomedicine containing antibiotic-conjugated polyacrylate nanoparticles, which has shown activity against methicillin-resistant Staphylococcus aureus (MRSA) in vitro and no cytotoxicity toward human dermal cells. The water-based nanoparticle emulsion is capable of solubilizing lipophilic antibiotics for systemic administration, and the nanoparticle drug delivery vehicle has shown protective properties for antibiotics from hydrolytic cleavage by bacterial penicillinases, thus rejuvenating the drug's activity against resistant microbes such as MRSA. Here we report the first in vivo study of this penicillin-conjugated nanoparticle emulsion in determining toxicological responses initiated upon systemic and topical application in a murine model. Favorable results were observed in vivo upon both routes of administration and, when topically applied to a dermal abrasion model, the emulsion enhanced wound healing by an average of 3 to 5 days. This study suggests that polyacrylate nanoparticle-containing emulsions may afford promising opportunities for treating both skin and systemic infections. PMID:18824416

  11. In vivo evaluation of drug delivery after ultrasound application: A new use for the photoacoustic technique

    NASA Astrophysics Data System (ADS)

    Barja, P. R.; Acosta-Avalos, D.; Rompe, P. C. B.; Dos Anjos, F. H.; Marciano, F. R.; da Silva, M. D.

    2005-06-01

    Ultrasound application is a therapeutical resource widely employed in physiotherapy. One of its applications is the phonophoresis, a technique in which the ultrasound radiation is utilized to deliver drugs through the skin to soft tissues. The proposal of our study was to employ the Photoacoustic Technique to evaluate the efficacy of such treatment, analyzing if phonophoresis could enhance drug delivery through skin when compared to the more traditional method of manual massage. The configuration of the system employed was such that it was possible to perform in vivo measurements, which is a pre-requisite for this kind of study. The changes observed in the photoacoustic signal amplitude after each form of drug application were attributed to changes in the thermal effusivity of the system, due to penetration of the drug. The technique was able to detect differences in drug delivery between the specified physiotherapy treatments, indicating that phonophoresis enhances drug absorption by tissue.

  12. Fabricated micro-nano devices for in vivo and in vitro biomedical applications.

    PubMed

    Barkam, Swetha; Saraf, Shashank; Seal, Sudipta

    2013-01-01

    In recent years, the innovative use of microelectromechanical systems (MEMSs) and nanoelectromechanical systems (NEMSs) in biomedical applications has opened wide opportunities for precise and accurate human diagnostics and therapeutics. The introduction of nanotechnology in biomedical applications has facilitated the exact control and regulation of biological environments. This ability is derived from the small size of the devices and their multifunctional capabilities to operate at specific sites for selected durations of time. Researchers have developed wide varieties of unique and multifunctional MEMS/NEMS devices with micro and nano features for biomedical applications (BioMEMS/NEMS) using the state of the art microfabrication techniques and biocompatible materials. However, the integration of devices with the biological milieu is still a fundamental issue to be addressed. Devices often fail to operate due to loss of functionality, or generate adverse toxic effects inside the body. The in vitro and in vivo performance of implantable BioMEMS such as biosensors, smart stents, drug delivery systems, and actuation systems are researched extensively to understand the interaction of the BioMEMS devices with physiological environments. BioMEMS developed for drug delivery applications include microneedles, microreservoirs, and micropumps to achieve targeted drug delivery. The biocompatibility of BioMEMS is further enhanced through the application of tissue and smart surface engineering. This involves the application of nanotechnology, which includes the modification of surfaces with polymers or the self-assembly of monolayers of molecules. Thereby, the adverse effects of biofouling can be reduced and the performance of devices can be improved in in vivo and in vitro conditions. PMID:23894041

  13. Description and application of instrumented staples for measuring in vivo bone strain.

    PubMed

    Buttermann, G R; Janevic, J T; Lewis, J L; Lindquist, C M; Wood, K B; Schendel, M J

    1994-08-01

    In vivo bone strain measurements using strain gages cemented to bony surfaces with cyanoacrylate polymers are limited in duration due to debonding of the gages from bone. As an alternative to the bone bonded strain gages, a technique was developed in which strain gages were first bonded to miniature staples and then the staples embedded into bone. The instrumented staples may be calibrated so that staple strain is directly proportional to bone strain. The method was first validated by comparing the staple output with cemented surface strain gages. Comparison of instrumented staples to cemented strain gages revealed only a 3% deviation from linearity during longitudinal bending; the staples were insensitive to transverse loading. The instrumented staples were then applied to the in vitro canine lumbar spine to determine L2-3 facet loads. Load testing, repeatibility of facet calibration, and validity testing of the in vitro instrumented staples were found to be comparable to that of the previous cemented strain gage techniques. In vivo facet joint application of the instrumented staples for periods of greater than 5 weeks gave load measurements comparable to our previous short-term in vivo studies obtained with cemented strain gages. The advantages of the instrumented staples are a more secure bonding to the bone, and less traumatic surgery for fixation. PMID:8089163

  14. Applications of nuclear techniques for in vivo body composition studies at Brookhaven National Laboratory

    SciTech Connect

    Cohn, S.H.; Ellis, K.J.; Vartsky, D.; Vaswani, A.N.; Wielopolski, L.

    1981-01-01

    A series of technical developments and their clinical applications in various nuclear technologies at Brookhaven National Laboratory is described. These include the development of a portable neutron activation facility for measuring cadmium in vivo in kidney and liver, a technique for the measurement of body iron utilizing nuclear resonant scattering of gamma rays, a non-invasive measure of the skeletal levels of lead by an x-ray fluorescence technique, and the development of a pulsed Van de Graaff generator as a source of pulsed neutrons for the measurement of lung silicon. (ACR)

  15. Non invasive in vivo investigation of hepatobiliary structure and function in STII medaka (Oryzias latipes): methodology and applications

    PubMed Central

    Hardman, Ron C; Kullman, Seth W; Hinton, David E

    2008-01-01

    Background A novel transparent stock of medaka (Oryzias latipes; STII), recessive for all pigments found in chromatophores, permits transcutaneous imaging of internal organs and tissues in living individuals. Findings presented describe the development of methodologies for non invasive in vivo investigation in STII medaka, and the successful application of these methodologies to in vivo study of hepatobiliary structure, function, and xenobiotic response, in both 2 and 3 dimensions. Results Using brightfield, and widefield and confocal fluorescence microscopy, coupled with the in vivo application of fluorescent probes, structural and functional features of the hepatobiliary system, and xenobiotic induced toxicity, were imaged at the cellular level, with high resolution (< 1 ?m), in living individuals. The findings presented demonstrate; (1) phenotypic response to xenobiotic exposure can be investigated/imaged in vivo with high resolution (< 1 ?m), (2) hepatobiliary transport of solutes from blood to bile can be qualitatively and quantitatively studied/imaged in vivo, (3) hepatobiliary architecture in this lower vertebrate liver can be studied in 3 dimensions, and (4) non invasive in vivo imaging/description of hepatobiliary development in this model can be investigated. Conclusion The non-invasive in vivo methodologies described are a unique means by which to investigate biological structure, function and xenobiotic response with high resolution in STII medaka. In vivo methodologies also provide the future opportunity to integrate molecular mechanisms (e.g., genomic, proteomic) of disease and toxicity with phenotypic changes at the cellular and system levels of biological organization. While our focus has been the hepatobiliary system, other organ systems are equally amenable to in vivo study, and we consider the potential for discovery, within the context of in vivo investigation in STII medaka, as significant. PMID:18838008

  16. Polymer-based, flexible glutamate and lactate microsensors for in vivo applications.

    PubMed

    Weltin, Andreas; Kieninger, Jochen; Enderle, Barbara; Gellner, Anne-Kathrin; Fritsch, Brita; Urban, Gerald A

    2014-11-15

    We present a flexible microsensor, based on a polymer substrate, for multiparametric, electrochemical in vivo monitoring. The sensor strip with a microelectrode array at the tip was designed for insertion into tissue, for fast and localized online monitoring of physiological parameters. The microsystem fabrication on a wafer-level is based on a polyimide substrate and includes the patterning of platinum microelectrodes as well as epoxy and dry-film-resist insulation in a cost-effective thin-film and laminate process. A stable, electrodeposited silver/silver chloride reference electrode on-chip and a perm-selective membrane as an efficient interference rejection scheme are integrated on a wafer-level. Amperometric, electrochemical, enzyme-based biosensors for the neurotransmitter L-glutamate and the energy metabolite L-lactate have been developed. Hydrogel membranes or direct cross-linking as stable concepts for the enzyme immobilization are shown. Sensor performance including high selectivity, tailoring of sensitivity and long-term stability is discussed. For glutamate, a high sensitivity of 2.16 nAmm(-2) µM(-1) was found. For lactate, a variation in sensitivity between 2.6 and 32 nAmm(-2)mM(-1) was achieved by different membrane compositions. The in vivo application in an animal model is demonstrated by glutamate measurements in the brain of rats. Local glutamate alterations in the micromolar range and in nanoliter-range volumes can be detected and quantified with high reproducibility and temporal resolution. A novel, versatile platform for the integration of various electrochemical sensors on a small, flexible sensor strip for a variety of in vivo applications is presented. PMID:24880657

  17. Sensitive single-layered oxygen-sensing systems: polypyridyl ruthenium(II) complexes covalently attached or deposited as langmuir-blodgett monolayer on glass surfaces.

    PubMed

    Chu, Ben Wai-Kin; Yam, Vivian Wing-Wah

    2006-08-15

    Oxygen-sensing elements containing single-layered structures of luminescent indicators of ruthenium(II) bipyridyl complexes on glass surfaces prepared by covalent attachment and LB deposition are described. They are capable of detecting gaseous oxygen concentration by luminescence quenching of the indicator with reproducible and large quenching efficiencies that are comparable to the best quenching efficiencies obtained by other ruthenium(II) polypyridine based complexes immobilized in matrixes. The large quenching efficiencies for both films imply that the probe complexes are effectively quenched by oxygen, which is probably due to the thin single-layered structures with large surface-to-area ratio and short distance between the probe complexes and oxygen. PMID:16893250

  18. Transoesophageal ultrasound applicator for sector-based thermal ablation: first in vivo experiments

    PubMed Central

    Melodelima, David; Lafon, Cyril; Prat, Frédéric; Theillère, Yves; Arefiev, Alexei; Cathignol, Dominique

    2003-01-01

    New curative and palliative treatments must be proposed in order to respond to the bad long-term prognosis of esophageal cancers. It has been demonstrated that High Intensity Ultrasound (HIU) can induce rapid, complete and well-defined coagulation necrosis. For the treatment of this cancer, we designed an applicator that uses an intraductal approach. The active part is an air-backed plane transducer. It has an external water-cooling system and operates at 10 MHz. Ex vivo experiments conducted on pig liver demonstrated the ability of this applicator to generate, by rotating the transducer, circular or sector-based coagulation necroses at predetermined depths up to 13 mm with an excellent angular precision. The treatment of sector-based esophageal tumour may be critical where both malignant and healthy tissues are covered by the ultrasound beam. Thus, in vivo trials were conducted on five healthy pig esophaguses in order to determine the maximal thermal dose that will not induce a perforation of the esophagus or surrounding tissues. From the results of previous studies, this dose is high enough in order to treat pathological tissues. These promising results indicate that this ultrasound system represents a safe and effective tool for the clinical treatment of esophageal tumours. PMID:12659916

  19. In vivo Raman spectroscopy of biochemical changes in human skin by cosmetic application

    NASA Astrophysics Data System (ADS)

    Tosato, Maira Gaspar; dos Santos, Edson Pereira; Alves, Rani de Souza; Raniero, Leandro; Menezes, Priscila Fernanda C.; Kruger, Odivânia; Praes, Carlos Eduardo O.; Martin, Airton Abrahão

    2010-02-01

    The skin aging process is mainly accelerated by external agents such as sunlight, air humidity and surfactants action. Changes in protein structures and hydration during the aging process are responsible for skin morphological variations. In this work the human skin was investigated by in vivo Raman spectroscopy before and after the topical applications of a cosmetic on 17 healthy volunteers (age 60 to 75). In vivo Raman spectra of the skin were obtained with a Spectrometer SpectraPro- 2500i (Pi-Acton), CCD detector and a 785 nm laser excitation source, collected at the beginning of experiment without cream (T0), after 30 (T30) and 60 (T60) days using the product. The primary changes occurred in the following spectral regions: 935 cm-1 (?CC), 1060 cm-1 (lipids), 1174 to 1201 cm-1 (tryptofan, phenylalanine and tyrosine), 1302 cm-1 (phospholipids), 1520 to 1580 cm-1 (C=C) and 1650 cm-1 (amide I). These findings indicate that skin positive effects were enhanced by a continuous cream application.

  20. An Ex Vivo Toe Model Used to Assess Applicators for the Iontophoretic Ungual Delivery of Terbinafine

    Microsoft Academic Search

    Anroop B. Nair; Hyun D. Kim; Shawn P. Davis; Robert Etheredge; Michael Barsness; Phillip M. Friden; S. Narasimha Murthy

    2009-01-01

    Purpose  An ex vivo intact toe model was developed to assess two different applicator designs for iontophoretic delivery of terbinafine into\\u000a the nail only or the nail and surrounding skin.\\u000a \\u000a \\u000a \\u000a Methods  Iontophoretic permeation studies were carried out on intact cadaver toes using nail-only and nail\\/skin applicators with a\\u000a current dose of 10 mA*min (0.5 mA for 20 min).\\u000a \\u000a \\u000a \\u000a Results  Iontophoresis enhanced drug permeation and tissue loading

  1. Qualichem In Vivo: A Tool for Assessing the Quality of In Vivo Studies and Its Application for Bisphenol A

    PubMed Central

    Maxim, Laura; van der Sluijs, Jeroen P.

    2014-01-01

    In regulatory toxicology, quality assessment of in vivo studies is a critical step for assessing chemical risks. It is crucial for preserving public health studies that are considered suitable for regulating chemicals are robust. Current procedures for conducting quality assessments in safety agencies are not structured, clear or consistent. This leaves room for criticism about lack of transparency, subjective influence and the potential for insufficient protection provided by resulting safety standards. We propose a tool called “Qualichem in vivo” that is designed to systematically and transparently assess the quality of in vivo studies used in chemical health risk assessment. We demonstrate its use here with 12 experts, using two controversial studies on Bisphenol A (BPA) that played an important role in BPA regulation in Europe. The results obtained with Qualichem contradict the quality assessments conducted by expert committees in safety agencies for both of these studies. Furthermore, they show that reliance on standardized guidelines to ensure scientific quality is only partially justified. Qualichem allows experts with different disciplinary backgrounds and professional experiences to express their individual and sometimes divergent views—an improvement over the current way of dealing with minority opinions. It provides a transparent framework for expressing an aggregated, multi-expert level of confidence in a study, and allows a simple graphical representation of how well the study integrates the best available scientific knowledge. Qualichem can be used to compare assessments of the same study by different health agencies, increasing transparency and trust in the work of expert committees. In addition, it may be used in systematic evaluation of in vivo studies submitted by industry in the dossiers that are required for compliance with the REACH Regulation. Qualichem provides a balanced, common framework for assessing the quality of studies that may or may not be following standardized guidelines. PMID:24489958

  2. Highly purified mussel adhesive protein to secure biosafety for in vivo applications

    PubMed Central

    2014-01-01

    Background Unique adhesive and biocompatibility properties of mussel adhesive proteins (MAPs) are known for their great potential in many tissue engineering and biomedical applications. Previously, it was successfully demonstrated that redesigned hybrid type MAP, fp-151, mass-produced in Gram-negative bacterium Escherichia coli, could be utilized as a promising adhesive biomaterial. However, purification of recombinant fp-151 has been unsatisfactory due to its adhesive nature and polarity which make separation of contaminants (especially, lipopolysaccharide, a toxic Gram-negative cell membrane component) very difficult. Results In the present work, we devised a high resolution purification approach to secure safety standards of recombinant fp-151 for the successful use in in vivo applications. Undesirable impurities were remarkably eliminated as going through sequential steps including treatment with multivalent ion and chelating agent for cell membrane washing, mechanical cell disruption, non-ionic surfactant treatment for isolated inclusion body washing, acid extraction of washed inclusion body, and ion exchange chromatography purification of acid extracted sample. Through various analyses, such as high performance liquid chromatographic purity assay, limulus amoebocyte lysate endotoxin assay, and in vitro mouse macrophage cell tests on inflammation, viability, cytotoxicity, and apoptosis, we confirmed the biological safety of bacterial-derived purified recombinant fp-151. Conclusions Through this purification design, recombinant fp-151 achieved 99.90% protein purity and 99.91% endotoxin reduction that nearly no inflammation response was observed in in vitro experiments. Thus, the highly purified recombinant MAP would be successfully used as a safety-secured in vivo bioadhesive for tissue engineering and biomedical applications. PMID:24725543

  3. A feasibility study of in vivo applications of single beam acoustic tweezers

    NASA Astrophysics Data System (ADS)

    Li, Ying; Lee, Changyang; Chen, Ruimin; Zhou, Qifa; Shung, K. Kirk

    2014-10-01

    Tools that are capable of manipulating micro-sized objects have been widely used in such fields as physics, chemistry, biology, and medicine. Several devices, including optical tweezers, atomic force microscope, micro-pipette aspirator, and standing surface wave type acoustic tweezers have been studied to satisfy this need. However, none of them has been demonstrated to be suitable for in vivo and clinical studies. Single beam acoustic tweezers (SBAT) is a technology that uses highly focused acoustic beam to trap particles toward the beam focus. Its feasibility was first theoretically and experimentally demonstrated by Lee and Shung several years ago. Since then, much effort has been devoted to improving this technology. At present, the tool is capable of trapping a microparticle as small as 1 ?m, as well as a single red blood cell. Although in comparing to other microparticles manipulating technologies, SBAT has advantages of providing stronger trapping force and deeper penetration depth in tissues, and producing less tissue damage, its potential for in vivo applications has yet been explored. It is worth noting that ultrasound has been used as a diagnostic tool for over 50 years and no known major adverse effects have been observed at the diagnostic energy level. This paper reports the results of an initial attempt to assess the feasibility of single beam acoustic tweezers to trap microparticles in vivo inside of a blood vessel. The acoustic intensity of SBAT under the trapping conditions that were utilized was measured. The mechanical index and thermal index at the focus of acoustic beam were found to be 0.48 and 0.044, respectively, which meet the standard of commercial diagnostic ultrasound system.

  4. The application of quantum dots for the melanoma tumor in vivo imaging

    NASA Astrophysics Data System (ADS)

    Feng, Yayi; Zhai, Peng; Wang, Xiaomei; Ying, Ming; Wu, Jinbo; Zhu, Xiaomei; Lin, Guimiao; Chen, Qiang; Xu, Gaixia

    2014-09-01

    Objective: Over the past decade, fluorescent semiconductor nanocrystals, also known as quantum dots (QDs), have been applied in biomedical imaging in vitro and in vivo because of their fascinating optical properties. In this work, we investigated the application of CdTe QDs for tumor fluorescence in vivo imaging. Methods: The transparent dorsal skin fold window chamber (DSFC) was constructed on the 4~6 week-old BALB/c mice. The melanoma cells stably expressing green fluorescent protein ---ZsGreen were transplanted into the chamber and the melanoma DSFC model was established successfully. The water soluble CdTe QDs were synthesized and then administrated in the model through the tail intravenous injection. The fluorescent expression of B16 cells were assayed by fluorescent microscopy, the tumor growth, the blood capillaries distributions and its dynamic changes were observed by stereomicroscopy and laser scanning confocal microscopy. Results: The results demonstrated that the expression efficiency of ZsGreen was 41%, which met the experimental requirement. The tumors was visible inside the chamber after implantation of melanoma cells for 5~6 days, while no obvious changes in mice behaviors were found. After injection of the QDs, CdTe QDs accumulated at the invading edge of a range of solid tumor. We could also observe the tumor cells growth near the blood vessels, the angiogenesis occurred inside the tumor and the local blood capillaries increased. Conclusions: This work provided a new strategy for the tumor in vivo imaging and the development of targeted antineoplastic drugs.

  5. A feasibility study of in vivo applications of single beam acoustic tweezers

    SciTech Connect

    Li, Ying, E-mail: yli582@usc.edu; Lee, Changyang; Chen, Ruimin; Zhou, Qifa; Shung, K. Kirk [NIH Transducer Resource Center and Department of Biomedical Engineering, University of Southern California, Los Angeles, California 90089-1111 (United States)

    2014-10-27

    Tools that are capable of manipulating micro-sized objects have been widely used in such fields as physics, chemistry, biology, and medicine. Several devices, including optical tweezers, atomic force microscope, micro-pipette aspirator, and standing surface wave type acoustic tweezers have been studied to satisfy this need. However, none of them has been demonstrated to be suitable for in vivo and clinical studies. Single beam acoustic tweezers (SBAT) is a technology that uses highly focused acoustic beam to trap particles toward the beam focus. Its feasibility was first theoretically and experimentally demonstrated by Lee and Shung several years ago. Since then, much effort has been devoted to improving this technology. At present, the tool is capable of trapping a microparticle as small as 1 ?m, as well as a single red blood cell. Although in comparing to other microparticles manipulating technologies, SBAT has advantages of providing stronger trapping force and deeper penetration depth in tissues, and producing less tissue damage, its potential for in vivo applications has yet been explored. It is worth noting that ultrasound has been used as a diagnostic tool for over 50 years and no known major adverse effects have been observed at the diagnostic energy level. This paper reports the results of an initial attempt to assess the feasibility of single beam acoustic tweezers to trap microparticles in vivo inside of a blood vessel. The acoustic intensity of SBAT under the trapping conditions that were utilized was measured. The mechanical index and thermal index at the focus of acoustic beam were found to be 0.48 and 0.044, respectively, which meet the standard of commercial diagnostic ultrasound system.

  6. In vivo study of nanostructured akermanite/PEO coating on biodegradable magnesium alloy for biomedical applications.

    PubMed

    Razavi, Mehdi; Fathi, Mohammadhossein; Savabi, Omid; Vashaee, Daryoosh; Tayebi, Lobat

    2015-05-01

    The major issue for biodegradable magnesium alloys is the fast degradation and release of hydrogen gas. In this article, we aim to overcome these disadvantages by using a surface modified magnesium implant. We have recently coated AZ91 magnesium implants by akermanite (Ca2 MgSi2 O7 ) through the combined electrophoretic deposition (EPD) and plasma electrolytic oxidation (PEO) methods. In this work, we performed the in vitro and in vivo examinations of these coated implants using L-929 cell line and rabbit animal model. The in vitro study confirmed the higher cytocompatibility of the coated implants compare to the uncoated ones. For the in vivo experiment, the rod samples were implanted into the greater trochanter of rabbits and monitored for two months. The results indicated a noticeable biocompatibility improvement of the coated implants which includes slower implant weight loss, reduction in Mg ion released from the coated samples in the blood plasma, lower release of hydrogen bubbles, increase in the amount of bone formation and ultimately lower bone inflammation after the surgery according to the histological images. Our data exemplifies that the proper surface treatment of the magnesium implants can improve their biocompatibility under physiological conditions to make them applicable in clinical uses. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 1798-1808, 2015. PMID:25203515

  7. The synthesis, characterisation and in vivo study of a bioceramic for potential tissue regeneration applications

    PubMed Central

    Poinern, Gérrard Eddy Jai; Brundavanam, Ravi Krishna; Thi Le, Xuan; Nicholls, Philip K.; Cake, Martin A.; Fawcett, Derek

    2014-01-01

    Hydroxyapatite (HAP) is a biocompatible ceramic that is currently used in a number of current biomedical applications. Recently, nanometre scale forms of HAP have attracted considerable interest due to their close similarity to the inorganic mineral component of the bone matrix found in humans. In this study ultrafine nanometre scale HAP powders were prepared via a wet precipitation method under the influence of ultrasonic irradiation. The resulting powders were compacted and sintered to form a series of ceramic pellets with a sponge-like structure with varying density and porosity. The crystalline structure, size and morphology of the powders and the porous ceramic pellets were investigated using advanced characterization techniques. The pellets demonstrated good biocompatibility, including mixed cell colonisation and matrix deposition, in vivo following surgical implantation into sheep M. latissimus dorsi. PMID:25168046

  8. In vivo application of a small molecular weight antifungal protein of Penicillium chrysogenum (PAF)

    SciTech Connect

    Palicz, Zoltán; Jenes, Ágnes; Gáll, Tamás [Department of Physiology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Miszti-Blasius, Kornél [Department of Clinical Biochemistry and Molecular Pathology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Kollár, Sándor; Kovács, Ilona [Department of Pathology, Kenézy Hospital LTD, Debrecen (Hungary); Emri, Miklós; Márián, Teréz [Department of Nuclear Medicine, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Leiter, Éva; Pócsi, István [Department of Microbial Biotechnology and Cell Biology, Faculty of Science and Technology, Centre of Arts, Humanities and Sciences, University of Debrecen, Debrecen (Hungary); Cs?sz, Éva; Kalló, Gerg? [Proteomics Core Facility, Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Heged?s, Csaba; Virág, László [Department of Medical Chemistry, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Csernoch, László [Department of Physiology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary); Szentesi, Péter, E-mail: szentesi.peter@med.unideb.hu [Department of Physiology, Medical and Health Science Center, University of Debrecen, Debrecen (Hungary)

    2013-05-15

    The antifungal protein of Penicillium chrysogenum (PAF) inhibits the growth of important pathogenic filamentous fungi, including members of the Aspergillus family and some dermatophytes. Furthermore, PAF was proven to have no toxic effects on mammalian cells in vitro. To prove that PAF could be safely used in therapy, experiments were carried out to investigate its in vivo effects. Adult mice were inoculated with PAF intranasally in different concentrations, up to 2700 ?g·kg{sup ?1} daily, for 2 weeks. Even at the highest concentration – a concentration highly toxic in vitro for all affected molds – used, animals neither died due to the treatment nor were any side effects observed. Histological examinations did not find pathological reactions in the liver, in the kidney, and in the lungs. Mass spectrometry confirmed that a measurable amount of PAF was accumulated in the lungs after the treatment. Lung tissue extracts from PAF treated mice exerted significant antifungal activity. Small-animal positron emission tomography revealed that neither the application of physiological saline nor that of PAF induced any inflammation while the positive control lipopolysaccharide did. The effect of the drug on the skin was examined in an irritative dermatitis model where the change in the thickness of the ears following PAF application was found to be the same as in control and significantly less than when treated with phorbol-12-myristate-13-acetate used as positive control. Since no toxic effects of PAF were found in intranasal application, our result is the first step for introducing PAF as potential antifungal drug in therapy. - Highlights: • PAF, the antifungal protein of Penicillium chrysogenum, was not toxic in mice. • Its intranasal application didn't induce pathological reactions in the lung. • PAF retained its antifungal activity in lung extracts. • Its application on the skin did not cause inflammation.

  9. Characterization of cardiovascular liver motion for the eventual application of elasticity imaging to the liver in vivo

    Microsoft Academic Search

    Alexander F Kolen; Naomi R Miller; Eltayeb E Ahmed; Jeffrey C Bamber

    2004-01-01

    Elastography, which uses ultrasound to image the tissue strain that results from an applied displacement, can display tumours and heat-ablated tissue with high contrast. However, its application to liver in vivo may be problematic due to the presence of respiratory and cardiovascular sources of displacement. The aim of this study was to measure the cardiovascular-induced component of natural liver motion

  10. Application of FRET Technology to the In Vivo Evaluation of Therapeutic Nucleic Acids (ANTs)

    NASA Astrophysics Data System (ADS)

    Benítez-Hess, María Luisa; Alvarez-Salas, Luis Marat

    2007-02-01

    Developing applications for therapeutic nucleic acids (TNAs) (i.e. ribozymes, antisense oligodeoxynucleotides (AS-ODNs), siRNA and aptamers) requires a reporter system designed to rapidly evaluate their in vivo effect. To this end we designed a reporter system based on the fluorescence resonance energy transfer (FRET) engineered to release the FRET effect produced by two green fluorescent protein (GFP) variants linked by a TNA target site. Because the FRET effect occurs instantaneously when two fluorophores are very close to each other (>100nm) stimulating emission of the acceptor fluorophore by the excitation of the donor fluorophore it has been widely use to reveal interactions between molecules. The present system (FRET2) correlates the FRET effect with the in vivo activity of distinct types of TNAs based on a model consisting of RNA from human papillomavirus type 16 (HPV-16) previously shown accessible to TNAs. HPV-16 is the most common papillomavirus associated with cervical cancer, the leading cause of death by cancer in México. The FRET2 system was first tested in vitro and then used in bacteria in which transcription is linked to translation allowing controlled expression and rapid evaluation of the FRET2 protein. To assure accessibility of the target mRNA to TNAs, the FRET2 mRNA was probed by RNaseH assays prior FRET testing. The fluorescence features of the FRET2 system was tested with different FRET-producing GFP donor-acceptor pairs leading to selection of green (donor) and yellow (acceptor) variants of GFP as the most efficient. Modifications in aminoacid composition and linker length of the target sequence did not affect FRET efficiency. In vivo AS-ODN-mediated destruction of the chimerical FRET2 reporter mRNA resulted in the recovery of GFP fluorescent spectrum in a concentration and time dependent manner. Reported anti-HPV ribozymes were also tested with similar results. Therefore, we conclude that the FRET effect can be a useful tool in the development of TNAs.

  11. A novel scaffold geometry for chondral applications: theoretical model and in vivo validation.

    PubMed

    Scaglione, Silvia; Ceseracciu, Luca; Aiello, Maurizio; Coluccino, Luca; Ferrazzo, Federica; Giannoni, Paolo; Quarto, Rodolfo

    2014-10-01

    A theoretical model of the 3D scaffold internal architecture has been implemented with the aim to predict the effects of some geometrical parameters on total porosity, Young modulus, buckling resistance and permeability of the graft. This model has been adopted to produce porous poly-caprolacton based grafts for chondral tissue engineering applications, best tuning mechanical and functional features of the scaffolds. Material prototypes were produced with an internal geometry with parallel oriented cylindrical pores of 200??m of radius (r) and an interpore distance/pores radius (d/r) ratio of 1. The scaffolds have been then extensively characterized; progenitor cells were then used to test their capability to support cartilaginous matrix deposition in an ectopic model. Scaffold prototypes fulfill both the chemical-physical requirements, in terms of Young's modulus and permeability, and the functional needs, such as surface area per volume and total porosity, for an enhanced cellular colonization and matrix deposition. Moreover, the grafts showed interesting chondrogenic potential in vivo, besides offering adequate mechanical performances in vitro, thus becoming a promising candidate for chondral tissues repair. Finally, a very good agreement was found between the prediction of the theoretical model and the experimental data. Many assumption of this theoretical model, hereby applied to cartilage, may be transposed to other tissue engineering applications, such as bone substitutes. PMID:25073412

  12. Direct optic nerve sheath (DONS) application of Schwann cells prolongs retinal ganglion cell survival in vivo

    PubMed Central

    Guo, L; Davis, B; Nizari, S; Normando, E M; Shi, H; Galvao, J; Turner, L; Shi, J; Clements, M; Parrinello, S; Cordeiro, M F

    2014-01-01

    Cell-based therapies are increasingly recognized as a potential strategy to treat retinal neurodegenerative disease. Their administration, however, is normally indirect and complex, often with an inability to assess in real time their effects on cell death and their migration/integration into the host retina. In the present study, using a partial optic nerve transection (pONT) rat model, we describe a new method of Schwann cell (SC) delivery (direct application to injured optic nerve sheath, SC/DONS), which was compared with intravitreal SC delivery (SC/IVT). Both SC/DONS and SC/IVT were able to be assessed in vivo using imaging to visualize retinal ganglion cell (RGC) apoptosis and SC retinal integration. RGC death in the pONT model was best fitted to the one-phase exponential decay model. Although both SC/DONS and SC/IVT altered the temporal course of RGC degeneration in pONT, SC/DONS resulted in delayed but long-lasting effects on RGC protection, compared with SC/IVT treatment. In addition, their effects on primary and secondary degeneration, and axonal regeneration, were also investigated, by histology, whole retinal counting, and modelling of RGC loss. SC/DONS was found to significantly reduce RGC apoptosis in vivo and significantly increase RGC survival by targeting secondary rather than primary degeneration. Both SC/DONS and SC/IVT were found to promote RGC axonal regrowth after optic nerve injury, with evidence of GAP-43 expression in RGC somas and axons. SC/DONS may have the potential in the treatment of optic neuropathies, such as glaucoma. We show that SC transplantation can be monitored in real time and that the protective effects of SCs are associated with targeting secondary degeneration, with implications for translating cell-based therapies to the clinic. PMID:25321467

  13. Direct optic nerve sheath (DONS) application of Schwann cells prolongs retinal ganglion cell survival in vivo.

    PubMed

    Guo, L; Davis, B; Nizari, S; Normando, E M; Shi, H; Galvao, J; Turner, L; Shi, J; Clements, M; Parrinello, S; Cordeiro, M F

    2014-01-01

    Cell-based therapies are increasingly recognized as a potential strategy to treat retinal neurodegenerative disease. Their administration, however, is normally indirect and complex, often with an inability to assess in real time their effects on cell death and their migration/integration into the host retina. In the present study, using a partial optic nerve transection (pONT) rat model, we describe a new method of Schwann cell (SC) delivery (direct application to injured optic nerve sheath, SC/DONS), which was compared with intravitreal SC delivery (SC/IVT). Both SC/DONS and SC/IVT were able to be assessed in vivo using imaging to visualize retinal ganglion cell (RGC) apoptosis and SC retinal integration. RGC death in the pONT model was best fitted to the one-phase exponential decay model. Although both SC/DONS and SC/IVT altered the temporal course of RGC degeneration in pONT, SC/DONS resulted in delayed but long-lasting effects on RGC protection, compared with SC/IVT treatment. In addition, their effects on primary and secondary degeneration, and axonal regeneration, were also investigated, by histology, whole retinal counting, and modelling of RGC loss. SC/DONS was found to significantly reduce RGC apoptosis in vivo and significantly increase RGC survival by targeting secondary rather than primary degeneration. Both SC/DONS and SC/IVT were found to promote RGC axonal regrowth after optic nerve injury, with evidence of GAP-43 expression in RGC somas and axons. SC/DONS may have the potential in the treatment of optic neuropathies, such as glaucoma. We show that SC transplantation can be monitored in real time and that the protective effects of SCs are associated with targeting secondary degeneration, with implications for translating cell-based therapies to the clinic. PMID:25321467

  14. Synthesis and surface modification of magnetic nanoparticles for in vivo biomedical applications

    NASA Astrophysics Data System (ADS)

    Sun, Conroy Ghin Chee

    Magnetic nanoparticles (MNPs) possess unique magnetic properties and the ability to function at the cellular and molecular level of biological interactions making them an attractive platform to serve as contrast agents for magnetic resonance imaging (MRI) and as carriers for drug delivery. Recent advances in nanotechnology have improved the ability to engineer the features and properties of MNPs allowing them to be tailored specifically for these biomedical applications. MNPs composed of metallic, oxide, and nanoalloy cores and a variety of protective coatings are being investigated for applications in the detection, diagnosis, and treatment of malignant tumors, cardiovascular disease, and neurological disease. To better address specific clinical needs, MNPs with higher magnetic moments, non-fouling surfaces, and increased functionalities are now being developed. The goal of this interdisciplinary research is to develop novel superparamagnetic nanoprobes for non-invasive cancer diagnosis and treatment. This strategy utilizes iron oxide nanoparticles coated with various biocompatible polymers, such as poly(ethylene glycol) (PEG) and chitosan, to serve as both a contrast agent for MRI and a carrier for drug delivery. In this project, we have conjugated various targeting agents, such as folic acid (FA) and chlorotoxin (CTX), to these iron oxide nanoparticles to improve their tumor specific accumulation. The folate receptor is known to be overexpressed on the surfaces of many human tumor cells, including ovarian, lung, breast, endometrial, renal, and colon cancers, while CTX binds with high affinity to gliomas, medulloblastomas, and other tumors of the neuroectodermal origin. To evaluate its effectiveness as a targeted drug carrier, methotrexate (MTX), a convention chemotherapeutic agent, was conjugated to iron oxide nanoparticles in combination with CTX. Specific tumor cell targeting of our nanoparticle system has been demonstrated through increased contrast enhancement both in vitro and in vivo in MRI experiments. The successful application of such smart molecular imaging probes will have a significant clinical impact on improved diagnosis and treatment of malignant tumors.

  15. In vivo pH monitoring using boron doped diamond microelectrode and silver needles: Application to stomach disorder diagnosis

    PubMed Central

    Fierro, Stéphane; Seishima, Ryo; Nagano, Osamu; Saya, Hideyuki; Einaga, Yasuaki

    2013-01-01

    This study presents the in vivo electrochemical monitoring of pH using boron doped diamond (BDD) microelectrode and silver needles for potential application in medical diagnosis. Accurate calibration curve for pH determination were obtained through in vitro electrochemical measurements. The increase induced in stomach pH by treatment with pantoprazole was used to demonstrate that it is possible to monitor the pH in vivo using the simple and noninvasive system proposed herein. Using the results of the in vivo and in vitro experiments, a quantitative analysis of the increase in stomach pH is also presented. It is proposed that the catheter-free pH monitoring system presented in this study could be potentially employed in any biological environment. PMID:24247214

  16. In vivo pH monitoring using boron doped diamond microelectrode and silver needles: Application to stomach disorder diagnosis

    NASA Astrophysics Data System (ADS)

    Fierro, Stéphane; Seishima, Ryo; Nagano, Osamu; Saya, Hideyuki; Einaga, Yasuaki

    2013-11-01

    This study presents the in vivo electrochemical monitoring of pH using boron doped diamond (BDD) microelectrode and silver needles for potential application in medical diagnosis. Accurate calibration curve for pH determination were obtained through in vitro electrochemical measurements. The increase induced in stomach pH by treatment with pantoprazole was used to demonstrate that it is possible to monitor the pH in vivo using the simple and noninvasive system proposed herein. Using the results of the in vivo and in vitro experiments, a quantitative analysis of the increase in stomach pH is also presented. It is proposed that the catheter-free pH monitoring system presented in this study could be potentially employed in any biological environment.

  17. Three-Photon Luminescence of Gold Nanorods and Its Applications for High Contrast Tissue and Deep In Vivo Brain Imaging

    PubMed Central

    Wang, Shaowei; Xi, Wang; Cai, Fuhong; Zhao, Xinyuan; Xu, Zhengping; Qian, Jun; He, Sailing

    2015-01-01

    Gold nanoparticles can be used as contrast agents for bio-imaging applications. Here we studied multi-photon luminescence (MPL) of gold nanorods (GNRs), under the excitation of femtosecond (fs) lasers. GNRs functionalized with polyethylene glycol (PEG) molecules have high chemical and optical stability, and can be used as multi-photon luminescent nanoprobes for deep in vivo imaging of live animals. We have found that the depth of in vivo imaging is dependent upon the transmission and focal capability of the excitation light interacting with the GNRs. Our study focused on the comparison of MPL from GNRs with two different aspect ratios, as well as their ex vivo and in vivo imaging effects under 760 nm and 1000 nm excitation, respectively. Both of these wavelengths were located at an optically transparent window of biological tissue (700-1000 nm). PEGylated GNRs, which were intravenously injected into mice via the tail vein and accumulated in major organs and tumor tissue, showed high image contrast due to distinct three-photon luminescence (3PL) signals upon irradiation of a 1000 nm fs laser. Concerning in vivo mouse brain imaging, the 3PL imaging depth of GNRs under 1000 nm fs excitation could reach 600 ?m, which was approximately 170 ?m deeper than the two-photon luminescence (2PL) imaging depth of GNRs with a fs excitation of 760 nm. PMID:25553113

  18. Experimental model to measure the increase of dental pulp temperature in vivo during laser application

    NASA Astrophysics Data System (ADS)

    Nicola, Ester M. D.; Junqueira, Silvio L. M.; Busato, Mara S.

    1994-09-01

    Carbon dioxide laser has been used in dental surgery. The existence of healthy teeth, which have pulp vitality needing to be preserved, is observed in a great number of cases. In this work we describe an experimental model which provides the measurement of temperature in pulp chamber `in vivo,' during oral surgeries in which the CO2 laser beam is applied to gingival tissue. The problems met during the search for the best way to place the thermal probe regarding the diameter and depth of pulp chamber and the thickness of the tissue layer formed by gum and maxillary bone are discussed. We use a thermocouple placed in the pulp chamber of superior canine teeth in dogs. After that, the probe was also placed between gum and dental root. Since the temperature at gingival surface was known, it was easy to determine the rise in temperature at pulp chamber and also to observe the thermal gradient from gum to tissue to bone, thus avoiding pulp damage during laser applications.

  19. Segmented surface coil resonator for in vivo EPR applications at 1.1 GHz

    PubMed Central

    Petryakov, Sergey; Samouilov, Alexandre; Chzhan-Roytenberg, Michael; Kesselring, Eric; Sun, Ziqi; Zweier, Jay L.

    2010-01-01

    A four-loop segmented surface coil resonator (SSCR) with electronic frequency and coupling adjustments was constructed with 18 mm aperture and loading capability suitable for in vivo Electron Paramagnetic Resonance (EPR) spectroscopy and imaging applications at L-band. Increased sample volume and loading capability were achieved by employing a multi-loop three-dimensional surface coil structure. Symmetrical design of the resonator with coupling to each loop resulted in high homogeneity of RF magnetic field. Parallel loops were coupled to the feeder cable via balancing circuitry containing varactor diodes for electronic coupling and tuning over a wide range of loading conditions. Manually adjusted high Q trimmer capacitors were used for initial tuning with subsequent tuning electronically controlled using varactor diodes. This design provides transparency and homogeneity of magnetic field modulation in the sample volume, while matching components are shielded to minimize interference with modulation and ambient RF fields. It can accommodate lossy samples up to 90% of its aperture with high homogeneity of RF and modulation magnetic fields and can function as a surface loop or a slice volume resonator. Along with an outer coaxial NMR surface coil, the SSCR enabled EPR/NMR co-imaging of paramagnetic probes in living rats to a depth of 20 mm. PMID:19268615

  20. In-vivo measurement of the human soft tissues constitutive laws. Applications to Computer Aided Surgery

    E-print Network

    Schiavone, Patrick; Ohayon, J; Payan, Y

    2007-01-01

    In the 80's, biomechanicians were asked to work on Computer Aided Surgery applications since orthopaedic surgeons were looking for numerical tools able to predict risks of fractures. More recently, biomechanicians started to address soft tissues arguing that most of the human body is made of such tissues that can move as well as deform during surgical gestures [1]. An intra-operative use of a continuous Finite Element (FE) Model of a given tissue mainly faces two problems: (1) the numerical simulations have to be "interactive", i.e. sufficiently fast to provide results during surgery (which can be a strong issue in the context of hyperelastic models for example) and (2) during the intervention, the surgeon needs a device that can be used to provide to the model an estimation of the patient-specific constitutive behaviour of the soft tissues. This work proposes an answer to the second point, with the design of a new aspiration device aiming at characterizing the in vivo constitutive laws of human soft tissues....

  1. A Freehand Ultrasound Elastography System with Tracking for In-vivo Applications

    PubMed Central

    Foroughi, Pezhman; Kang, Hyun-Jae; Carnegie, Daniel A.; van Vledder, Mark G.; Choti, Michael A.; Hager, Gregory D.; Boctor, Emad M.

    2012-01-01

    Ultrasound transducers are commonly tracked in modern ultrasound navigation/guidance systems. In this paper, we demonstrate the advantages of incorporating tracking information into ultrasound elastography for clinical applications. First, we address a common limitation of freehand palpation: speckle decorrelation due to out-of-plane probe motion. We show that by automatically selecting pairs of radio frequency (RF) frames with minimal lateral and out-of-plane motions combined with a fast and robust displacement estimation technique greatly improves in-vivo elastography results. We also use tracking information and image quality measure to fuse multiple images with similar strain that are taken roughly from the same location to obtain a high quality elastography image. Finally, we show that tracking information can be used to give the user partial control over the rate of compression. Our methods are tested on tissue mimicking phantom and experiments have been conducted on intra-operative data acquired during animal and human experiments involving liver ablation. Our results suggest that in challenging clinical conditions, our proposed method produces reliable strain images and eliminates the need for a manual search through the ultrasound data in order to find RF pairs suitable for elastography. PMID:23257351

  2. In vivo selection to identify bacterial strains with enhanced ecological performance in synbiotic applications.

    PubMed

    Krumbeck, Janina A; Maldonado-Gomez, María X; Martínez, Inés; Frese, Steven A; Burkey, Thomas E; Rasineni, Karuna; Ramer-Tait, Amanda E; Harris, Edward N; Hutkins, Robert W; Walter, Jens

    2015-04-01

    One strategy for enhancing the establishment of probiotic bacteria in the human intestinal tract is via the parallel administration of a prebiotic, which is referred to as a synbiotic. Here we present a novel method that allows a rational selection of putative probiotic strains to be used in synbiotic applications: in vivo selection (IVS). This method consists of isolating candidate probiotic strains from fecal samples following enrichment with the respective prebiotic. To test the potential of IVS, we isolated bifidobacteria from human subjects who consumed increasing doses of galactooligosaccharides (GOS) for 9 weeks. A retrospective analysis of the fecal microbiota of one subject revealed an 8-fold enrichment in Bifidobacterium adolescentis strain IVS-1 during GOS administration. The functionality of GOS to support the establishment of IVS-1 in the gastrointestinal tract was then evaluated in rats administered the bacterial strain alone, the prebiotic alone, or the synbiotic combination. Strain-specific quantitative real-time PCR showed that the addition of GOS increased B. adolescentis IVS-1 abundance in the distal intestine by nearly 2 logs compared to rats receiving only the probiotic. Illumina 16S rRNA sequencing not only confirmed the increased establishment of IVS-1 in the intestine but also revealed that the strain was able to outcompete the resident Bifidobacterium population when provided with GOS. In conclusion, this study demonstrated that IVS can be used to successfully formulate a synergistic synbiotic that can substantially enhance the establishment and competitiveness of a putative probiotic strain in the gastrointestinal tract. PMID:25616794

  3. Skin imaged by femtosecond laser irradiation: a risk assessment for in vivo applications

    NASA Astrophysics Data System (ADS)

    Fischer, F.; Volkmer, B.; Puschmann, S.; Greinert, R.; Breitbart, W.; Kiefer, J.; Wepf, R.

    2006-04-01

    During the last couple of years new imaging techniques using femtosecond lasers (fs-lasers) in the near infrared spectral range evolved for a variety of in vitro applications. We wanted to know, whether fs-lasers have a non-invasive imaging potential for in vivo applications for human skin. So far, little is known about possible risks of this irradiation type. To estimate the risk of irradiation damage in human skin we used a "biological dosimeter" in this investigation. We irradiated fresh human skin samples with both an fs-laser and a solar simulator (UV-source) for comparison. DNA damage introduced in the epidermis was evaluated using fluorescent antibodies against cyclobutane-pyrimidin-dimers (CPDs) in combination with immuno-fluorescence image analysis. Various fs-irradiation regimes were evaluated differing in laser power and step width of horizontal irradiation scans. When using 15 mW or 30 mW fs-laser power combined with horizontal irradiation scans applied every 5 mm in depth around the epidermal-dermal junction no induction of CPDs was found. However, induction of CPDs could be seen using 60 mW laser power and 5 ?m step width. Narrowing the step width to 1 mm and using increasing laser power (up to 35 mW) from the surface of the skin to the epidermis led to CPD formation, too. Quantitative comparison of CPD production at various laser regimes with CPD production using a solar simulator was done. We could show that the number of CPDs formed by the 60 mW laser irradiation regime is comparable to an UV-irradiation giving 0.6 MED (minimal erythemal dose). The smaller step width laser irradiation regime (1 ?m step width and up to 35 mW) was comparable to a UV-irradiation regime resulting in 0.45 MED.

  4. Combining whispering gallery mode lasers and microstructured optical fibers for in-vivo biosensing applications

    NASA Astrophysics Data System (ADS)

    François, A.; Rowland, K. J.; Reynolds, T.; Nicholls, S. J.; Monro, T. M.

    2013-10-01

    Whispering Gallery Modes (WGMs) have been widely studied for the past 20 years for various applications, including biological sensing. While the different WGM-based sensing approaches reported in the literature enable useful sensor characteristics, at present this technology is not yet mature, mainly for practical reasons. Our work has been focused on developing a simple, yet efficient, WGM-based sensing platform capable of being used as a dip sensor for in-vivo biosensing applications. We recently demonstrated that a dye-doped polymer microresonator, supporting WGMs, positioned onto the tip of a suspended core Microstructured Optical Fiber can be used as a dip sensor. In this architecture, the resonator is located on an air hole next to the fiber core at the fiber's tip, enabling a significant portion of the sphere to overlap with the guided light emerging from the fiber tip. This architecture offers significant benefits that have never been reported in the literature in terms of radiation efficiency, compared to the standard freestanding resonators, which arise from breaking the symmetry of the resonator. In addition to providing the remote excitation and collection of the WGMs' signal, the fiber also allows easy manipulation of the microresonator and the use this sensor in a dip sensing architecture, alleviating the need for a complex microfluidic interface. Here, we present our recent results on the microstructured fiber tip WGM-based sensor, including its lasing behavior and enhancement of the radiation efficiency as a function of the position of the resonator on the fiber tip. We also show that this platform can be used for clinical diagnostics and applying this technology to the detection of Troponin T, an acute myocardial infarction biomarker, down to a concentration of 7.4 pg/mL.

  5. Synthesis of Luminescent Near-Infrared AgInS2 Nanocrystals as Optical Probes for In Vivo Applications

    PubMed Central

    Liu, Liwei; Hu, Rui; Roy, Indrajit; Lin, Guimiao; Ye, Ling; Reynolds, Jessica L.; Liu, Jianwei; Liu, Jing; Schwartz, Stanley A.; Zhang, Xihe; Yong, Ken-Tye

    2013-01-01

    Near infrared quantum dots have been receiving great attention as fluorescent optical probes for in vivo imaging applications. In this contribution, we report the synthesis and surface functionalization of cadmium free ternary AgInS2 nanocrystals emitting in the near infrared range for successful in vitro and in vivo bioimaging applications. The FDA approved triblock copolymer Pluronic F127 was used to encapsulate the nanocrystals and made them dispersible in aqueous solution. By employing a whole body small animal optical imaging setup, we were able to use the AgInS2 nanocrystals formulation for passive targeted delivery to the tumor site. The ultra-small crystal size, near-infrared emitting luminescence, and high quantum yield make the AgInS2 nanocrystals an attractive candidate as a biological contrast agent for cancer sensing and imaging. PMID:23422953

  6. A Multimode Optical Imaging System for Preclinical Applications In Vivo: Technology Development, Multiscale Imaging, and Chemotherapy Assessment

    PubMed Central

    Hwang, Jae Youn; Wachsmann-Hogiu, Sebastian; Ramanujan, V. Krishnan; Ljubimova, Julia; Gross, Zeev; Gray, Harry B.; Medina-Kauwe, Lali K.; Farkas, Daniel L.

    2012-01-01

    Purpose Several established optical imaging approaches have been applied, usually in isolation, to preclinical studies; however, truly useful in vivo imaging may require a simultaneous combination of imaging modalities to examine dynamic characteristics of cells and tissues. We developed a new multimode optical imaging system designed to be application-versatile, yielding high sensitivity, and specificity molecular imaging. Procedures We integrated several optical imaging technologies, including fluorescence intensity, spectral, lifetime, intravital confocal, two-photon excitation, and bioluminescence, into a single system that enables functional multiscale imaging in animal models. Results The approach offers a comprehensive imaging platform for kinetic, quantitative, and environmental analysis of highly relevant information, with micro-to-macroscopic resolution. Applied to small animals in vivo, this provides superior monitoring of processes of interest, represented here by chemo-/nanoconstruct therapy assessment. Conclusions This new system is versatile and can be optimized for various applications, of which cancer detection and targeted treatment are emphasized here. PMID:21874388

  7. Selective perturbation of in vivo linear energy transfer using high- Z vaginal applicators for Cf-252 brachytherapy

    NASA Astrophysics Data System (ADS)

    Rivard, M. J.; Evans, K. E.; Leal, L. C.; Kirk, B. L.

    2004-01-01

    Californium-252 ( 252Cf) brachytherapy sources emit both neutrons and photons, and have the potential to vastly improve the current standard-of-practice for brachytherapy. While hydrogenous materials readily attenuate the 252Cf fission energy neutrons, high- Z materials are utilized to attenuate the 252Cf gamma-rays. These differences in shielding materials may be exploited when treating with a vaginal applicator to possibly improve patient survival through perturbation of the in vivo linear energy transfer radiation.

  8. Informatics approach using metabolic reactivity classifiers to link in vitro to in vivo data in application to the ToxCast Phase I dataset

    EPA Science Inventory

    Strategic combinations and tiered application of alternative testing methods to replace or minimize the use of animal models is attracting much attention. With the advancement of high throughput screening (HTS) assays and legacy databases providing in vivo testing results, suffic...

  9. Application of electrical stimulation for functional tissue engineering in vitro and in vivo

    NASA Technical Reports Server (NTRS)

    Radisic, Milica (Inventor); Park, Hyoungshin (Inventor); Langer, Robert (Inventor); Freed, Lisa (Inventor); Vunjak-Novakovic, Gordana (Inventor)

    2013-01-01

    The present invention provides new methods for the in vitro preparation of bioartificial tissue equivalents and their enhanced integration after implantation in vivo. These methods include submitting a tissue construct to a biomimetic electrical stimulation during cultivation in vitro to improve its structural and functional properties, and/or in vivo, after implantation of the construct, to enhance its integration with host tissue and increase cell survival and functionality. The inventive methods are particularly useful for the production of bioartificial equivalents and/or the repair and replacement of native tissues that contain electrically excitable cells and are subject to electrical stimulation in vivo, such as, for example, cardiac muscle tissue, striated skeletal muscle tissue, smooth muscle tissue, bone, vasculature, and nerve tissue.

  10. Preparation of 2 nm gold nanoparticles for in vitro and in vivo applications

    PubMed Central

    Moyano, Daniel F.; Duncan, Bradley; Rotello, Vincent M.

    2014-01-01

    Summary Gold nanoparticles have been a versatile tool in recent years for the exploration of biological systems. However, challenges with purification and adequate surface coverage limit the biocompatibility of gold nanoparticles. Here, we describe a detailed procedure for the synthesis, purification, and functionalization of biologically compatible gold nanoparticles for in vitro and in vivo studies. PMID:23918325

  11. Development of in vivo Constitutive Models for Liver: Application to Surgical Simulation

    PubMed Central

    Lister, Kevin; Gao, Zhan; Desai, Jaydev P.

    2011-01-01

    Advancements in real-time surgical simulation techniques have provided the ability to utilize more complex nonlinear constitutive models for biological tissues which result in increased haptic and graphic accuracy. When developing such a model, verification is necessary to determine the accuracy of the force response as well as the magnitude of tissue deformation for tool-tissue interactions. In this study, we present an experimental device which provides the ability to obtain force-displacement information as well as surface deformation of porcine liver for in vivo probing tasks. In addition, the system is capable of accurately determining the geometry of the liver specimen. These combined attributes provide the context required to simulate the experiment with accurate boundary conditions, whereby the only variable in the analysis is the material properties of the liver specimen. During the simulation, effects of settling due to gravity have been taken into account by a technique which incorporates the proper internal stress conditions in the model without altering the geometry. Initially, an Ogden model developed from ex vivo tension and compression experimentation is run through the simulation to determine the efficacy of utilizing an ex vivo model for simulation of in vivo probing tasks on porcine liver. Subsequently, a method for improving upon the ex vivo model was developed using different hyperelastic models such that increased accuracy could be achieved for the force characteristics compared to the displacement characteristics, since changes in the force variation would be more perceptible to a user in the simulation environment, while maintaining a high correlation with the surface displacement data. Furthermore, this study also presents the probing simulation which includes the capsule surrounding the liver. PMID:21161684

  12. A new strategy for in vivo spectral editing. Application to GABA editing using selective homonuclear polarization transfer spectroscopy

    NASA Astrophysics Data System (ADS)

    Shen, Jun; Yang, Jehoon; Choi, In-Young; Li, Shizhe Steve; Chen, Zhengguang

    2004-10-01

    A novel single-shot in vivo spectral editing method is proposed in which the signal to be detected, is regenerated anew from the thermal equilibrium magnetization of a source to which it is J-coupled. The thermal equilibrium magnetization of the signal to be detected together with those of overlapping signals are suppressed by single-shot gradient dephasing prior to the signal regeneration process. Application of this new strategy to in vivo GABA editing using selective homonuclear polarization transfer allows complete suppression of overlapping creatine and glutathione while detecting the GABA-4 methylene resonance at 3.02 ppm with an editing yield similar to that of conventional editing methods. The NAA methyl group at 2.02 ppm was simultaneously detected and can be used as an internal navigator echo for correcting the zero order phase and frequency shifts and as an internal reference for concentration. This new method has been demonstrated for robust in vivo GABA editing in the rat brain and for study of GABA synthesis after acute vigabatrin administration.

  13. Brief overview of control of genetic expression by antisense oligonucleotides and in vivo applications

    Microsoft Academic Search

    Gerald Zon

    1995-01-01

    Over the past several years, the use of synthetic oligonucleotides and functional analogs thereof as a possibly general means\\u000a of controlling genetic expression has received widespread attention. Following a brief overview of some of the basic principles\\u000a and strategies for this approach, attention is focused here on summarizing some recent reports of in vitro and, in particular,\\u000a in vivo investigations

  14. In vivo elemental analysis by counting neutron-induced gamma rays for medical and biological applications

    NASA Astrophysics Data System (ADS)

    Kehayias, Joseph J.; Ma, Ruimei; Zhuang, Hong; Moore, Robert; Dowling, Lisa

    1995-03-01

    Non-invasive in vivo elemental analysis is a technique used to assess human body composition which is indicative of nutritional status and health condition. The in vivo measurement of the body's major elements is used for a variety of medical studies requiring the determination of the body's compartments (protein, fat, water, bone). Whole body gamma-ray counters, consisting of Nal(Tl) crystal detectors in a shielded room, are used for measuring in vivo the body's Ca, Cl, Na and P by delayed neutron activation analysis. Thermal neutrons from a moderated 238Pu-Be source are used for the measurement of total body nitrogen (and thus protein) and chlorine at low radiation exposure (0.80 mSv). The resulting high energy prompt gamma-rays from nitrogen (10.83 MeV) and chlorine (6.11 MeV) are detected simultaneously with the irradiation. Body fat (the main energy store) and fat distribution (which relates to risk for cardiovascular disease) are measured by detecting C and O in vivo through fast neutron inelastic scattering. A small sealed D-T neutron generator is used for the pulsed (4 - 8 KHz) production of fast neutrons. Carbon and oxygen are detected by counting the 4.44 and 6.13 MeV gamma-rays resulting from the inelastic scattering of the fast neutrons from the 12C and 16O nuclei, respectively. One use of this method is the systematic study of the mechanisms driving the age-associated depletion of the metabolizing, oxygen-consuming cellular compartment of the body. The understanding of this catabolism may suggest ways to maintain lean tissue and thus to preserve quality of life for the very old.

  15. In Vivo Activities of Amoxicillin and Amoxicillin-Clavulanate against Streptococcus pneumoniae: Application to Breakpoint Determinations

    Microsoft Academic Search

    D. ANDES; W. A. CRAIG

    1998-01-01

    The in vivo activities of amoxicillin and amoxicillin-clavulanate against 17 strains of Streptococcus pneu- moniae with penicillin MICs of 0.12-8.0 mg\\/liter were assessed in a cyclophosphamide-induced neutropenic murine thigh infection model. Renal impairment was produced by administration of uranyl nitrate to prolong the amoxicillin half-life in the mice from 21 to 65 min, simulating human pharmacokinetics. Two hours after thigh

  16. Applications of RNA interference: current state and prospects for siRNA-based strategies in vivo

    Microsoft Academic Search

    Achim Aigner

    2007-01-01

    Within the recent years, RNA interference (RNAi) has become an almost-standard method for in vitro knockdown of any target\\u000a gene of interest. Now, one major focus is to further explore its potential in vivo, including the development of novel therapeutic\\u000a strategies. From the mechanism, it becomes clear that small interfering RNAs (siRNAs) play a pivotal role in triggering RNAi.\\u000a Thus,

  17. Two-photon excited fluorescence microscopy application for ex vivo investigation of ocular fundus samples

    NASA Astrophysics Data System (ADS)

    Peters, Sven; Hammer, Martin; Schweitzer, Dietrich

    2011-07-01

    Two-photon excited fluorescence (TPEF) imaging of ocular tissue has recently become a promising tool in ophthalmology for diagnostic and research purposes. The feasibility and the advantages of TPEF imaging, namely deeper tissue penetration and improved high-resolution imaging of microstructures, have been demonstrated lately using human ocular samples. The autofluorescence properties of endogenous fluorophores in ocular fundus tissue are well known from spectrophotometric analysis. But fluorophores, especially when it comes to fluorescence lifetime, typically display a dependence of their fluorescence properties on local environmental parameters. Hence, a more detailed investigation of ocular fundus autofluorescence ideally in vivo is of utmost interest. The aim of this study is to determine space-resolved the stationary and time-resolved fluorescence properties of endogenous fluorophores in ex vivo porcine ocular fundus samples by means of two-photon excited fluorescence spectrum and lifetime imaging microscopy (FSIM/FLIM). By our first results, we characterized the autofluorescence of individual anatomical structures of porcine retina samples excited at 760 nm. The fluorescence properties of almost all investigated retinal layers are relatively homogenous. But as previously unknown, ganglion cell bodies show a significantly shorter fluorescence lifetime compared to the adjacent mueller cells. Since all retinal layers exhibit bi-exponential autofluorescence decays, we were able to achieve a more precise characterization of fluorescence properties of endogenous fluorophores compared to a present in vivo FLIM approach by confocal scanning laser ophthalmoscope (cSLO).

  18. Windows on the Human Body – in Vivo High-Field Magnetic Resonance Research and Applications in Medicine and Psychology

    PubMed Central

    Moser, Ewald; Meyerspeer, Martin; Fischmeister, Florian Ph. S.; Grabner, Günther; Bauer, Herbert; Trattnig, Siegfried

    2010-01-01

    Analogous to the evolution of biological sensor-systems, the progress in “medical sensor-systems”, i.e., diagnostic procedures, is paradigmatically described. Outstanding highlights of this progress are magnetic resonance imaging (MRI) and spectroscopy (MRS), which enable non-invasive, in vivo acquisition of morphological, functional, and metabolic information from the human body with unsurpassed quality. Recent achievements in high and ultra-high field MR (at 3 and 7 Tesla) are described, and representative research applications in Medicine and Psychology in Austria are discussed. Finally, an overview of current and prospective research in multi-modal imaging, potential clinical applications, as well as current limitations and challenges is given. PMID:22219684

  19. A multi-fiber handling device for in vivo solid phase microextraction-liquid chromatography-mass spectrometry applications.

    PubMed

    Cudjoe, Erasmus; Pawliszyn, Janusz

    2012-04-01

    Solid phase microextraction, an in vivo and ex vivo sample preparation method, continues to capture growing interest among researchers for bioanalytical applications. When coupled with liquid chromatography mass spectrometry, the procedure often involves large numbers of fibers in, for example, both pharmacokinetic and pharmadynamic studies as well as other bioapplications. In this regard, appropriate and adequate precaution will be critical in preventing the fibers firstly from any possible external contamination and damage to maintain high analytical data integrity. In addition, improving the offline desorption of fibers specifically for in vivo SPME will not only help in improving data quality, but will also significantly decrease the overall analysis time. This article introduces a prototype multi-fiber handling device capable of simultaneous extraction/desorption of multiple solid phase microextraction (SPME) fibers on a 96-deep well plate format. This device thus provides an alternative approach to improving higher sample throughput for in vivo SPME liquid chromatography mass spectrometry applications. The portable design of the device ensures effective protection and prevention of fibers against damage and possible contamination and thus maintains analytical data reliability. To ensure its suitability for parallel extraction/desorption, the device was carefully evaluated using four benzodiazepines (diazepam, nordiazepam, oxazepam and lorazepam) as model drugs by monitoring inter- and intra-well variability. The effect of agitation speed on data precision and accuracy, effect of device weight on data precision, and comparison of the overall performance of the device with traditional manual desorption approach were also assessed. Results obtained from evaluation of the device with particular focus on the desorption process indicated that the weight of the device has no effect on the reliability and reproducibility of data acquired using the device. The average amount of diazepam obtained for 20 selected wells with and without device was 48.8pg and 49.4pg, respectively. Intra-, inter-well, and inter fiber variations recorded were all ?13% indicating an excellent precision and reproducibility can be attained with the device. PMID:22078236

  20. A fiber tip label free biological sensing platform for in vivo applications

    NASA Astrophysics Data System (ADS)

    François, A.; Rowland, K. J.; Monro, T. M.

    2013-03-01

    The novel platform presented in this paper is design to answer the unmet challenge of real time label free in-vivo sensing by bringing together a refractive index transducing mechanism based on Whispering Gallery Modes in dye doped microspheres combined with Microstructured Optical Fibers. In addition to providing the provides the remote excitation and collection of the WGM signal, the fibre also allows easily manipulation of the microresonator and the use this sensor in a dip sensing architecture, alleviating the need for a complex microfluidic interface. Here, we demonstrate the ability of this sensing platform to be operated above its lasing threshold, enabling enhanced performance.

  1. Application of laser scan microscopy in vivo for wound healing characterization

    NASA Astrophysics Data System (ADS)

    Czaika, V.; Alborova, A.; Sterry, W.; Lademann, J.; Koch, S.

    2010-09-01

    Considering the advancing age of the population, wound healing disturbances are becoming increasingly important in clinical routine. The development of wound healing creams and lotions as well as therapy control require an objective evaluation of the wound healing process, which represents the destruction of the barrier. Therefore, transepidermal water loss measurements are often carried out. These measurements have the disadvantage that they are disturbed by the interstitial fluid, which is located on the surface of chronic wounds and also by water components of the creams and lotions. Additionally, the TEWL measurements are very sensitive to temperature changes and to the anxiety of the volunteers. In the present study, in vivo laser scanning microscopy was used to analyze the reepithelialization and barrier recovery of standardized wounds produced by the suction blister technique. It was demonstrated that this non-invasive, on-line spectroscopic method allows the evaluation of the wound healing process, without any disturbances. It was found that the wound healing starts not only from the edges of the wound, but also out of the hair follicles. The in vivo laser scanning microscopy is well suited to evaluate the efficacy of wound healing creams and for therapy control.

  2. Monoclonal antibodies reactive with human breast or ovarian carcinoma: In vivo applications

    SciTech Connect

    Thor, A.D.; Edgerton, S.M. (Harvard Medical School, Boston, MA (USA))

    1989-10-01

    Monoclonal antibodies (MoAbs) are unique and useful bioprobes that allow in vivo targeting of membrane-associated or circulating antigens. Most of the clinical trials to date have used low dosages of radiolabeled MoAb given in a single dose. Newer studies have included antibody fragments, repeated injections, intraperitoneal (IP) administration, and other labels such as 90Y. Clinical MoAb trials are often arduous, expensive, and time-consuming to perform. Before human use, animal studies and extensive MoAb characterization are required. The production of pharmaceutical grade, radiolabeled MoAb is technically difficult and costly. Clinical trials require administrative and patient consent as well as extensive written protocols. These studies necessitate interdepartmental and intradepartmental cooperation and coordination. Furthermore, the use of in vivo radiolabeled probes impacts many levels of health care providers from janitorial, nursing, and technical staff to laboratories and physicians. Simple blood tests or disposal of body excretions may concern nursing or technical staff with the possibility of radiation exposure. The responsibility for study design, personnel involvement, and prospective use in patients without a definitive cancer diagnosis ultimately rests with the physician. While many issues have been addressed, additional clinical trials, consideration of safety issues, and standardization between institutions will be necessary before the use of radiolabeled MoAb for diagnosis, management, or therapy of human tumors becomes routine. Continued cooperation and funding should ensure its achievement. 136 references.

  3. In vitro & in vivo studies on lornoxicam loaded nanoemulsion gels for topical application.

    PubMed

    Dasgupta, Sandipan; Ghosh, Surajit K; Ray, Subhabrata; Kaurav, Surendra Singh; Mazumder, Bhaskar

    2014-01-01

    The objective of this work was to increase the solubility, in vitro skin permeability of lornoxicam from semisolid topical formulations and also to investigate the in vivo potential of nanoemulsion formulation. Optimized lornoxicam loaded nanoemulsion was prepared successfully by spontaneous self-emulsification method and the size of the stable formulations was found within the range of 102 to 200 nm. The stable nanoemulsion formulations characterized for viscosity, droplet size, transmission electron microscopy (TEM) and refractive index. In vitro permeation rate of nanoemulsion and conventional gel of lornoxicam (LX) were determined. Prmeability parameters like steady-state flux (Jss), permeability coefficient (Kp), and enhancement ratio (Er) were significantly increased in nanoemulsion NE8 and the nanogel NG8 as compared to conventional gel (LG). In vivo studies revealed a significant increase in anti-inflammatory effects as compared with conventional gel of LX. The anti-inflammatory effects of formulation NG8 showed a significant increase in percent inhibition value when compared with control, this difference was found to be highly significant (p<0.001). This work shows for the first time that lornoxicam can be formulated into nanoemulsions and may show promise in enhancing solubility and permeation. PMID:24266509

  4. Luminescent magnetic quantum dots for in vitro/in vivo imaging and applications in therapeutics.

    PubMed

    Acharya, Amitabha

    2013-06-01

    The quest for design of newer/advanced methods for medical diagnosis and targeted therapeutics are of utmost interest and challenging too, because of its importance in clinical diagnosis. Currently available diagnosis methodologies have their own disadvantages. These shortcomings can be overcome by using multimodal imaging systems where two or more imaging modalities may be coupled. Nanoparticles being widely studied for targeted drug delivery and as biological contrasting agents, might play a decisive role in such findings. This review is focused towards the ongoing research in the area of hybrid nanocomposites which can be used for both as MRI contrasting agent (magnetic nanoparticles) and molecular imaging studies (using fluorescent quantum dots) at in vitro and in vivo level. Though several reports are available in literature for such bimodal imaging systems, their clinical trials are very restricted, possibly because of the lack of communication between the in vitro and in vivo studies. This review is expected to bridge the gap between such studies. PMID:23862405

  5. Application of a new high-speed magnetic deformable mirror for in-vivo retinal imaging

    NASA Astrophysics Data System (ADS)

    Balderas-Mata, Sandra E.; Jones, Steven M.; Zawadzki, Robert J.; Werner, John S.

    2011-08-01

    Nowadays in ophthalmologic practice several commercial instruments are available to image patient retinas in vivo. Many modern fundus cameras and confocal scanning laser ophthalmoscopes allow acquisition of two dimensional en face images of the retina with both back reflected as well as fluorescent light. Additionally, optical coherence tomography systems allow non-invasive probing of three-dimensional retinal morphology. For all of these instruments the available lateral resolution is limited by optical quality of the human eye used as the imaging objective. To improve lateral resolution and achieve diffraction-limited imaging, adaptive optics (AO) can be implemented with any of these imaging systems to correct both static and dynamic aberrations inherent in human eyes. Most of the wavefront correctors used previously in AO systems have limited dynamic range and an insufficient number of actuators to achieve diffraction-limited correction of most human eyes. Thus, additional corrections were necessary, either by trial lenses or additional deformable mirrors (DMs). The UC Davis AO flood-illuminated fundus camera system described in this paper has been previously used to acquire in vivo images of the photoreceptor mosaic and for psychophysical studies on normal and diseased retinas. These results were acquired using a DM manufactured by Litton ITEK (DM109), which has 109 actuators arranged in a hexagonal array below a continuous front-surface mirror. It has an approximate surface actuator stroke of +/-2?m. Here we present results with a new hi-speed magnetic DM manufactured by ALPAO (DM97, voice coil technology), which has 97 actuators and similar inter-actuator stroke (>3?m, mirror surface) but much higher low-order aberration correction (defocus stroke of at least +/-30?m) than the previous one. In this paper we report results of testing performance of the ALPAO DM for the correction of human eye aberrations. Additionally changes made to our AO flood illuminated system are presented along with images of the model eye retina and in-vivo human retina acquired with this system.

  6. New labeled derivatives of the neuroprotective peptide colivelin: synthesis, characterization, and first in vitro and in vivo applications.

    PubMed

    Kostomoiri, Myrta; Zikos, Christos; Benaki, Dimitra; Triantis, Charalampos; Sagnou, Marina; Paravatou-Petsotas, Maria; Papadaki, Amalia; Boleti, Haralabia; Papadopoulos, Minas; Pirmettis, Ioannis; Pelecanou, Maria; Livaniou, Evangelia

    2015-02-01

    Colivelin (CL), first reported in 2005, is the most potent member of the humanin family of neuroprotective peptides with in vitro and in vivo rescuing action against insults associated with Alzheimer's disease (AD). The objective of the present work is the design, synthesis and characterization of specific CL derivatives that can be used as molecular probes in the investigation of the unknown mechanism of CL action. Within this framework, three CL derivatives bearing suitable tags, i.e., the fluorescent moiety FITC, the streptavidin-counterpart biotinyl-group, and the (99m)Tc-radiometal chelating unit dimethylGly-Ser-Cys, were developed and subsequently applied in biological evaluation experiments. Specifically, the FITC-labeled derivative of CL was used in confocal microscopy, where specific binding at the periphery of F11 cells was observed; the biotin-labeled derivative of CL was used in an in-house developed ELISA-type assay, where specific and concentration-dependent binding with the ?-amyloid peptide of AD was shown; finally, the (99m)Tc-radiolabeled derivative of CL was used in in vivo biodistribution studies in healthy Swiss Albino mice, where 0.58% of the radioactivity administered was measured in the mouse brain 2min after injection. The above first successful applications of the CL probes demonstrate their potential to contribute in the field of neuroprotective peptides. PMID:25575783

  7. Medical applications of in vivo neutron inelastic scattering and neutron activation analysis: Technical similarities to detection of explosives and contraband

    NASA Astrophysics Data System (ADS)

    Kehayias, J. J.

    2001-07-01

    Nutritional status of patients can be evaluated by monitoring changes in elemental body composition. Fast neutron activation (for N and P) and neutron inelastic scattering (for C and O) are used in vivo to assess elements characteristic of specific body compartments. There are similarities between the body composition techniques and the detection of hidden explosives and narcotics. All samples have to be examined in depth and the ratio of elements provides a "signature" of the chemical of interest. The N/H and C/O ratios measure protein and fat content in the body. Similarly, a high C/O ratio is characteristic of narcotics and a low C/O together with a strong presence of N is a signature of some explosives. The available time for medical applications is about 20 min—compared to a few seconds for the detection of explosives—but the permitted radiation exposure is limited. In vivo neutron analysis is used to measure H, O, C, N, P, Na, Cl, and Ca for the study of the mechanisms of lean tissue depletion with aging and wasting diseases, and to investigate methods of preserving function and quality of life in the elderly.

  8. Near-infrared luminescent cubic silicon carbide nanocrystals for in vivo biomarker applications: an ab initio Study

    NASA Astrophysics Data System (ADS)

    Gali, Adam; Zólyomi, Viktor; Somogyi, Bálint

    2013-03-01

    Small molecule-sized fluorescent emitters are needed as probes to image and track the locations of targeted nano-sized objects with minimal perturbation, and are much sought-after to probe biomolecules in living cells. For in vivo biological imaging, fluorescent biomarkers have to meet the following stringent requirements: (i) they should be non-toxic and bioinert, (ii) their hydrodynamical size should be sufficiently small for clearance, (iii) they should be photo-stable. Furthermore, it is highly desirable that (iv) they have intense, stable emission in the near-infrared range, and (v) they can be produced in relatively large amount for biological studies. Here we report time-density functional calculations on SiC-based QDs in the aspect of in vivo biological imaging applications. We find that Si-vacancy, divacancy, as well as single metal dopants such as Vanadium (V), Molybdenum (Mo) and Tungsten (W) in molecule-sized (1-2 nm) SiC QDs emit light efficiently in the near-infrared range. Furthermore, their emission wavelength varies on the size of host SiC QDs at less extent than that of pristine SiC QDs, thus sharper emission spectrum is expected even in a disperse size distribution of these QDs. These fluorescent SiC QDs are paramagnetic in the ground state.

  9. Intelligent spectral signature bio-imaging in vivo for surgical applications

    NASA Astrophysics Data System (ADS)

    Jeong, Jihoon; Frykman, Philip K.; Gaon, Mark; Chung, Alice P.; Lindsley, Erik H.; Hwang, Jae Y.; Farkas, Daniel L.

    2007-02-01

    Multi-spectral imaging provides digital images of a scene or object at a large, usually sequential number of wavelengths, generating precise optical spectra at every pixel. We use the term "spectral signature" for a quantitative plot of optical property variations as a function of wavelengths. We present here intelligent spectral signature bio-imaging methods we developed, including automatic signature selection based on machine learning algorithms and database search-based automatic color allocations, and selected visualization schemes matching these approaches. Using this intelligent spectral signature bio-imaging method, we could discriminate normal and aganglionic colon tissue of the Hirschsprung's disease mouse model with over 95% sensitivity and specificity in various similarity measure methods and various anatomic organs such as parathyroid gland, thyroid gland and pre-tracheal fat in dissected neck of the rat in vivo.

  10. Application Of Micro-Highspeed Flow Visualization In Study Of Blood Cells Rheology In Vivo

    NASA Astrophysics Data System (ADS)

    Gui-shah, Li; Ni, Liang; Yu-ju, Lin; Jian, Zhang; Qiang, Wang

    1990-01-01

    A new experimental method has been developed in study of rheological behaviour of single red blood cell (RBC) in passing through the capillaries in vivo, using the technique of micro-highspeed cinecamera and micro-highspeed video system. It is one of the most important topics in the study of microcirculatory theories that fur-ther understand the deformability of RBC, flow states, velocities and dynamic mechanimi. A micro-highspeed flow visualization system consisted of essential elements: a biological microscope, a highspeed cinecmera with 35 mm film, a highspeed motion analysis system SP2000 (Kodak U.S.A) and a cold-light source etc. We have investigated the rheological parameters of single RBC in vivo in single capillaries which are about 3.3 to 6.9 um in diameters. The RBCs velocities are 0.1 to 0.25 mm/sec, and maximum shear stress on the outside surface of RBC is 13.8 dyn/cml, and maximum extension of RBC is 10.3 um. In aforementioned experiment, the highspeed flow visualization system frequency at 530 frames/sec and 200 frames/sec were used respectively. In addition, the vasomotion of precapillary sphincters have been measured and a complicated coupling phenomena between the RBC and sphincter have also been recorded and analysed. The experiment were performed with intravital hamsters and frogs. The results obtained by this system shown that the method designed by us are an effective tool in the study of rheological behaviour of single RBC in passing through the blood capillaries in vivoz.

  11. In vivo determination of the diclofenac skin reservoir: comparison between passive, occlusive, and iontophoretic application

    PubMed Central

    Clijsen, Ron; Baeyens, Jean Pierre; Barel, André Odilon; Clarys, Peter

    2015-01-01

    Aim There is scarce information concerning the pharmacodynamic behavior of topical substances used in the physiotherapy setting. The aim of the present study was to estimate the formation and emptying of the diclofenac (DF) skin reservoir after passive, semiocclusive, and electrically assisted applications of DF. Subjects and methods Five different groups of healthy volunteers (ntotal=60, 23 male and 37 female), participated in this study. A 1% DF (Voltaren Emulgel) formulation (12 mg) was applied on the volar forearms on randomized defined circular skin areas of 7 cm2. DF was applied for 20 minutes under three different conditions at the same time. The presence of DF in the skin results in a reduction of the methyl nicotinate (MN) response. To estimate the bioavailability of DF in the skin, MN responses at different times following initial DF application (1.5, 6, 24, 32, 48, 72, 96, and 120 hours) were analyzed. Results At 1.5 hours after the initial DF application, a significant decrease in MN response was detected for the occluded and iontophoretic delivery. Passive application resulted in a decrease of the MN response from 6 hours post-DF application. The inhibition remained up to 32 hours post-DF application for the iontophoretic delivery, 48 hours for the occluded application, and 72 hours for the passive delivery. At 96 and 120 hours post-DF application none of the MN responses was inhibited. Conclusion The formation and emptying of a DF skin reservoir was found to be dependent on the DF-application mode. Penetration-enhanced delivery resulted in a faster emptying of the reservoir. PMID:25709408

  12. 21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 false Tissue culture media for human ex vivo tissue and cell culture...Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture...a) Identification. Tissue culture media for human ex vivo tissue and cell...

  13. Ketorolac tromethamine floating beads for oral application: Characterization and in vitro/in vivo evaluation

    PubMed Central

    Abou el Ela, Amal El Sayeh F.; Hassan, Maha A.; El- Maraghy, Dalia A.

    2013-01-01

    The floating beads have been employed to make a sustained release of the drug in the stomach and to decrease the dose of the drug and hence overcome its side effects. The common benefits of the floating beads were it is easy preparation, without the need of a high temperature, and high percentage of the drug entrapment. In the present work, the Ketorolac tromethamine (KT) floating beads were prepared by extrusion congealing method utilizing calcium carbonate as a gas forming agent. The physical characters of the produced beads were investigated such as KT yield, KT loading, and entrapment efficiency of the drug. In addition, floating behavior, swelling, particle size, morphology and KT stability were also evaluated. In vitro drug release study was carried out, and the kinetics of the release was evaluated using the linear regression method. Furthermore, the in vivo analgesic effect of KT after oral administration of the selected formula of floating beads (F10) was carried out using hot plate and tail flick methods. Oral commercial KT tablets and KT solution were used for the comparison. The prepared beads remained floated for more than 8 h. The optimized formulation (F10) exhibited prolonged drug release (more than 8 h) and the drug release follows the Higuchi kinetic model, with a Fickian diffusion mechanism according to Korsmeyer-Peppas (n = 0.466). Moreover, F10 showed a sustained analgesic effect as compared to the commercial tablet. PMID:25161380

  14. In vivo studies of the ceramic coated titanium alloy for enhanced osseointegration in dental applications.

    PubMed

    Alzubaydi, Thair L; Alameer, Shatha S; Ismaeel, Thekra; Alhijazi, Athraa Y; Geetha, M

    2009-12-01

    This paper reports the effect of the various ceramic coatings viz., hydroxyapatite (HA) and partially stabilized zirconia (PSZ) on the bond strength between the bone and implant, and cell compatibility of screw-shaped Ti-6Al-7Nb dental implants. Electrophoretic deposition technique (EPD) was used to obtain a uniform coating of one of the three types of ceramic layers (HA, PSZ and 50%HA + 50%PSZ) on the screws. Structural investigations were carried out on the prepared HA powder and the modified surfaces of the Ti-6Al-7Nb alloy using different techniques, namely X-ray diffraction (XRD), and Fourier transform infrared spectroscopy (FTIR). The in vivo studies were performed by the implantation of screw-shaped uncoated and coated implants in the tibia of white New Zealand rabbits. To understand the bone-implant interface, biomechanical test was carried out after 2, 6 and 18 weeks healing periods. There was increased mechanical strength (torque value) of bone-implant interface with time, and the highest increment in the bond strength was recorded for implants coated with a 50% HA and 50% PSZ. Histological results show that the coated Ti-6Al-7Nb screws after 18 weeks of the implantation seem to be well-tolerated by the bone since no adverse tissue reaction was evident. However, there was a faster reaction of bone towards the coated implants compared to the uncoated one. The histochemical stain studies shows higher cellular activity and mature bone formation on all the samples. PMID:18592351

  15. In Vivo Application of Short-Lag Spatial Coherence Imaging in Human Liver

    PubMed Central

    Jakovljevic, Marko; Trahey, Gregg E.; Nelson, Rendon C.; Dahl, Jeremy J.

    2013-01-01

    We present the results of a patient study conducted to assess the performance of two novel imaging methods, namely Short-Lag Spatial Coherence (SLSC) and Harmonic Spatial Coherence Imaging (HSCI), in in vivo liver environment. Similar in appearance to the B-mode images, SLSC and HSCI images are based solely on the spatial coherence of fundamental and harmonic echo data, respectively, and do not depend on the echo magnitude. SLSC and HSCI suppress incoherent echo signals and thus tend to reduce clutter. The SLSC and HSCI images of 17 patients demonstrate sharper delineation of blood vessel walls, suppressed clutter inside the vessel lumen, and show reduced speckle in surrounding tissue compared to matched B-modes. Target contrast and contrast-to-noise ratio (CNR) show statistically significant improvements between fundamental B-mode and SLSC imaging and between harmonic B-mode and HSCI imaging (in all cases p < 0.01). The magnitude of improvement in contrast and CNR increases as the overall quality of B-mode images decreases. Poor quality fundamental B-mode images (where image quality classification is based on both contrast and CNR) exhibit the highest improvements in both contrast and CNR (288 % improvement in contrast and 533 % improvement in CNR). PMID:23347642

  16. Optical clearing of skin tissue produced by application of glucose solution: in vivo study

    NASA Astrophysics Data System (ADS)

    Bashkatov, Alexey N.; Korolevich, Alexander N.; Stolnitz, Mikhail M.; Genina, Elina A.; Tuchin, Valery V.; Sinichkin, Yuri P.; Dubina, Nataly S.; Vecherinski, Sergey I.; Belsley, Michael S.

    2006-08-01

    We present experimental results on optical properties of the human skin controlled by administration ofthe 40%-glucose solution. In vivo reflectance spectra of the human skin were measured. Results of the experimental study of influence of the 40%-glucose solution on reflectance spectra of the human skin are presented. A significant decrease of reflectance of the human skin under action of the osmotic agent is demonstrated. The experiments show that administration of the glucose solution allows for effective control of tissue optical characteristics, that makes skin more transparent, thereby increasing the ability of light penetration through the tissue. Laser Doppler flowmetry has been used for study of skin blood microcirculation under the action of the glucose solution. Results of the experiments demonstrated that at the action of the glucose solution blood perfusion and blood concentration increase, however the mean blood velocity does not change. The presented results can be used in developing functional imaging techniques, including OCT and reflectance spectroscopy. A potential benefit of the optical clearing technique is the improvement of laser therapeutic techniques that rely on sufficient light penetration to a target embedded in tissue.

  17. Ketorolac tromethamine floating beads for oral application: Characterization and in vitro/in vivo evaluation.

    PubMed

    Abou El Ela, Amal El Sayeh F; Hassan, Maha A; El-Maraghy, Dalia A

    2014-09-01

    The floating beads have been employed to make a sustained release of the drug in the stomach and to decrease the dose of the drug and hence overcome its side effects. The common benefits of the floating beads were it is easy preparation, without the need of a high temperature, and high percentage of the drug entrapment. In the present work, the Ketorolac tromethamine (KT) floating beads were prepared by extrusion congealing method utilizing calcium carbonate as a gas forming agent. The physical characters of the produced beads were investigated such as KT yield, KT loading, and entrapment efficiency of the drug. In addition, floating behavior, swelling, particle size, morphology and KT stability were also evaluated. In vitro drug release study was carried out, and the kinetics of the release was evaluated using the linear regression method. Furthermore, the in vivo analgesic effect of KT after oral administration of the selected formula of floating beads (F10) was carried out using hot plate and tail flick methods. Oral commercial KT tablets and KT solution were used for the comparison. The prepared beads remained floated for more than 8 h. The optimized formulation (F10) exhibited prolonged drug release (more than 8 h) and the drug release follows the Higuchi kinetic model, with a Fickian diffusion mechanism according to Korsmeyer-Peppas (n = 0.466). Moreover, F10 showed a sustained analgesic effect as compared to the commercial tablet. PMID:25161380

  18. Preliminary experimental investigation of in vivo magnetic manipulation: results and potential application in hyperthermia.

    PubMed

    Grady, M S; Howard, M A; Molloy, J A; Ritter, R C; Quate, E G; Gillies, G T

    1989-01-01

    The first in vivo experiments in support of a new technique for delivering stereotaxic hyperthermia have been conducted at the Experimental Surgery Facility of the University of Virginia's Medical Center. We call this technique the "Video Tumor Fighter." In each of twelve trials a single, small permanent magnet or train of small permanent magnets was implanted on the brain surface of adult canine models. In three of the trials, this "seed" (typically 6-mm diameter X 6-mm long) was moved by magnetic manipulation to different locations within the brain. In two other trials, the seed moved along the interface between the brain and the inner vault of the skull. The noncontact magnetic manipulation was accomplished by coupling the permanently magnetized seed to the large dc magnetic field gradient created by a water-cooled coil surrounding the animal's head. The seed's motions were monitored with x-ray fluoroscopy; its rate of movement was found to be approximately 0.8 mm s-1. The forces required to produce these motions were on the order of 0.07 N. We document here the instrumentation used in these trials, describe the experimental procedures employed, and discuss the technical aspects of the results. PMID:2654597

  19. A naphthalimide-based fluorescent probe for highly selective detection of histidine in aqueous solution and its application in in vivo imaging.

    PubMed

    Un, Hio-Ieng; Wu, Shuai; Huang, Chang-Bo; Xu, Zheng; Xu, Lin

    2015-02-01

    A naphthalimide-based fluorescent ensemble probe NPC for selectively detecting His in aqueous solution (10 mM HEPES, pH 7.4) has been reported. Moreover, the application of NPC in in vivo (both in Hela cells and in C. elegans) fluorescence imaging was carried out as well. PMID:25605588

  20. Multi-transmit beam forming for fast cardiac imaging--experimental validation and in vivo application.

    PubMed

    Tong, Ling; Ramalli, Alessandro; Jasaityte, Ruta; Tortoli, Piero; D'hooge, Jan

    2014-06-01

    High frame rate (HFR) echocardiography may be of benefit for functional analysis of the heart. In current clinical equipment, HFR is obtained using multi-line acquisition (MLA) which typically requires broadening of transmit beams. As this may result in a significant degradation of spatial resolution and signal-to-noise ratio (SNR), the capacity of MLA to obtain high quality HFR images remains limited. As an alternative, we have demonstrated by computer simulation that simultaneously transmitting multiple focused beams into different directions [multi-line transmit (MLT)], can increase the frame rate without significantly compromising the spatial resolution or SNR. This study aimed to experimentally verify these theoretical predictions both in vitro and in vivo to demonstrate, for the first time, that cardiac MLT imaging is feasible. Hereto, the ultrasound advanced open platform, equipped with a 2.0 MHz phased array, was programmed to interleave MLT and conventional single line transmit (SLT) beam forming. Using these two beam forming methods, images of phantoms and healthy volunteers were acquired and investigated both qualitatively and quantitatively. The results confirmed our simulations that image quality of a 4MLT imaging system with a Tukey apodization scheme is very competitive to that of SLT while providing a 4 times higher frame rate. It is also demonstrated that MLT can be combined with MLA to provide images at 12- to 16-fold frame rate (about 340-450 Hz) without significantly compromising spatial resolution and SNR. This is thus the first study to demonstrate that this new ultrasound imaging paradigm is viable which could have significant impact on future cardiac ultrasound systems. PMID:24893253

  1. The biodegradation of zein in vitro and in vivo and its application in implants.

    PubMed

    Lin, Teng; Lu, Chuanhua; Zhu, Ligen; Lu, Tao

    2011-03-01

    A unique polymer-based sustained-release implant drug delivery system was prepared by using biocompatible and biodegradable Zein as the skeleton material. After preparing Zein colloids, the Zein-loaded implant rods were formulated by injection molding followed by evaporating the solvent, and being coated with poly(lactic-co-glycolic) acid (PLGA) solution. Drug release kinetics was examined by using Fluorouracil (5-FU) as model drug. Nearly zero-order release was achieved for the model drugs for a period of 0-25 days when the implants were incubated in distilled water at 37 °C. And then the degradation kinetics of the rods in vivo and in vitro were evaluated, which indicated that Zein could be absorbed by body and has good degradation property. The effects of different ratios of Zein/5-FU and the rods' diameter on drug release were studied, respectively. The plasma concentration of 5-FU in the implants were determined by HPLC after implanting a single dose of the implants in rats. All data were subsequently processed by using the computer program 3P97, and the values were showed as follows: the area under the plasma concentration-time curve (AUC) value was 321.88 (?g/ml) × day, and the mean residence time (MRT) value was 23.05 days. The sustained-release implants of Zein/5-FU were successfully formulated. The uniqueness of the article is that Zein has been used as a skeleton material in implant delivery system for the first time and zero-order release kinetics has been obtained successfully. PMID:21184205

  2. Sphene ceramics for orthopedic coating applications: an in vitro and in vivo study.

    PubMed

    Ramaswamy, Yogambha; Wu, Chengtie; Dunstan, Colin R; Hewson, Benjamin; Eindorf, Tanja; Anderson, Gail I; Zreiqat, Hala

    2009-10-01

    The host response to titanium alloy (Ti-6Al-4V) is not always favorable as a fibrous layer may form at the skeletal tissue-device interface, causing aseptic loosening. Recently, sphene (CaTiSiO(5)) ceramics were developed by incorporating Ti in the Ca-Si system, and found to exhibit improved chemical stability. The aim of this study is to evaluate the in vitro response of human osteoblast-like cells, human osteoclasts and human microvascular endothelial cells to sphene ceramics and determine whether coating Ti-6Al-4V implants with sphene enhances anchorage to surrounding bone. The study showed that sphene ceramics support human osteoblast-like cell attachment with organized cytoskeleton structure and express increased mRNA levels of osteoblast-related genes. Sphene ceramics were able to induce the differentiation of monocytes to form functional osteoclasts with the characteristic features of f-actin and alpha(v)beta(3) integrin, and express osteoclast-related genes. Human endothelial cells were also able to attach and express the endothelial cell markers ZO-1 and VE-Cadherin when cultured on sphene ceramics. Histological staining, enzyme histochemistry and immunolabelling were used for identification of mineralized bone and bone remodelling around the coated implants. Ti-6Al-4V implants coated with sphene showed new bone formation and filled the gap between the implants and existing bone in a manner comparable to that of the hydroxyapatite coatings used as control. The new bone was in direct contact with the implants, whereas fibrous tissue formed between the bone and implant with uncoated Ti-6Al-4V. The in vivo assessment of sphene-coated implants supports our in vitro observation and suggests that they have the ability to recruit osteogenic cells, and thus support bone formation around the implants and enhance osseointegration. PMID:19457458

  3. Nanomiemgel - A Novel Drug Delivery System for Topical Application - In Vitro and In Vivo Evaluation

    PubMed Central

    Somagoni, Jaganmohan; Boakye, Cedar H. A.; Godugu, Chandraiah; Patel, Apurva R.; Mendonca Faria, Henrique Antonio; Zucolotto, Valtencir; Singh, Mandip

    2014-01-01

    Aim The objective of this study was to formulate and evaluate a unique matrix mixture (nanomiemgel) of nanomicelle and nanoemulsion containing aceclofenac and capsaicin using in vitro and in vivo analyses and to compare it to a marketed formulation (Aceproxyvon). Methods Nanomicelles were prepared using Vitamin E TPGS by solvent evaporation method and nanoemulsion was prepared by high-pressure homogenization method. In vitro drug release and human skin permeation studies were performed and analyzed using HPLC. The efficiency of nanomiemgel as a delivery system was investigated using an imiquimod-induced psoriatic like plaque model developed in C57BL/6 mice. Results Atomic Force Microscopy images of the samples exhibited a globular morphology with an average diameter of 200, 250 and 220 nm for NMI, NEM and NMG, respectively. Nanomiemgel demonstrated a controlled release drug pattern and induced 2.02 and 1.97-fold more permeation of aceclofenac and capsaicin, respectively than Aceproxyvon through dermatomed human skin. Nanomiemgel also showed 2.94 and 2.09-fold greater Cmax of aceclofenac and capsaicin, respectively than Aceproxyvon in skin microdialysis study in rats. The PASI score, ear thickness and spleen weight of the imiquimod-induced psoriatic-like plaque model were significantly (p<0.05) reduced in NMG treated mice compared to free drug, NEM, NMI & Aceproxyvon. Conclusion Using a new combination of two different drug delivery systems (NEM+NMI), the absorption of the combined system (NMG) was found to be better than either of the individual drug delivery systems due to the utilization of the maximum possible paths of absorption available for that particular drug. PMID:25546392

  4. Applicability of human dental follicle cells to bone regeneration without dexamethasone: an in vivo pilot study.

    PubMed

    Takahashi, K; Ogura, N; Tomoki, R; Eda, T; Okada, H; Kato, R; Iwai, S; Ito, K; Kuyama, K; Kondoh, T

    2015-05-01

    The aim of this study was to investigate the capacity of human dental follicle cells (hDFCs) for bone formation in vivo. hDFCs were obtained from wisdom teeth extracted from patients aged 14 and 22 years. hDFCs from the 5th to 8th passages were grown in three-dimensional (3D) culture using gelatin sponges. Cells were transplanted onto the calvaria of F344/NJcl-rnu/rnu male rats (immunodeficient rats). Haematoxylin and eosin (HE) staining and immunohistochemistry were performed, and newly formed bone was evaluated by micro-computed tomography (micro-CT). HE staining showed newly formed bone in 3D culture. Immunohistochemistry showed bone morphogenetic protein 2 (BMP-2), runt-related transcription factor 2 (RUNX2), and osterix staining in areas with newly formed bone. Furthermore, micro-CT showed that, in comparison to controls, transplanted hDFCs promoted better bone quality and bone mineral density (BMD 582±131.1 vs. 300.5±77.7mg/cm(3); P=0.039), bone mineral content (BMC 5.6±1.1 vs. 2.1±0.4mg; P=0.006), bone volume (BV 9.7±0.5×10(-3) vs. 7.0±0.4×10(-3)cm(3); P=0.002), BMC/total volume (TV) (399.9±76.3 vs. 147.7±30.8mg/cm(3); P=0.006), and BV/TV (69.1±3.6% vs. 49.6±3.1%; P=0.002). This suggests that human dental follicles are potentially useful for regenerative therapy. PMID:25496849

  5. In vitro and in vivo application of PLGA nanofiber for artificial blood vessel

    Microsoft Academic Search

    Mi Jin Kim; Ji-Heung Kim; Gijong Yi; Sang-Hyun Lim; You Sun Hong; Dong June Chung

    2008-01-01

    Poly(lactic-co-glycolic acid) (PLGA) tubes (5 mm in diameter) were fabricated using an electro spinning method and used as a scaffold for\\u000a artificial blood vessels through the hybridization of smooth muscle cells (SMCs) and endothelial cells (ECs) differentiated\\u000a from canine bone marrow under previously reported conditions. The potential clinical applications of these artificial blood\\u000a vessels were investigated using a canine model.

  6. {ital In vivo} local determination of tissue optical properties: applications to human brain

    SciTech Connect

    Bevilacqua, F.; Piguet, D.; Marquet, P.; Depeursinge, C. [Department of Micro-Engineering, institute of Applied Optics, Swiss Federal Institute of Technology Lausanne, 1015 Lausanne (Switzerland); Gross, J.D.; Tromberg, B.J. [Laser Microbeam and Medical Program, Beckman Laser Institute and Medical Clinic, University of California, Irvine, 1002 Health Sciences Road East, Irvine, California 92612 (United States)

    1999-08-01

    Local and superficial near-infrared (NIR) optical-property characterization of turbid biological tissues can be achieved by measurement of spatially resolved diffuse reflectance at small source{endash}detector separations ({lt}1.4 mm). However, in these conditions the inverse problem, i.e., calculation of localized absorption and the reduced scattering coefficients, is necessarily sensitive to the scattering phase function. This effect can be minimized if a new parameter of the phase function {gamma}, which depends on the first and the second moments of the phase function, is known. If {gamma} is unknown, an estimation of this parameter can be obtained by the measurement, but the uncertainty of the absorption coefficient is increased. A spatially resolved reflectance probe employing multiple detector fibers (0.3{endash}1.4 mm from the source) is described. Monte Carlo simulations are used to determine {gamma}, the reduced scattering and absorption coefficients from reflectance data. Probe performance is assessed by measurements on phantoms, the optical properties of which were measured by other techniques [frequency domain photon migration (FDPM) and spatially resolved transmittance]. Our results show that changes in the absorption coefficient, the reduced scattering coefficient, and {gamma} can be measured to within {plus_minus}0.005 mm{sup {minus}1}, {plus_minus}0.05 mm{sup {minus}1}, and {plus_minus}0.2, respectively. {ital In vivo} measurements performed intraoperatively on a human skull and brain are reported for four NIR wavelengths (674, 811, 849, 956 nm) when the spatially resolved probe and FDPM are used. The spatially resolved probe shows optimum measurement sensitivity in the measurement volume immediately beneath the probe (typically 1 mm{sup 3} in tissues), whereas FDPM typically samples larger regions of tissues. Optical-property values for human skull, white matter, scar tissue, optic nerve, and tumors are reported that show distinct absorption and scattering differences between structures and a dependence on the phase-function parameter {gamma}. {copyright} 1999 Optical Society of America

  7. Modulating Gold Nanoparticle in vivo Delivery for Photothermal Therapy Applications Using a T Cell Delivery System

    NASA Astrophysics Data System (ADS)

    Kennedy, Laura Carpin

    This thesis reports new gold nanoparticle-based methods to treat chemotherapy-resistant and metastatic tumors that frequently evade conventional cancer therapies. Gold nanoparticles represent an innovative generation of diagnostic and treatment agents due to the ease with which they can be tuned to scatter or absorb a chosen wavelength of light. One area of intensive investigation in recent years is gold nanoparticle photothermal therapy (PTT), in which gold nanoparticles are used to heat and destroy cancer. This work demonstrates the utility of gold nanoparticle PTT against two categories of cancer that are currently a clinical challenge: trastuzumab-resistant breast cancer and metastatic cancer. In addition, this thesis presents a new method of gold nanoparticle delivery using T cells that increases gold nanoparticle tumor accumulation efficiency, a current challenge in the field of PTT. I ablated trastuzumab-resistant breast cancer in vitro for the first time using anti-HER2 labeled silica-gold nanoshells, demonstrating the potential utility of PTT against chemotherapy-resistant cancers. I next established for the first time the use of T cells as gold nanoparticle vehicles in vivo. When incubated with gold nanoparticles in culture, T cells can internalize up to 15000 nanoparticles per cell with no detrimental effects to T cell viability or function (e.g. migration and cytokine secretion). These AuNP-T cells can be systemically administered to tumor-bearing mice and deliver gold nanoparticles four times more efficiently than by injecting free nanoparticles. In addition, the biodistribution of AuNP-T cells correlates with the normal biodistribution of T cell carrier, suggesting the gold nanoparticle biodistribution can be modulated through the choice of nanoparticle vehicle. Finally, I apply gold nanoparticle PTT as an adjuvant treatment for T cell adoptive transfer immunotherapy (Hyperthermia-Enhanced Immunotherapy or HIT) of distant tumors in a melanoma mouse model. The results presented in this thesis expand the potential of gold nanoparticle PTT from only chemotherapy-sensitive or localized cancers to chemotherapy-resistant non-localized cancers that currently defy conventional therapies.

  8. Transurethral ultrasound applicators with dynamic multi-sector control for prostate thermal therapy: In vivo evaluation under MR guidance

    SciTech Connect

    Kinsey, Adam M.; Diederich, Chris J.; Rieke, Viola; Nau, William H.; Pauly, Kim Butts; Bouley, Donna; Sommer, Graham [Thermal Therapy Research Group, Department of Radiation Oncology, University of California, San Francisco, California 94143 (United States) and Joint Graduate Group in Bioengineering, University of California, Berkeley and San Francisco, California 94158 (United States); Department of Radiology, Stanford University Medical Center, Stanford, California 94305 (United States); Thermal Therapy Research Group, Department of Radiation Oncology, University of California, San Francisco, California 94143 (United States); Department of Radiology, Stanford University Medical Center, Stanford, California 94305 (United States); Department of Comparative Medicine, Stanford University, Stanford, California 94305 (United States); Department of Radiology, Stanford University Medical Center, Stanford, California 94305 (United States)

    2008-05-15

    The purpose of this study was to explore the feasibility and performance of a multi-sectored tubular array transurethral ultrasound applicator for prostate thermal therapy, with potential to provide dynamic angular and length control of heating under MR guidance without mechanical movement of the applicator. Test configurations were fabricated, incorporating a linear array of two multi-sectored tubular transducers (7.8-8.4 MHz, 3 mm OD, 6 mm length), with three 120 deg. independent active sectors per tube. A flexible delivery catheter facilitated water cooling (100 ml min{sup -1}) within an expandable urethral balloon (35 mm longx10 mm diameter). An integrated positioning hub allows for rotating and translating the transducer assembly within the urethral balloon for final targeting prior to therapy delivery. Rotational beam plots indicate {approx}90 deg. - 100 deg. acoustic output patterns from each 120 deg. transducer sector, negligible coupling between sectors, and acoustic efficiencies between 41% and 53%. Experiments were performed within in vivo canine prostate (n=3), with real-time MR temperature monitoring in either the axial or coronal planes to facilitate control of the heating profiles and provide thermal dosimetry for performance assessment. Gross inspection of serial sections of treated prostate, exposed to TTC (triphenyl tetrazolium chloride) tissue viability stain, allowed for direct assessment of the extent of thermal coagulation. These devices created large contiguous thermal lesions (defined by 52 deg. C maximum temperature, t{sub 43}=240 min thermal dose contours, and TTC tissue sections) that extended radially from the applicator toward the border of the prostate ({approx}15 mm) during a short power application ({approx}8-16 W per active sector, 8-15 min), with {approx}200 deg. or 360 deg. sector coagulation demonstrated depending upon the activation scheme. Analysis of transient temperature profiles indicated progression of lethal temperature and thermal dose contours initially centered on each sector that coalesced within {approx}5 min to produce uniform and contiguous zones of thermal destruction between sectors, with smooth outer boundaries and continued radial propagation in time. The dimension of the coagulation zone along the applicator was well-defined by positioning and active array length. Although not as precise as rotating planar and curvilinear devices currently under development for MR-guided procedures, advantages of these multi-sectored transurethral applicators include a flexible delivery catheter and that mechanical manipulation of the device using rotational motors is not required during therapy. This multi-sectored tubular array transurethral ultrasound technology has demonstrated potential for relatively fast and reasonably conformal targeting of prostate volumes suitable for the minimally invasive treatment of BPH and cancer under MR guidance, with further development warranted.

  9. Recent advances in in vivo applications of intein-mediated protein splicing

    PubMed Central

    2014-01-01

    Intein-mediated protein splicing has become an essential tool in modern biotechnology. Fundamental progress in the structure and catalytic strategies of cis- and trans-splicing inteins has led to the development of modified inteins that promote efficient protein purification, ligation, modification and cyclization. Recent work has extended these in vitro applications to the cell or to whole organisms. We review recent advances in intein-mediated protein expression and modification, post-translational processing and labeling, protein regulation by conditional protein splicing, biosensors, and expression of trans-genes. PMID:24490831

  10. Stabilization of Collagen-Model, Triple-Helical Peptides for In Vitro and In Vivo Applications

    PubMed Central

    Bhowmick, Manishabrata; Fields, Gregg B.

    2014-01-01

    The triple-helical structure of collagen has been accurately reproduced in numerous chemical and recombinant model systems. Triple-helical peptides and proteins have found application for dissecting collagen-stabilizing forces, isolating receptor- and protein-binding sites in collagen, mechanistic examination of collagenolytic proteases, and development of novel biomaterials. Introduction of native-like sequences into triple-helical constructs can reduce the thermal stability of the triple-helix to below that of the physiological environment. In turn, incorporation of nonnative amino acids and/or templates can enhance triple-helix stability. We presently describe approaches by which triple-helical structure can be modulated for use under physiological or near-physiological conditions. PMID:24014440

  11. First in vivo application and evaluation of a novel method for non-invasive estimation of cardiac output.

    PubMed

    Papaioannou, Theodore G; Soulis, Dimitrios; Vardoulis, Orestis; Protogerou, Athanase; Sfikakis, Petros P; Stergiopulos, Nikolaos; Stefanadis, Christodoulos

    2014-10-01

    Surgical or critically ill patients often require continuous assessment of cardiac output (CO) for diagnostic purposes or for guiding therapeutic interventions. A new method of non-invasive CO estimation has been recently developed, which is based on pressure wave analysis. However, its validity has been examined only in silico. Aim of this study was to evaluate in vivo the reproducibility and accuracy of the "systolic volume balance" method (SVB). Twenty two subjects underwent 2-D transthoracic echocardiography for CO measurement (reference value of CO). The application of SVB method required aortic pressure wave analysis and estimation of total arterial compliance. Aortic pulses were derived by mathematical transformation of radial pressure waves recorded by applanation tonometry. Total compliance was estimated by the "pulse pressure" method. The agreement, association, variability, bias and precision between Doppler and SVB measures of CO were evaluated by intraclass correlation coefficient (ICC), mean difference, SD of differences, percentage error (PR) and Bland-Altman analysis. SVB yielded very reproducible CO estimates (ICC=0.84, mean difference 0.27 ± 0.73 L/min, PR = 16.7%). SVB-derived CO was comparable with Doppler measurements, indicating a good agreement and accuracy (ICC = 0.74, mean difference = -0.22 ± 0.364 L/min, PR ? 15). The basic mathematical and physical principles of the SVB method provide highly reproducible and accurate estimates of CO compared with echocardiography. PMID:25108554

  12. The Development and Application of Magnetic Resonance Elastography of the Normal and Pathological Thyroid Gland in vivo

    PubMed Central

    Bahn, Mark M.; Brennan, Michael D.; Bahn, Rebecca S.; Dean, Diana S.; Kugel, Jennifer L.; Ehman, Richard L.

    2010-01-01

    Purpose To non-invasively assess the shear stiffness of the thyroid gland in vivo in order to determine whether Magnetic Resonance Elastography (MRE) might hold clinical utility in the diagnosis of thyroid disease. Materials and Methods Quantitative parametric images of thyroid stiffness in normal volunteers and patients were produced and quantitative stiffness values measured. Average gland stiffness was determined by region of interest analysis of the parametric images. This technique was used to assess stiffness of the thyroid in normal individuals (n=12), patients with Hashimoto's thyroiditis (HT; n=5), and patients with a solitary benign (n=8) or malignant (n=2) thyroid nodule. Results Mean shear modulus of normal thyroid glands was 1.9 ± 0.6 kPa at 100 Hz and 1.3 ± 0.5 kPa at 80 Hz, while that of HT glands was 2.8 ± 0.6 kPa and 1.8 ± 0.6 kPa at 80 Hz, respectively (p=0.004 at 100 Hz). Elastographic parameters could not differentiate benign from malignant thyroid nodules in these small sample sizes. Conclusion We have developed a method for the application of MRE to the study of thyroid gland pathology. Results show that the HT gland can be differentiated from normal thyroid. The clinical utility of this imaging modality in the diagnosis and management of thyroid disease awaits further study. PMID:19856448

  13. In vitro and in vivo degradation evaluation of novel iron-bioceramic composites for bone implant applications.

    PubMed

    Ulum, M F; Arafat, A; Noviana, D; Yusop, A H; Nasution, A K; Abdul Kadir, M R; Hermawan, H

    2014-03-01

    Biodegradable metals such as magnesium, iron and their alloys have been known as potential materials for temporary medical implants. However, most of the studies on biodegradable metals have been focusing on optimizing their mechanical properties and degradation behavior with no emphasis on improving their bioactivity behavior. We therefore investigated the possibility of improving iron biodegradation rate and bioactivity by incorporating various bioactive bioceramics. The iron-based bioceramic (hydroxyapatite, tricalcium phosphate and biphasic calcium phosphate) composites were prepared by mechanical mixing and sintering process. Degradation studies indicated that the addition of bioceramics lowered the corrosion potential of the composites and slightly increased their corrosion rate compared to that of pure iron. In vitro cytotoxicity results showed an increase of cellular activity when rat smooth muscle cells interacted with the degrading composites compared to pure iron. X-ray radiogram analysis showed a consistent degradation progress with that found in vivo and positive tissue response up to 70 days implantation in sheep animal model. Therefore, the iron-based bioceramic composites have the potential to be used for biodegradable bone implant applications. PMID:24433920

  14. Diagnostic applications of gastric carcinoma cell aptamers in vitro and in vivo.

    PubMed

    Ding, Fei; Guo, Shan; Xie, Min; Luo, Wei; Yuan, Chunhui; Huang, Weihua; Zhou, Yan; Zhang, Xiao-Lian; Zhou, Xiang

    2015-03-01

    Gastric carcinoma is the most malignant tumor. Due to lacking of efficient means to diagnose the cancer at the early stage, it is necessary to develop effective molecular probes for early diagnosis and treatment. We have selected aptamers with high specificity and affinity against SGC7901 cells by cell-SELEX (Systematic Evolution of Ligands by Exponential Enrichment) method, which shown important clinical applications: (1) Specific recognize human gastric tumor tissues compared to the normal tissues. (2)When used to capture cancerous cells, the aptamer-functionalized fluorescent-magnetic nanospheres (FMNS) could specifically capture 93% target cancer cells and about 70% target cells can be released. (3) The aptamer probe displayed a quenched fluorescence in the absence of target cancer cells and went through a conformational transformation upon binding to target cancer cells that induced fluorescence. (4) The aptamer probe could target gastric tumors transplanted into mice with obvious fluorescence. The newly generated aptamers hold great potential in early cancer diagnosis. PMID:25618637

  15. A USPL functional system with articulated mirror arm for in-vivo applications in dentistry

    NASA Astrophysics Data System (ADS)

    Schelle, Florian; Meister, Jörg; Dehn, Claudia; Oehme, Bernd; Bourauel, Christoph; Frentzen, Mathias

    Ultra-short pulsed laser (USPL) systems for dental application have overcome many of their initial disadvantages. However, a problem that has not yet been addressed and solved is the beam delivery into the oral cavity. The functional system that is introduced in this study includes an articulated mirror arm, a scanning system as well as a handpiece, allowing for freehand preparations with ultra-short laser pulses. As laser source an Nd:YVO4 laser is employed, emitting pulses with a duration of tp < 10 ps at a repetition rate of up to 500 kHz. The centre wavelength is at 1064 nm and the average output power can be tuned up to 9 W. The delivery system consists of an articulated mirror arm, to which a scanning system and a custom made handpiece are connected, including a 75 mm focussing lens. The whole functional system is compact in size and moveable. General characteristics like optical losses and ablation rate are determined and compared to results employing a fixed setup on an optical table. Furthermore classical treatment procedures like cavity preparation are being demonstrated on mammoth ivory. This study indicates that freehand preparation employing an USPL system is possible but challenging, and accompanied by a variety of side-effects. The ablation rate with fixed handpiece is about 10 mm3/min. Factors like defocussing and blinding affect treatment efficiency. Laser sources with higher average output powers might be needed in order to reach sufficient preparation speeds.

  16. Preparation, characterization, and in vitro testing of poly(lactide-co-glycolide) and dextran magnetic microspheres for in vivo applications

    NASA Astrophysics Data System (ADS)

    Leamy, Patrick J.

    Many research groups are investigating degradable magnetic particles for magnetic resonance imaging (MRI) contrast agents and as carriers for magnetic drug guidance. These particles are composite materials with a degradable polymer matrix and iron oxide nanoparticles for magnetic properties. The degradable polymer matrix acts to provide colloidal stability and, for drug delivery applications, provides a reservoir for the storage and release of drugs. Natural polymers, like albumin and dextran, which degrade by the action of enzymes; have been used for the polymer matrix. Iron oxide nanoparticles are used for magnetic properties since they can be digested in vivo and have low toxicities. Polylactic acid (PLA) and its copolymers with polyglycolic acid (PLGA) are versatile polymers that degrade by simple hydrolysis without the aid of enzymes. Microspheres are easily formed using the solvent extraction/evaporation method and a wide range of drugs can be encapsulated in them. Magnetic PLGA microspheres suitable for applications were synthesized for the first time in this dissertation. This was accomplished by coating iron oxide nanoparticles with oleic acid to make them dispersible in the organic solvents used in the extraction/evaporation microsphere preparation method. In addition to the magnetic PLGA microspheres, a novel all-aqueous method for preparing crosslinked dextran magnetic microspheres was developed in this dissertation. This method uses free radical polymerization for crosslinking and does not require the use of flammable and harmful solvents. For efficient MRI contrast and magnetic drug guidance, maximized iron oxide content of microspheres is desirable. The two different microsphere preparation methods were optimized for iron oxide content. The effect of iron oxide content on microsphere size and morphology was studied. In addition, an in vitro circulation model was used to evaluate the ability of magnetic microspheres to be guided at physiologic blood flow velocities. The MRI contrast effect was studied as a function of microsphere concentration.

  17. Design and application of an in vivo reporter assay for phenylalanine ammonia-lyase.

    PubMed

    Wang, Siyuan; Zhang, Shuwei; Zhou, Tong; Zeng, Jia; Zhan, Jixun

    2013-09-01

    Phenylalanine ammonia-lyase (PAL) is an important enzyme that links primary metabolism to secondary metabolism. Its efficiency is often a critical factor that affects the overall flux of a related metabolic pathway, the titer of the final products, and the efficacy of PAL-based therapies. Thus, PAL is a common target for metabolic engineering, and it is of significant interest to screen efficient PALs for industrial and medical applications. In this study, a novel and efficient visible reporter assay for screening of PAL efficiency in Escherichia coli was established based on a plant type III polyketide biosynthetic pathway. The candidate PALs were co-expressed with a 4-coumarate:CoA ligase 4CL1 from Arabidopsis thaliana and curcuminoid synthase (CUS) from Oryza sativa in E. coli BL21(DE3) to form a dicinnamoylmethane biosynthetic pathway. Taking advantage of the yellow color of the product, a microplate-based assay was designed to measure the titer of dicinnamoylmethane, which was validated by HPLC analysis. The different titers of the product reflect the overall performance (expression level and enzymatic activity) of the individual PALs in E. coli. Using this system, we have screened three PALs (PAL1, PAL3, and PAL4) from Trifolium pratense, among which PAL1 showed the best performance in E. coli. The engineered E. coli strain containing PAL1, 4CL1, and CUS led to the production of dicinnamoylmethane at a high level of 0.36 g/l. Supplement of 2-fluoro-phenylalanine yielded two fluorinated dicinnamoylmethane derivatives, 6,6'-difluoro-dicinnamoylmethane and 6-fluoro-dicinnamoylmethane, of which the latter is a new curcuminoid. PMID:23907258

  18. Probe depth matters in dermal microdialysis sampling of benzoic acid after topical application: an ex vivo study in human skin.

    PubMed

    Holmgaard, R; Benfeldt, E; Bangsgaard, N; Sorensen, J A; Brosen, K; Nielsen, F; Nielsen, J B

    2012-01-01

    Microdialysis (MD) in the skin - dermal microdialysis (DMD) - is a unique technique for sampling of topically as well as systemically administered drugs at the site of action, e.g. sampling of dermatological drug concentrations in the dermis. Debate has concerned the existence of a correlation between the depth of the sampling device - the probe - in the dermis and the amount of drug sampled following topical drug administration. This study evaluates the relation between probe depth and drug sampling using dermal DMD sampling ex vivo in human skin. We used superficial (<1 mm), intermediate (1-2 mm) and deep (>2 mm) positioning of the linear MD probe in the dermis of human abdominal skin, followed by topical application of 4 mg/ml of benzoic acid (BA) in skin chambers overlying the probes. Dialysate was sampled every hour for 12 h and analysed for BA content by high-performance liquid chromatography. Probe depth was measured by 20-MHz ultrasound scanning. The area under the time-versus-concentration curve (AUC) describes the drug exposure in the tissue during the experiment and is a relevant parameter to compare for the 3 dermal probe depths investigated. The AUC(0-12) were: superficial probes: 3,335 ± 1,094 ?g·h/ml (mean ± SD); intermediate probes: 2,178 ± 1,068 ?g·h/ml, and deep probes: 1,159 ± 306 ?g·h/ml. AUC(0-12) sampled by the superficial probes was significantly higher than that of samples from the intermediate and deeply positioned probes (p value <0.05). There was a significant inverse correlation between probe depth and AUC(0-12) sampled by the same probe (p value <0.001, r(2) value = 0.5). The mean extrapolated lag-times (±SD) for the superficial probes were 0.8 ± 0.1 h, for the intermediate probes 1.7 ± 0.5 h, and for the deep probes 2.7 ± 0.5 h, which were all significantly different from each other (p value <0.05). In conclusion, this paper demonstrates that there is an inverse relationship between the depth of the probe in the dermis and the amount of drug sampled following topical penetration ex vivo. The result is of relevance to the in vivo situation, and it can be predicted that the differences in sampling at different probe depths will have a more significant impact in the beginning of a study or in studies of short duration. Based on this study it can be recommended that studies of topical drug penetration using DMD sampling should include measurements of probe depth and that efforts should be made to minimize probe depth variability. PMID:21849814

  19. Influence of formulation type on the deposition of glycolic acid and glycerol in hairless mouse skin following topical in vivo application

    Microsoft Academic Search

    M. OHTA; C. RAMACHANDRAN; N. D. WEINER

    Synopsis The kinetics and extent of uptake of glycolic acid and glycerol in various strata of hairless mouse skin following topical in vivo application of several glycolic acid and glycerol formulations were determined. The formulations tested included an aqueous solution, a 30% propylene glycol (PG) aqueous solution, an oil-in-water (O\\/W) emulsion, a water-in-oil (W\\/O) emulsion, and two nonionic liposomal systems.

  20. Preparation and in vitro/in vivo evaluation of cyclosporin a-loaded nanodecorated ocular implants for subconjunctival application.

    PubMed

    Pehlivan, Sibel Bozda?; Yavuz, Burçin; Çalamak, Semih; Ulubayram, Kezban; Kaffashi, Abbas; Vural, ?mran; Çakmak, Hasan Basri; Durgun, Meltem Ezgi; Denkba?, Emir Baki; Ünlü, Nur?en

    2015-05-01

    In terms of ocular drug delivery, biodegradable implant systems have several advantages including the ability to provide constant drug concentration at the target site, no necessity for surgical removal, and minimum systemic side effects. Cyclosporin A (CsA) is a neutral, hydrophobic, cyclic peptide of amino acids that frequently used for dry eye disease treatment. The aim of this study was to develop a nanoparticle-loaded implant system for sustained-release CsA delivery following subconjunctival implantation. Poly(lactide-co-glycolide) (85:15) or poly-?-caprolactone (PCL) were used to prepare two different nanoparticle formulations. These nanoparticles loaded into PCL or poly(lactide-co-caprolactone) implant formulations were prepared by two different methods, which were molding and electrospinning. Size and zeta potential of nanoparticles were determined and the morphology of the formulations were investigated by scanning electron microscopy. CsA-loading efficiencies were calculated and the in vitro degradation and in vitro release studies were performed. MTT test was also performed using L929 fibroblast cells to evaluate the cytotoxicity of the formulations. PCL-PCL-NP-I formulation was implanted to Swiss Albino mice with induced dry eye syndrome to evaluate the efficacy. In vitro release studies showed that the release from the formulations continues between 30 and 60 days, and the cell viability was found to be 77.4%-99.0%. In vivo studies showed that healing is significantly faster in the presence of the selected implant formulation. Results indicated that nanodecorated implants are promising ocular carriers for controlled-release CsA application. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 104:1709-1720, 2015. PMID:25716582

  1. In vitro and in vivo corrosion properties of new iron-manganese alloys designed for cardiovascular applications.

    PubMed

    Drynda, Andreas; Hassel, Thomas; Bach, Friedrich Wilhelm; Peuster, Matthias

    2015-04-01

    The principle of biodegradation for the production of temporary implant materials (e.g. stents) plays an important role in the treatment of congenital heart defects. In the last decade several attempts have been made with different alloy materials-mainly based on iron and magnesium. None of the currently available materials in this field have demonstrated satisfying results and have therefore not found entry into broad clinical practice. While magnesium or magnesium alloy systems corrode too fast, the corrosion rate of pure iron-stents is too slow for cardiovascular applications. In the last years FeMn alloy systems were developed with the idea that galvanic effects, caused by different electrochemical properties of Fe and Mn, would increase the corrosion rate. In vitro tests with alloys containing up to 30% Mn showed promising results in terms of biocompatibility. This study deals with the development of new FeMn alloy systems with lower Mn concentrations (FeMn 0.5 wt %, FeMn 2.7 wt %, FeMn 6.9 wt %) to avoid Mn toxicity. Our results show, that these alloys exhibit good mechanical features as well as suitable in vitro biocompatibility and corrosion properties. In contrast, the evaluation of these alloys in a mouse model led to unexpected results-even after 9 months no significant corrosion was detectable. Preliminary SEM investigations showed that passivation layers (FeMn phosphates) might be the reason for corrosion resistance. If this can be proved in further experiments, strategies to prevent or dissolve those layers need to be developed to expedite the in vivo corrosion of FeMn alloys. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 103B: 649-660, 2015. PMID:24976236

  2. 21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture processing...

  3. 21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture processing...

  4. 21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture processing...

  5. 21 CFR 876.5885 - Tissue culture media for human ex vivo tissue and cell culture processing applications.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES GASTROENTEROLOGY-UROLOGY DEVICES Therapeutic Devices § 876.5885 Tissue culture media for human ex vivo tissue and cell culture processing...

  6. Laguerre-based method for analysis of time-resolved fluorescence data: application to in-vivo characterization and diagnosis of atherosclerotic lesions

    PubMed Central

    Jo, Javier A.; Fang, Qiyin; Papaioannou, Thanassis; Baker, J. Dennis; Dorafshar, Amir H.; Reil, Todd; Qiao, Jian-Hua; Fishbein, Michael C.; Freischlag, Julie A.; Marcu, Laura

    2007-01-01

    We report the application of the Laguerre deconvolution technique (LDT) to the analysis of in-vivo time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) data and the diagnosis of atherosclerotic plaques. TR-LIFS measurements were obtained in vivo from normal and atherosclerotic aortas (eight rabbits, 73 areas), and subsequently analyzed using LDT. Spectral and time-resolved features were used to develop four classification algorithms: linear discriminant analysis (LDA), stepwise LDA (SLDA), principal component analysis (PCA), and artificial neural network (ANN). Accurate deconvolution of TR-LIFS in-vivo measurements from normal and atherosclerotic arteries was provided by LDT. The derived Laguerre expansion coefficients reflected changes in the arterial biochemical composition, and provided a means to discriminate lesions rich in macrophages with high sensitivity (>85%) and specificity (>95%). Classification algorithms (SLDA and PCA) using a selected number of features with maximum discriminating power provided the best performance. This study demonstrates the potential of the LDT for in-vivo tissue diagnosis, and specifically for the detection of macrophages infiltration in atherosclerotic lesions, a key marker of plaque vulnerability. PMID:16674179

  7. Comparative in vitro and in vivo pharmacological investigation of platinum(IV) complexes as novel anticancer drug candidates for oral application

    PubMed Central

    Theiner, Sarah; Varbanov, Hristo P.; Galanski, Markus; Egger, Alexander E.; Berger, Walter; Heffeter, Petra; Keppler, Bernhard K.

    2015-01-01

    Platinum(IV) complexes are promising candidates as prodrugs for oral application in anticancer chemotherapy. However, only a few Pt(IV) compounds entered (pre)clinical trials, e.g. satraplatin, while most of the others were only tested in vitro. Aim of the study was investigation of the in vivo pharmacological behavior as well as the anticancer activity of two novel platinum(IV) complexes vs. satraplatin. The drugs were selected due to significantly different in vitro cytotoxicity while sharing some physicochemical properties (e.g. lipophilicity). Initial experiments indicated that the highly in vitro cytotoxic compound 1 ((OC-6-33)-dichloridobis((4-ethoxy)-4-oxobutanoato)-bis(ethylamine)platinum(IV)) was also characterized by high drug absorption and tissue platinum levels after oral application. Interestingly, analysis of serum samples using SEC-ICP-MS revealed that the administered drugs have completely been metabolized and/or bound to proteins in serum within 2 h after treatment. With regard to the activity in vivo, the outcomes were rather unexpected: although potent anticancer effect of 1 was observed in cell culture, the effects in vivo were rather minor. Nevertheless, 1 was superior to 2 ((OC-6-33)-diammine(cyclobutane-1,1-dicarboxylato)-bis((4-cyclopentylamino)-4-oxobutanoato)platinum(IV)) after i.p. administration, which was, at least to some extent, in accordance to the cell culture experiments. After oral gavage, both compounds exhibited comparable activity. This is remarkable considering the distinctly lower activity of 2 in cell culture as well as the low platinum levels detected both in serum and tissues after oral application. Consequently, our data indicate that the prediction of in vivo anticancer activity by cell culture experiments is not trivial, especially for orally applied drugs. PMID:25413442

  8. Comparative in vitro and in vivo pharmacological investigation of platinum(IV) complexes as novel anticancer drug candidates for oral application.

    PubMed

    Theiner, Sarah; Varbanov, Hristo P; Galanski, Markus; Egger, Alexander E; Berger, Walter; Heffeter, Petra; Keppler, Bernhard K

    2015-01-01

    Platinum(IV) complexes are promising candidates as prodrugs for oral application in anticancer chemotherapy. However, only a few Pt(IV) compounds entered (pre)clinical trials, e.g. satraplatin, while most of the others were only tested in vitro. Aim of the study was investigation of the in vivo pharmacological behavior as well as the anticancer activity of two novel platinum(IV) complexes vs. satraplatin. The drugs were selected due to significantly different in vitro cytotoxicity while sharing some physicochemical properties (e.g. lipophilicity). Initial experiments indicated that the highly in vitro cytotoxic compound 1 ((OC-6-33)-dichloridobis((4-ethoxy)-4-oxobutanoato)-bis(ethylamine)platinum(IV)) was also characterized by high drug absorption and tissue platinum levels after oral application. Interestingly, analysis of serum samples using SEC-ICP-MS revealed that the administered drugs have completely been metabolized and/or bound to proteins in serum within 2 h after treatment. With regard to the activity in vivo, the outcomes were rather unexpected: although potent anticancer effect of 1 was observed in cell culture, the effects in vivo were rather minor. Nevertheless, 1 was superior to 2 ((OC-6-33)-diammine(cyclobutane-1,1-dicarboxylato)-bis((4-cyclopentylamino)-4-oxobutanoato)platinum(IV)) after i.p. administration, which was, at least to some extent, in accordance to the cell culture experiments. After oral gavage, both compounds exhibited comparable activity. This is remarkable considering the distinctly lower activity of 2 in cell culture as well as the low platinum levels detected both in serum and tissues after oral application. Consequently, our data indicate that the prediction of in vivo anticancer activity by cell culture experiments is not trivial, especially for orally applied drugs. PMID:25413442

  9. A volume birdcage coil with an adjustable sliding tuner ring for neuroimaging in high field vertical magnets: Ex and in vivo applications at 21.1 T

    NASA Astrophysics Data System (ADS)

    Qian, Chunqi; Masad, Ihssan S.; Rosenberg, Jens T.; Elumalai, Malathy; Brey, William W.; Grant, Samuel C.; Gor'kov, Peter L.

    2012-08-01

    A tunable 900 MHz transmit/receive volume coil was constructed for 1H MR imaging of biological samples in a 21.1 T vertical bore magnet. To accommodate a diverse range of specimen and RF loads at such a high frequency, a sliding-ring adaptation of a low-pass birdcage was implemented through simultaneous alteration of distributed capacitance. To make efficient use of the constrained space inside the vertical bore, a modular probe design was implemented with a bottom-adjustable tuning and matching apparatus. The sliding ring coil displays good homogeneity and sufficient tuning range for different samples of various dimensions representing large span of RF loads. High resolution in vivo and ex vivo images of large rats (up to 350 g), mice and human postmortem tissues were obtained to demonstrate coil functionality and to provide examples of potential applications at 21.1 T.

  10. A volume birdcage coil with an adjustable sliding tuner ring for neuroimaging in high field vertical magnets: ex and in vivo applications at 21.1 T

    PubMed Central

    Qian, Chunqi; Masad, Ihssan S.; Rosenberg, Jens T.; Elumalai, Malathy; Brey, William W.; Grant, Samuel C.; Gor’kov, Peter L.

    2012-01-01

    A tunable 900 MHz transmit/receive volume coil was constructed for 1H MR imaging of biological samples in a 21.1 T vertical bore magnet. To accommodate a diverse range of specimen and RF loads at such a high frequency, a sliding-ring adaptation of a low-pass birdcage was implemented through simultaneous alteration of distributed capacitance. To make efficient use of the constrained space inside the vertical bore, a modular probe design was implemented with a bottom-adjustable tuning and matching apparatus. The sliding ring coil displays good homogeneity and sufficient tuning range for different samples of various dimensions representing large span of RF loads. High resolution in vivo and ex vivo images of large rats (up to 350 g), mice and human postmortem tissues were obtained to demonstrate coil functionality and to provide examples of potential applications at 21.1 T. PMID:22750638

  11. The application of statistical moment theory to the evaluation of in vivo dissolution time and absorption time

    Microsoft Academic Search

    Sidney Riegelman; Paul Collier

    1980-01-01

    Moments analysis has been applied to the calculation of mean (in vivo)dissolution time (MDT) and mean absorption time (MAT) from plasma level of drug versus time data. Methods for accurately estimating the MDT under varying conditions, limitations of the methods, and interpretation of the data are presented. The importance of accurate estimates of the terminal rate constant (?z) and the

  12. Characteristics and Applications of the ToxRefDB In Vivo Datasets from Chronic, Reproductive and Developmental Assays

    EPA Science Inventory

    ToxRefDB was developed to store data from in vivo animal toxicity studies. The initial focus was populating ToxRefDB with pesticide registration toxicity data that has been historically stored as hard-copy and scanned documents by the Office of Pesticide Programs. A significant p...

  13. Application of a partial-thickness human ex vivo skin culture model in cutaneous wound healing study.

    PubMed

    Xu, Wei; Jong Hong, Seok; Jia, Shengxian; Zhao, Yanan; Galiano, Robert D; Mustoe, Thomas A

    2012-04-01

    A number of in vivo and ex vivo skin models have been applied to human wound healing studies. A reliable skin model, which recapitulates the features of human wound repair, is essential for the clinical and mechanical investigation of human cutaneous wound healing. Full-skin ex vivo culture systems have been used in wound healing studies. However, important structures of the skin, such as the differentiation of keratinocytes and epidermis-dermis junction, are poorly characterized in this model. This study aims to develop an optimized partial-thickness human ex vivo skin culture (HESC) model to maintain human skin characteristics in vitro. During our culture, the basal layer, suprabasal layer, and stratum granulosum layer of epidermis were preserved until day 8. Analyses of hemidesmosome proteins, bullous pemphigoid antigen 1 (BP180) and 2 (BP230), showed that the integrity of the basement membrane of the epidermis was well preserved in the HESC model. In contrast, an organotypic culture with human keratinocytes and fibroblasts failed to show an integrated basement membrane. Maintenance of skin structure by histological analysis and proliferation of epidermal keratinocytes by Ki67 staining were observed in our model for 12 days. Complete re-epithelialization of the wounding area was observed at day 6 post wounding when a superficial incisional wound was created. The expression of Ki-67 and keratin 6, indicators of activated keratinocytes in epidermis, was significantly upregulated and new collagen synthesis was found in the dermis during the wound healing process. As control, we also used organotypic culture in studying the differentiation of the keratinocyte layers and incisional wound repair. It turned out that our model has advantage in these study fields. The results suggest that our HESC model retains important elements of in vivo skin and has significant advantages for the wound healing studies in vitro. PMID:22231737

  14. ?TCP ceramic doped with dicalcium silicate for bone regeneration applications prepared by powder metallurgy method: in vitro and in vivo studies.

    PubMed

    Velasquez, Pablo; Luklinska, Zofia B; Meseguer-Olmo, Luis; Mate-Sanchez de Val, Jose E; Delgado-Ruiz, Rafael A; Calvo-Guirado, Jose L; Ramirez-Fernandez, Ma P; de Aza, Piedad N

    2013-07-01

    This study reports on the in vitro and in vivo behavior of ?-tricalcium phosphate (?TCP) and also ?TCP doped with either 1.5 or 3.0 wt % of dicalcium silicate (C2 S). The ceramics were successfully prepared by powder metallurgy method combined with homogenization and heat treatment procedures. All materials were composed of a single-phase, ?TCP in the case of a pure material, or solid solution of C2 S in ?TCP for the doped ?TCP, which were stable at room temperature. The ceramics were tested for bioactivity in simulated body fluid, cell culture medium containing adult mesenchymal stem cells of human origin, and in animals. Analytical scanning electron microscopy combined with chemical elemental analysis was used and Fourier transform infrared and conventional histology methods. The in vivo behavior of the ceramics matched the in vitro results, independently of the C2 S content in ?TCP. Carbonated hydroxyapatite (CHA) layer was formed on the surface and within the inner parts of the specimens in all cases. A fully mineralized new bone growing in direct contact with the implants was found under the in vivo conditions. The bioactivity and biocompatibility of the implants increased with the C2 S content in ?TCP. The C2 S doped ceramics also favoured a phase transformation of ?TCP into CHA, important for full implant integration during the natural bone healing processes. ?TCP ceramic doped with 3.0 wt % C2 S showed the best bioactive in vitro and in vivo properties of all the compositions and hence could be of interest in specific applications for bone restorative purposes. PMID:23225787

  15. Oxygen Sensing Strategies in Mammals and Bacteria

    PubMed Central

    Taabazuing, Cornelius Y.; Hangasky, John A.; Knapp, Michael J.

    2014-01-01

    The ability to sense and adapt to changes in pO2 is crucial for basic metabolism in most organisms, leading to elaborate pathways for sensing hypoxia (low pO2). This review focuses on the mechanisms utilized by mammals and bacteria to sense hypoxia. While responses to acute hypoxia in mammalian tissues lead to altered vascular tension, the molecular mechanism of signal transduction is not well understood. In contrast, chronic hypoxia evokes cellular responses that lead to transcriptional changes mediated by the hypoxia inducible factor (HIF), which is directly controlled by post-translational hydroxylation of HIF by the non-heme Fe(II)/?KG-dependent enzymes FIH and PHD2. Research on PHD2 and FIH is focused on developing inhibitors and understanding the links between HIF binding and the O2 reaction in these enzymes. Sulfur speciation is a putative mechanism for acute O2-sensing, with special focus on the role of H2S. This sulfur-centered model is discussed, as are some of the directions for further refinement of this model. In contrast to mammals, bacterial O2-sensing relies on protein cofactors that either bind O2 or oxidatively decompose. The sensing modality for bacterial O2-sensors is either via altered DNA binding affinity of the sensory protein, or else due to the actions of a two-component signaling cascade. Emerging data suggests that proteins containing a hemerythrin-domain, such as FBXL5, may serve to connect iron sensing to O2-sensing in both bacteria and humans. As specific molecular machinery becomes identified, these hypoxia sensing pathways present therapeutic targets for diseases including ischemia, cancer, or bacterial infection. PMID:24468676

  16. Rapid response oxygen-sensing nanofibers

    PubMed Central

    Xue, Ruipeng; Behera, Prajna; Viapiano, Mariano S.; Lannutti, John J.

    2014-01-01

    Molecular oxygen has profound effects on cell and tissue viability. Relevant sensor forms that can rapidly determine dissolved oxygen levels under biologically relevant conditions provide critical metabolic information. Using 0.5 ?m diameter electrospun polycaprolactone (PCL) fiber containing an oxygen-sensitive probe, tris (4,7-diphenyl-1,10-phenanthroline) ruthenium(II) dichloride, we observed a response time of 0.9±0.12 seconds – 4–10 times faster than previous reports – while the t95 for the corresponding film was more than two orders of magnitude greater. Interestingly, the response and recovery times of slightly larger diameter PCL fibers were 1.79±0.23 s and 2.29±0.13 s, respectively, while the recovery time was not statistically different likely due to the more limited interactions of nitrogen with the polymer matrix. A more than 10-fold increase in PCL fiber diameter reduces oxygen sensitivity while having minor effects on response time; conversely, decreases in fiber diameter to less than 0.5 ?m would likely decrease response times even further. In addition, a 50°C heat treatment of the electrospun fiber resulted in both increased Stern-Volmer slope and linearity likely due to secondary recrystallization that further homogenized the probe microenvironment. At exposure times up to 3600 s in length, photobleaching was observed but was largely eliminated by the use of either polyethersulfone (PES) or a PES-PCL core-shell composition. However, this resulted in 2- and 3-fold slower response times. Finally, even the non-core shell compositions containing the Ru oxygen probe result in no apparent cytotoxicity in representative glioblastoma cell populations. PMID:23706233

  17. TRANSCRIPTION: Oxygen Sensing Gets a Second Wind

    NSDL National Science Digital Library

    Richard K. Bruick (University of Texas Southwestern Medical Center; Department of Biochemistry)

    2002-02-01

    Access to the article is free, however registration and sign-in are required. The recent discovery of molecules that mediate the hypoxia response pathway demonstrates that mammalian cells are supremely well adapted to dealing with conditions of low oxygen. In their Perspective, Bruick and McKnight detail new findings (Lando et al.) that describe an additional way in which the transcription factor HIF, a central player in the hypoxia response pathway, can be regulated.

  18. In Vivo Imaging of Brain Development: Technologies, Models, Applications, and Impact on Understanding the Etiology of Mental Retardation

    Microsoft Academic Search

    Vicko Gluncic

    \\u000a Development of the mammalian brain proceeds in a precisely controlled sequence of cell divisions, migration, differentiation,\\u000a and synaptogenesis. It is a process of precise dynamic assembly, and time lapse in vivo imaging of these processes is fundamental\\u000a for the multidisciplinary endeavor to merge and understand the morphological, physiological, and regulatory processes of neurogenesis.\\u000a \\u000a \\u000a Modern optical and non-optical imaging technologies enable

  19. Application of Ultrasound on Monitoring the Evolution of the Collagen Fiber Reinforced nHAC/CS Composites In Vivo

    PubMed Central

    Chen, Yan; Yan, Yuting; Li, Xiaoming; Tan, Huiting; Li, Huajun; Zhu, Yanwen; Niemeyer, Philipp; Yaega, Matin; Yu, Bo

    2014-01-01

    To date, fiber reinforce scaffolds have been largely applied to repair hard and soft tissues. Meanwhile, monitoring the scaffolds for long periods in vivo is recognized as a crucial issue before its wide use. As a consequence, there is a growing need for noninvasive and convenient methods to analyze the implantation remolding process in situ and in real time. In this paper, diagnostic medical ultrasound was used to monitor the in vivo bone formation and degradation process of the novel mineralized collagen fiber reinforced composite which is synthesized by chitosan (CS), nanohydroxyapatite (nHA), and collagen fiber (Col). To observe the impact of cells on bone remodeling process, the scaffolds were planted into the back of the SD rats with and without rat bone mesenchymal stem cells (rBMSCs). Systematic data of scaffolds in vivo was extracted from ultrasound images. Significant consistency between the data from the ultrasound and DXA could be observed (P < 0.05). This indicated that ultrasound may serve as a feasible alternative for noninvasive monitoring the evolution of scaffolds in situ during cell growth. PMID:24822206

  20. Metabolic engineering applications of in vivo sup 31 P and sup 13 C NMR studies of Saccharomyces cerevisiae

    SciTech Connect

    Shanks, J.V.

    1989-01-01

    With intent to quantify NMR measurements as much as possible, analysis techniques of the in vivo {sup 31}P NMR spectrum are developed. A systematic procedure is formulated for estimating the relative intracellular concentrations of the sugar phosphates in S. cerevisiae from the {sup 31}P NMR spectrum. In addition, in vivo correlation of inorganic phosphate chemical shift with the chemical shifts of 3-phosphoglycerate, {beta}-fructose 1,6-diphosphate, fructose 6-phosphate, and glucose 6-phosphate are determined. Also, a method was developed for elucidation of the cytoplasmic and vacuolar components of inorganic phosphate in the {sup 31}P NMR spectrum of S. cerevisiae. An in vivo correlation relating the inorganic phosphate chemical shift of the vacuole with the chemical shift of the resonance for pyrophosphate and the terminal phosphate of polyphosphate (PP{sub 1}) is established. Transient measurements provided by {sup 31}P NMR are applied to reg1 mutant and standard strains. {sup 31}P and {sup 13}C NMR measurements are used to analyze the performance of recombinant strains in which the glucose phosphorylation step had been altered.

  1. Feasibility Study of Glass Dosimeter for In Vivo Measurement: Dosimetric Characterization and Clinical Application in Proton Beams

    SciTech Connect

    Rah, Jeong-Eun; Oh, Do Hoon; Kim, Jong Won [Department of Radiation Oncology, Myongji Hospital, Kwandong University College of Medicine, Goyang (Korea, Republic of)] [Department of Radiation Oncology, Myongji Hospital, Kwandong University College of Medicine, Goyang (Korea, Republic of); Kim, Dae-Hyun; Suh, Tae-Suk [Department of Biomedical Engineering, Catholic University of Korea, Seoul (Korea, Republic of)] [Department of Biomedical Engineering, Catholic University of Korea, Seoul (Korea, Republic of); Ji, Young Hoon [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of)] [Korea Institute of Radiological and Medical Sciences, Seoul (Korea, Republic of); Shin, Dongho; Lee, Se Byeong; Kim, Dae Yong [Proton Therapy Center, National Cancer Center, Goyang (Korea, Republic of)] [Proton Therapy Center, National Cancer Center, Goyang (Korea, Republic of); Park, Sung Yong, E-mail: Sung.Park@mclaren.org [Proton Therapy Center, McLaren Cancer Institute, Flint, Michigan (United States)

    2012-10-01

    Purpose: To evaluate the suitability of the GD-301 glass dosimeter for in vivo dose verification in proton therapy. Methods and Materials: The glass dosimeter was analyzed for its dosimetrics characteristic in proton beam. Dosimeters were calibrated in a water phantom using a stairlike holder specially designed for this study. To determine the accuracy of the glass dosimeter in proton dose measurements, we compared the glass dosimeter and thermoluminescent dosimeter (TLD) dose measurements using a cylindrical phantom. We investigated the feasibility of the glass dosimeter for the measurement of dose distributions near the superficial region for proton therapy plans with a varying separation between the target volume and the surface of 6 patients. Results and Discussion: Uniformity was within 1.5%. The dose-response has good linearity. Dose-rate, fading, and energy dependence were found to be within 3%. The beam profile measured using the glass dosimeter was in good agreement with the profile obtained from the ionization chamber. Depth-dose distributions in nonmodulated and modulated proton beams obtained with the glass dosimeter were estimated to be within 3%, which was lower than those with the ionization chamber. In the phantom study, the difference of isocenter dose between the delivery dose calculated by the treatment planning system and that measured by the glass dosimeter was within 5%. With in vivo dosimetry, the calculated surface doses overestimated measurements by 4%-16% using glass dosimeter and TLD. Conclusion: It is recommended that bolus be added for these clinical cases. We also believe that the glass dosimeter has considerable potential for use with in vivo patient proton dosimetry.

  2. Application of wide-field optical coherence tomography to monitoring of viability of rat brain in vivo

    NASA Astrophysics Data System (ADS)

    Sato, Manabu; Nishidate, Izumi

    2014-05-01

    We investigated the feasibility of OCT in monitoring the viability of the brain. It was confirmed that after an overdose of pentobarbital sodium salt for an euthanasia, the OCT signal intensity increased before cardiac arrest and finally became 2.7 times, and by periodically changing the tissue temperature from 20 to 32 °C in vivo, average correlation coefficients between the ratio of signal intensity (RSI) and temperature were determined to be -0:42 to -0:50. RSI reversibly changed with subsequent variations of temperatures and finally increased rapidly just before cardiac arrest. These results indicate that RSI could correspond to decreases in viability.

  3. Near-infrared emission Ba3(PO4)2:Mn5+ phosphor and potential application in vivo fluorescence imaging.

    PubMed

    Cao, Renping; Yu, Xiaoguang; Sun, Xinyuan; Cao, Chunyan; Qiu, Jianrong

    2014-07-15

    Fluorescence imaging in the second near-infrared window (NIR-II, 1000-1350 nm) is attracting attention due to negligible tissue scattering and lower tissue autofluorescence, etc. Here, Ba3(PO4)2:Mn(5+) phosphor is prepared via solid state reaction method in air, and NIR emission band peaking at ?1191 nm in the NIR-II region is observed. According to experiment results, Ba3(PO4)2:Mn(5+) phosphor has a great potential for the study of the NIR-II fluorescence imaging in vivo. PMID:24691378

  4. Observation of Multiphoton-induced Fluorescence from Nano Graphene Oxide and Its Applications in In vitro and In vivo Bioimaging

    E-print Network

    Qian, Jun; Peng, Li; Xi, Wang; Cai, Fu-Hong; Zhu, Zhen-Feng; He, Hao; Hu, Ming-Lie; He, Sailing

    2012-01-01

    In the present paper, we observed both two-photon and three-photon induced distinct photoluminescence from GO nanoparticles under fs laser excitation. Conjugated with PEG molecules, GO nanoparticles exhibited high chemical stability, and could effectively label HeLa cells. Imaged with a two-photon scanning microscope, GO nanoparticles were observed to localize in the mitochondria, endoplasmic reticulum, Golgi and lysosome of HeLa cells. Furthermore, GO nanoparticles were micro-injected into the brain of a black mouse, and in vivo two-photon luminescence imaging illustrated that GO nanoparticles located at 300 {\\mu}m depth in the brain could be clearly distinguished.

  5. In vivo application of an optical segment tracking approach for bone loading regimes recording in humans: a reliability study.

    PubMed

    Yang, Peng-Fei; Sanno, Maximilian; Ganse, Bergita; Koy, Timmo; Brüggemann, Gert-Peter; Müller, Lars Peter; Rittweger, Jörn

    2014-08-01

    This paper demonstrates an optical segment tracking (OST) approach for assessing the in vivo bone loading regimes in humans. The relative movement between retro-reflective marker clusters affixed to the tibia cortex by bone screws was tracked and expressed as tibia loading regimes in terms of segment deformation. Stable in vivo fixation of bone screws was tested by assessing the resonance frequency of the screw-marker structure and the relative marker position changes after hopping and jumping. Tibia deformation was recorded during squatting exercises to demonstrate the reliability of the OST approach. Results indicated that the resonance frequencies remain unchanged prior to and after all exercises. The changes of Cardan angle between marker clusters induced by the exercises were rather minor, maximally 0.06°. The reproducibility of the deformation angles during squatting remained small (0.04°/m-0.65°/m). Most importantly, all surgical and testing procedures were well tolerated. The OST method promises to bring more insights of the mechanical loading acting on bone than in the past. PMID:24907129

  6. In vitro and in vivo investigations of upconversion and NIR emitting Gd?O?:Er³?,Yb³? nanostructures for biomedical applications.

    PubMed

    Hemmer, Eva; Takeshita, Hiroyuki; Yamano, Tomoyoshi; Fujiki, Takanori; Kohl, Yvonne; Löw, Karin; Venkatachalam, Nallusamy; Hyodo, Hiroshi; Kishimoto, Hidehiro; Soga, Kohei

    2012-10-01

    The use of an "over 1000-nm near-infrared (NIR) in vivo fluorescence bioimaging" system based on lanthanide containing inorganic nanostructures emitting in the visible and NIR range under 980-nm excitation is proposed. It may overcome problems of currently used biomarkers including color fading, phototoxicity and scattering. Gd(2)O(3):Er(3+),Yb(3+) nanoparticles and nanorods showing upconversion and NIR emission are synthesized and their cytotoxic behavior is investigated by incubation with B-cell hybridomas and macrophages. Surface modification with PEG-b-PAAc provides the necessary chemical durability reducing the release of toxic Gd(3+) ions. NIR fluorescence microscopy is used to investigate the suitability of the nanostructures as NIR-NIR biomarkers. The in vitro uptake of bare and modified nanostructures by macrophages is investigated by confocal laser scanning microscopy. In vivo investigations revealed nanostructures in liver, lung, kidneys and spleen a few hours after injection into mice, while most of the nanostructures have been removed from the body after 24 h. PMID:22588504

  7. Carotid artery wall motion analysis from B-mode ultrasound using adaptive block matching: in silico evaluation and in vivo application

    NASA Astrophysics Data System (ADS)

    Gastounioti, A.; Golemati, S.; Stoitsis, J. S.; Nikita, K. S.

    2013-12-01

    Valid risk stratification for carotid atherosclerotic plaques represents a crucial public health issue toward preventing fatal cerebrovascular events. Although motion analysis (MA) provides useful information about arterial wall dynamics, the identification of motion-based risk markers remains a significant challenge. Considering that the ability of a motion estimator (ME) to handle changes in the appearance of motion targets has a major effect on accuracy in MA, we investigated the potential of adaptive block matching (ABM) MEs, which consider changes in image intensities over time. To assure the validity in MA, we optimized and evaluated the ABM MEs in the context of a specially designed in silico framework. ABMFIRF2, which takes advantage of the periodicity characterizing the arterial wall motion, was the most effective ABM algorithm, yielding a 47% accuracy increase with respect to the conventional block matching. The in vivo application of ABMFIRF2 revealed five potential risk markers: low movement amplitude of the normal part of the wall adjacent to the plaques in the radial (RMAPWL) and longitudinal (LMAPWL) directions, high radial motion amplitude of the plaque top surface (RMAPTS), and high relative movement, expressed in terms of radial strain (RSIPL) and longitudinal shear strain (LSSIPL), between plaque top and bottom surfaces. The in vivo results were reproduced by OFLK(WLS) and ABMKF-K2, MEs previously proposed by the authors and with remarkable in silico performances, thereby reinforcing the clinical values of the markers and the potential of those MEs. Future in vivo studies will elucidate with confidence the full potential of the markers.

  8. Carotid artery wall motion analysis from B-mode ultrasound using adaptive block matching: in silico evaluation and in vivo application.

    PubMed

    Gastounioti, A; Golemati, S; Stoitsis, J S; Nikita, K S

    2013-12-21

    Valid risk stratification for carotid atherosclerotic plaques represents a crucial public health issue toward preventing fatal cerebrovascular events. Although motion analysis (MA) provides useful information about arterial wall dynamics, the identification of motion-based risk markers remains a significant challenge. Considering that the ability of a motion estimator (ME) to handle changes in the appearance of motion targets has a major effect on accuracy in MA, we investigated the potential of adaptive block matching (ABM) MEs, which consider changes in image intensities over time. To assure the validity in MA, we optimized and evaluated the ABM MEs in the context of a specially designed in silico framework. ABM(FIRF2), which takes advantage of the periodicity characterizing the arterial wall motion, was the most effective ABM algorithm, yielding a 47% accuracy increase with respect to the conventional block matching. The in vivo application of ABM(FIRF2) revealed five potential risk markers: low movement amplitude of the normal part of the wall adjacent to the plaques in the radial (RMA(PWL)) and longitudinal (LMA(PWL)) directions, high radial motion amplitude of the plaque top surface (RMA(PTS)), and high relative movement, expressed in terms of radial strain (RSI(PL)) and longitudinal shear strain (LSSI(PL)), between plaque top and bottom surfaces. The in vivo results were reproduced by OF(LK(WLS)) and ABM(KF-K2), MEs previously proposed by the authors and with remarkable in silico performances, thereby reinforcing the clinical values of the markers and the potential of those MEs. Future in vivo studies will elucidate with confidence the full potential of the markers. PMID:24256708

  9. Photoacoustic flow cytometry: principle and application for real-time detection of circulating single nanoparticles, pathogens, and contrast dyes in vivo.

    PubMed

    Zharov, Vladimir P; Galanzha, Ekaterina I; Shashkov, Evgeny V; Kim, Jin-Woo; Khlebtsov, Nikolai G; Tuchin, Valery V

    2007-01-01

    The goal of this work is to develop in vivo photoacoustic (PA) flow cytometry (PAFC) for time-resolved detection of circulating absorbing objects, either without labeling or with nanoparticles as PA labels. This study represents the first attempt, to our knowledge, to demonstrate the capability of PAFC with tunable near-infrared (NIR) pulse lasers for real-time monitoring of gold nanorods, Staphylococcus aureus and Escherichia coli labeled with carbon nanotubes (CNTs), and contrast dye Lymphazurin in the microvessels of mouse and rat ears and mesenteries. PAFC shows the unprecedented threshold sensitivity in vivo as one gold nanoparticle in the irradiated volume and as one bacterium in the background of 10(8) of normal blood cells. The CNTs are demonstrated to serve as excellent new NIR high-PA contrast agents. Fast Lymphazurin diffusion in live tissue is observed with rapid blue coloring of a whole animal body. The enhancement of the thermal and acoustic effects is obtained with clustered, multilayer, and exploded nanoparticles. This novel combination of PA microscopy/spectroscopy and flow cytometry may be considered as a new powerful tool in biological research with the potential of quick translation to humans, providing ultrasensitive diagnostics of pathogens (e.g., bacteria, viruses, fungi, protozoa, parasites, helminthes), metastatic, infected, inflamed, stem, and dendritic cells, and pharmacokinetics of drug, liposomes, and nanoparticles in deep vessels (with focused transducers) among other potential applications. PMID:17994867

  10. Potential application of in vivo imaging of impaired lymphatic duct to evaluate the severity of pressure ulcer in mouse model

    NASA Astrophysics Data System (ADS)

    Kasuya, Akira; Sakabe, Jun-Ichi; Tokura, Yoshiki

    2014-02-01

    Ischemia-reperfusion (IR) injury is a cause of pressure ulcer. However, a mechanism underlying the IR injury-induced lymphatic vessel damage remains unclear. We investigated the alterations of structure and function of lymphatic ducts in a mouse cutaneous IR model. And we suggested a new method for evaluating the severity of pressure ulcer. Immunohistochemistry showed that lymphatic ducts were totally vanished by IR injury, while blood vessels were relatively preserved. The production of harmful reactive oxygen species (ROS) was increased in injured tissue. In vitro study showed a high vulnerability of lymphatic endothelial cells to ROS. Then we evaluated the impaired lymphatic drainage using an in vivo imaging system for intradermally injected indocyanine green (ICG). The dysfunction of ICG drainage positively correlated with the severity of subsequent cutaneous changes. Quantification of the lymphatic duct dysfunction by this imaging system could be a useful strategy to estimate the severity of pressure ulcer.

  11. In Vivo Noninvasive Analysis of Human Forearm Muscle Function and Fatigue: Applications to EVA Operations and Training Maneuvers

    NASA Technical Reports Server (NTRS)

    Fotedar, L. K.; Marshburn, T.; Quast, M. J.; Feeback, D. L.

    1999-01-01

    Forearm muscle fatigue is one of the major limiting factors affecting endurance during performance of deep-space extravehicular activity (EVA) by crew members. Magnetic resonance (MR) provides in vivo noninvasive analysis of tissue level metabolism and fluid exchange dynamics in exercised forearm muscles through the monitoring of proton magnetic resonance imaging (MRI) and phosphorus magnetic resonance spectroscopy (P-31-MRS) parameter variations. Using a space glove box and EVA simulation protocols, we conducted a preliminary MRS/MRI study in a small group of human test subjects during submaximal exercise and recovery and following exhaustive exercise. In assessing simulated EVA-related muscle fatigue and function, this pilot study revealed substantial changes in the MR image longitudinal relaxation times (T2) as an indicator of specific muscle activation and proton flux as well as changes in spectral phosphocreatine-to-phosphate (PCr/Pi) levels as a function of tissue bioenergetic potential.

  12. Non-contact respiration monitoring for in-vivo murine micro computed tomography: characterization and imaging applications

    NASA Astrophysics Data System (ADS)

    Burk, Laurel M.; Lee, Yueh Z.; Wait, J. Matthew; Lu, Jianping; Zhou, Otto Z.

    2012-09-01

    A cone beam micro-CT has previously been utilized along with a pressure-tracking respiration sensor to acquire prospectively gated images of both wild-type mice and various adult murine disease models. While the pressure applied to the abdomen of the subject by this sensor is small and is generally without physiological effect, certain disease models of interest, as well as very young animals, are prone to atelectasis with added pressure, or they generate too weak a respiration signal with this method to achieve optimal prospective gating. In this work we present a new fibre-optic displacement sensor which monitors respiratory motion of a subject without requiring physical contact. The sensor outputs an analogue signal which can be used for prospective respiration gating in micro-CT imaging. The device was characterized and compared against a pneumatic air chamber pressure sensor for the imaging of adult wild-type mice. The resulting images were found to be of similar quality with respect to physiological motion blur; the quality of the respiration signal trace obtained using the non-contact sensor was comparable to that of the pressure sensor and was superior for gating purposes due to its better signal-to-noise ratio. The non-contact sensor was then used to acquire in-vivo micro-CT images of a murine model for congenital diaphragmatic hernia and of 11-day-old mouse pups. In both cases, quality CT images were successfully acquired using this new respiration sensor. Despite the presence of beam hardening artefacts arising from the presence of a fibre-optic cable in the imaging field, we believe this new technique for respiration monitoring and gating presents an opportunity for in-vivo imaging of disease models which were previously considered too delicate for established animal handling methods.

  13. Dynamic in vivo imaging of dual-triggered microspheres for sustained release applications: synthesis, characterization and cytotoxicity study.

    PubMed

    Efthimiadou, Eleni K; Tapeinos, Christos; Chatzipavlidis, Alexandros; Boukos, Nikos; Fragogeorgi, Eirini; Palamaris, Lazaros; Loudos, George; Kordas, George

    2014-01-30

    This paper deals with the synthesis, characterization and property evaluation of drug-loaded magnetic microspheres with pH-responsive cross-linked polymer shell. The synthetic procedure consists of 3 steps, of which the first two comprise the synthesis of a poly methyl methacrylate (PMMA) template and the synthesis of a shell by using acrylic acid (AA) and methyl methacrylate (MMA) as monomers, and divinyl benzene (DVB) as cross-linker. The third step of the procedure refers to the formation of magnetic nanoparticles on the microsphere's surface. AA that attaches pH-sensitivity in the microspheres and magnetic nanoparticles in the inner and the outer surface of the microspheres, enhance the efficacy of this intelligent drug delivery system (DDS), which constitutes a promising approach toward cancer therapy. A number of experimental techniques were used to characterize the resulting microspheres. In order to investigate the in vitro controlled release behavior of the synthesized microspheres, we studied the Dox release percentage under different pH conditions and under external magnetic field. Hyperthermia caused by an alternating magnetic field (AFM) is used in order to study the doxorubicin (Dox) release behavior from microspheres with pH functionality. The in vivo fate of these hybrid-microspheres was tracked by labeling them with the ?-emitting radioisotope (99m)Tc after being intravenously injected in normal mice. According to our results, microsphere present a pH depending and a magnetic heating, release behavior. As expected, labeled microspheres were mainly found in the mononuclear phagocyte system (MPS). The highlights of the current research are: (i) to illustrate the advantages of controlled release by combining hyperthermia and pH-sensitivity and (ii) to provide noninvasive, in vivo information on the spatiotemporal biodistribution of these microsphere by dynamic ?-imaging. PMID:24286923

  14. Non-contact respiration monitoring for in-vivo murine micro computed tomography: characterization and imaging applications

    PubMed Central

    Burk, Laurel M; Lee, Yueh Z; Wait, J Matthew; Lu, Jianping; Zhou, Otto Z

    2012-01-01

    A cone beam micro-CT has previously been utilized along with a pressure-tracking respiration sensor to acquire prospectively gated images of both wild-type mice and various adult murine disease models. While the pressure applied to the abdomen of the subject by this sensor is small and is generally without physiological effect, certain disease models of interest, as well as very young animals, are prone to atelectasis with added pressure, or they generate too weak of a respiration signal with this method to achieve optimal prospective gating. In this work we present a new fiber-optic displacement sensor which monitors respiratory motion of a subject without requiring physical contact. The sensor outputs an analog signal which can be used for prospective respiration gating in micro-CT imaging. The device was characterized and compared against a pneumatic air chamber pressure sensor for the imaging of adult wild-type mice. The resulting images were found to be of similar quality with respect to physiological motion blur; the quality of the respiration signal trace obtained using the non-contact sensor was comparable to that of the pressure sensor and was superior for gating purposes due to its better signal-to-noise ratio. The non-contact sensor was then used to acquire in-vivo micro-CT images of a murine model for congenital diaphragmatic hernia and of 11-day-old mouse pups. In both cases, quality CT images were successfully acquired using this new respiration sensor. Despite the presence of beam hardening artifact arising from the presence of a fiber-optic cable in the imaging field, we believe this new technique for respiration monitoring and gating presents an opportunity for in-vivo imaging of disease models which were previously considered too delicate for established animal handling methods. PMID:22948192

  15. Measured and Modeled Toxicokinetics in Cultured Fish Cells and Application to In Vitro - In Vivo Toxicity Extrapolation

    PubMed Central

    Stadnicka-Michalak, Julita; Tanneberger, Katrin; Schirmer, Kristin; Ashauer, Roman

    2014-01-01

    Effect concentrations in the toxicity assessment of chemicals with fish and fish cells are generally based on external exposure concentrations. External concentrations as dose metrics, may, however, hamper interpretation and extrapolation of toxicological effects because it is the internal concentration that gives rise to the biological effective dose. Thus, we need to understand the relationship between the external and internal concentrations of chemicals. The objectives of this study were to: (i) elucidate the time-course of the concentration of chemicals with a wide range of physicochemical properties in the compartments of an in vitro test system, (ii) derive a predictive model for toxicokinetics in the in vitro test system, (iii) test the hypothesis that internal effect concentrations in fish (in vivo) and fish cell lines (in vitro) correlate, and (iv) develop a quantitative in vitro to in vivo toxicity extrapolation method for fish acute toxicity. To achieve these goals, time-dependent amounts of organic chemicals were measured in medium, cells (RTgill-W1) and the plastic of exposure wells. Then, the relation between uptake, elimination rate constants, and log KOW was investigated for cells in order to develop a toxicokinetic model. This model was used to predict internal effect concentrations in cells, which were compared with internal effect concentrations in fish gills predicted by a Physiologically Based Toxicokinetic model. Our model could predict concentrations of non-volatile organic chemicals with log KOW between 0.5 and 7 in cells. The correlation of the log ratio of internal effect concentrations in fish gills and the fish gill cell line with the log KOW was significant (r>0.85, p?=?0.0008, F-test). This ratio can be predicted from the log KOW of the chemical (77% of variance explained), comprising a promising model to predict lethal effects on fish based on in vitro data. PMID:24647349

  16. Folic acid-functionalized up-conversion nanoparticles: toxicity studies in vivo and in vitro and targeted imaging applications

    NASA Astrophysics Data System (ADS)

    Sun, Lining; Wei, Zuwu; Chen, Haige; Liu, Jinliang; Guo, Jianjian; Cao, Ming; Wen, Tieqiao; Shi, Liyi

    2014-07-01

    Folate receptors (FRs) are overexpressed on a variety of human cancer cells and tissues, including cancers of the breast, ovaries, endometrium, and brain. This over-expression of FRs can be used to target folate-linked imaging specifically to FR-expressing tumors. Fluorescence is emerging as a powerful new modality for molecular imaging in both the diagnosis and treatment of disease. Combining innovative molecular biology and chemistry, we prepared three kinds of folate-targeted up-conversion nanoparticles as imaging agents (UCNC-FA: UCNC-Er-FA, UCNC-Tm-FA, and UCNC-Er,Tm-FA). In vivo and in vitro toxicity studies showed that these nanoparticles have both good biocompatibility and low toxicity. Moreover, the up-conversion luminescence imaging indicated that they have good targeting to HeLa cells and can therefore serve as potential fluorescent contrast agents.Folate receptors (FRs) are overexpressed on a variety of human cancer cells and tissues, including cancers of the breast, ovaries, endometrium, and brain. This over-expression of FRs can be used to target folate-linked imaging specifically to FR-expressing tumors. Fluorescence is emerging as a powerful new modality for molecular imaging in both the diagnosis and treatment of disease. Combining innovative molecular biology and chemistry, we prepared three kinds of folate-targeted up-conversion nanoparticles as imaging agents (UCNC-FA: UCNC-Er-FA, UCNC-Tm-FA, and UCNC-Er,Tm-FA). In vivo and in vitro toxicity studies showed that these nanoparticles have both good biocompatibility and low toxicity. Moreover, the up-conversion luminescence imaging indicated that they have good targeting to HeLa cells and can therefore serve as potential fluorescent contrast agents. Electronic supplementary information (ESI) available: Up-conversion luminescence spectra of UCNC-Er and UCNC-Er-FA, UCNC-Tm and UCNC-Tm-FA. Confocal luminescence imaging data collected as a series along the Z optical axis. See DOI: 10.1039/c4nr02312a

  17. Preparation and In Vitro/Ex Vivo Evaluation of Moxifloxacin-Loaded PLGA Nanosuspensions for Ophthalmic Application

    PubMed Central

    Mudgil, Meetali; Pawar, Pravin K.

    2013-01-01

    The aim of the present investigation was to prepare a colloidal ophthalmic formulation to improve the residence time of moxifloxacin. Moxifloxacin-loaded poly(dl-lactide-co-glycolide) (PLGA) nanosuspensions were prepared by using the solvent evaporation technique. The nanosuspensions were characterised physically by using different techniques like particle size, zeta potential, FTIR, DSC, and XRD analysis. In vitro and ex vivo studies of nanosuspensions were carried out using a modified USP dissolution apparatus and all-glass Franz diffusion cells, respectively. The antibacterial activities of the nanosuspension and marketed formulations were performed against S. aureus and P. aeroginosa. The moxifloxacin-loaded PLGA nanosuspensions showed uniform particle size, ranging between 164–490 nm with negative zeta potential for all batches. The percentage entrapment efficiency of the drug-loaded nano-suspension was found to be between 84.09 to 92.05%. In vitro drug release studies suggest that all of the formulations showed extended drug release profiles and follow Korsemeyer-Peppas release kinetics. In vitro corneal permeability was found to be comparable with that of the marketed formulation across isolated goat cornea, indicating the suitability of the nanosuspension formulation in the ophthalmic delivery of moxifloxacin. The optimised nano-suspension was found to be more active against S. aureus and P. aeruginosa compared to the marketed eye drops. PMID:23833723

  18. Automated Segmentation and Object Classification of CT Images: Application to In Vivo Molecular Imaging of Avian Embryos

    PubMed Central

    Schmidt, Jana; Zimmermann, Johannes; Saluz, Hans Peter

    2013-01-01

    Background. Although chick embryogenesis has been studied extensively, there has been growing interest in the investigation of skeletogenesis. In addition to improved poultry health and minimized economic loss, a greater understanding of skeletal abnormalities can also have implications for human medicine. True in vivo studies require noninvasive imaging techniques such as high-resolution microCT. However, the manual analysis of acquired images is both time consuming and subjective. Methods. We have developed a system for automated image segmentation that entails object-based image analysis followed by the classification of the extracted image objects. For image segmentation, a rule set was developed using Definiens image analysis software. The classification engine was implemented using the WEKA machine learning tool. Results. Our system reduces analysis time and observer bias while maintaining high accuracy. Applying the system to the quantification of long bone growth has allowed us to present the first true in ovo data for bone length growth recorded in the same chick embryos. Conclusions. The procedures developed represent an innovative approach for the automated segmentation, classification, quantification, and visualization of microCT images. MicroCT offers the possibility of performing longitudinal studies and thereby provides unique insights into the morpho- and embryogenesis of live chick embryos. PMID:23997760

  19. In vivo biocompatibility assessment of (PTFE–PVDF–PP) terpolymer-based membrane with potential application for glaucoma treatment

    PubMed Central

    Leszczynski, Rafa?; Stodolak, Ewa; Wieczorek, Jaros?aw; Orlowska-Heitzman, Jolanta; Gumula, Teresa

    2010-01-01

    The aim of the work was to evaluate the in vivo biological behaviour of polymeric membrane materials for glaucoma implants. The base material was biostable synthetic terpolymer (PTFE–PVDF–PP) with proved biocompability (PN-EN ISO 10993). The samples manufactured in the form a membrane were subjected to chemical and physical treatment to create an open pore system within the polymer matrix. As a porogenic phase biodegradable natrium alginate in a fibrous form was employed. The non-perforating deep sclerectomy technique was performed in a rabbit model. The clinical observations were made after 14 and 30 days. During the study clinical symptoms of a moderate degree were observed, and histopathological changes were typical for foreign body implantation. At the end stage of the study no significant difference in histopathological assessment was found between control and experimental group. Similarities observed in both groups and relatively mild histopathological changes in the tissue surrounding the implant indicate that the observed symptoms come from a deep scleral trauma caused by surgery, and not by the presence of the implant itself. PMID:20652824

  20. EX VIVO APPLICATION OF CARBON MONOXIDE IN UW SOLUTION PREVENTS TRANSPLANT-INDUCED RENAL ISCHEMIA/REPERFUSION INJURY IN PIGS

    PubMed Central

    Yoshida, Junichi; Ozaki, Kikumi S.; Nalesnik, Michael A.; Ueki, Shinya; Castillo-Rama, Marcela; Faleo, Gaetano; Ezzelarab, Mohamed; Nakao, Atsunori; Ekser, Burcin; Echeverri, Gabriel J.; Ross, Mark A.; Stolz, Donna B.; Murase, Noriko

    2010-01-01

    I/R injury is a major deleterious factor of successful kidney transplantation (KTx). Carbon monoxide (CO) is an endogenous gaseous regulatory molecule, and exogenously delivered CO in low concentrations provides potent cytoprotection. This study evaluated efficacies of CO exposure to excised kidney grafts to inhibit I/R injury in the pig KTx model. Porcine kidneys were stored for 48 hrs in control UW or UW supplemented with CO (CO-UW) and autotransplanted in a 14-day follow-up study. In the control UW group, animal survival was 80% (4/5) with peak serum creatinine levels of 12.0±5.1 mg/dl. CO-UW showed potent protection, and peak creatinine levels were reduced to 6.9±1.4 mg/dl with 100% (5/5) survival without any noticeable adverse event or abnormal COHb value. Control grafts at 14d showed significant tubular damages, focal fibrotic changes, and numerous infiltrates. The CO-UW group showed significantly less severe histopathological changes with less TGF-? and p-Smad3 expression. Grafts in CO-UW also showed significantly lower early mRNA levels for proinflammatory cytokines and less lipid peroxidation. CO in UW provides significant protection against renal I/R injury in the porcine KTx model. Ex vivo exposure of kidney grafts to CO during cold storage may therefore be a safe strategy to reduce I/R injury. PMID:20199500

  1. Clinical Application of In-Room Positron Emission Tomography for In Vivo Treatment Monitoring in Proton Radiation Therapy

    SciTech Connect

    Min, Chul Hee [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States)] [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States); Zhu, Xuping [Center for Advanced Radiological Sciences, Nuclear Medicine and Molecular Imaging, Radiology Department, Massachusetts General Hospital, Boston, Massachusetts (United States)] [Center for Advanced Radiological Sciences, Nuclear Medicine and Molecular Imaging, Radiology Department, Massachusetts General Hospital, Boston, Massachusetts (United States); Winey, Brian A. [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States)] [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States); Grogg, Kira [Center for Advanced Radiological Sciences, Nuclear Medicine and Molecular Imaging, Radiology Department, Massachusetts General Hospital, Boston, Massachusetts (United States)] [Center for Advanced Radiological Sciences, Nuclear Medicine and Molecular Imaging, Radiology Department, Massachusetts General Hospital, Boston, Massachusetts (United States); Testa, Mauro [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States)] [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States); El Fakhri, Georges [Center for Advanced Radiological Sciences, Nuclear Medicine and Molecular Imaging, Radiology Department, Massachusetts General Hospital, Boston, Massachusetts (United States)] [Center for Advanced Radiological Sciences, Nuclear Medicine and Molecular Imaging, Radiology Department, Massachusetts General Hospital, Boston, Massachusetts (United States); Bortfeld, Thomas R.; Paganetti, Harald [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States)] [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States); Shih, Helen A., E-mail: hshih@partners.org [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts (United States)

    2013-05-01

    Purpose: The purpose of this study is to evaluate the potential of using in-room positron emission tomography (PET) for treatment verification in proton therapy and for deriving suitable PET scan times. Methods and Materials: Nine patients undergoing passive scattering proton therapy underwent scanning immediately after treatment with an in-room PET scanner. The scanner was positioned next to the treatment head after treatment. The Monte Carlo (MC) method was used to reproduce PET activities for each patient. To assess the proton beam range uncertainty, we designed a novel concept in which the measured PET activity surface distal to the target at the end of range was compared with MC predictions. The repositioning of patients for the PET scan took, on average, approximately 2 minutes. The PET images were reconstructed considering varying scan times to test the scan time dependency of the method. Results: The measured PET images show overall good spatial correlations with MC predictions. Some discrepancies could be attributed to uncertainties in the local elemental composition and biological washout. For 8 patients treated with a single field, the average range differences between PET measurements and computed tomography (CT) image-based MC results were <5 mm (<3 mm for 6 of 8 patients) and root-mean-square deviations were 4 to 11 mm with PET-CT image co-registration errors of approximately 2 mm. Our results also show that a short-length PET scan of 5 minutes can yield results similar to those of a 20-minute PET scan. Conclusions: Our first clinical trials in 9 patients using an in-room PET system demonstrated its potential for in vivo treatment monitoring in proton therapy. For a quantitative range prediction with arbitrary shape of target volume, we suggest using the distal PET activity surface.

  2. A novel application of maleimide for advanced drug delivery: in vitro and in vivo evaluation of maleimide-modified pH-sensitive liposomes

    PubMed Central

    Li, Tianshu; Takeoka, Shinji

    2013-01-01

    Maleimide is a stable and easy-to-handle moiety that rapidly and covalently conjugates thiol groups of cysteine residues in proteins or peptides. Herein, we use maleimide to modify the surface of liposomes in order to obtain an advanced drug delivery system. Employing a small amount (0.3 mol%) of maleimide-polyethylene glycol (PEG) to modify the surface of the liposomes M-GGLG-liposomes, composed of 1,5-dihexadecyl N,N-diglutamyl-lysyl-L-glutamate (GGLG)/cholesterol/poly(ethylene glycol) 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (PEG5000-DSPE)/maleimide-PEG5000-Glu2C18 at a molar ratio of 5:5:0.03:0.03, drug delivery efficiency was remarkably improved both in vitro and in vivo compared to unmodified liposomes (GGLG-liposomes, composed of GGLG/cholesterol/PEG5000-DSPE/PEG5000-Glu2C18 at a molar ratio of 5:5:0.03:0.03). Moreover, this modification did not elicit any detectable increase in cytotoxicity. The maleimide-modification did not alter the physical characteristics of the liposomes such as size, zeta potential, pH sensitivity, dispersibility and drug encapsulation efficiency. However, M-GGLG-liposomes were more rapidly (?2-fold) internalized into HeLa, HCC1954, and MDA-MB-468 cells compared to GGLG-liposomes. In vivo, M-GGLG-liposomes encapsulating doxorubicin (M-GGLG-DOX-liposomes) also showed a more potent antitumor effect than GGLG-DOX-liposomes and the widely used 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC)-DOX-liposomes after two subcutaneous injections around breast cancer tissue in mice. The biodistribution of liposomes in this model was observed using an in vivo imaging system, which showed that M-GGLG-liposomes were present for significantly longer at the injection site compared to GGLG-liposomes. The outstanding biological functions of the maleimide-modified liposomes as a novel drug delivery system make them ideally suited to a wide range of applications. PMID:24143089

  3. Thermal increase in the oral mucosa and in the jawbone during Nd:YAG laser applications. Ex vivo study

    PubMed Central

    Vescovi, Paolo; Fornaini, Carlo; Rocca, Jean P.; Nammour, Samir

    2012-01-01

    Objective: Literature reports bactericidal and biostimulant effects for Nd:YAG laser procedures on bone and oral mucosa but the possible overheating can cause damage to anatomical structures. The aim of the study is the evaluation of thermal increase in different levels of oral tissues: mucosa, periosteum and bone during defocused application of Nd:YAG laser at different parameters. Study Design: Superficial thermal evaluation was performed in pig jaws with a thermal camera device; deep thermal evaluation was realized by 4 thermocouples placed at a subperiosteal level and at 1,2 and 4 mm depth in the jaw bone. Laser applications of 1 minute were performed 5 times (with a pause of 1 minute) on a surface of 4 cm2 with a Nd:YAG laser (VSP mode, 320 micrometer fiber, defocused mode) with different parameters. Temperatures were recorded before and after laser applications and after each pause in order to evaluate also the thermal relaxation of tissues. Results: At submucosal level, mean thermal increase was between 1.1°C and 13.2°C, at 1 mm depth between 1.1°C and 8.5°C, at 2 mm depth between 1.1°C and 6.8°C, at 4 mm depth between 1.0°C and 5.3°C. Temperature decrease during the rest time period was variable between 0°C and 2.5°C. Conclusions: Temperatures reached during clinical procedures with parameters reported in the literature in biostimulation protocols (1.25-2 Watts) for the five minutes of application are not dangerous for biological structures. The decrease in temperature during the rest time period is less considerable in the bone in comparison to oral mucosa. Key words:Nd:YAG laser, thermal increase, thermocouple, thermal camera, low level laser therapy. PMID:22322506

  4. Application of laser-induced autofluorescence spectra detection system in human colorectal cancer in-vivo screening

    NASA Astrophysics Data System (ADS)

    Chia, Teck Chee; Fu, Sheng; Chia, Yee Hong; Kwek, Leong Chuan; Tang, Choong Leong

    2005-09-01

    This study aimed at applying Laser induced-autofluorescence (LIAF) diagnostics method as an in-vivo screening of colorectal polyplcancer. The spectrum algorithm based on the ratio of autofluorescence intensity was used to identify the diseased tissues from the normal tissues as it was generally performed better than an algorithm based only simply on the intensity of the spectrum. Histopathological biopsy results were compared with the detected AF spectra characteristics for different kinds of polyps. 73 patients had been examined via the LIAF spectroscopy detection system during their colonoscopy screening in Endoscopy Center, Singapore General Hospital. The autofluorescence from the surface of the colorectal tissues under 405 nm laser light excitation was detected using our detecting system. In the experimental investigation two groups of patients were involved. One group was "abnormal" group. There were 25 patients belonging to this group since polyps or carcinoma was found in their colorectal tract during colonoscopy. The histopathology reports confirm the group classification. Total 36 polyps' AF spectra and 9 carcinoma' AF spectra were detected from 25 patients of the abnormal group during their regular endoscopy examination. The intensity ratios RI-680/I-500 and RI-630/I-500 of polyps/cancerous AF spectra and intensity ratios of corresponding normal colorectal AF spectra were calculated. Two critical intensity ratios for separating the AF intensity ratios RI-680/I-500 and RI-630/I-500 of normal and abnormal colorectal tissues were defined as 0.5 and 0.6 respectively. Using the critical intensity ratio values, 48 "normal" group patients' rectums were checked via the LIAF detection system. There were 20 patients (41.7%) whose AF spectra of colorectal tract mucosa belonging to abnormal spectra. However, these 20 patients had not been found under white light via traditional endoscopy. For small diseased area like small plat polyp disease and carcinoma, it was very difficult to identify under white light by endoscopy. However, the LIAF spectra technique and AF intensity ratio algorithm was able to detect these kinds of abnormal area earlier than traditional endoscopy. Using this algorithm, it is able to identify the onset of abnormal tissue growth during real-time clinical endoscope examination.

  5. The application of micro-CT in monitoring bone alterations in tail-suspended rats in vivo

    NASA Astrophysics Data System (ADS)

    Luan, Hui-Qin; Sun, Lian-Wen; Huang, Yun-Fei; Wang, Ying; McClean, Colin J.; Fan, Yu-Bo

    2014-06-01

    Osteopenia is a pathological process that affects human skeletal health not only on earth but also in long-time spaceflight. Micro-computed tomography (micro-CT) is a nondestructive method for assessing both bone quantity and bone quality. To investigate the characteristics of micro-CT on evaluating the microgravity-induced osteopenia (e.g. early detection time and the sensitive parameters), the bone loss process of tail-suspended rats was monitored by micro-CT in this study. 8-Week-old female Sprague Dawley rats were divided into two groups: tail suspension (TS) and control (CON). Volumetric bone mineral density (vBMD) and microstructure of the femur and tibia were evaluated in vivo by micro-CT at 0, 7, 14, 22 days. Biomechanical properties of the femur and tibia were determined by three-point bending test. The ash weight of bone was also investigated. The results showed that (1) bone loss in the proximal tibia appeared earlier than in the distal femur. (2) On day 7, the percent bone volume (BV/TV) of the tibia 15.44% decreased significantly, and the trabecular separation (Tb.Sp) 30.29% increased significantly in TS group, both of which were detected earlier than other parameters. (3) Biomechanical properties (e.g. femur, -22.4% maximum load and -23.75% Young’s modulus vs. CON) and ash weight of the femur and tibia decreased significantly in the TS group in comparison to CON group. (4) vBMD of the femur and tibia were clearly related to bone ash and dry weight (r = 0.75-0.87, p < 0.05). (5) BV/TV of both femur and tibia were clearly related to maximum load and Young’s modulus (r = 0.66-0.87, p < 0.05). Similarly, trabecular vBMD and BV/TV of the femur and tibia were clearly related to Young’s modulus (r = 0.73-0.89, p < 0.05). These indicated that BV/TV and Tb.Sp were more sensitive than other parameters for evaluating bone loss induced by tail suspension, moreover, trabecular vBMD and other parameters might be used to evaluate bone strength. Therefore, micro-CT is a reliable and sensitive method for predicting unloading-induced bone loss in small animals.

  6. Resistance after Chronic Application of the HDAC-Inhibitor Valproic Acid Is Associated with Elevated Akt Activation in Renal Cell Carcinoma In Vivo

    PubMed Central

    Juengel, Eva; Makarevi?, Jasmina; Tsaur, Igor; Bartsch, Georg; Nelson, Karen; Haferkamp, Axel; Blaheta, Roman A.

    2013-01-01

    Targeted drugs have significantly improved the therapeutic options for advanced renal cell carcinoma (RCC). However, resistance often develops, negating the benefit of these agents. In the present study, the molecular mechanisms of acquired resistance towards the histone deacetylase (HDAC) inhibitor valproic acid (VPA) in a RCC in vivo model were investigated. NMRI:nu/nu mice were transplanted with Caki-1 RCC cells and then treated with VPA (200 mg/kg/day). Controls remained untreated. Based on tumor growth dynamics, the mice were divided into “responders” and “non-responders” to VPA. Histone H3 and H4 acetylation and expression of cell signaling and cell cycle regulating proteins in the RCC mouse tumors were evaluated by Western blotting. Tumor growth of VPA responders was significantly diminished, whereas that of VPA non-responders even exceeded control values. Cdk1, 2 and 4 proteins were strongly enhanced in the non-responders. Importantly, Akt expression and activity were massively up-regulated in the tumors of the VPA non-responders. Chronic application (12 weeks) of VPA to Caki-1 cells in vitro evoked a distinct elevation of Akt activity and cancer cells no longer responded with cell growth reduction, compared to the short 2 week treatment. We assume that chronic use of an HDAC-inhibitor is associated with (re)-activation of Akt, which may be involved in resistance development. Consequently, combined blockade of both HDAC and Akt may delay or prevent drug resistance in RCC. PMID:23372654

  7. In vivo assessments of bioabsorbable AZ91 magnesium implants coated with nanostructured fluoridated hydroxyapatite by MAO/EPD technique for biomedical applications.

    PubMed

    Razavi, Mehdi; Fathi, Mohammadhossein; Savabi, Omid; Vashaee, Daryoosh; Tayebi, Lobat

    2015-03-01

    Although magnesium (Mg) is a unique biodegradable metal which possesses mechanical property similar to that of the natural bone and can be an attractive material to be used as orthopedic implants, its quick corrosion rate restricts its actual clinical applications. To control its rapid degradation, we have modified the surface of magnesium implant using fluoridated hydroxyapatite (FHA: Ca10(PO4)6OH2-xFx) through the combined micro-arc oxidation (MAO) and electrophoretic deposition (EPD) techniques, which was presented in our previous paper. In this article, the biocompatibility examinations were conducted on the coated AZ91 magnesium alloy by implanting it into the greater trochanter area of rabbits. The results of the in vivo animal test revealed a significant enhancement in the biocompatibility of FHA/MAO coated implant compared to the uncoated one. By applying the FHA/MAO coating on the AZ91 implant, the amount of weight loss and magnesium ion release in blood plasma decreased. According to the histological results, the formation of the new bone increased and the inflammation decreased around the implant. In addition, the implantation of the uncoated AZ91 alloy accompanied by the release of hydrogen gas around the implant; this release was suppressed by applying the coated implant. Our study exemplifies that the surface coating of magnesium implant using a bioactive ceramic such as fluoridated hydroxyapatite may improve the biocompatibility of the implant to make it suitable as a commercialized biomedical product. PMID:25579892

  8. Synthesis of chemically pure, luminescent Eu3+ doped HAp nanoparticles: a promising fluorescent probe for in vivo imaging applications.

    PubMed

    Hasna, K; Kumar, S Sasanka; Komath, Manoj; Varma, Manoj Raama; Jayaraj, M K; Kumar, K Rajeev

    2013-06-01

    The poor solubility, poor biocompatibility and disposal issues make fluorescent quantum dots such as CdSe, CdS, ZnS, InP, InAs, etc. impractical for imaging tissues or intercellular structures. As calcium phosphate is the main inorganic component of human bone and teeth, hydroxyapatite (Ca10(PO4)6(OH)2, HAp) is highly biocompatible and bioactive. Since HAp nanoparticles are not luminescent, a novel inorganic biocompatible fluorescent probe was suggested by doping HAp with lanthanides. Here we report the growth of chemically pure fluorescent HAp nanoparticles synthesized by a new methodology, liquid phase pulsed laser ablation using third harmonics (355 nm) of Nd-YAG laser. Europium doped HAp nanoparticles show emission with prominent peaks at 531 nm, 572 nm, 601 nm and 627 nm upon excitation at a wavelength of 325 nm. The red luminescence could also be observed under visible excitation at 459 nm and is suitable for living cell applications. PMID:23580129

  9. LC analysis of benzophenone-3: II application to determination of ‘in vitro’ and ‘in vivo’ skin penetration from solvents, coarse and submicron emulsions

    Microsoft Academic Search

    C. Fernandez; G. Marti-Mestres; J. Ramos; H. Maillols

    2000-01-01

    The aim of this study was to determine the skin penetration of benzophenone-3 in vitro and in vivo in order to investigate a possible influence of formulation. Six different vehicles, three solvents and three different emulsion types were evaluated in vitro and in vivo. Each vehicle was applied to the skin model at 2 mg cm?2. First, histological studies on

  10. Application of in vivo microdialysis for studying the efficacy of protective preparations against sulfur mustard penetrating the skin.

    PubMed

    Karvaly, G; Gachályi, A; Furész, J

    2008-01-01

    Subcutaneous microdialysis was employed for monitoring thiodiglycol (2,2'-thiodiethanol, TDG) levels with the aim of characterizing the transdermal penetration of topically applied liquid sulfur mustard (2,2'-dichlorodiethyl sulfide, SM) in rats. TDG levels, evaluated in 20 min dialysates collected over a 6 h sampling period, were plotted against time after pooling. Linear correlation was identified between the SM dose and the mean areas under the 0-60 min or the whole curve (AUC(0-60) and AUC, respectively) as well as mean peak concentrations (C(max)) in the range of 1.0-3.0 microl applied volume (7.2-21.7 nmol).A commercially available barrier cream, a perfluoropolyether oil and a vaseline based ointment containing zinc oxide were subsequently tested as topical protectants. Each preparation was layered on the skin surface prior to the application of 2.0 microl SM. The evaluation of the efficacy of the preparations was based on obtained AUC(0-60), AUC and C(max) values. A statistical comparison of these parameters with those obtained when 2.0 microl SM was applied without pretreatment indicated that the barrier cream and the perfluoropolyether oil significantly (P < 0.01) reduced the amount of penetrating SM within the sampling period. In addition, the perfluoropolyether oil almost completely prevented the penetration of SM for 20 min. Pretreatment with the ointment did not prove to be an effective countermeasure as its administration resulted in no significant reduction in AUC(0-60), AUC and C(max) values. PMID:17429799

  11. Theory and application of optimal linear resolution to MRI truncation artifacts, multiexponential decays and in vivo multiple sclerosis pathology

    NASA Astrophysics Data System (ADS)

    Cover, Keith S.

    It is widely believed that one of the best way to proceed when analysing data is to generate estimates which fit the data. However, when the relationship between the unknown model and data is linear for highly underdetermined systems, is it common practice to find estimates with good linear resolution with no regard for fitting the data. For example, windowed Fourier transforms produces estimates that have good linear resolution but do not fit the data. Surprisingly, many researchers do not seem to be explicitly aware of this fact. This thesis presents a theoretical basis for the linear resolution which demonstrates that, for a wide range of problems, algorithms which produce estimates with good linear resolution can be a more powerful and convenient way of presenting the information in the data, than models that fit the data. Linear resolution was also applied to two outstanding problems in linear inverse theory. The first was the problem of truncation artifacts in magnetic resonance imaging (MRI). Truncation artifacts were heavily suppressed or eliminated by the choice of one of two novel Fourier transform windows. Complete elimination of truncation artifacts generally led to unexpectedly blurry images. Heavy suppression seemed to be the best compromise between truncation artifacts and blurriness. The second problem was estimating the relaxation distribution of a multiexponential system from its decay curve. This is an example where hundreds of papers have been written on the subject, yet almost no one has made a substantial effort to apply linear resolution. I found the application to be very successful. As an example, the algorithm was applied to the decay of MRI data from multiple sclerosis patients in an attempt to differentiate between various pathologies.

  12. In vivo application of short-lag spatial coherence and harmonic spatial coherence imaging in fetal ultrasound.

    PubMed

    Kakkad, Vaibhav; Dahl, Jeremy; Ellestad, Sarah; Trahey, Gregg

    2015-04-01

    Fetal scanning is one of the most common applications of ultrasound imaging and serves as a source of vital information about maternal and fetal health. Visualization of clinically relevant structures, however, can be severely compromised in difficult-to-image patients due to poor resolution and the presence of high levels of acoustical noise or clutter. We have developed novel coherence-based beamforming methods called Short-Lag Spatial Coherence (SLSC) imaging and Harmonic Spatial Coherence imaging (HSCI), and applied them to suppress the effects of clutter in fetal imaging. This method is used to create images of the spatial coherence of the backscattered ultrasound as opposed to images of echo magnitude. We present the results of a patient study to assess the benefits of coherence-based beamforming in the context of first trimester fetal exams. Matched fundamental B-mode, SLSC, harmonic B-mode, and HSCI images were generated using raw radio frequency data collected on 11 volunteers in the first trimester of pregnancy. The images were compared for qualitative differences in image texture and target conspicuity as well as using quantitative imaging metrics such as signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and contrast. SLSC and HSCI showed statistically significant improvements across all imaging metrics compared with B-mode and harmonic B-mode, respectively. These improvements were greatest for poor quality B-mode images where contrast of anechoic targets was improved from 15 dB in fundamental B-mode to 27 dB in SLSC and 17 dB in harmonic B-mode to 30 dB in HSCI. CNR improved from 1.4 to 2.5 in the fundamental images and 1.4 to 3.1 in the harmonic case. These results exhibit the potential of coherence-based beamforming to improve image quality and target detectability, especially in high noise environments. PMID:25116292

  13. Ex vivo lung perfusion

    PubMed Central

    Machuca, Tiago N.

    2014-01-01

    Lung transplantation (LTx) is an established treatment option for eligible patients with end-stage lung disease. Nevertheless, the imbalance between suitable donor lungs available and the increasing number of patients considered for LTx reflects in considerable waitlist mortality. Among potential alternatives to address this issue, ex vivo lung perfusion (EVLP) has emerged as a modern preservation technique that allows for more accurate lung assessment and also improvement of lung function. Its application in high-risk donor lungs has been successful and resulted in safe expansion of the donor pool. This article will: (I) review the technical details of EVLP; (II) the rationale behind the method; (III) report the worldwide clinical experience with the EVLP, including the Toronto technique and others; (IV) finally, discuss the growing literature on EVLP application for donation after cardiac death (DCD) lungs. PMID:25132972

  14. ITRAQ MASS SPECTROMETRIC PROTEOMIC APPLICATIONS FOR IN VIVO TOXICOLOGY STUDIES OF AMPHIBIAN SPECIES: DATA HANDLING AND INTERPRETATION USING PEPTIDE-TAGGING SOFTWARE

    EPA Science Inventory

    This addresses the USEPA's need for a cost effective, non-mammalian screening assay for thyroid axis disrupting chemicals; a multi-endpoint strategy combining molecular and in vivo protocols in an amphibian model is being applied at MED Duluth....

  15. In vivo imaging flow cytometer

    NASA Astrophysics Data System (ADS)

    Lee, Ho; Alt, Clemens; Pitsillides, Costas M.; Puoris'haag, Mehron; Lin, Charles P.

    2006-08-01

    We introduce an in vivo imaging flow cytometer, which provides fluorescence images simultaneously with quantitative information on the cell population of interest in a live animal. As fluorescent cells pass through the slit of light focused across a blood vessel, the excited fluorescence is confocally detected. This cell signal triggers a strobe beam and a high sensitivity CCD camera that captures a snapshot image of the cell as it moves down-stream from the slit. We demonstrate that the majority of signal peaks detected in the in vivo flow cytometer arise form individual cells. The instrument’s capability to image circulating T cells and measure their speed in the blood vessel in real time in vivo is demonstrated. The cell signal irradiance variation, clustering percentage, and potential applications in biology and medicine are discussed.

  16. In vivo methods for drug absorption - comparative physiologies, model selection, correlations with in vitro methods (IVIVC), and applications for formulation/API/excipient characterization including food effects.

    PubMed

    Sjögren, Erik; Abrahamsson, Bertil; Augustijns, Patrick; Becker, Dieter; Bolger, Michael B; Brewster, Marcus; Brouwers, Joachim; Flanagan, Talia; Harwood, Matthew; Heinen, Christian; Holm, René; Juretschke, Hans-Paul; Kubbinga, Marlies; Lindahl, Anders; Lukacova, Viera; Münster, Uwe; Neuhoff, Sibylle; Nguyen, Mai Anh; Peer, Achiel van; Reppas, Christos; Hodjegan, Amin Rostami; Tannergren, Christer; Weitschies, Werner; Wilson, Clive; Zane, Patricia; Lennernäs, Hans; Langguth, Peter

    2014-06-16

    This review summarizes the current knowledge on anatomy and physiology of the human gastrointestinal tract in comparison with that of common laboratory animals (dog, pig, rat and mouse) with emphasis on in vivo methods for testing and prediction of oral dosage form performance. A wide range of factors and methods are considered in addition, such as imaging methods, perfusion models, models for predicting segmental/regional absorption, in vitro in vivo correlations as well as models to investigate the effects of excipients and the role of food on drug absorption. One goal of the authors was to clearly identify the gaps in today's knowledge in order to stimulate further work on refining the existing in vivo models and demonstrate their usefulness in drug formulation and product performance testing. PMID:24637348

  17. Clinical applications of a real-time scanning-slit confocal microscope designed for real-time observations of the in-vivo human cornea

    NASA Astrophysics Data System (ADS)

    Masters, Barry R.

    1995-05-01

    We describe a new, real-time, flying slit confocal microscope, that has unique features and imaging characteristics for in vivo human ocular imaging. In vivo real-time confocal microscopy is currently used to investigate the tear film, renewal of the ocular surface, the role of epithelial innervation in epithelial cell proliferation, wound healing, kinetics of drug penetration, the effects of laser refractive surgery on the keratocyte activation and distribution in the stroma, and the nature of endothelial defects. The following clinical examples will be presented and discussed: confocal microscopy of normal human basal and wing cells in the epithelium, confocal microscopy of lamellar and penetrating corneal grafts, confocal microscopy of corneal ulcer, confocal microscopy of scar formation after herpes keratitis, and confocal microscopy of corneal innervation. The use of scanning slit confocal microscopes has unique advantages over other instrumental systems based on pinhole-containing Nipkow disks (tandem-scanning confocal microscopes) for clinical in vivo confocal microscopy.

  18. APPLICATION OF THE EPR SPIN-TRAPPING TECHNIQUE TO THE DETECTION OF RADICALS PRODUCED IN VIVO DURING INHALATION EXPOSURE OF RATS TO OZONE

    EPA Science Inventory

    Ozone is known to induce lipid peroxidation of lung tissue, although no direct evidence of free radical formation has been reported. e have use the electron paramagnetic resonance (EPR) spin-trapping technique to search for free radicals produced in vivo by ozone exposure. he spi...

  19. Simultaneous Measurement of Monoamine, Amino Acid, and Drug Levels, Using High Performance Liquid Chromatography and Coulometric Array Technology: Application to In Vivo Microdialysis Perfusate Analysis

    Microsoft Academic Search

    Ian N. Acworth; Jian Yu; Elizabeth Ryan; Kathleen Cox Gaiuepy; Paul Gamache; Keith Hull; Timothy Maher

    1994-01-01

    An automated HPLC coulometric array-ECD method is described for the simultaneous analysis of monoamines, their metabolites, derivatized amino acids, and pharmacological agents. This method has been used with in vivo microdialysis in urethane-anesthetized animals to examine extracellular fluid levels of endogenous and exogenous analytes after the peripheral administration of drugs. An aliquot of dialysate was initially analyzed for the monoamines,

  20. Feasibility and application of a retronasal aroma-trapping device to study in vivo aroma release during the consumption of model wine-derived beverages

    PubMed Central

    Muñoz-González, Carolina; Rodríguez-Bencomo, Juan José; Moreno-Arribas, Maria Victoria; Pozo-Bayón, Maria Ángeles

    2014-01-01

    New types of wine-derived beverages are now in the market. However, little is known about the impact of ingredient formulation on aroma release during consumption, which is directly linked to consumer preferences and liking. In this study, the optimization and validation of a retronasal aroma-trapping device (RATD) for the in vivo monitoring of aroma release was carried out. This device was applied to assess the impact of two main ingredients (sugar and ethanol) in these types of beverages on in vivo aroma release. Two aroma-trapping materials (Lichrolut and Tenax) were firstly assayed. Tenax provided higher recovery and lower intra- and inter-trap variability. In in vivo conditions, RATD provided an adequate linear range (R2 > 0.91) between 0 and 50 mg L?1 of aroma compounds. Differences in the total aroma release were observed in equally trained panelists. It was proven that the addition of sugar (up to 150 mg kg?1) did not have effect on aroma release, while ethanol (up to 40 mg L?1) enhanced the aroma release during drinking. The RATD is a useful tool to collect real in vivo data to extract reliable conclusions about the effect of beverage components on aroma release during consumption. The concentration of ethanol should be taken into consideration for the formulation of wine-derived beverages. PMID:25473493

  1. Establishment of an Allo-Transplantable Hamster Cholangiocarcinoma Cell Line and Its Application for In Vivo Screening of Anti-Cancer Drugs

    PubMed Central

    Puthdee, Nattapong; Seubwai, Wunchana; Wonkchalee, Orasa; Kaewkong, Worasak; Juasook, Amornrat; Pinlaor, Somchai; Pairojkul, Chawalit; Wongkham, Chaisiri; Okada, Seiji; Boonmars, Thidarut; Wongkham, Sopit

    2013-01-01

    Opisthorchis viverrini (O. viverrini) is a well-known causative agent of cholangiocarcinoma (CCA) in humans. CCA is very resistant to chemotherapy and is frequently fatal. To understand the pathogenesis of CCA in humans, a rodent model was developed. However, the development of CCA in rodents is time-consuming and the xenograft-transplantation model of human CCA in immunodeficient mice is costly. Therefore, the establishment of an in vivo screening model for O. viverrini-associated CCA treatment was of interest. We developed a hamster CCA cell line, Ham-1, derived from the CCA tissue of O. viverrini-infected and N-nitrosodimethylamine-treated Syrian golden hamsters. Ham-1 has been maintained in Dulbecco's Modified Essential Medium supplemented with 10% fetal bovine serum for more than 30 subcultures. These cells are mostly diploid (2n=44) with some being polyploid. Tumorigenic properties of Ham-1 were demonstrated by allograft transplantation in hamsters. The transplanted tissues were highly proliferative and exhibited a glandular-like structure retaining a bile duct marker, cytokeratin 19. The usefulness of this for in vivo model was demonstrated by berberine treatment, a traditional medicine that is active against various cancers. Growth inhibitory effects of berberine, mainly by an induction of G1 cell cycle arrest, were observed in vitro and in vivo. In summary, we developed the allo-transplantable hamster CCA cell line, which can be used for chemotherapeutic drug testing in vitro and in vivo. PMID:24516278

  2. Behavior of Tip-Steerable Needles in ex vivo and in vivo Tissue

    PubMed Central

    Majewicz, Ann; Marra, Steven P.; van Vledder, Mark G.; Lin, MingDe; Choti, Michael A.; Song, Danny Y.; Okamura, Allison M.

    2012-01-01

    Robotic needle steering is a promising technique to improve the effectiveness of needle-based clinical procedures, such as biopsies and ablation, by computer-controlled, curved insertions of needles within solid organs. In this paper, we explore the capabilities, challenges, and clinical relevance of asymmetric-tip needle steering though experiments in ex vivo and in vivo tissue. We evaluate the repeatability of needle insertion in inhomogeneous biological tissue and compare ex vivo and in vivo needle curvature and insertion forces. Steerable needles curved more in kidney than in liver and prostate, likely due to differences in tissue properties. Pre-bent needles produced higher insertion forces in liver and more curvature in vivo than ex vivo. When compared to straight stainless steel needles, steerable needles did not cause a measurable increase in tissue damage and did not exert more force during insertion. The minimum radius of curvature achieved by pre-bent needles was 5.23 cm in ex vivo tissue, and 10.4 cm in in vivo tissue. The curvatures achieved by bevel tip needles were negligible for in vivo tissue. The minimum radius of curvature for bevel tip needles in ex vivo tissue was 16.4 cm; however, about half of the bevel tip needles had negligible curvatures. We also demonstrate a potential clinical application of needle steering by targeting and ablating overlapping regions of cadaveric canine liver. PMID:22711767

  3. Enhanced priming of antigen-specific CTLs in vivo by embryonic stem cell-derived dendritic cells expressing chemokine along with antigenic protein: application to antitumor vaccination.

    PubMed

    Matsuyoshi, Hidetake; Senju, Satoru; Hirata, Shinya; Yoshitake, Yoshihiro; Uemura, Yasushi; Nishimura, Yasuharu

    2004-01-15

    Dendritic cell (DC)-based immunotherapy is regarded as a promising means for anti-cancer therapy. The efficiency of T cell-priming in vivo by transferred DCs should depend on their encounter with T cells. In the present study, we attempted to improve the capacity of DCs to prime T cells in vivo by genetic modification to express chemokine with a T cell-attracting property. For genetic modification of DCs, we used a recently established method to generate DCs from mouse embryonic stem cells. We generated double-transfectant DCs expressing a chemokine along with a model Ag (OVA) by sequential transfection of embryonic stem cells, and then induced differentiation to DCs. We comparatively evaluated the effect of three kinds of chemokines; secondary lymphoid tissue chemokine (SLC), monokine induced by IFN-gamma (Mig), and lymphotactin (Lptn). All three types of double transfectant DCs primed OVA-specific CTLs in vivo more efficiently than did DCs expressing only OVA, and the coexpression of SLC or Lptn was more effective than that of Mig. Immunization with DCs expressing OVA plus SLC or Mig provided protection from OVA-expressing tumor cells more potently than did immunization with OVA alone, and SLC was more effective than Mig. In contrast, coexpression of Lptn gave no additive effect on protection from the tumor. Collectively, among the three chemokines, expression of SLC was the most effective in enhancing antitumor immunity by transferred DCs in vivo. The findings provide useful information for the development of a potent DC-based cellular immunotherapy. PMID:14707047

  4. Motion-artifact-robust, polarization-resolved second-harmonic-generation microscopy based on rapid polarization switching with electro-optic Pockells cell and its application to in vivo visualization of collagen fiber orientation in human facial skin

    PubMed Central

    Tanaka, Yuji; Hase, Eiji; Fukushima, Shuichiro; Ogura, Yuki; Yamashita, Toyonobu; Hirao, Tetsuji; Araki, Tsutomu; Yasui, Takeshi

    2014-01-01

    Polarization-resolved second-harmonic-generation (PR-SHG) microscopy is a powerful tool for investigating collagen fiber orientation quantitatively with low invasiveness. However, the waiting time for the mechanical polarization rotation makes it too sensitive to motion artifacts and hence has hampered its use in various applications in vivo. In the work described in this article, we constructed a motion-artifact-robust, PR-SHG microscope based on rapid polarization switching at every pixel with an electro-optic Pockells cell (PC) in synchronization with step-wise raster scanning of the focus spot and alternate data acquisition of a vertical-polarization-resolved SHG signal and a horizontal-polarization-resolved one. The constructed PC-based PR-SHG microscope enabled us to visualize orientation mapping of dermal collagen fiber in human facial skin in vivo without the influence of motion artifacts. Furthermore, it implied the location and/or age dependence of the collagen fiber orientation in human facial skin. The robustness to motion artifacts in the collagen orientation measurement will expand the application scope of SHG microscopy in dermatology and collagen-related fields. PMID:24761292

  5. Application of in vivo microdialysis for investigation of unbound drug concentrations of intravenously administered sulfadimidine in the paranasal sinus mucosa of horses.

    PubMed

    Bienert-Zeit, Astrid; Gietz, Caroline; Staszyk, Carsten; Kietzmann, Manfred; Stahl, Jessica; Ohnesorge, Bernhard

    2015-04-01

    OBJECTIVE To monitor concentrations of sulfadimidine in the paranasal sinus mucosa (PSM) of unsedated horses following IV administration of trimethoprim-sulfadimidine via in vivo microdialysis. ANIMALS 10 healthy adult horses. PROCEDURES Concentric microdialysis probes were implanted into the subepithelial layers of the frontal sinus mucosa of standing sedated horses. Four hours after implantation, trimethoprim-sulfadimidine (30 mg/kg) was administered IV every 24 hours for 2 days; dialysate and plasma samples were collected at intervals during that 48-hour period and analyzed for concentrations of sulfadimidine. The dialysate concentration and relative loss of sulfadimidine from the perfusate were used to calculate the PSM concentration. RESULTS Microdialysis probe implantation and subsequent in vivo microdialysis were successfully performed for all 10 horses. Following the first and second administration of trimethoprim-sulfadimidine, mean ± SD peak concentrations of sulfadimidine were 55.3 ± 10.3 ?g/mL and 51.5 ± 8.7 ?g/mL, respectively, in plasma and 9.6 ± 4.5 ?g/mL and 7.0 ± 3.3 ?g/mL, respectively, in the PSM. Peak sulfadimidine concentrations in the PSM were detected at 5.9 ± 2.7 hours and 5.4 ± 2.3 hours following the first and second drug administrations, respectively. For 12 hours, mean PSM sulfadimidine concentration remained greater than the minimum inhibitory concentration indicative of sulfonamide susceptibility of equine bacterial isolates (4.75 ?g/mL). CONCLUSIONS AND CLINICAL RELEVANCE In vivo microdialysis for continuous monitoring of PSM sulfadimidine concentrations in unsedated horses was feasible. Intravenous administration of trimethoprim (5 mg/kg) and sulfadimidine (25 mg/kg) proved likely to be efficient for treating sinusitis caused by highly susceptible pathogens, providing that the dosing interval is 12 hours. PMID:25815573

  6. Novel application of human periodontal ligament stem cells and water-soluble chitin for collagen tissue regeneration: in vitro and in vivo investigations.

    PubMed

    Jung, Im Hee; Park, Jung Chul; Kim, Jane C; Jeon, Dong Won; Choi, Seong Ho; Cho, Kyoo Sung; Im, Gun Il; Kim, Byung Soo; Kim, Chang Sung

    2012-03-01

    Human periodontal ligament stem cells (hPDLSCs) have been proposed as an alternative to conventional cosmetic fillers because they display an innate ability to synthesize collagen. The aims of this study were to determine the effects of water-soluble chitin (WSC) on the proliferation and migration of hPDLSCs, and to quantify collagen synthesis in vitro and in vivo compared with human adipose-derived stem cell (hADSC)s. hPDLSCs were isolated from healthy extracted teeth, and the cell proliferation and cell migration capacities of untreated hPDLSCs (control group) and WSC-treated hPDLSCs (test group) were compared. Insoluble/soluble collagen synthesis were also assessed, and collagen related markers were evaluated including lysyl oxidase (LOX), lysyl oxidase like (LOXL)1, LOXL2, and hydroxyproline. In vivo collagen formation was examined by transplanting hyaluronic acid as a cell carrier into the subcutaneous pockets of immunocompromised mice in the control and test groups; histology and immunohistochemistry analyses were performed 4 (n=4) and 8 (n=4) weeks later. There was a dose-dependent enhancement of hPDLSCs proliferation in the test group, and a concomitant reduction in cell migration. The amount of insoluble collagen formed was greater in the test group than in the control group (p<0.05), whereas soluble collagen formation was significantly reduced in the test group (p<0.05). The histology and immunohistochemistry results revealed that the amount of collagen formed in vivo was greater in WSC-treated hPDLSCs than in the control cells at 4 and 8 weeks (p<0.05), and histometric analysis at 8 weeks revealed that enhancement of collagen formation by hPDLSCs was greater than by hADSCs. These results indicate that WSC modulates the properties of hPDLSCs, rendering them more suitable for cosmetic soft-tissue augmentation. PMID:21981356

  7. Terahertz pulsed imaging in vivo

    NASA Astrophysics Data System (ADS)

    Pickwell-MacPherson, E.

    2011-03-01

    Terahertz (1012 Hz) pulsed imaging is a totally non-destructive and non-ionising imaging modality and thus potential applications in medicine are being investigated. In this paper we present results using our hand-held terahertz probe that has been designed for in vivo use. In particular, we use the terahertz probe to perform reflection geometry in vivo measurements of human skin. The hand-held terahertz probe gives more flexibility than a typical flat-bed imaging system, but it also results in noisier data and requires existing processing methods to be improved. We describe the requirements and limitations of system geometry, data acquisition rate, image resolution and penetration depth and explain how various factors are dependent on each other. We show how some of the physical limitations can be overcome using novel data processing methods.

  8. EDITORIAL: Nanotechnology in vivo Nanotechnology in vivo

    NASA Astrophysics Data System (ADS)

    Demming, Anna

    2010-04-01

    Since the development of x-rays the ability to image inside our bodies has provided medicine with a potent diagnostic tool, as well as fascinating us with the eerie evidence of our mechanistic mortality. In December 2008 Osamu Shimomura, Martin Chalfie and Roger Y Tsien received a Nobel Prize for the discovery and development of the green fluorescent protein. The award recognised a new discovery that further facilitated our abilities to follow cellular activities and delve deeper into the workings of living organisms. Since the first observation of green fluorescent protein in jelly fish over thirty years ago, quantum dots have emerged as a potential alternative tool for imaging [1]. The advantages of quantum dots over organic dyes and fluorescent proteins include intense luminescence, high molar extinction coefficient, resistance to photobleaching, and broad excitation with narrow emission bands. However, one drawback for biological applications has been the layer of hydrophobic organic ligands often present at the surface as a result of the synthesis procedures. One solution to improve the solubility of quantum dots has been to conjugate them with a hydrophilic substance, as reported by Nie et al [2]. Chitosan is a hydrophilic, non-toxic, biocompatible and biodegradable substance and has been conjugated with quantum dots such as CdSe-ZnS [2] for bioassays and intracellular labelling. As well as luminescence, different nanoparticles present a variety of exceptional properties that render them useful in a range of bio applications, including MRI, drug delivery and cancer hyperthermia therapy. The ability to harness these various attributes in one system was reported by researchers in China, who incorporated magnetic nanoparticles, fluorescent quantum dots and pharmaceutical drugs into chitosan nanoparticles for multifunctional smart drug delivery systems [3]. More recently silicon quantum dots have emerged as a less cytotoxic alternative to CdSe for bio-imaging labels [4]. A surface hydroxyl group renders silicon quantum dots soluble in water and the photoluminescence can be made stable with oxygen-passivation. In addition, researchers in Japan have demonstrated how the initially modest yield in the preparation of silicon quantum dots can be improved to tens of milligrams per batch, thus further promoting their application in bio-imaging [5]. In the search for non-toxic quantum dots, researchers at the Amrita Centre for Nanoscience in India have prepared heavy metal-free quantum dot bio-probes based on single phase ZnS [6]. The quantum dots are selectively doped with metals, transition metals and halides to provide tuneable luminescence properties, and they are surface conjugated with folic acid for cancer targeting. The quantum dots were demonstrated to be water-soluble, non-toxic in normal and cancer cell lines, and have bright, tuneable luminescence. So far most of the quantum dots developed for bio-imaging have had excitation and emission wavelengths in the visible spectrum, which is highly absorbed by tissue. This limits imaging with these quantum dots to superficial tissues. This week, researchers in China and the US reported work developing functionalized dots for in vivo tumour vasculature in the infrared part of the spectrum [7]. In addition the quantum dots were functionalised with glycine-aspartic acid (RGD) peptides, which target the vasculature of almost all types of growing tumours, unlike antibody- or aptamer-mediated targeting strategies that are specific to a particular cancer type. In this issue, researchers in China and the US demonstrate a novel type of contrast agent for ultrasonic tumour imaging [8]. Contrast-enhanced ultrasonic tumour imaging extends the diagnostic and imaging capabilities of traditional techniques. The use of nanoparticles as ultrasound contrast agents exploits the presence of open pores in the range of 380 to 780 nm in tumour blood vessels, which enhance the permeability and retention of nanoparticles in the tumour vasculature. However, previous reports on techniques to generate

  9. Application of time-resolved autofluorescence to label-free in vivo optical mapping of changes in tissue matrix and metabolism associated with myocardial infarction and heart failure

    PubMed Central

    Lagarto, João; Dyer, Benjamin T.; Talbot, Clifford; Sikkel, Markus B.; Peters, Nicholas S.; French, Paul M. W.; Lyon, Alexander R.; Dunsby, Chris

    2015-01-01

    We investigate the potential of an instrument combining time-resolved spectrofluorometry and diffuse reflectance spectroscopy to measure structural and metabolic changes in cardiac tissue in vivo in a 16 week post-myocardial infarction heart failure model in rats. In the scar region, we observed changes in the fluorescence signal that can be explained by increased collagen content, which is in good agreement with histology. In areas remote from the scar tissue, we measured changes in the fluorescence signal (p < 0.001) that cannot be explained by differences in collagen content and we attribute this to altered metabolism within the myocardium. A linear discriminant analysis algorithm was applied to the measurements to predict the tissue disease state. When we combine all measurements, our results reveal high diagnostic accuracy in the infarcted area (100%) and border zone (94.44%) as well as in remote regions from the scar (> 77%). Overall, our results demonstrate the potential of our instrument to characterize structural and metabolic changes in a failing heart in vivo without using exogenous labels. PMID:25780727

  10. Application of time-resolved autofluorescence to label-free in vivo optical mapping of changes in tissue matrix and metabolism associated with myocardial infarction and heart failure.

    PubMed

    Lagarto, João; Dyer, Benjamin T; Talbot, Clifford; Sikkel, Markus B; Peters, Nicholas S; French, Paul M W; Lyon, Alexander R; Dunsby, Chris

    2015-02-01

    We investigate the potential of an instrument combining time-resolved spectrofluorometry and diffuse reflectance spectroscopy to measure structural and metabolic changes in cardiac tissue in vivo in a 16 week post-myocardial infarction heart failure model in rats. In the scar region, we observed changes in the fluorescence signal that can be explained by increased collagen content, which is in good agreement with histology. In areas remote from the scar tissue, we measured changes in the fluorescence signal (p < 0.001) that cannot be explained by differences in collagen content and we attribute this to altered metabolism within the myocardium. A linear discriminant analysis algorithm was applied to the measurements to predict the tissue disease state. When we combine all measurements, our results reveal high diagnostic accuracy in the infarcted area (100%) and border zone (94.44%) as well as in remote regions from the scar (> 77%). Overall, our results demonstrate the potential of our instrument to characterize structural and metabolic changes in a failing heart in vivo without using exogenous labels. PMID:25780727

  11. Implanted, inductively-coupled, radiofrequency coils fabricated on flexible polymeric material: Application to in vivo rat brain MRI at 7 T

    NASA Astrophysics Data System (ADS)

    Ginefri, J.-C.; Rubin, A.; Tatoulian, M.; Woytasik, M.; Boumezbeur, F.; Djemaï, B.; Poirier-Quinot, M.; Lethimonnier, F.; Darrasse, L.; Dufour-Gergam, E.

    2012-11-01

    Combined with high-field MRI scanners, small implanted coils allow for high resolution imaging with locally improved SNR, as compared to external coils. Small flexible implantable coils dedicated to in vivo MRI of the rat brain at 7 T were developed. Based on the Multi-turn Transmission Line Resonator design, they were fabricated with a Teflon substrate using copper micromolding process and a specific metal-polymer adhesion treatment. The implanted coils were made biocompatible by PolyDimethylSiloxane (PDMS) encapsulation. The use of low loss tangent material achieves low dielectric losses within the substrate and the use of the PDMS layer reduces the parasitic coupling with the surrounding media. An implanted coil was implemented in a 7 T MRI system using inductive coupling and a dedicated external pick-up coil for signal transmission. In vivo images of the rat brain acquired with in plane resolution of (150 ?m)2 thanks to the implanted coil revealed high SNR near the coil, allowing for the visualization of fine cerebral structures.

  12. Development of a liquid chromatographic method for the quantification of paromomycin. Application to in vitro release and ex vivo permeation studies

    NASA Astrophysics Data System (ADS)

    Pujol-Brugués, A.; Calpena-Campmany, A. C.; Riera-Lizandra, C.; Halbaut-Bellowa, L.; Clares-Naveros, B.

    2014-12-01

    We have developed a reversed phase high performance liquid chromatography pulsed amperometric detection (RPHPLC-PAD) method for the determination of paromomycin. It is sensitive, repeatable, and selective without the pretreatment step. Trifluoroacetic acid-water was utilized as the eluent and detected by PAD under NaOH alkaline conditions. The paromomycin detection limit (S/N = 3.3) was 2 ?g mL-1 and the quantification limit (S/N = 10) was 6 ?g mL-1. Coefficients of linear regression were higher than 0.99 for concentrations between 6.25 and 200 ?g mL-1. The intra and inter-day precision (RSD) was less than 6.5%. The average recoveries were 97.53-102.01%. The proposed HPLC-PAD method presented advantageous performance characteristics and it can be considered suitable for the evaluation of paromomycin loaded nanogel formulation in ex vivo permeation and in vitro release studies.

  13. Patient-derived Models of Human Breast Cancer: Protocols for In vitro and In vivo Applications in Tumor Biology and Translational Medicine

    PubMed Central

    DeRose, Yoko S.; Gligorich, Keith M.; Wang, Guoying; Georgelas, Ann; Bowman, Paulette; Courdy, Samir J.; Welm, Alana L.; Welm, Bryan E.

    2013-01-01

    Research models that replicate the diverse genetic and molecular landscape of breast cancer are critical for developing the next generation therapeutic entities that can target specific cancer subtypes. Patient-derived tumorgrafts, generated by transplanting primary human tumor samples into immune-compromised mice, are a valuable method to model the clinical diversity of breast cancer in mice, and are a potential resource in personalized medicine. Primary tumorgrafts also enable in vivo testing of therapeutics and make possible the use of patient cancer tissue for in vitro screens. Described in this unit are a variety of protocols including tissue collection, biospecimen tracking, tissue processing, transplantation, and 3-dimensional culturing of xenografted tissue, that enable use of bona fide uncultured human tissue in designing and validating cancer therapies. PMID:23456611

  14. Immobilized Cytochrome P450 2C9 (CYP2C9): Applications for Metabolite Generation, Monitoring Protein-Protein Interactions, and Improving In-vivo Predictions Using Enhanced In-vitro Models

    NASA Astrophysics Data System (ADS)

    Wollenberg, Lance A.

    Cytochrome P450 (P450) enzymes are a family of oxoferroreductase enzymes containing a heme moiety and are well known to be involved in the metabolism of a wide variety of endogenous and xenobiotic materials. It is estimated that roughly 75% of all pharmaceutical compounds are metabolized by these enzymes. Traditional reconstituted in-vitro incubation studies using recombinant P450 enzymes are often used to predict in-vivo kinetic parameters of a drug early in development. However, in many cases, these reconstituted incubations are prone to aggregation which has been shown to affect the catalytic activity of an enzyme. Moreover, the presence of other isoforms of P450 enzymes present in a metabolic incubation, as is the case with microsomal systems, may affect the catalytic activity of an enzyme through isoform-specific protein-protein interactions. Both of these effects may result in inaccurate prediction of in-vivo drug metabolism using in-vitro experiments. Here we described the development of immobilized P450 constructs designed to elucidate the effects of aggregation and protein-protein interactions between P450 isoforms on catalytic activities. The long term objective of this project is to develop a system to control the oligomeric state of Cytochrome P450 enzymes to accurately elucidate discrepancies between in vitro reconstituted systems and actual in vivo drug metabolism for the precise prediction of metabolic activity. This approach will serve as a system to better draw correlations between in-vivo and in-vitro drug metabolism data. The central hypothesis is that Cytochrome P450 enzymes catalytic activity can be altered by protein-protein interactions occurring between Cytochrome P450 enzymes involved in drug metabolism, and is dependent on varying states of protein aggregation. This dissertation explains the details of the construction and characterization of a nanostructure device designed to control the state of aggregation of a P450 enzyme. Moreover, applications of immobilized P450 enzyme constructs will also be used for monitoring protein-protein interaction and metabolite production with the use of immobilized-P450 bioreactor constructs. This work provides insight into the effect on catalytic activity caused by both P450 aggregation as well as isoform-specific protein-protein interactions and provides insight in the production of biosynthetically produced drug metabolites

  15. Design, synthesis and binding properties of a fluorescent ????/???? integrin antagonist and its application as an in vivo probe for bone marrow haemopoietic stem cells.

    PubMed

    Cao, Benjamin; Hutt, Oliver E; Zhang, Zhen; Li, Songhui; Heazlewood, Shen Y; Williams, Brenda; Smith, Jessica A; Haylock, David N; Savage, G Paul; Nilsson, Susan K

    2014-02-14

    The ?9?1 and ?4?1 integrin subtypes are expressed on bone marrow haemopoietic stem cells and have important roles in stem cell regulation and trafficking. Although the roles of ?4?1 integrin have been thoroughly investigated with respect to HSC function, the role of ?9?1 integrin remains poorly characterised. Small molecule fluorescent probes are useful tools for monitoring biological processes in vivo, to determine cell-associated protein localisation and activation, and to elucidate the mechanism of small molecule mediated protein interactions. Herein, we report the design, synthesis and integrin-dependent cell binding properties of a new fluorescent ?9?1 integrin antagonist (R-BC154), which was based on a series of N-phenylsulfonyl proline dipeptides and assembled using the Cu(I)-catalyzed azide alkyne cycloaddition (CuAAC) reaction. Using transfected human glioblastoma LN18 cells, we show that R-BC154 exhibits high nanomolar binding affinities to ?9?1 integrin with potent cross-reactivity against ?4?1 integrin under physiological mimicking conditions. On-rate and off-rate measurements revealed distinct differences in the binding kinetics between ?9?1 and ?4?1 integrins, which showed faster binding to ?4?1 integrin relative to ?9?1, but more prolonged binding to the latter. Finally, we show that R-BC154 was capable of binding rare populations of bone marrow haemopoietic stem and progenitor cells when administered to mice. Thus, R-BC154 represents a useful multi-purpose fluorescent integrin probe that can be used for (1) screening small molecule inhibitors of ?9?1 and ?4?1 integrins; (2) investigating the biochemical properties of ?9?1 and ?4?1 integrin binding and (3) investigating integrin expression and activation on defined cell phenotypes in vivo. PMID:24363056

  16. Measuring Proteome Dynamics in Vivo

    PubMed Central

    Rachdaoui, Nadia; Austin, Leanne; Kramer, Eric; Previs, Michael J.; Anderson, Vernon E.; Kasumov, Takhar; Previs, Stephen F.

    2009-01-01

    Proteomics investigations typically yield information regarding static gene expression profiles. The central issues that limit the study of proteome dynamics include how to (i) administer a labeled amino acid in vivo, (ii) measure the isotopic labeling of a protein(s) (which may be low), and (iii) reliably interpret the precursor/product labeling relationships. In this study, we demonstrate the potential of quantifying proteome dynamics by coupling the administration of stable isotopes with mass spectrometric assays. Although the direct administration of a labeled amino acid(s) is typically used to measure protein synthesis, we explain the application of labeled water, comparing 2H2O versus H218O for measuring albumin biosynthesis in vivo. This application emphasizes two distinct advantages of using labeled water over a labeled amino acid(s). First, in long term studies (e.g. days or weeks), it is not practical to continuously administer a labeled amino acid(s); however, in the presence of labeled water, organisms will generate labeled amino acids. Second, to calculate rates of protein synthesis in short term studies (e.g. hours), one must utilize a precursor/product labeling ratio; when using labeled water it is possible to reliably identify and easily measure the precursor labeling (i.e. water). We demonstrate that labeled water permits studies of protein synthesis (e.g. albumin synthesis in mice) during metabolic “steady-state” or “non-steady-state” conditions, i.e. integrating transitions between the fed and fasted state or during an acute perturbation (e.g. following a meal), respectively. We expect that the use of labeled water is applicable to wide scale investigations of proteome dynamics and can therein be used to obtain a functional image of gene expression in vivo. PMID:19724074

  17. Ex vivo optical metabolic measurements from cultured tissue reflect in vivo tissue status

    NASA Astrophysics Data System (ADS)

    Walsh, Alex J.; Poole, Kristin M.; Duvall, Craig L.; Skala, Melissa C.

    2012-11-01

    Optical measurements of metabolism are ideally acquired in vivo; however, intravital measurements are often impractical. Accurate ex vivo assessments would greatly broaden the applicability of optical measurements of metabolism. We investigate the use of live tissue culture experiments to serve as a surrogate for in vivo metabolic measurements. To validate this approach, NADH and FAD fluorescence intensity and lifetime images were acquired with a two-photon microscope from hamster cheek pouch epithelia in vivo, from biopsies maintained in live tissue culture up to 48 h, and from flash-frozen and thawed biopsies. We found that the optical redox ratio (fluorescence intensity of NADH/FAD) of the cultured biopsy was statistically identical to the in vivo measurement until 24 h, while the redox ratio of the frozen-thawed samples decreased by 15% (p<0.01). The NADH mean fluorescence lifetime (?m) remained unchanged (p>0.05) during the first 8 h of tissue culture, while the NADH ?m of frozen-thawed samples increased by 13% (p<0.001). Cellular morphology did not significantly change between in vivo, cultured, and frozen-thawed tissues (p>0.05). All results were consistent across multiple depth layers in this stratified squamous epithelial tissue. Histological markers for proliferation and apoptosis also confirm the viability of tissues maintained in culture. This study suggests that short-term ex vivo tissue culture may be more appropriate than frozen-thawed tissue for optical metabolic and morphologic measurements that approximate in vivo status.

  18. Supramolecular quantum dot-porphyrin assemblies for biological oxygen sensing

    E-print Network

    Lemon, Christopher M. (Christopher Michael)

    2013-01-01

    Generating metabolic profiles of tumors provides a spatiotemporal map of the concentration of key species to assess and quantify tumor growth, metabolism, and response to therapy. Because the tumor microenvironment is ...

  19. FRET excited ratiometric oxygen sensing in living tissue

    PubMed Central

    Ingram, Justin M.; Zhang, Chunfeng; Xu, Jian; Schiff, Steven J.

    2013-01-01

    Dynamic analysis of oxygen (O2) has been limited by the lack of a real-time, quantitative, and biocompatible sensor. To address these demands, we designed a ratiometric optode matrix consisting of the phosphorescence quenching dye platinum (II) octaethylporphine ketone (PtOEPK) and nanocystal quantum dots (NQDs), which when embedded within an inert polymer matrix allows long-term pre-designed excitation through fluorescence resonance energy transfer (FRET). Depositing this matrix on various glass substrates allowed the development of a series of optical sensors able to measure interstitial oxygen concentration [O2] with several hundred millisecond temporal resolution in varying biological microdomains of active brain tissue. PMID:23333398

  20. TOR signaling couples oxygen sensing to lifespan in C. elegans

    PubMed Central

    Schieber, Michael; Chandel, Navdeep S.

    2014-01-01

    Summary Metazoans adapt to a low oxygen environment (hypoxia) through activation of stress response pathways. Here we report that transient hypoxia exposure extends lifespan in C. elegans through mitochondrial ROS-dependent regulation of the nutrient sensing kinase TOR and its upstream activator RHEB-1. The increase in lifespan during hypoxia requires the intestinal GATA-type transcription factor, ELT-2, downstream of TOR signaling. Using RNA-Sequencing, we describe an ELT-2-dependent hypoxia response that includes an intestinal glutathione S-transferase, GSTO-1, and uncover that GSTO-1 is required for lifespan under hypoxia. These results indicate mitochondrial ROS-dependent TOR signaling integrates metabolic adaptations to confer survival under hypoxia. PMID:25284791

  1. Molecular aspects of oxygen sensing in physiological adaptation to hypoxia.

    PubMed

    Czyzyk-Krzeska, M F

    1997-11-01

    Oxygen is an essential substrate in aerobic metabolism for most eukaryotic organisms. Thus organisms and cells have developed numerous immediate and long-term compensatory mechanisms for dealing with oxygen deprivation. Adaptation to hypoxia at the organismal level includes reflex hyperventilation, polycythemia and angiogenesis, which lead to increased O2 delivery to the tissues. Adaptation at the cellular level involves a shift from oxidative phosphorylation to anaerobic glycolysis, increased glucose metabolism, and expression of hypoxic stress-related proteins. Regulation of many proteins participating in adaptation to hypoxia occurs at the level of gene expression. The most widespread molecular mechanism of hypoxia-dependent regulation is transcriptional induction via the binding of a transcription factor, hypoxia-inducible factor-1 (Hif-1), to the specific sequences on the regulated genes. Long-term induction of many proteins also requires an increase in mRNA stability, which is mediated by the binding of regulatory proteins to specific sequences within the mRNAs. The current theories of coupling between the O2 sensor and mechanisms controlling gene expression are discussed. PMID:9407604

  2. Phosphorescent semiconductor nanocrystals and proteins for biological oxygen sensing

    E-print Network

    McLaurin, Emily J. (Emily Jane)

    2011-01-01

    Oxygen is required for cellular respiration by all complex life making it a key metabolic profiling factor in biological systems. Tumors are defined by hypoxia (low pO2), which has been shown to influence response to ...

  3. Multimodal Mn-doped I-III-VI quantum dots for near infrared fluorescence and magnetic resonance imaging: from synthesis to in vivo application

    NASA Astrophysics Data System (ADS)

    Sitbon, Gary; Bouccara, Sophie; Tasso, Mariana; Francois, Aurélie; Bezdetnaya, Lina; Marchal, Frédéric; Beaumont, Marine; Pons, Thomas

    2014-07-01

    The development of sensitive multimodal contrast agents is a key issue to provide better global, multi-scale images for diagnostic or therapeutic purposes. Here we present the synthesis of Zn-Cu-In-(S, Se)/Zn1-xMnxS core-shell quantum dots (QDs) that can be used as markers for both near-infrared fluorescence imaging and magnetic resonance imaging (MRI). We first present the synthesis of Zn-Cu-In-(S, Se) cores coated with a thick ZnS shell doped with various proportions of Mn. Their emission wavelengths can be tuned over the NIR optical window suitable for deep tissue imaging. The incorporation of manganese ions (up to a few thousand ions per QD) confers them a paramagnetic character, as demonstrated by structural analysis and electron paramagnetic resonance spectroscopy. These QDs maintain their optical properties after transfer to water using ligand exchange. They exhibit T1-relaxivities up to 1400 mM-1 [QD] s-1 at 7 T and 300 K. We finally show that these QDs are suitable multimodal in vivo probes and demonstrate MRI and NIR fluorescence detection of regional lymph nodes in mice.The development of sensitive multimodal contrast agents is a key issue to provide better global, multi-scale images for diagnostic or therapeutic purposes. Here we present the synthesis of Zn-Cu-In-(S, Se)/Zn1-xMnxS core-shell quantum dots (QDs) that can be used as markers for both near-infrared fluorescence imaging and magnetic resonance imaging (MRI). We first present the synthesis of Zn-Cu-In-(S, Se) cores coated with a thick ZnS shell doped with various proportions of Mn. Their emission wavelengths can be tuned over the NIR optical window suitable for deep tissue imaging. The incorporation of manganese ions (up to a few thousand ions per QD) confers them a paramagnetic character, as demonstrated by structural analysis and electron paramagnetic resonance spectroscopy. These QDs maintain their optical properties after transfer to water using ligand exchange. They exhibit T1-relaxivities up to 1400 mM-1 [QD] s-1 at 7 T and 300 K. We finally show that these QDs are suitable multimodal in vivo probes and demonstrate MRI and NIR fluorescence detection of regional lymph nodes in mice. Electronic supplementary information (ESI) available: Determination of Mn content; magnetization measurements; additional TEM and spectroscopic data; additional NIR fluorescence image; MTT assay results. See DOI: 10.1039/c4nr02239d

  4. Fluorescent Multiblock ?-Conjugated Polymer Nanoparticles for In Vivo Tumor Targeting

    PubMed Central

    Ahmed, Eilaf; Morton, Stephen W.

    2014-01-01

    Highly fluorescent multiblock conjugated polymer nanoparticles with folic acid surface ligands are highly effective for bioimaging and in vivo tumor targeting. The targeted nanoparticles were preferentially localized in tumor cells in vivo, thereby illustrating their potential for diagnostic and therapeutic applications. PMID:23794490

  5. Volumetric tomography of fluorescent proteins through small animals in vivo

    E-print Network

    Lorenzo, Jorge Ripoll

    Volumetric tomography of fluorescent proteins through small animals in vivo Giannis Zacharakis of the art and should find wide in vivo imaging applications in drug discovery, immunology, and cancer research. fluorescence whole-body imaging imaging gene expression gene transfer multispectral imaging

  6. In vivo application of chitosan to improve bioavailability of cyanocobalamin, a form of vitamin B12, following intraintestinal administration in rats.

    PubMed

    Goto, Yuko; Masuda, Ayumi; Aiba, Tetsuya

    2015-04-10

    The effect of chitosan on the intestinal absorption of cyanocobalamin (VB12), a stable form of vitamin B12, was investigated in vivo in rats, with the aim of improving the oral bioavailability of VB12 for anemia treatment in patients with gastrectomy. The bioavailability was evaluated based on the plasma concentration profile of VB12 following intraintestinal administration of the VB12 solution containing chitosan at various concentrations. The bioavailability of VB12 was 0.6±0.2% when the chitosan-free VB12 solution was administered, while it increased to 10.5±3.3% when chitosan was dissolved in the VB12 solution at a concentration of 1%. The bioavailability of VB12 increases with the chitosan concentration, in which chitosan seems to augment the amount of VB12 absorbed without affecting the absorption rate constant of VB12. It was also shown that the bioavailability of VB12 does not increase further when the degree of chitosan deacetylation is increased from 83 to 100% by substitutively employing the fully deacetylated chitosan. These findings suggest that the oral administration of VB12 with readily available chitosan may be a practical approach for anemia treatment in patients with gastrectomy. PMID:25681732

  7. Determination of Oxycodone, Noroxycodone and Oxymorphone by High-Performance Liquid Chromatography–Electrospray Ionization-Tandem Mass Spectrometry in Human Matrices: In vivo and In vitro Applications

    PubMed Central

    Fang, Wenfang B.; Lofwall, Michelle R.; Walsh, Sharon L.; Moody, David E.

    2013-01-01

    The opioid analgesic oxycodone is widely abused and increasingly associated with overdose deaths. A sensitive analytical method was developed for oxycodone and its metabolites, noroxycodone and oxymorphone, in human plasma, urine (±enzymatic hydrolysis at 50°C for 16 h) and liver microsomes (HLMs). Liquid–liquid extraction was followed by high-performance liquid chromatography–electrospray ionization-tandem mass spectrometry. The calibration range was 0.2–250 ng/mL for plasma and HLM and 10–5000 ng/mL for urine. Intra- and interrun accuracies were within 13.3% of target; precisions were within 12.8% for all matrices. Recoveries from plasma were: oxycodone, 75.6%; noroxycodone, 37.4% and oxymorphone, 18.2%. Analytes exhibited room temperature stability in plasma and urine up to 24 h, and freeze–thaw stability in plasma up to three cycles. In 24-h hydrolyzed urine from subjects administered intranasal oxycodone (30 mg/70 kg, n = 5), mean concentrations (ng/mL) and % daily doses excreted were: oxycodone, 1150, 6.53%; noroxycodone, 1330, 7.81% and oxymorphone, 3000, 17.1%. Oxycodone incubated with HLM produced more noroxycodone than oxymorphone. With a panel of recombinant human cytochrome P450s (CYPs), CYP2C18 and CYP3A4 produced the most noroxycodone, whereas CYP2D6 produced the most oxymorphone. These results demonstrate a new method suitable for both in vivo and in vitro metabolism and pharmacokinetic studies of oxycodone. PMID:23743505

  8. Cultivation of keratinocytes and fibroblasts in a three-dimensional bovine collagen-elastin matrix (Matriderm®) and application for full thickness wound coverage in vivo.

    PubMed

    Killat, Jasper; Reimers, Kerstin; Choi, Claudia Y; Jahn, Sabrina; Vogt, Peter M; Radtke, Christine

    2013-01-01

    New skin substitutes for burn medicine or reconstructive surgery pose an important issue in plastic surgery. Matriderm® is a clinically approved three-dimensional bovine collagen-elastin matrix which is already used as a dermal substitute of full thickness burn wounds. The drawback of an avital matrix is the limited integration in full thickness skin defects, depending on the defect size. To further optimize this process, Matriderm® has also been studied as a matrix for tissue engineering of skin albeit long-term cultivation of the matrix with cells has been difficult. Cells have generally been seeded onto the matrix with high cell loss and minimal time-consuming migration. Here we developed a cell seeded skin equivalent after microtransfer of cells directly into the matrix. First, cells were cultured, and microinjected into Matriderm®. Then, cell viability in the matrix was determined by histology in vitro. As a next step, the skin substitute was applied in vivo into a full thickness rodent wound model. The wound coverage and healing was observed over a period of two weeks followed by histological examination assessing cell viability, proliferation and integration into the host. Viable and proliferating cells could be found throughout the entire matrix. The presented skin substitute resembles healthy skin in morphology and integrity. Based on this study, future investigations are planned to examine behaviour of epidermal stem cells injected into a collagen-elastin matrix under the aspects of establishment of stem cell niches and differentiation. PMID:23852021

  9. Localizing the chemical forms of sulfur in vivo using X-ray fluorescence spectroscopic imaging: application to onion (Allium cepa) tissues.

    PubMed

    Pickering, Ingrid J; Sneeden, Eileen Yu; Prince, Roger C; Block, Eric; Harris, Hugh H; Hirsch, Gregory; George, Graham N

    2009-07-28

    Sulfur has a particularly rich biochemistry and fills a number of important roles in biology. In situ information on sulfur biochemistry is generally difficult to obtain because of a lack of biophysical techniques that have sufficient sensitivity to molecular form. We have recently reported that sulfur K-edge X-ray absorption spectroscopy can be used as a direct probe of the sulfur biochemistry of living mammalian cells [Gnida, M., et al. (2007) Biochemistry 46, 14735-14741]. Here we report an extension of this work and develop sulfur K-edge X-ray fluorescence spectroscopic imaging as an in vivo probe of sulfur metabolism in living cells. For this work, we have chosen onion (Allium cepa) as a tractable model system with well-developed sulfur biochemistry and present evidence of the localization of a number of different chemical forms. X-ray absorption spectroscopy of onion sections showed increased levels of lachrymatory factor (LF) and thiosulfinate and decreased levels of sulfoxide (LF precursor) following cell breakage. In intact cells, X-ray fluorescence spectroscopic imaging showed elevated levels of sulfoxides in the cytosol and elevated levels of reduced sulfur in the central transport vessels and bundle sheath cells. PMID:19463015

  10. Novel estimation of the electrical bioimpedance using the local polynomial method. Application to in vivo real-time myocardium tissue impedance characterization during the cardiac cycle.

    PubMed

    Sanchez, Benjamin; Schoukens, Johan; Bragos, Ramon; Vandersteen, Gerd

    2011-12-01

    Classical measurements of myocardium tissue electrical impedance for characterizing the morphology of myocardium cells, as well as cell membranes integrity and intra/extra cellular spaces, are based on the frequency-sweep electrical impedance spectroscopy (EIS) technique. In contrast to the frequency-sweep EIS approach, measuring with broadband signals, i.e., multisine excitations, enables to collect, simultaneously, multiple myocardium tissue impedance data in a short measuring time. However, reducing the measuring time makes the measurements to be prone to the influence of the transients introduced by noise and the dynamic time-varying properties of tissue. This paper presents a novel approach for the impedance-frequency-response estimation based on the local polynomial method (LPM). The fast LPM version presented rejects the leakage error's influence on the impedance frequency response when measuring electrical bioimpedance in a short time. The theory is supported by a set of validation measurements. Novel preliminary experimental results obtained from real-time in vivo healthy myocardium tissue impedance characterization within the cardiac cycle using multisine excitation are reported. PMID:21878408

  11. Measurement of glycine in the human brain in vivo by 1H-MRS at 3T: Application in brain tumors

    PubMed Central

    Choi, Changho; Ganji, Sandeep K.; DeBerardinis, Ralph J.; Dimitrov, Ivan E.; Pascual, Juan M.; Bachoo, Robert; Mickey, Bruce E.; Malloy, Craig R.; Maher, Elizabeth A.

    2011-01-01

    Glycine (Gly) is a key metabolic intermediate required for the synthesis of proteins, nucleic acids and other molecules, and its detection in cancer could therefore provide biologically relevant information about the growth of the tumor. Here we report measurement of Gly in human brain and gliomas by an optimized point-resolved spectroscopy (PRESS) sequence at 3T. Echo time dependence of the major obstacle, myo-inositol (mI) multiplet, was investigated with numerical simulations, incorporating the 3D volume localization. The simulations indicated that a subecho pair (TE1, TE2) = (60, 100) ms permits detection of both Gly and mI with optimum selectivity. In vivo validation of the optimized PRESS was conducted on the right parietal cortex of five healthy volunteers. Metabolite signals estimated from LCModel were normalized with respect to the brain water signal and the concentrations were evaluated assuming the total creatine concentration at 8 mM. The Gly concentration was estimated as 0.6±0.1 mM (mean±SD, n=5), with a mean Cramér-Rao lower bound of 9±1%. The PRESS sequence was applied to measure the Gly levels in patients with glioblastoma multiforme. Metabolite concentrations were obtained using the water signal from the tumor mass. The study revealed that a subset of human gliomas contains glycine levels elevated 1.5–8 fold relative to normal. PMID:21394775

  12. The application of hyaluronic acid-derivatized carbon nanotubes in hematoporphyrin monomethyl ether-based photodynamic therapy for in vivo and in vitro cancer treatment.

    PubMed

    Shi, Jinjin; Ma, Rourou; Wang, Lei; Zhang, Jing; Liu, Ruiyuan; Li, Lulu; Liu, Yan; Hou, Lin; Yu, Xiaoyuan; Gao, Jun; Zhang, Zhenzhong

    2013-01-01

    Carbon nanotubes (CNTs) have shown great potential in both photothermal therapy and drug delivery. In this study, a CNT derivative, hyaluronic acid-derivatized CNTs (HA-CNTs) with high aqueous solubility, neutral pH, and tumor-targeting activity, were synthesized and characterized, and then a new photodynamic therapy agent, hematoporphyrin monomethyl ether (HMME), was adsorbed onto the functionalized CNTs to develop HMME-HA-CNTs. Tumor growth inhibition was investigated both in vivo and in vitro by a combination of photothermal therapy and photodynamic therapy using HMME-HA-CNTs. The ability of HMME-HA-CNT nanoparticles to combine local specific photodynamic therapy with external near-infrared photothermal therapy significantly improved the therapeutic efficacy of cancer treatment. Compared with photodynamic therapy or photothermal therapy alone, the combined treatment demonstrated a synergistic effect, resulting in higher therapeutic efficacy without obvious toxic effects to normal organs. Overall, it was demonstrated that HMME-HA-CNTs could be successfully applied to photodynamic therapy and photothermal therapy simultaneously in future tumor therapy. PMID:23843694

  13. The application of hyaluronic acid-derivatized carbon nanotubes in hematoporphyrin monomethyl ether-based photodynamic therapy for in vivo and in vitro cancer treatment

    PubMed Central

    Shi, Jinjin; Ma, Rourou; Wang, Lei; Zhang, Jing; Liu, Ruiyuan; Li, Lulu; Liu, Yan; Hou, Lin; Yu, Xiaoyuan; Gao, Jun; Zhang, Zhenzhong

    2013-01-01

    Carbon nanotubes (CNTs) have shown great potential in both photothermal therapy and drug delivery. In this study, a CNT derivative, hyaluronic acid-derivatized CNTs (HA-CNTs) with high aqueous solubility, neutral pH, and tumor-targeting activity, were synthesized and characterized, and then a new photodynamic therapy agent, hematoporphyrin monomethyl ether (HMME), was adsorbed onto the functionalized CNTs to develop HMME-HA-CNTs. Tumor growth inhibition was investigated both in vivo and in vitro by a combination of photothermal therapy and photodynamic therapy using HMME-HA-CNTs. The ability of HMME-HA-CNT nanoparticles to combine local specific photodynamic therapy with external near-infrared photothermal therapy significantly improved the therapeutic efficacy of cancer treatment. Compared with photodynamic therapy or photothermal therapy alone, the combined treatment demonstrated a synergistic effect, resulting in higher therapeutic efficacy without obvious toxic effects to normal organs. Overall, it was demonstrated that HMME-HA-CNTs could be successfully applied to photodynamic therapy and photothermal therapy simultaneously in future tumor therapy. PMID:23843694

  14. In vivo detection of human papilloma virus-induced lesions of anogenital area after application of acetic acid: a novel and accurate approach to a trivial method

    Microsoft Academic Search

    Irene M. Stefanaki; Androniki D. Tosca; George C. Themelis; Eleftheria M. Vazgiouraki; Despina N. Dokianakis; John G. Panayiotidis; Demetrios A. Spandidos; Costas J. Balas

    2001-01-01

    Human papilloma virus infection is increasing at an alarming rate. The ability of the virus to establish a subclinical infection and its association with malignancy of the lower genital tract make the statistics even more worrisome. Topical application of acetic acid solution provokes temporal alterations of the light-scattering properties of human papilloma virus-induced lesions of anogenital area. For the in

  15. In Vivo skin hydration and anti-erythema effects of Aloe vera, Aloe ferox and Aloe marlothii gel materials after single and multiple applications

    PubMed Central

    Fox, Lizelle T.; du Plessis, Jeanetta; Gerber, Minja; van Zyl, Sterna; Boneschans, Banie; Hamman, Josias H.

    2014-01-01

    Objective: To investigate the skin hydrating and anti-erythema activity of gel materials from Aloe marlothii A. Berger and A. ferox Mill. in comparison to that of Aloe barbadensis Miller (Aloe vera) in healthy human volunteers. Materials and Methods: Aqueous solutions of the polisaccharidic fractions of the selected aloe leaf gel materials were applied to the volar forearm skin of female subjects. The hydration effect of the aloe gel materials were measured with a Corneometer® CM 825, Visioscan® VC 98 and Cutometer® dual MPA 580 after single and multiple applications. The Mexameter® MX 18 was used to determine the anti-erythema effects of the aloe material solutions on irritated skin areas. Results: The A. vera and A. marlothii gel materials hydrated the skin after a single application, whereas the A. ferox gel material showed dehydration effects compared to the placebo. After multiple applications all the aloe materials exhibited dehydration effects on the skin. Mexameter® readings showed that A. vera and A. ferox have anti-erythema activity similar to that of the positive control group (i.e. hydrocortisone gel) after 6 days of treatment. Conclusion: The polysaccharide component of the gel materials from selected aloe species has a dehydrating effect on the skin after multiple applications. Both A. vera and A. ferox gel materials showed potential to reduce erythema on the skin similar to that of hydrocortisone gel. PMID:24991119

  16. Optimization of In Vivo Confocal Autofluorescence Imaging of the Ocular Fundus in Mice and Its Application to Models of Human Retinal Degeneration

    PubMed Central

    Issa, Peter Charbel; Singh, Mandeep S.; Lipinski, Daniel M.; Chong, Ngaihang V.; Delori, François C.; Barnard, Alun R.; MacLaren, Robert E.

    2012-01-01

    Purpose. To investigate the feasibility and to identify sources of experimental variability of quantitative and qualitative fundus autofluorescence (AF) assessment in mice. Methods. Blue (488 nm) and near-infrared (790 nm) fundus AF imaging was performed in various mouse strains and disease models (129S2, C57Bl/6, Abca4?/?, C3H-Pde6brd1/rd1, Rho?/?, and BALB/c mice) using a commercially available scanning laser ophthalmoscope. Gray-level analysis was used to explore factors influencing fundus AF measurements. Results. A contact lens avoided cataract development and resulted in consistent fundus AF recordings. Fundus illumination and magnification were sensitive to changes of the camera position. Standardized adjustment of the recorded confocal plane and consideration of the pupil area allowed reproducible recording of fundus AF from the retinal pigment epithelium with an intersession coefficient of repeatability of ±22%. Photopigment bleaching occurred during the first 1.5 seconds of exposure to 488 nm blue light (?10 mW/cm2), resulting in an increase of fundus AF. In addition, there was a slight decrease in fundus AF during prolonged blue light exposure. Fundus AF at 488 nm was low in animals with an absence of a normal visual cycle, and high in BALB/c and Abca4?/? mice. Degenerative alterations in Pde6brd1/rd1 and Rho?/? were reminiscent of findings in human retinal disease. Conclusions. Investigation of retinal phenotypes in mice is possible in vivo using standardized fundus AF imaging. Correlation with postmortem analysis is likely to lead to further understanding of human disease phenotypes and of retinal degenerations in general. Fundus AF imaging may be useful as an outcome measure in preclinical trials, such as for monitoring effects aimed at lowering lipofuscin accumulation in the retinal pigment epithelium. PMID:22169101

  17. Ocular Distribution, Spectrum of Activity, and In Vivo Viral Neutralization of a Fully Humanized Anti-Herpes Simplex Virus IgG Fab Fragment following Topical Application

    PubMed Central

    Berdugo, Marianne; Larsen, Inna V.; Abadie, Claire; Deloche, Catherine; Kowalczuk, Laura; Touchard, Elodie; Dubielzig, Richard; Brandt, Curtis R.; Combette, Jean-Marc

    2012-01-01

    Herpes simplex ocular infection is a major cause of corneal blindness. Local antiviral treatments exist but are associated with corneal toxicity, and resistance has become an issue. We evaluated the biodistribution and efficacy of a humanized anti-herpes simplex virus (anti-HSV) IgG FAb fragment (AC-8; 53 kDa) following repeated topical administration. AC-8 was found in the corneal epithelium, anterior stroma, subepithelial stromal cells, and retinal glial cells, with preferential entry through the ocular limbus. AC-8 was active against 13 different strains of HSV-1, with 50% and 90% mean effective concentrations (MEC50 and MEC90, respectively) ranging from 0.03 to 0.13 ?g/ml, indicating broad-spectrum activity. The in vivo efficacy of AC-8 was evaluated in a mouse model of herpes-induced ocular disease. Treatment with low-dose AC-8 (1 mg/ml) slightly reduced the ocular disease scores. A greater reduction of the disease scores was observed in the 10-mg/ml AC-8-treated group, but not as much as with trifluridine (TFT). AC-8 treatment reduced viral titers but less than trifluridine. AC-8 did not display any toxicity to the cornea or other structures in the eye. In summary, topical instillation of an anti-HSV FAb can be used on both intact and ulcerated corneas. It is well tolerated and does not alter reepithelialization. Further studies to improve the antiviral effect are needed for AC-8 to be considered for therapeutic use. PMID:22203590

  18. In vitro and in vivo experiments with iron oxide nanoparticles functionalized with DEXTRAN or polyethylene glycol for medical applications: magnetic targeting.

    PubMed

    Mojica Pisciotti, M L; Lima, E; Vasquez Mansilla, M; Tognoli, V E; Troiani, H E; Pasa, A A; Creczynski-Pasa, T B; Silva, A H; Gurman, P; Colombo, L; Goya, G F; Lamagna, A; Zysler, R D

    2014-05-01

    In this research work, DEXTRAN- and polyethylene glycol (PEG)-coated iron-oxide superparamagnetic nanoparticles were synthetized and their cytotoxicity and biodistribution assessed. Well-crystalline hydrophobic Fe3 O4 SPIONs were formed by a thermal decomposition process with d = 18 nm and ? = 2 nm; finally, the character of SPIONs was changed to hydrophilic by a post-synthesis procedure with the functionalization of the SPIONs with PEG or DEXTRAN. The nanoparticles present high saturation magnetization and superparamagnetic behavior at room temperature, and the hydrodynamic diameters of DEXTRAN- and PEG-coated SPIONs were measured as 170 and 120 nm, respectively. PEG- and DEXTRAN-coated SPIONs have a Specific Power Absorption SPA of 320 and 400 W/g, respectively, in an ac magnetic field with amplitude of 13 kA/m and frequency of 256 kHz. In vitro studies using VERO and MDCK cell lineages were performed to study the cytotoxicity and cell uptake of the SPIONs. For both cell lineages, PEG- and DEXTRAN-coated nanoparticles presented high cell viability for concentrations as high as 200 ?g/mL. In vivo studies were conducted using BALB/c mice inoculating the SPIONs intravenously and exposing them to the presence of an external magnet located over the tumour. It was observed that the amount of PEG-coated SPIONs in the tumor increased by up to 160% when using the external permanent magnetic as opposed to those animals that were not exposed to the external magnetic field. PMID:24458920

  19. Elastography Using Multi-Stream GPU: An Application to Online Tracked Ultrasound Elastography, In-Vivo and the da Vinci Surgical System

    PubMed Central

    Deshmukh, Nishikant P.; Kang, Hyun Jae; Billings, Seth D.; Taylor, Russell H.; Hager, Gregory D.; Boctor, Emad M.

    2014-01-01

    A system for real-time ultrasound (US) elastography will advance interventions for the diagnosis and treatment of cancer by advancing methods such as thermal monitoring of tissue ablation. A multi-stream graphics processing unit (GPU) based accelerated normalized cross-correlation (NCC) elastography, with a maximum frame rate of 78 frames per second, is presented in this paper. A study of NCC window size is undertaken to determine the effect on frame rate and the quality of output elastography images. This paper also presents a novel system for Online Tracked Ultrasound Elastography (O-TRuE), which extends prior work on an offline method. By tracking the US probe with an electromagnetic (EM) tracker, the system selects in-plane radio frequency (RF) data frames for generating high quality elastograms. A novel method for evaluating the quality of an elastography output stream is presented, suggesting that O-TRuE generates more stable elastograms than generated by untracked, free-hand palpation. Since EM tracking cannot be used in all systems, an integration of real-time elastography and the da Vinci Surgical System is presented and evaluated for elastography stream quality based on our metric. The da Vinci surgical robot is outfitted with a laparoscopic US probe, and palpation motions are autonomously generated by customized software. It is found that a stable output stream can be achieved, which is affected by both the frequency and amplitude of palpation. The GPU framework is validated using data from in-vivo pig liver ablation; the generated elastography images identify the ablated region, outlined more clearly than in the corresponding B-mode US images. PMID:25541954

  20. Transdermal delivery system for zidovudine: in vitro, ex vivo and in vivo evaluation.

    PubMed

    Narishetty, Sunil Thomas Kumar; Panchagnula, Ramesh

    2004-01-01

    The objective of this study was to prepare a transdermal delivery system (TDS) for zidovudine (AZT) with a combination of menthol and oleic acid as penetration enhancers incorporated in hydroxypropyl methylcellulose, and to evaluate ex vivo as well as in vivo permeation across rat skin. It was found that AZT in gel formulation was stable in both refrigerated as well as accelerated stability conditions for 3 months and further, the gel did not significantly retard the permeability of AZT across the skin in comparison with solution formulation. Ex vivo steady state flux of AZT across rat skin from gel was 2.26 mg cm(-2) h(-1), which is sufficient to achieve therapeutic plasma concentrations. Intravenous pharmacokinetic parameters of AZT in rats were determined and used together with ex vivo flux data to generate theoretical plasma profiles of AZT and compared with plasma concentrations achieved after application of TDS. Further, steady state plasma concentrations of drug following multiple applications of TDS were determined and good correlations between ex vivo and in vivo data were observed. In addition, the combination of penetration enhancers used at 2.5% w/w in this study proved efficient in achieving sufficient enhancement in the transdermal permeability of AZT across rat skin with reduced skin irritation potential when compared with individual penetration enhancers at higher concentrations. PMID:14716748

  1. Quantitative determination of zolmitriptan in rat blood and cerebrospinal fluid by reversed phase HPLC-ESI-MS/MS analysis: application to in vivo preclinical pharmacokinetic study.

    PubMed

    Dalpiaz, Alessandro; Marchetti, Nicola; Cavazzini, Alberto; Pasti, Luisa; Velaga, Sitaram; Gavini, Elisabetta; Beggiato, Sarah; Ferraro, Luca

    2012-07-15

    A fast HPLC-ESI-MS/MS method has been developed and validated for the quantification of the potent and selective antimigraine zolmitriptan in rat blood and cerebrospinal fluid (CSF). The assay has been then applied for in vivo preclinical studies. The analytical determination has been used to obtain pharmacokinetics of zolmitriptan in the two biological matrices after its intravenous or nasal administration. Liquid-liquid extraction of zolmitriptan was performed from 100 ?L rat blood samples in the presence of N(6)-cyclopentyladenosine (internal standard) with the employment of ethyl acetate. Calibration standards were prepared by using blood matrix and following the same liquid-liquid extraction procedure. CSF samples were analyzed without any pre-treatment steps and by using an external calibration method in pure water matrix. Chromatographic separation was performed under reversed phase and a gradient elution condition on a C18 packed column (100 × 2.0 mm, 2.5 ?m particles diameter). The mobile phase was a mixture between acetonitrile, water and formic acid (0.1% v/v). The applied HPLC-MS/MS method allowed low limits of detection, as calculated from calibration curves, of 6.6 and 24.4 ng/mL for water matrix and rat blood extracts, respectively. Linearity of the calibration curves was established up to 5 ?M (1.44 ?g/mL), as well as good assay accuracy. The intravenous infusion of 20 ?g zolmitriptan to male Sprague-Dawley rats produced blood concentrations ranging from 9.4±0.7 to 1.24±0.07 ?g/mL within 10 h, with a terminal half-life of 3.4±0.2h. The nasal administration of a water suspension of 20 ?g zolmitriptan produced blood concentrations ranging from 2.92±0.21 to 0.85±0.07 ?g/mL within 6h. One hour after zolmitriptan intravenous infusion or nasal administration, its CSF concentrations were 0.0539±0.0016 and 0.0453±0.0012 ?g/mL, respectively. This study determined the suitability of the herein proposed method to investigate the pharmacokinetics of zolmitriptan after its administration by means of novel formulations and, hence, to evaluate the efficacy of innovative nose-to-brain drug delivery in preclinical studies. PMID:22743338

  2. One-pot hydrothermal synthesis of lanthanide ions doped one-dimensional upconversion submicrocrystals and their potential application in vivo CT imaging.

    PubMed

    Gao, Guo; Zhang, Chunlei; Zhou, Zhijun; Zhang, Xin; Ma, Jiebing; Li, Chao; Jin, Weilin; Cui, Daxiang

    2013-01-01

    Multi-functional rare-earth Yb(3+) and Ln(3+) (Ln = Er, Tm and Ho) ions doped one-dimensional (1-D) upconversion submicrocrystals (NaYF(4) and NaGdF(4)) possessing upconversion luminescence, biocompatibility and magnetic properties have been synthesized by a one-pot hydrothermal method. Rare-earth Yb(3+) and Ln(3+) ions doped NaYF(4) microrods (~1 ?m in diameter, 3-5 ?m in length) exhibit porous properties, and the average pore sizes are ~28.2 nm. They show paramagnetism in the magnetic range of -60 to -2 kOe and 2 to 60 kOe at 300 K, and exhibit near superparamagnetic behaviour at the magnetic range of -2 to 2 kOe. Saturation magnetization was ~12.1 emu g(-1) at 2 K. The Yb(3+) and Ln(3+) ions doped NaGdF(4) submicrocrystals (~100 nm in diameter, 200-300 nm in length) show paramagnetism at 300 K, and exhibit superparamagnetic behaviour with a saturation magnetization of 129.2 emu g(-1) at 2 K. The magnetic properties of Yb(3+) and Ln(3+) ions doped 1-D upconversion submicrocrystals indicate they can be used for drug targeting under a magnetic field. Their unique upconversion emission (green for Yb(3+)/Er(3+) and blue for Yb(3+)/Tm(3+)) under 980 nm laser excitation indicate that they could be used for specific luminescent immunolabeling and imaging. MTT assays reveal that 1-D upconversion submicrocrystals have satisfactory bio-affinity, where the viability keeps in good state even at a concentration of 500 ?g mL(-1), which is much higher than the concentration usually used in cell labelling. Luminescent microscopy images show that the morphologies of the cytoskeleton and cell nucleus are well maintained after incubating different concentrations of 1-D upconversion submicrocrystals. After injecting upconversion submicrocrystals into the mice (tumor sites or back normal tissue), a clearly distinguished CT signal was observed, indicating the synthesized 1-D submicrocrystals are effective for CT imaging in vivo. PMID:23168841

  3. In vivo microautoradiography of [ 3 H]1,24(OH) 2 D 3 (tacalcitol) following topical application to normal rats and in vitro metabolism in human keratinocytes

    Microsoft Academic Search

    Tomohiro Ohta; Kohichiro Okabe; Yoshiaki Azuma; Mamoru Kiyoki

    1996-01-01

    This study was conducted to investigate the mechanism of topical absorption of [3H]1,24(OH)2D3 (1,24-dihydroxyvitamin D3; tacalcitol) by applying an ointment containing 4 ?g\\/g [3H]1,24(OH)2D3 to the skin of rats using an occlusion method. Microautoradiography of the skin at the application site 1 h after topical\\u000a treatment showed a high concentration of radiolabel in the stratum corneum, the epidermis and around

  4. Application of the Principles of Systems Biology and Wiener’s Cybernetics for Analysis of Regulation of Energy Fluxes in Muscle Cells in Vivo

    PubMed Central

    Guzun, Rita; Saks, Valdur

    2010-01-01

    The mechanisms of regulation of respiration and energy fluxes in the cells are analyzed based on the concepts of systems biology, non-equilibrium steady state kinetics and applications of Wiener’s cybernetic principles of feedback regulation. Under physiological conditions cardiac function is governed by the Frank-Starling law and the main metabolic characteristic of cardiac muscle cells is metabolic homeostasis, when both workload and respiration rate can be changed manifold at constant intracellular level of phosphocreatine and ATP in the cells. This is not observed in skeletal muscles. Controversies in theoretical explanations of these observations are analyzed. Experimental studies of permeabilized fibers from human skeletal muscle vastus lateralis and adult rat cardiomyocytes showed that the respiration rate is always an apparent hyperbolic but not a sigmoid function of ADP concentration. It is our conclusion that realistic explanations of regulation of energy fluxes in muscle cells require systemic approaches including application of the feedback theory of Wiener’s cybernetics in combination with detailed experimental research. Such an analysis reveals the importance of limited permeability of mitochondrial outer membrane for ADP due to interactions of mitochondria with cytoskeleton resulting in quasi-linear dependence of respiration rate on amplitude of cyclic changes in cytoplasmic ADP concentrations. The system of compartmentalized creatine kinase (CK) isoenzymes functionally coupled to ANT and ATPases, and mitochondrial-cytoskeletal interactions separate energy fluxes (mass and energy transfer) from signalling (information transfer) within dissipative metabolic structures – intracellular energetic units (ICEU). Due to the non-equilibrium state of CK reactions, intracellular ATP utilization and mitochondrial ATP regeneration are interconnected by the PCr flux from mitochondria. The feedback regulation of respiration occurring via cyclic fluctuations of cytosolic ADP, Pi and Cr/PCr ensures metabolic stability necessary for normal function of cardiac cells. PMID:20479996

  5. Application of Carbon-Ion Beams or Gamma-Rays on Primary Tumors Does Not Change the Expression Profiles of Metastatic Tumors in an In Vivo Murine Model

    SciTech Connect

    Tamaki, Tomoaki [RadGenomics Research Group, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba (Japan); Department of Radiation Oncology, Gunma University Graduate School of Medicine, Maebashi, Gunma (Japan); Iwakawa, Mayumi [RadGenomics Research Group, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba (Japan)], E-mail: mayumii@nirs.go.jp; Ohno, Tatsuya; Imadome, Kaori; Nakawatari, Miyako; Sakai, Minako; Tsujii, Hirohiko [RadGenomics Research Group, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba (Japan); Nakano, Takashi [Department of Radiation Oncology, Gunma University Graduate School of Medicine, Maebashi, Gunma (Japan); Imai, Takashi [RadGenomics Research Group, Research Center for Charged Particle Therapy, National Institute of Radiological Sciences, Chiba (Japan)

    2009-05-01

    Purpose: To clarify how carbon-ion radiotherapy (C-ion) on primary tumors affects the characteristics of subsequently arising metastatic tumor cells. Methods and Materials: Mouse squamous cell carcinomas, NR-S1, in synergic C3H/HeMsNrs mice were irradiated with a single dose of 5-50 Gy of C-ion (290 MeV per nucleon, 6-cm spread-out Bragg peak) or {gamma}-rays ({sup 137}Cs source) as a reference beam. The volume of the primary tumors and the number of metastatic nodules in lung were studied, and histologic analysis and microarray analysis of laser-microdissected tumor cells were also performed. Results: Including 5 Gy of C-ion and 8 Gy of {gamma}-rays, which did not inhibit the primary tumor growth, all doses used in this study inhibited lung metastasis significantly. Pathologic findings showed no difference among the metastatic tumor nodules in the nonirradiated, C-ion-irradiated, and {gamma}-ray-irradiated groups. Clustering analysis of expression profiles among metastatic tumors and primary tumors revealed a single cluster consisting of metastatic tumors different from their original primary tumors, indicating that the expression profiles of the metastatic tumor cells were not affected by the local application of C-ion or {gamma}-ray radiotherapy. Conclusion: We found no difference in the incidence and histology, and only small differences in expression profile, of distant metastasis between local C-ion and {gamma}-ray radiotherapy. The application of local radiotherapy per se or the type of radiotherapy applied did not influence the transcriptional changes caused by metastasis in tumor cells.

  6. Ex vivo DNA Assembly

    PubMed Central

    Fisher, Adam B.; Canfield, Zachary B.; Hayward, Laura C.; Fong, Stephen S.; McArthur, George H.

    2013-01-01

    Even with decreasing DNA synthesis costs there remains a need for inexpensive, rapid, and reliable methods for assembling synthetic DNA into larger constructs or combinatorial libraries. Advances in cloning techniques have resulted in powerful in vitro and in vivo assembly of DNA. However, monetary and time costs have limited these approaches. Here, we report an ex vivo DNA assembly method that uses cellular lysates derived from a commonly used laboratory strain of Escherichia coli for joining double-stranded DNA with short end homologies embedded within inexpensive primers. This method concurrently shortens the time and decreases costs associated with current DNA assembly methods. PMID:25024067

  7. In-vivo optical investigation of psoriasis

    NASA Astrophysics Data System (ADS)

    Kapsokalyvas, Dimitrios; Cicchi, Riccardo; Bruscino, Nicola; Alfieri, Domenico; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2011-03-01

    Psoriasis is an autoimmune disease of the skin characterized by hyperkeratosis, hyperproliferation of the epidermis, inflammatory cell accumulation and increased dilatation of dermal papillary blood vessels. Cases of psoriasis were investigated in vivo with optical means in order to evaluate the potential of in vivo optical biopsy. A Polarization Multispectral Dermoscope was employed for the macroscopic observation. Features such as the 'dotted' blood vessels pattern was observed with high contrast. The average size of dot vessels in Psoriasis was measured to be 974 ?m2 which is much higher compared to healthy skin. High resolution image sections of the epidermis and the dermis were produced with a custom made Multiphoton Microscope. Imaging extended from the surface of the lesion down to the papillary dermis, at a depth of 200 ?m. In the epidermis, a characteristic morphology of the stratum corneum found only in Psoriasis was revealed. Additionally, the cytoplasmic area of the cells in the stratum spinosum layer was found to be smaller than normal. In the dermis the morphological features were more pronounced, where the elongated dermal papillae dominated the papillary layer. Their length exceeds 100?m, which is a far greater value compared to that of healthy skin. These in vivo observations are consistent with the ex vivo histopathological observations, supporting both the applicability and potentiality of multispectral dermoscopy and multiphoton microscopy in the field of in vivo optical investigation and biopsy of skin.

  8. In Vivo Tumor Cell Targeting with “Click” Nanoparticles

    PubMed Central

    von Maltzahn, Geoffrey; Ren, Yin; Park, Ji-Ho; Min, Dal-Hee; Kotamraju, Venkata Ramana; Jayakumar, Jayanthi; Fogel, Valentina; Sailor, Michael J.; Ruoslahti, Erkki; Bhatia, Sangeeta N.

    2008-01-01

    The in vivo fate of nanomaterials strongly determines their biomedical efficacy. Accordingly, much effort has been invested into the development of library screening methods to select targeting ligands for a diversity of sites in vivo. Still, broad application of chemical and biological screens to the in vivo targeting of nanomaterials requires ligand attachment chemistries that are generalizable, efficient, covalent, orthogonal to diverse biochemical libraries, applicable under aqueous conditions, and stable in in vivo environments. To date, the copper(I)-catalyzed Huisgen 1,3-dipolar cycloaddition or “click” reaction has shown considerable promise as a method for developing targeted nanomaterials in vitro. Here, we investigate the utility of “click” chemistry for the in vivo targeting of inorganic nanoparticles to tumors. We find that “click” chemistry allows cyclic LyP-1 targeting peptides to be specifically linked to azido-nanoparticles and to direct their binding to p32-expressing tumor cells in vitro. Moreover, “click” nanoparticles are able to stably circulate for hours in vivo following intravenous administration (>5h circulation time), extravasate into tumors, and penetrate the tumor interstitium to specifically bind p32-expressing cells in tumors. In the future, in vivo use of “click” nanomaterials should expedite the progression from ligand discovery to in vivo evaluation and diversify approaches toward multifunctional nanoparticle development. PMID:18611045

  9. A ruthenium(II) complex as turn-on Cu(II) luminescent sensor based on oxidative cyclization mechanism and its application in vivo

    PubMed Central

    Zhang, Yunfei; Liu, Zonglun; Yang, Kui; Zhang, Yi; Xu, Yongqian; Li, Hongjuan; Wang, Chaoxia; Lu, Aiping; Sun, Shiguo

    2015-01-01

    Copper ions play a vital role in a variety of fundamental physiological processes not only in human beings and plants, but also for extensive insects and microorganisms. In this paper, a novel water-soluble ruthenium(II) complex as a turn-on copper(II) ions luminescent sensor based on o-(phenylazo)aniline was designed and synthesized. The azo group would undergo a specific oxidative cyclization reaction with copper(II) ions and turn into high luminescent benzotriazole, triggering significant luminescent increasements which were linear to the concentrations of copper(II) ions. The sensor distinguished by its high sensitivity (over 80-fold luminescent switch-on response), good selectivity (the changes of the emission intensity in the presence of other metal ions or amino acids were negligible) and low detection limit (4.42?nM) in water. Moreover, the copper(II) luminescent sensor exhibited good photostability under light irradiation. Furthermore, the applicability of the proposed sensor in biological samples assay was also studied and imaged copper(II) ions in living pea aphids successfully. PMID:25640000

  10. A ruthenium(II) complex as turn-on Cu(II) luminescent sensor based on oxidative cyclization mechanism and its application in vivo

    NASA Astrophysics Data System (ADS)

    Zhang, Yunfei; Liu, Zonglun; Yang, Kui; Zhang, Yi; Xu, Yongqian; Li, Hongjuan; Wang, Chaoxia; Lu, Aiping; Sun, Shiguo

    2015-02-01

    Copper ions play a vital role in a variety of fundamental physiological processes not only in human beings and plants, but also for extensive insects and microorganisms. In this paper, a novel water-soluble ruthenium(II) complex as a turn-on copper(II) ions luminescent sensor based on o-(phenylazo)aniline was designed and synthesized. The azo group would undergo a specific oxidative cyclization reaction with copper(II) ions and turn into high luminescent benzotriazole, triggering significant luminescent increasements which were linear to the concentrations of copper(II) ions. The sensor distinguished by its high sensitivity (over 80-fold luminescent switch-on response), good selectivity (the changes of the emission intensity in the presence of other metal ions or amino acids were negligible) and low detection limit (4.42 nM) in water. Moreover, the copper(II) luminescent sensor exhibited good photostability under light irradiation. Furthermore, the applicability of the proposed sensor in biological samples assay was also studied and imaged copper(II) ions in living pea aphids successfully.

  11. A ruthenium(II) complex as turn-on Cu(II) luminescent sensor based on oxidative cyclization mechanism and its application in vivo.

    PubMed

    Zhang, Yunfei; Liu, Zonglun; Yang, Kui; Zhang, Yi; Xu, Yongqian; Li, Hongjuan; Wang, Chaoxia; Lu, Aiping; Sun, Shiguo

    2015-01-01

    Copper ions play a vital role in a variety of fundamental physiological processes not only in human beings and plants, but also for extensive insects and microorganisms. In this paper, a novel water-soluble ruthenium(II) complex as a turn-on copper(II) ions luminescent sensor based on o-(phenylazo)aniline was designed and synthesized. The azo group would undergo a specific oxidative cyclization reaction with copper(II) ions and turn into high luminescent benzotriazole, triggering significant luminescent increasements which were linear to the concentrations of copper(II) ions. The sensor distinguished by its high sensitivity (over 80-fold luminescent switch-on response), good selectivity (the changes of the emission intensity in the presence of other metal ions or amino acids were negligible) and low detection limit (4.42?nM) in water. Moreover, the copper(II) luminescent sensor exhibited good photostability under light irradiation. Furthermore, the applicability of the proposed sensor in biological samples assay was also studied and imaged copper(II) ions in living pea aphids successfully. PMID:25640000

  12. In vitro and in vivo evaluation of porous PCL-PLLA 3D polymer scaffolds fabricated via salt leaching method for bone tissue engineering applications.

    PubMed

    Sadiasa, Alexander; Nguyen, Thi Hiep; Lee, Byong-Taek

    2014-01-01

    Three dimensional porous scaffolds composed of various ratios of polycaprolactone and poly(L-lactic acid) (PLLA) were prepared using salt leaching method for bone regeneration applications. Surfaces of the scaffolds were visualized using scanning electron microscope (SEM) and the combination of the polymers was confirmed by FT-IR. Addition of PLLA increased the porosity and pore sizes of the scaffolds and also the scaffolds' compressive strength initially. Osteoblast-like cells were used and it was found that the samples' cell biocompatibility was further promoted with the increase in PLLA content as observed via cell proliferation assays using MTT, gene expression with RT-PCR, and micrographs from SEM and confocal microscopy. Samples were then implanted into male rabbits for 2 months, and histological staining and micro-CT histomorphometry show that new bone formations were detected in the site containing the implants of the scaffolds and that bone regeneration was further promoted with the increased concentration of PLLA in the scaffold. PMID:24138179

  13. Compensatory inter vivos gifts

    Microsoft Academic Search

    Stefan Hochguertel; Henry Ohlsson

    2000-01-01

    Empirical studies of intergenerational transfers usually find that bequests are equally divided among heirs while inter vivos gifts tend to be compensatory. Using the 1992 and 1994 waves of the Health and Retirement Study, we find that only 4% of parents who give, divide their gifts equally among their children. Estimating probit models, using family panels, we find that gifts

  14. Compensatory Inter Vivos Gifts

    Microsoft Academic Search

    Stefan Hochguertel; Henry Ohlsson

    2001-01-01

    Empirical studies of intergenerational transfers usually find that bequests are equally divided among heirs while inter vivos gifts tend to be compensatory. Using the 1992 and 1994 waves of the Health and Retirement Study, we find that only 4 percent of parents who give divide their gifts equally among their children. Estimating probit models using family panels, we find that

  15. Apoptosis In Vivo

    Microsoft Academic Search

    L. C. Stephens; L. Milas; K. K. Ang; K. A. Mason; R. E. Meyn

    \\u000a Apoptosis is a complex and highly regulated process with numerous and varied biological consequences, it is typically described\\u000a as a sequence of morphological events that can be easily recognized histologically. In fact, the initial identification and\\u000a subsequent characterization of apoptosis were based on microscopic observations of its occurrence in vivo. In the early 1970’s, an experimental pathologist recognized variations in

  16. Bi-layer composite dressing of gelatin nanofibrous mat and poly vinyl alcohol hydrogel for drug delivery and wound healing application: in-vitro and in-vivo studies.

    PubMed

    Jaiswal, Maneesh; Gupta, Asheesh; Agrawal, Ashwini K; Jassal, Manjeet; Dinda, Amit Kr; Koul, Veena

    2013-09-01

    Present investigation involves the development of a bi-layer dressing of gelatin nanofibrous mat loaded with epigallocatechin gallate (EGCG)/poly vinyl alcohol (PVA) hydrogel and its in-vivo evaluation on full-thickness excision wounds in experimental Wistar rats. Nanomorphological observation, porosity, effect of crosslinking on tensile strength, physical stability and drug release profile in phosphate buffer and biocompatibility aspects of electrospun nanomat were investigated by various physico-chemical tools. EGCGa release profile was found to increase from 2-4 days with decreasing crosslinking time from 15 to 5 min. PVA hydrogels were prepared by freeze-thaw method and has been utilized as a protective and hydrating outer layer of the bi-layer dressing. Topical application of bi-layer composite dressing loaded with EGCG improve the healing rate in experimental rats as acute wounds model which was evidenced by significant increase in DNA (approximately 42%), total protein (approximately 32%), hydroxyproline (approximately 26%) and hexosamine approximately 24%) contents. A faster wound contraction was observed in wounds treated with composite dressing from approximately 14% to 47%. Histopathological examination revealed significant improvement in angiogenesis, re-epithelialization and less inflammatory response in comparison to control. Van-Gieson's collagen stains revealed matured, compact and parallel deposition of collagen fibrils on day 12. These results were supported by up-regulated expressions of matrix metalloproteinase (MMPs-2 and 9) by gelatin zymography. Control release of EGCG, 3D porous architecture of nanofibrous scaffolds as well as moist microenvironment provides ideal conditions for uninterrupted wound healing. PMID:23980498

  17. In vivo Noninvasive Small Animal Molecular Imaging

    PubMed Central

    Youn, Hyewon; Hong, Kee-Jong

    2012-01-01

    The remarkable efforts that are made on molecular imaging technologies demonstrate its potential importance and range of applications. The generation of disease-specific animal models, and the developments of target-specific probes and genetically encoded reporters are another important component. Continued improvements in the instrumentation, the identification of novel targets and genes, and the availability of improved imaging probes should be made. Multimodal imaging probes should provide easier transitions between laboratory studies, including small animal studies and clinical applications. Here, we reviewed basic strategies of noninvasive in vivo imaging methods in small animals to introducing the concept of molecular imaging. PMID:24159487

  18. Protocol: ex vivo culture of mouse embryonic mammary buds.

    PubMed

    Voutilainen, Maria; Lindfors, Päivi H; Mikkola, Marja L

    2013-06-01

    The explant culture techniques of embryonic tissues allow continuous monitoring of organ growth and morphogenesis ex vivo. The effect of growth factors and other soluble molecules can be examined by applying them to the culture medium. Relatively few studies have reported application of tissue culture techniques to analysis of embryonic mammary glands. Here we describe a protocol for murine mammary rudiments that permits ex vivo development up to branching stage. PMID:23674216

  19. In vivo dosimetry for IMRT

    SciTech Connect

    Vial, Philip [Department of Medical Physics, Liverpool Cancer Therapy Centre (Australia); Institute of Medical Physics, School of Physics, University of Sydney (Australia)

    2011-05-05

    In vivo dosimetry has a well established role in the quality assurance of 2D radiotherapy and 3D conformal radiotherapy. The role of in vivo dosimetry for IMRT is not as well established. IMRT introduces a range of technical issues that complicate in vivo dosimetry. The first decade or so of IMRT implementation has largely relied upon pre-treatment phantom based dose verification. During that time, several new devices and techniques for in vivo dosimetry have emerged with the promise of providing the ultimate form of IMRT dose verification. Solid state dosimeters continue to dominate the field of in vivo dosimetry in the IMRT era. In this report we review the literature on in vivo dosimetry for IMRT, with an emphasis on clinical evidence for different detector types. We describe the pros and cons of different detectors and techniques in the IMRT setting and the roles that they are likely to play in the future.

  20. In Vivo Imaging of Quantum Dots

    NASA Astrophysics Data System (ADS)

    Texier, Isabelle; Josser, Véronique

    Noninvasive whole-body near-infrared fluorescence imaging is now acknowledged as a powerful method for the molecular mapping of biological events in live small animals such as mouse models. With outstanding optical properties such as high fluorescence quantum yields and low photobleaching rates, quantum dots (QDs) are labels of choice in the near-infrared domain. The main applications described in the literature for in vivo imaging of mice after injection of QDs encompass imaging of lymph nodes and tumors and cell tracking. Standard methods for the preparation, the purification, and the in vivo fluorescence whole-body imaging of QDs in the live mouse are described. Nanoparticles coated by PEG chains of different sizes and terminal groups are prepared using 705-nm-emitting commercial QDs. Their biodistribution after intravenous or intradermal injections in tumor-bearing mice is reported here.

  1. Electrodeposition of platinum-iridium coatings and nanowires for neurostimulating applications: Fabrication, characterization and in-vivo retinal stimulation/recording. EIS studies of hexavalent and trivalent chromium based military coating systems

    NASA Astrophysics Data System (ADS)

    Petrossians, Artin

    The studies presented in this thesis are composed of two different projects demonstrated in two different parts. The first part of this thesis represents an electrochemical approach to possible improvements of the interface between an implantable device and biological tissue. The second part of this thesis represents electrochemical impedance spectroscopy (EIS) studies on the corrosion resistance behavior of different types of polymer coated Al2024 alloys. In the first part of this thesis, a broad range of investigations on the development of an efficient and reproducible electrochemical deposition method for fabrication of thin-film platinum-iridium alloys were performed. The developed method for production of dense films was then modified to produce very high surface area coatings with ultra-low electrochemical impedance characteristics. The high-surface area platinum-iridium coating was applied on microelectrode arrays for chronic in-vitro stimulation. Using the same method of producing dense films, platinum-iridium nanowires were fabricated using Anodized Aluminum Oxide (AAO) templates for hermetic packaging applications to be used in implantable microelectronics. The implantable microelectronics will be used to perform data reception and transmission management, power recovery, digital processing and analog output of stimulus current. Finally, in-vivo electrical stimulation tests were performed on an animal retina using high surface-area platinum-iridium coated single microelectrodes to verify the charge transfer characteristics of the coatings. In the second part of this thesis, three different sets of samples with different combinations of pretreatments, primers with the same type of topcoat were tested in 0.5 N NaCl for period of 30 days. The surface changes measured by EIS as a function of time were then analyzed. The analysis of the fit parameters of the impedance spectra showed that the different primers had the most effect on the corrosion protection properties of the coatings in which the primers with hexavalent chromium ions (Cr6+) provided better corrosion protection compared to primers with trivalent chromium ions (Cr3+). After 30 days of the exposure of the samples in 0.5 N NaCl, one sample from each set of samples was scribed and exposed to 0.5 N NaCl for 3 days. Analysis of the impedance spectra revealed that the samples with chromium conversion coating pretreatment and hexavalent chromium primer showed "self healing" characteristics and provided better corrosion protection on the scribed areas compared to the scribed samples with trivalent chromium pretreatment and non-hexavalent chromium primer.

  2. Reactive polymer enables efficient in vivo bioorthogonal chemistry

    PubMed Central

    Devaraj, Neal K.; Thurber, Greg M.; Keliher, Edmund J.; Marinelli, Brett; Weissleder, Ralph

    2012-01-01

    There has been intense interest in the development of selective bioorthogonal reactions or “click” chemistry that can proceed in live animals. Until now however, most reactions still require vast surpluses of reactants because of steep temporal and spatial concentration gradients. Using computational modeling and design of pharmacokinetically optimized reactants, we have developed a predictable method for efficient in vivo click reactions. Specifically, we show that polymer modified tetrazines (PMT) are a key enabler for in vivo bioorthogonal chemistry based on the very fast and catalyst-free [4 + 2] tetrazine/trans-cyclooctene cycloaddition. Using fluorescent PMT for cellular resolution and 18F labeled PMT for whole animal imaging, we show that cancer cell epitopes can be easily reacted in vivo. This generic strategy should help guide the design of future chemistries and find widespread use for different in vivo bioorthogonal applications, particularly in the biomedical sciences. PMID:22411831

  3. Study on advancement of in vivo counting using mathematical simulation

    E-print Network

    Kinase, S

    2003-01-01

    To obtain an assessment of the committed effective dose, individual monitoring for the estimation of intakes of radionuclides is required. For individual monitoring of exposure to intakes of radionuclides, direct measurement of radionuclides in the body - in vivo counting- is very useful. To advance in a precision in vivo counting which fulfills the requirements of ICRP 1990 recommendations, some problems, such as the investigation of uncertainties in estimates of body burdens by in vivo counting, and the selection of the way to improve the precision, have been studied. In the present study, a calibration technique for in vivo counting application using Monte Carlo simulation was developed. The advantage of the technique is that counting efficiency can be obtained for various shapes and sizes that are very difficult to change for phantoms. To validate the calibration technique, the response functions and counting efficiencies of a whole-body counter installed in JAERI were evaluated using the simulation and m...

  4. In vivo imaging of hydrogen peroxide with chemiluminescent nanoparticles

    NASA Astrophysics Data System (ADS)

    Lee, Dongwon; Khaja, Sirajud; Velasquez-Castano, Juan C.; Dasari, Madhuri; Sun, Carrie; Petros, John; Taylor, W. Robert; Murthy, Niren

    2007-10-01

    The overproduction of hydrogen peroxide is implicated in the development of numerous diseases and there is currently great interest in developing contrast agents that can image hydrogen peroxide in vivo. In this report, we demonstrate that nanoparticles formulated from peroxalate esters and fluorescent dyes can image hydrogen peroxide in vivo with high specificity and sensitivity. The peroxalate nanoparticles image hydrogen peroxide by undergoing a three-component chemiluminescent reaction between hydrogen peroxide, peroxalate esters and fluorescent dyes. The peroxalate nanoparticles have several attractive properties for in vivo imaging, such as tunable wavelength emission (460-630nm), nanomolar sensitivity for hydrogen peroxide and excellent specificity for hydrogen peroxide over other reactive oxygen species. The peroxalate nanoparticles were capable of imaging hydrogen peroxide in the peritoneal cavity of mice during a lipopolysaccharide-induced inflammatory response. We anticipate numerous applications of peroxalate nanoparticles for in vivo imaging of hydrogen peroxide, given their high specificity and sensitivity and deep-tissue-imaging capability.

  5. Ultrasound biomicroscopy permits in vivo

    E-print Network

    Cai, Long

    Ultrasound biomicroscopy permits in vivo characterization of zebrafish liver tumors Wolfram that tumors could be readily detected in vivo using high-resolution microscopic ultrasound in zebrafish. We and response to treatment. Ultrasound biomicroscopy allows longitudinal studies of tumor development and real

  6. Fluorescence Tomography and Magnetic Resonance Imaging of Myocardial Macrophage Infiltration in Infarcted Myocardium In Vivo

    Microsoft Academic Search

    David E. Sosnovik; Matthias Nahrendorf; Nikolaos Deliolanis; Mikhail Novikov; Elena Aikawa; Lee Josephson; Anthony Rosenzweig; Ralph Weissleder; Vasilis Ntziachristos

    2010-01-01

    Background—Fluorescence imaging of the heart is currently limited to invasive ex vivo or in vitro applications. We hypothesized that the adaptation of advanced transillumination and tomographic techniques would allow noninvasive fluorescence images of the heart to be acquired in vivo and be coregistered with in vivo cardiac magnetic resonance images. Methods and Results—The uptake of the magnetofluorescent nanoparticle CLIO-Cy5.5 by

  7. In vivo hybridization of technetium-99m-labeled peptide nucleic acid (PNA)

    Microsoft Academic Search

    G. Mardirossian; K. Lei; M. Rusckowski

    1997-01-01

    Hybridization of a radiolabeled single-stranded DNA oligonucleotide with its single-stranded complement in vivo has not yet been convincingly demonstrated. A contributing factor may be unfavorable in vivo properties of the phosphodiester and phosphorothioate DNAs. Peptide nucleic acid (PNA) oligomers have been reported to possess in vivo properties more suitable for radiopharmaceutical applications. We have radiolabeled an amine-derivatized 15-base PNA oligomer

  8. Quantification of carbon nanomaterials in vivo.

    PubMed

    Wang, Haifang; Yang, Sheng-Tao; Cao, Aoneng; Liu, Yuanfang

    2013-03-19

    A diverse array of carbon nanomaterials (NMs), including fullerene, carbon nanotubes (CNTs), graphene, nanodiamonds, and carbon nanoparticles, have been discovered and widely applied in a variety of industries. Carbon NMs have been detected in the environment and have a strong possibility of entering the human body. The safety of carbon NMs has thus become a serious concern in academia and society. To achieve strict biosafety assessments, researchers need to fully understand the effects and fates of NMs in the human body, including information about absorption, distribution, metabolism, excretion, and toxicity (ADME/T). To acquire the ADME data, researchers must quantify NMs, but carbon NMs are very difficult to quantify in vivo. The carbon background in a typical biological system is high, particularly compared with the much lower concentration of carbon NMs. Moreover, carbon NMs lack a specific detection signal. Therefore, isotopic labeling, with its high sensitivity and specificity, is the first choice to quantify carbon NMs in vivo. Previously, researchers have used many isotopes, including ¹³C, ¹?C, ¹²?I, ¹³¹I, ³H, ??Cu, ¹¹¹In, ??Y, 99mTc, and ??Ga, to label carbon NMs. We used these isotopic labeling methods to study the ADME of carbon NMs via different exposure pathways in animal models. Except for the metabolism of carbon NMs, which has seldom been investigated, significant amounts of data have been reported on the in vivo absorption, distribution, excretion, and toxicity of carbon NMs, which have revealed characteristic behaviors of carbon NMs, such as reticuloendothelial system (RES) capture. However, the complexity of the biological systems and diverse preparation and functionalization of the same carbon NMs have led to inconsistent results across different studies. Therefore, the data obtained so far have not provided a compatible and systematic profile of biosafety. Further efforts are needed to address these problems. In this Account, we review the in vivo quantification methods of carbon NMs, focusing on isotopic labeling and tracing methods, and summarize the related labeling, purification, bio-sampling, and detection of carbon NMs. We also address the advantages, applicable situations, and limits of various labeling and tracing methods and propose guidelines for choosing suitable labeling methods. A collective analysis of the ADME information on various carbon NMs in vivo would provide general principles for understanding the fate of carbon NMs and the effects of chemical functionalization and aggregation of carbon NMs on their ADME/T in vivo and their implications in nanotoxicology and biosafety evaluations. PMID:23035715

  9. Type I cell ROS kinetics under hypoxia in the intact mouse carotid body ex vivo: a FRET-based study.

    PubMed

    Bernardini, A; Brockmeier, U; Metzen, E; Berchner-Pfannschmidt, U; Harde, E; Acker-Palmer, A; Papkovsky, D; Acker, H; Fandrey, J

    2015-01-01

    Reactive oxygen species (ROS) mainly originating from NADPH oxidases have been shown to be involved in the carotid body (CB) oxygen-sensing cascade. For measuring ROS kinetics, type I cells of the mouse CB in an ex vivo preparation were transfected with the ROS sensor construct FRET-HSP33. After 2 days of tissue culture, type I cells expressed FRET-HSP33 as shown by immunohistochemistry. In one population of CBs, 5 min of hypoxia induced a significant and reversible decrease of type I cell ROS levels (n = 9 CBs; P < 0.015), which could be inhibited by 4-(2-aminoethyl)benzensulfonylfluorid (AEBSF), a highly specific inhibitor of the NADPH oxidase subunits p47(phox) and p67(phox). In another population of CBs, however, 5 min of hypoxia induced a significant and reversible increase of ROS levels in type I cells (n = 8 CBs; P < 0.05), which was slightly enhanced by administration of 3 mM AEBSF. These different ROS kinetics seemed to coincide with different mice breeding conditions. Type I cells of both populations showed a typical hypoxia-induced membrane potential (MP) depolarization, which could be inhibited by 3 mM AEBSF. ROS and MP closely followed the hypoxic decrease in CB tissue oxygen as measured with an O2-sensitive dye. We conclude that attenuated p47(phox) subunit activity of the NADPH oxidase under hypoxia is the physiological trigger for type I cell MP depolarization probably due to ROS decrease, whereas the observed ROS increase has no influence on type I cell MP kinetics under hypoxia. PMID:25318107

  10. Imaging schistosomes in vivo

    PubMed Central

    Krautz-Peterson, Greice; Ndegwa, David; Vasquez, Kristine; Korideck, Houari; Zhang, Jun; Peterson, Jeffrey D.; Skelly, Patrick J.

    2009-01-01

    Schistosomes are intravascular, parasitic helminths that cause a chronic, often debilitating disease afflicting over 200 million people in over 70 countries. Here we describe novel imaging methods that, for the first time, permit visualization of live schistosomes within their living hosts. The technology centers on fluorescent agent uptake and activation in the parasite’s gut, and subsequent detection and signal quantitation using fluorescence molecular tomography (FMT). There is a strong positive correlation between the signal detected and parasite number. Schistosoma mansoni parasites of both sexes recovered from infected experimental animals exhibit vivid fluorescence throughout their intestines. Likewise, the remaining important human schistosome parasites, S. japonicum and S. hematobium, also exhibit gut fluorescence when recovered from infected animals. Imaging has been used to efficiently document the decline in parasite numbers in infected mice treated with the antischistosome drug praziquantel. This technology will provide a unique opportunity both to help rapidly identify much-needed, novel antischistosome therapies and to gain direct visual insight into the intravascular lives of the major schistosome parasites of humans.—Krautz-Peterson, G., Ndegwa, D., Vasquez, K., Korideck, H., Zhang, J., Peterson, J. D., Skelly, P. J. Imaging schistosomes in vivo. PMID:19346298

  11. Nanocrystal targeting in vivo

    NASA Astrophysics Data System (ADS)

    Åkerman, Maria E.; Chan, Warren C. W.; Laakkonen, Pirjo; Bhatia, Sangeeta N.; Ruoslahti, Erkki

    2002-10-01

    Inorganic nanostructures that interface with biological systems have recently attracted widespread interest in biology and medicine. Nanoparticles are thought to have potential as novel intravascular probes for both diagnostic (e.g., imaging) and therapeutic purposes (e.g., drug delivery). Critical issues for successful nanoparticle delivery include the ability to target specific tissues and cell types and escape from the biological particulate filter known as the reticuloendothelial system. We set out to explore the feasibility of in vivo targeting by using semiconductor quantum dots (qdots). Qdots are small (<10 nm) inorganic nanocrystals that possess unique luminescent properties; their fluorescence emission is stable and tuned by varying the particle size or composition. We show that ZnS-capped CdSe qdots coated with a lung-targeting peptide accumulate in the lungs of mice after i.v. injection, whereas two other peptides specifically direct qdots to blood vessels or lymphatic vessels in tumors. We also show that adding polyethylene glycol to the qdot coating prevents nonselective accumulation of qdots in reticuloendothelial tissues. These results encourage the construction of more complex nanostructures with capabilities such as disease sensing and drug delivery.

  12. Application

    Cancer.gov

    Application To apply, either email, fax, or mail the following information postmarked by December 31, 2008 to the address below: Curriculum Vitae (that includes complete address, street address, telephone, fax and e-mail) or resume; Two (2) letters

  13. Image Stabilization for In Vivo Microscopy by High-Speed Visual Feedback Control

    Microsoft Academic Search

    Sungon Lee; Yoshihiko Nakamura; Katsu Yamane; Takeshi Toujo; Seiya Takahashi; Yoshihisa Tanikawa; Hajime Takahashi

    2008-01-01

    This paper presents image stabilization for microscopy using horizontal visual feedback control of the objective lens through a five-bar linkage and piezoelectric actuators, and its application to in vivo imaging. Even very small in vivo motion due to heartbeat and breathing makes microscopic observation difficult by blurring the microscope image or impossible by sending a region of interest out of

  14. Steady-state theory for quantitative microdialysis of solutes and water in vivo and in vitro

    Microsoft Academic Search

    P. M. Bungay; P. F. Morrison; R. L. Dedrick

    1990-01-01

    A mathematical framework was developed to provide a quantitative basis for either in vivo tissue or in vitro microdialysis. Established physiological and mass transport principles were employed to obtain explicit expressions relating dialysate concentration to tissue extracellular concentration for in vivo applications or external medium concentration in vitro probe characterization. Some of the important generalizations derived from the modeling framework

  15. In vivo Confocal Laser Scanning Microscopy and Micropuncture in Intact Rat

    Microsoft Academic Search

    Yoshio Ohno; Henrik Birn; Erik I. Christensen

    2005-01-01

    Background: Intravital microscopy theoretically provides the optimal conditions for studying specific organ functions. However, the application of microscopy in intact organs in vivo has been limited so far due to technical difficulties. The purpose of this study was to establish a method of in vivo confocal laser scanning microscopy (CLSM) for the study of endocytosis in proximal tubules of intact

  16. Survival of thefittest: in vivo selection and stem cell gene therapy

    Microsoft Academic Search

    Tobias Neff; Brian C. Beard; Hans-Peter Kiem

    2006-01-01

    human stem cells has long been the most prominent obstacle to widespread clinical application of stem cell gene therapy. A solution to this problem has been in vivo selection. In vivo selection increases the proportion of circulating gene-corrected cells by conferring a selective growth and\\/or survival advantage to the corrected cell population. While improvements in gene transfer technology did contribute,

  17. RESEARCH ARTICLE In Vivo Resolution of Multiexponential Decays of Multiple

    E-print Network

    Larson-Prior, Linda

    OF DISEASE BIOMARKERS and mole- cular targets for cancer application has been accelerated by advances molecular targets by the same tumor, occasionally dependent on the stage of disease development.2 Subsequent to in vivo optical molecular imaging because of concurrent development of narrowband laser light sources

  18. Gene-based cancer vaccines: an ex vivo approach

    Microsoft Academic Search

    VFI Van Tendeloo; C Van Broeckhoven; ZN Berneman

    2001-01-01

    The application of gene transfer techniques to immunotherapy has animated the field of gene-based cancer vaccine research. Gene transfer strategies were developed to bring about active immunization against tumor-associated antigens (TAA) through gene transfer technology. A wide variety of viral and nonviral gene transfer methods have been investigated for immunotherapeutic purposes. Ex vivo strategies include gene delivery into tumor cells

  19. In Vivo Monitoring of Fluorescently Labeled Cancer Cells

    Microsoft Academic Search

    Eva Barton; Angel Ang; Kevin Francis; Jae-Beom Kim

    Whole animal in vivo imaging has contributed significantly to the detection of disease progression and drug efficacy for the past several years (1). With the introduction of sensitive imaging instruments, applications in fluorescent imaging have expanded to monitoring tumor cell growth and gene expression. The new fluorescent proteins have improved brightness, photostability and better tissue penetration of signals at a

  20. Recommendations for conducting the in vivo alkaline Comet assay

    Microsoft Academic Search

    A. Hartmann; E. Agurell; C. Beevers; S. Brendler-Schwaab; B. Burlinson; P. Clay; A. Collins; A. Smith; G. Speit; V. Thybaud; R. R. Tice

    2003-01-01

    The in vivo alkaline single cell gel electrophoresis assay, hereafter the Comet assay, can be used to investigate the genotoxicity of industrial chemicals, biocides, agrochem- icals and pharmaceuticals. The major advantages of this assay include the relative ease of application to any tissue of interest, the detection of multiple classes of DNA damage and the generation of data at the

  1. Rapid and simultaneous measurement of midazolam, 1?-hydroxymidazolam and digoxin by liquid chromatography\\/tandem mass spectrometry: Application to an in vivo study to simultaneously measure P-glycoprotein and Cytochrome P450 3A activity

    Microsoft Academic Search

    Xinping Xue; Min Huang; Huanyu Xiao; Xiaoling Qin; Ling Huang; Guoping Zhong; Huichang Bi

    2011-01-01

    In order to simultaneously determine in vivo P-glycoprotein (P-gp) and Cytochrome P450 3A (CYP3A) activity, a new, rapid and sensitive liquid chromatography\\/tandem mass spectrometry (LC–MS\\/MS) method has been developed and fully validated to simultaneously determine midazolam (MDZ, as CYP3A substrate), 1?-hydroxymidazolam (1?-OHMDZ) and digoxin (DG, as P-gp substrate) in rat plasma using digitoxin as the internal standard (IS). After a

  2. 31P-MRS-Based Determination of Brain Intracellular and Interstitial pH: Its Application to In Vivo H+ Compartmentation and Cellular Regulation during Hypoxic\\/Ischemic Conditions

    Microsoft Academic Search

    D. B. Kintner; M. K. Anderson; J. H. Fitzpatrick; K. A. Sailor; D. D. Gilboe

    2000-01-01

    In the last decade, significant progress has been made in the characterization of pH regulation in nervous tissue in vitro. However, little work has been directed at understanding how pH regulatory mechanisms function in vivo. We are interested in how ischemic acidosis can effect pH regulation and modulate the extent of post-ischemic brain damage. We used 31P-MRS to determine normal

  3. Defining human mesenchymal stem cell efficacy in vivo.

    PubMed

    Bonfield, Tracey L; Nolan Koloze, Mary T; Lennon, Donald P; Caplan, Arnold I

    2010-01-01

    Allogeneic human mesenchymal stem cells (hMSCs) can suppress graft versus host disease (GvHD) and have profound anti-inflammatory and regenerative capacity in stroke, infarct, spinal cord injury, meniscus regeneration, tendinitis, acute renal failure, and heart disease in human and animal models of disease. There is significant clinical hMSC variability in efficacy and the ultimate response in vivo. The challenge in hMSC based therapy is defining the efficacy of hMSC in vivo. Models which may provide insight into hMSC bioactivity in vivo would provide a means to distinguish hMSCs for clinical utility. hMSC function has been described as both regenerative and trophic through the production of bioactive factors. The regenerative component involves the multi-potentiality of hMSC progenitor differentiation. The secreted factors generated by the hMSCs are milieu and injury specific providing unique niches for responses in vivo. These bioactive factors are anti-scarring, angiogenic, anti-apoptotic as well as regenerative. Further, from an immunological standpoint, hMSC's can avoid host immune response, providing xenographic applications. To study the in vivo immuno-regulatory effectiveness of hMSCs, we used the ovalbumin challenge model of acute asthma. This is a quick 3 week in vivo pulmonary inflammation model with readily accessible ways of measuring effectiveness of hMSCs. Our data show that there is a direct correlation between the traditional ceramic cube score to hMSCs attenuation of cellular recruitment due to ovalbumin challenge. The results from these studies verify the in vivo immuno-modulator effectiveness of hMSCs and support the potential use of the ovalbumin model as an in vivo model of hMSC potency and efficacy. Our data also support future directions toward exploring hMSCs as an alternative therapeutic for the treatment of airway inflammation associated with asthma. PMID:20974000

  4. Defining human mesenchymal stem cell efficacy in vivo

    PubMed Central

    2010-01-01

    Allogeneic human mesenchymal stem cells (hMSCs) can suppress graft versus host disease (GvHD) and have profound anti-inflammatory and regenerative capacity in stroke, infarct, spinal cord injury, meniscus regeneration, tendinitis, acute renal failure, and heart disease in human and animal models of disease. There is significant clinical hMSC variability in efficacy and the ultimate response in vivo. The challenge in hMSC based therapy is defining the efficacy of hMSC in vivo. Models which may provide insight into hMSC bioactivity in vivo would provide a means to distinguish hMSCs for clinical utility. hMSC function has been described as both regenerative and trophic through the production of bioactive factors. The regenerative component involves the multi-potentiality of hMSC progenitor differentiation. The secreted factors generated by the hMSCs are milieu and injury specific providing unique niches for responses in vivo. These bioactive factors are anti-scarring, angiogenic, anti-apoptotic as well as regenerative. Further, from an immunological standpoint, hMSC's can avoid host immune response, providing xenographic applications. To study the in vivo immuno-regulatory effectiveness of hMSCs, we used the ovalbumin challenge model of acute asthma. This is a quick 3 week in vivo pulmonary inflammation model with readily accessible ways of measuring effectiveness of hMSCs. Our data show that there is a direct correlation between the traditional ceramic cube score to hMSCs attenuation of cellular recruitment due to ovalbumin challenge. The results from these studies verify the in vivo immuno-modulator effectiveness of hMSCs and support the potential use of the ovalbumin model as an in vivo model of hMSC potency and efficacy. Our data also support future directions toward exploring hMSCs as an alternative therapeutic for the treatment of airway inflammation associated with asthma. PMID:20974000

  5. In Vivo Treg Suppression Assays

    PubMed Central

    Workman, Creg J.; Collison, Lauren W.; Bettini, Maria; Pillai, Meenu R.; Rehg, Jerold E.; Vignali, Dario A.A.

    2011-01-01

    To fully examine the functionality of a regulatory T cell (Treg) population, one needs to assess their ability to suppress in a variety of in vivo models. We describe five in vivo models that examine the suppressive capacity of Tregs upon different target cell types. The advantages and disadvantages of each model includ ing resources, time, and technical expertise required to execute each model are also described. PMID:21287333

  6. In vivo photoacoustic imaging of mouse embryos

    NASA Astrophysics Data System (ADS)

    Laufer, Jan; Norris, Francesca; Cleary, Jon; Zhang, Edward; Treeby, Bradley; Cox, Ben; Johnson, Peter; Scambler, Pete; Lythgoe, Mark; Beard, Paul

    2012-06-01

    The ability to noninvasively image embryonic vascular anatomy in mouse models is an important requirement for characterizing the development of the normal cardiovascular system and malformations in the heart and vascular supply. Photoacoustic imaging, which can provide high resolution non invasive images of the vasculature based upon optical absorption by endogenous hemoglobin, is well suited to this application. In this study, photoacoustic images of mouse embryos were obtained ex vivo and in vivo. The images show intricate details of the embryonic vascular system to depths of up to 10 mm, which allowed whole embryos to be imaged in situ. To achieve this, an all-optical photoacoustic scanner and a novel time reversal image reconstruction algorithm, which provide deep tissue imaging capability while maintaining high spatial resolution and contrast were employed. This technology may find application as an imaging tool for preclinical embryo studies in developmental biology as well as more generally in preclinical and clinical medicine for studying pathologies characterized by changes in the vasculature.

  7. Drug-Induced Secondary Cataract Prevention: Experimental ex vivo and in vivo Results with Disulfiram, Methotrexate and Actinomycin D

    Microsoft Academic Search

    Katrin Sternberg; Thom Terwee; Oliver Stachs; Rudolf Guthoff; Marian Löbler; Klaus-Peter Schmitz

    2010-01-01

    Background\\/Aims: A clinical approach to prevent secondary cataract after lens implantation involves the intraocular application of pharmacological agents. The goals of our study were to develop an ex vivo model to test the drug effectiveness for lens epithelial cell ablation from the basal membrane and to verify the data in rabbit intraocular lens implantation experiments. Methods: Human capsular rhexis specimens

  8. Bond failure patterns in vivo.

    PubMed

    Linklater, Rognvald A; Gordon, Peter H

    2003-05-01

    The aim of this study was to identify the presence and pattern of differences in bond failure between tooth types in vivo when bonding orthodontic brackets with the no-mix orthodontic composite adhesive Right-On. In vivo bond failure for a single operator was recorded for 108 consecutive patients undergoing fixed-appliance orthodontic treatment. The bond failure data were analyzed by survival analysis. Time to first failure or censorship was recorded for each bonded attachment. Overall failure in the sample matched previous clinical studies but conflicted with previous ex vivo bond strength data. Mandibular and posterior teeth had significantly higher rates of failure than did maxillary and anterior teeth. The type of attachment used had a significant effect on bond survival. The results of this study confirm that in vivo bond survival is not uniform for all teeth. Comparisons between the findings of this study and those of a previous ex vivo study by the same authors failed to validate ex vivo bond strength testing as clinically relevant. PMID:12750672

  9. Phase Diagrams in Vivo

    NSDL National Science Digital Library

    This activity uses three experiments for students to construct a phase diagram; the experiments have been videotaped and can be seen online. The purpose of this laboratory as designed is to gain familiarity with simple phase diagrams, their construction, and their applications to the understanding of geological and environmental problems. Subsidiary objectives include development of strategies for data processing including evaluation of assumptions and sources of errors, as well as honing of computer, spreadsheet, presentation (tabular and graphical), and report writing skills.

  10. Photoacoustic Tomography of a Rat Cerebral Cortex in vivo with Au

    E-print Network

    Wang, Lihong

    be better suited for in vivo applications. We sequentially injected Au nanocages into the circulatory system to dysfunctional anatomi- cal conditions such as localized leaky circulatory and lymphatic systems. This feature

  11. In vitro and in vivo two-photon luminescence imaging of single gold nanorods

    E-print Network

    Wei, Alexander

    In vitro and in vivo two-photon luminescence imaging of single gold nanorods Haifeng Wang*, Terry B that of the two-photon fluorescence signal from a single rhodamine molecule. The application of gold nanorods rhodamine molecule. These fe

  12. Application

    Microsoft Academic Search

    Michael Ströbel

    \\u000a This chapter serves two major purposes. First of all it outlines general usage domains for the enMedia framework that has\\u000a been presented in this book. The second purpose is to demonstrate in detail the application of the enMedia framework and its\\u000a prototype implementation, SilkRoad, through a sequence of electronic negotiation scenario cases. In these cases, specific emphasis is set on

  13. Progress Toward In Vivo Use of siRNAs-II

    PubMed Central

    Rettig, Garrett R; Behlke, Mark A

    2012-01-01

    RNA interference (RNAi) has been extensively employed for in vivo research since its use was first demonstrated in mammalian cells 10 years ago. Design rules have improved, and it is now routinely possible to obtain reagents that suppress expression of any gene desired. At the same time, increased understanding of the molecular basis of unwanted side effects has led to the development of chemical modification strategies that mitigate these concerns. Delivery remains the single greatest hurdle to widespread adoption of in vivo RNAi methods. However, exciting advances have been made and new delivery systems under development may help to overcome these barriers. This review discusses advances in RNAi biochemistry and biology that impact in vivo use and provides an overview of select publications that demonstrate interesting applications of these principles. Emphasis is placed on work with synthetic, small interfering RNAs (siRNAs) published since the first installment of this review which appeared in 2006. PMID:22186795

  14. In vivo modeling of biofilm-infected wounds: a review.

    PubMed

    Seth, Akhil K; Geringer, Matthew R; Hong, Seok J; Leung, Kai P; Mustoe, Thomas A; Galiano, Robert D

    2012-11-01

    Chronic wounds continue to represent a difficult and complex problem for both patients and healthcare providers. Bacterial biofilms represent a critical component of nonhealing wounds, utilizing several different mechanisms to inhibit innate inflammatory pathways and resist traditional therapeutics. Although in vitro biofilm systems have been well described and studied, understanding the intricacies of wound biofilm pathology requires appropriate in vivo models to understand the interactions between bacteria and host. In an effort to clarify the available literature, this review describes and critically evaluates all of the in vivo wound biofilm models currently published to-date, including model advantages and clinical applicability. We will also address the need for continued therapeutic development and testing using these currently available in vivo models. PMID:22835953

  15. RNA circularization strategies in vivo and in vitro.

    PubMed

    Petkovic, Sonja; Müller, Sabine

    2015-02-27

    In the plenitude of naturally occurring RNAs, circular RNAs (circRNAs) and their biological role were underestimated for years. However, circRNAs are ubiquitous in all domains of life, including eukaryotes, archaea, bacteria and viruses, where they can fulfill diverse biological functions. Some of those functions, as for example playing a role in the life cycle of viral and viroid genomes or in the maturation of tRNA genes, have been elucidated; other putative functions still remain elusive. Due to the resistance to exonucleases, circRNAs are promising tools for in vivo application as aptamers, trans-cleaving ribozymes or siRNAs. How are circRNAs generated in vivo and what approaches do exist to produce ring-shaped RNAs in vitro? In this review we illustrate the occurrence and mechanisms of RNA circularization in vivo, survey methods for the generation of circRNA in vitro and provide appropriate protocols. PMID:25662225

  16. RNA circularization strategies in vivo and in vitro

    PubMed Central

    Petkovic, Sonja; Müller, Sabine

    2015-01-01

    In the plenitude of naturally occurring RNAs, circular RNAs (circRNAs) and their biological role were underestimated for years. However, circRNAs are ubiquitous in all domains of life, including eukaryotes, archaea, bacteria and viruses, where they can fulfill diverse biological functions. Some of those functions, as for example playing a role in the life cycle of viral and viroid genomes or in the maturation of tRNA genes, have been elucidated; other putative functions still remain elusive. Due to the resistance to exonucleases, circRNAs are promising tools for in vivo application as aptamers, trans-cleaving ribozymes or siRNAs. How are circRNAs generated in vivo and what approaches do exist to produce ring-shaped RNAs in vitro? In this review we illustrate the occurrence and mechanisms of RNA circularization in vivo, survey methods for the generation of circRNA in vitro and provide appropriate protocols. PMID:25662225

  17. Tracking immune cells in vivo using magnetic resonance imaging

    PubMed Central

    Ahrens, Eric T.; Bulte, Jeff W. M.

    2013-01-01

    The increasing complexity of in vivo imaging technologies, coupled with the development of cell therapies, has fuelled a revolution in immune cell tracking in vivo. Powerful magnetic resonance imaging (MRI) methods are now being developed that use iron oxide- and 19F-based probes. These MRI technologies can be used for image-guided immune cell delivery and for the visualization of immune cell homing and engraftment, inflammation, cell physiology and gene expression. MRI-based cell tracking is now also being applied to evaluate therapeutics that modulate endogenous immune cell recruitment and to monitor emerging cellular immunotherapies. These recent uses show that MRI has the potential to be developed in many applications to follow the fate of immune cells in vivo. PMID:24013185

  18. Progress toward in vivo use of siRNAs-II.

    PubMed

    Rettig, Garrett R; Behlke, Mark A

    2012-03-01

    RNA interference (RNAi) has been extensively employed for in vivo research since its use was first demonstrated in mammalian cells 10 years ago. Design rules have improved, and it is now routinely possible to obtain reagents that suppress expression of any gene desired. At the same time, increased understanding of the molecular basis of unwanted side effects has led to the development of chemical modification strategies that mitigate these concerns. Delivery remains the single greatest hurdle to widespread adoption of in vivo RNAi methods. However, exciting advances have been made and new delivery systems under development may help to overcome these barriers. This review discusses advances in RNAi biochemistry and biology that impact in vivo use and provides an overview of select publications that demonstrate interesting applications of these principles. Emphasis is placed on work with synthetic, small interfering RNAs (siRNAs) published since the first installment of this review which appeared in 2006. PMID:22186795

  19. In vivo Raman spectroscopy of cervix cancers

    NASA Astrophysics Data System (ADS)

    Rubina, S.; Sathe, Priyanka; Dora, Tapas Kumar; Chopra, Supriya; Maheshwari, Amita; Krishna, C. Murali

    2014-03-01

    Cervix-cancer is the third most common female cancer worldwide. It is the leading cancer among Indian females with more than million new diagnosed cases and 50% mortality, annually. The high mortality rates can be attributed to late diagnosis. Efficacy of Raman spectroscopy in classification of normal and pathological conditions in cervix cancers on diverse populations has already been demonstrated. Our earlier ex vivo studies have shown the feasibility of classifying normal and cancer cervix tissues as well as responders/non-responders to Concurrent chemoradiotherapy (CCRT). The present study was carried out to explore feasibility of in vivo Raman spectroscopic methods in classifying normal and cancerous conditions in Indian population. A total of 182 normal and 132 tumor in vivo Raman spectra, from 63 subjects, were recorded using a fiberoptic probe coupled HE-785 spectrometer, under clinical supervision. Spectra were acquired for 5 s and averaged over 3 times at 80 mW laser power. Spectra of normal conditions suggest strong collagenous features and abundance of non-collagenous proteins and DNA in case of tumors. Preprocessed spectra were subjected to Principal Component-Linear Discrimination Analysis (PCLDA) followed by leave-one-out-cross-validation. Classification efficiency of ~96.7% and 100% for normal and cancerous conditions respectively, were observed. Findings of the study corroborates earlier studies and suggest applicability of Raman spectroscopic methods in combination with appropriate multivariate tool for objective, noninvasive and rapid diagnosis of cervical cancers in Indian population. In view of encouraging results, extensive validation studies will be undertaken to confirm the findings.

  20. Design and optimization of topical methotrexate loaded niosomes for enhanced management of psoriasis: Application of Box-Behnken design, in-vitro evaluation and in-vivo skin deposition study.

    PubMed

    Abdelbary, Aly A; AbouGhaly, Mohamed H H

    2015-05-15

    Psoriasis, a skin disorder characterized by impaired epidermal differentiation, is regularly treated by systemic methotrexate (MTX), an effective cytotoxic drug but with numerous side effects. The aim of this work was to design topical MTX loaded niosomes for management of psoriasis to avoid systemic toxicity. To achieve this goal, MTX niosomes were prepared by thin film hydration technique. A Box-Behnken (BB) design, using Design-Expert(®) software, was employed to statistically optimize formulation variables. Three independent variables were evaluated: MTX concentration in hydration medium (X1), total weight of niosomal components (X2) and surfactant: cholesterol ratio (X3). The encapsulation efficiency percent (Y1: EE%) and particle size (Y2: PS) were selected as dependent variables. The optimal formulation (F12) displayed spherical morphology under transmission electron microscopy (TEM), optimum particle size of 1375.00nm and high EE% of 78.66%. In-vivo skin deposition study showed that the highest value of percentage drug deposited (22.45%) and AUC0-10 (1.15mg.h/cm(2)) of MTX from niosomes were significantly greater than that of drug solution (13.87% and 0.49mg.h/cm(2), respectively). Moreover, in-vivo histopathological studies confirmed safety of topically applied niosomes. Concisely, the results showed that targeted MTX delivery might be achieved using topically applied niosomes for enhanced treatment of psoriasis. PMID:25773359

  1. Tailoring vessel morphology in vivo

    NASA Astrophysics Data System (ADS)

    Gould, Daniel Joseph

    Tissue engineering is a rapidly growing field which seeks to provide alternatives to organ transplantation in order to address the increasing need for transplantable tissues. One huge hurdle in this effort is the provision of thick tissues; this hurdle exists because currently there is no way to provide prevascularized or rapidly vascularizable scaffolds. To design thick, vascularized tissues, scaffolds are needed that can induce vessels which are similar to the microvasculature found in normal tissues. Angiogenic biomaterials are being developed to provide useful scaffolds to address this problem. In this thesis angiogenic and cell signaling and adhesion factors were incorporated into a biomimetic poly(ethylene glycol) (PEG) hydrogel system. The composition of these hydrogels was precisely tuned to induce the formation of differing vessel morphology. To sensitively measure induced microvascular morphology and to compare it to native microvessels in several tissues, this thesis developed an image-based tool for quantification of scale invariant and classical measures of vessel morphology. The tool displayed great utility in the comparison of native vessels and remodeling vessels in normal tissues. To utilize this tool to tune the vessel response in vivo, Flk1::myr-mCherry fluorescently labeled mice were implanted with Platelet Derived Growth Factor-BB (PDGF-BB) and basic Fibroblast Growth Factor (FGF-2) containing PEG-based hydrogels in a modified mouse corneal angiogenesis assay. Resulting vessels were imaged with confocal microscopy, analyzed with the image based tool created in this thesis to compare morphological differences between treatment groups, and used to create a linear relationship between space filling parameters and dose of growth factor release. Morphological parameters of native mouse tissue vessels were then compared to the linear fit to calculate the dose of growth factors needed to induce vessels similar in morphology to native vessels. Resulting induced vessels did match in morphology to the target vessels. Several other covalently bound signals were then analyzed in the assay and resulting morphology of vessels was compared in several studies which further highlighted the utility of the micropocket assay in conjunction with the image based tool for vessel morphological quantification. Finally, an alternative method to provide rapid vasculature to the constructs, which relied on pre-seeded hydrogels encapsulated endothelial cells was also developed and shown to allow anastamosis between induced host vessels and the implanted construct within 48 hours. These results indicate great promise in the rational design of synthetic, bioactive hydrogels, which can be used as a platform to study microvascular induction for regenerative medicine and angiogenesis research. Future applications of this research may help to develop therapeutic strategies to ameliorate human disease by replacing organs or correcting vessel morphology in the case of ischemic diseases and cancer.

  2. Measuring PLD activity in vivo.

    PubMed

    Munnik, Teun; Laxalt, Ana M

    2013-01-01

    Phospholipase D (PLD) hydrolyzes structural phospholipids like phosphatidylcholine (PC) and phosphatidylethanolamine (PE) into phosphatidic acid (PA) and free choline/ethanolamine. In plants, this activity can be stimulated by a wide variety of biotic and abiotic stresses (Li et al., Biochim Biophys Acta 1791:927-935, 2009; Testerink and Munnik, J Exp Bot 62(7):2349-2361, 2011). This chapter describes a protocol for the measurement of PLD activity in vivo. The protocol takes advantage of a unique property of PLD, i.e., its ability to substitute a primary alcohol, such as 1-butanol, for water in the hydrolytic reaction. This transphosphatidylation reaction results in the formation of phosphatidylbutanol (PBut), which is a specific and unique reporter for PLD activity. The assay is highly sensitive for detecting PLD activity in vivo, following stimulation of intact plant cells, seedlings, and tissues, being a valuable method for studying the regulation of plant PLD activity in vivo. PMID:23681537

  3. Experimental Cryosurgery Investigations In Vivo

    PubMed Central

    Gage, A.A.; Baust, J.M.; Baust, J.G.

    2009-01-01

    Cryosurgery is the use of freezing temperatures to elicit an ablative response in a targeted tissue. This review provides a global overview of experimentation in vivo which has been the basis of advancement of this widely applied therapeutic option. The cellular and tissue-related events that underlie the mechanisms of destruction, including direct cell injury (cryolysis), vascular stasis, apoptosis and necrosis, are described and are related to the optimal methods of technique of freezing to achieve efficacious therapy. In vivo experiments with major organs, including wound healing, the putative immunological response following thawing, and the use of cryoadjunctive strategies to enhance cancer cell sensitivity to freezing, are described. PMID:19833119

  4. In-vivo morphologic and spectroscopic investigation of Psoriasis

    NASA Astrophysics Data System (ADS)

    Kapsokalyvas, Dimitrios; Cicchi, Riccardo; Bruscino, Nicola; Alfieri, Domenico; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2011-07-01

    Psoriasis is an autoimmune disease of the skin characterized by hyperkeratosis, hyperproliferation of the epidermis, inflammatory cell accumulation and increased dilatation of dermal papillary blood vessels. Cases of psoriasis were investigated in vivo with optical means in order to evaluate the potential of in vivo optical biopsy. A Polarization Multispectral Dermoscope was employed for the macroscopic observation. Features such as the 'dotted' blood vessels pattern was observed with high contrast. High resolution image sections of the epidermis and the dermis were produced with a custom made Multiphoton Microscope. Imaging extended from the surface of the lesion down to the papillary dermis, at a depth of 200 ?m. In the epidermis, a characteristic morphology of the stratum corneum found only in Psoriasis was revealed. Additionally, the cytoplasmic area of the cells in the stratum spinosum layer was found to be smaller than normal. In the dermis the morphological features were more pronounced, where the elongated dermal papillae dominated the papillary layer. Their length exceeds 100?m, which is a far greater value compared to that of healthy skin. These in vivo observations are consistent with the ex vivo histopathological observations, supporting both the applicability and potentiality of multispectral dermoscopy and multiphoton microscopy in the field of in vivo optical investigation and biopsy of skin.

  5. The in vivo activation of persistent nanophosphors for optical imaging of vascularization, tumours and grafted cells

    NASA Astrophysics Data System (ADS)

    Maldiney, Thomas; Bessière, Aurélie; Seguin, Johanne; Teston, Eliott; Sharma, Suchinder K.; Viana, Bruno; Bos, Adrie J. J.; Dorenbos, Pieter; Bessodes, Michel; Gourier, Didier; Scherman, Daniel; Richard, Cyrille

    2014-04-01

    Optical imaging for biological applications requires more sensitive tools. Near-infrared persistent luminescence nanoparticles enable highly sensitive in vivo optical detection and complete avoidance of tissue autofluorescence. However, the actual generation of persistent luminescence nanoparticles necessitates ex vivo activation before systemic administration, which prevents long-term imaging in living animals. Here, we introduce a new generation of optical nanoprobes, based on chromium-doped zinc gallate, whose persistent luminescence can be activated in vivo through living tissues using highly penetrating low-energy red photons. Surface functionalization of this photonic probe can be adjusted to favour multiple biomedical applications such as tumour targeting. Notably, we show that cells can endocytose these nanoparticles in vitro and that, after intravenous injection, we can track labelled cells in vivo and follow their biodistribution by a simple whole animal optical detection, opening new perspectives for cell therapy research and for a variety of diagnosis applications.

  6. The in vivo activation of persistent nanophosphors for optical imaging of vascularization, tumours and grafted cells.

    PubMed

    Maldiney, Thomas; Bessière, Aurélie; Seguin, Johanne; Teston, Eliott; Sharma, Suchinder K; Viana, Bruno; Bos, Adrie J J; Dorenbos, Pieter; Bessodes, Michel; Gourier, Didier; Scherman, Daniel; Richard, Cyrille

    2014-04-01

    Optical imaging for biological applications requires more sensitive tools. Near-infrared persistent luminescence nanoparticles enable highly sensitive in vivo optical detection and complete avoidance of tissue autofluorescence. However, the actual generation of persistent luminescence nanoparticles necessitates ex vivo activation before systemic administration, which prevents long-term imaging in living animals. Here, we introduce a new generation of optical nanoprobes, based on chromium-doped zinc gallate, whose persistent luminescence can be activated in vivo through living tissues using highly penetrating low-energy red photons. Surface functionalization of this photonic probe can be adjusted to favour multiple biomedical applications such as tumour targeting. Notably, we show that cells can endocytose these nanoparticles in vitro and that, after intravenous injection, we can track labelled cells in vivo and follow their biodistribution by a simple whole animal optical detection, opening new perspectives for cell therapy research and for a variety of diagnosis applications. PMID:24651431

  7. Non-Invasive in vivo Mapping and Long-Term Monitoring of Magnetic Nanoparticles in Different Organs of Animals

    NASA Astrophysics Data System (ADS)

    Nikitin, Maxim; Yuriev, Mikhail; Brusentsov, Nikolai; Vetoshko, Petr; Nikitin, Petr

    2010-12-01

    Quantitative detection of magnetic nanoparticles (MP) in vivo is very important for various biomedical applications. Our original detection method based on non-linear MP magnetization has been modified for non-invasive in vivo mapping of the MP distribution among different organs of rats. A novel highly sensitive room-temperature device equipped with an external probe has been designed and tested for quantification of MP within 20-mm depth from the animal skin. Results obtained by external in vivo scanning of rats by the probe and ex vivo MP quantification in different organs of rats well correlated. The method allows long-term in vivo study of MP evolution, clearance and redistribution among different organs of the animal. Experiments showed that dynamics in vivo strongly depend on MP characteristics (size, material, coatings, etc.), site of injection and dose. The developed detection method combined with the magnetic nanolabels can substitute the radioactive labeling in many applications.

  8. In vivo static field perturbations in magnetic resonance

    NASA Astrophysics Data System (ADS)

    Koch, Kevin Matthew

    2007-12-01

    Fundamental magnetic resonance (MR) theory assumes the spatial homogeneity of a dominating static magnetic field B = B 0?. When this assumption is violated, a myriad of artifacts and compromising factors are introduced to MR spectra and images. Though in vivo nuclear magnetic resonance (NMR) is one of the most widely used scientific and diagnostic tools in medicine and biology, it remains haunted by the continual and persistant ghost of B0 inhomogeneity. An inclusive list of in vivo NMR applications severely impacted by B0 inhomogeneity could go on ad infinitum. Examples of such applications include neurosurgical utility in functional magnetic resonance imaging (fMRI), cerebral metabolic flux mapping, cerebral diffusion tractography, and abdominal diagnostic imaging. Given this wide impact on in vivo NMR, significant effort has been exerted in developing methods of compensating B0 inhomogeneity. Complicating this task is the sample-specific nature of in vivo B 0 inhomogeneity and its exacerbation with ever increasing B 0 field strengths. State of the art B 0 inhomogeneity compensation is currently at a critical juncture where homogenization demands are overwhelming the outer capabilities of existing technology and methods. This thesis addresses the B 0 inhomogeneity problem in the mammalian brain and presents novel solutions to the homogenization technology stalemate.

  9. Monitoring protein stability in vivo

    Microsoft Academic Search

    Zoya Ignatova

    2005-01-01

    Reduced protein stability in vivo is a prerequisite to aggregation. While this is merely a nuisance factor in recombinant protein production, it holds a serious impact for man. This review focuses on specific approaches to selectively determine the solubility and\\/or stability of a target protein within the complex cellular environment using different detection techniques. Noninvasive techniques mapping folding\\/misfolding events on

  10. Respiratory Rhythm Generation In Vivo

    PubMed Central

    Richter, Diethelm W.; Smith, Jeffrey C.

    2014-01-01

    The cellular and circuit mechanisms generating the rhythm of breathing in mammals have been under intense investigation for decades. Here, we try to integrate the key discoveries into an updated description of the basic neural processes generating respiratory rhythm under in vivo conditions. PMID:24382872

  11. In aqua vivo EPID dosimetry

    SciTech Connect

    Wendling, Markus; McDermott, Leah N.; Mans, Anton; Olaciregui-Ruiz, Igor; Pecharroman-Gallego, Raul; Sonke, Jan-Jakob; Stroom, Joep; Herk, Marcel J.; Mijnheer, Ben van [Department of Radiation Oncology, Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital, Plesmanlaan 121, 1066 CX Amsterdam (Netherlands)

    2012-01-15

    Purpose: At the Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital in vivo dosimetry using an electronic portal imaging device (EPID) has been implemented for almost all high-energy photon treatments of cancer with curative intent. Lung cancer treatments were initially excluded, because the original back-projection dose-reconstruction algorithm uses water-based scatter-correction kernels and therefore does not account for tissue inhomogeneities accurately. The aim of this study was to test a new method, in aqua vivo EPID dosimetry, for fast dose verification of lung cancer irradiations during actual patient treatment. Methods: The key feature of our method is the dose reconstruction in the patient from EPID images, obtained during the actual treatment, whereby the images have been converted to a situation as if the patient consisted entirely of water; hence, the method is termed in aqua vivo. This is done by multiplying the measured in vivo EPID image with the ratio of two digitally reconstructed transmission images for the unit-density and inhomogeneous tissue situation. For dose verification, a comparison is made with the calculated dose distribution with the inhomogeneity correction switched off. IMRT treatment verification is performed for each beam in 2D using a 2D {gamma} evaluation, while for the verification of volumetric-modulated arc therapy (VMAT) treatments in 3D a 3D {gamma} evaluation is applied using the same parameters (3%, 3 mm). The method was tested using two inhomogeneous phantoms simulating a tumor in lung and measuring its sensitivity for patient positioning errors. Subsequently five IMRT and five VMAT clinical lung cancer treatments were investigated, using both the conventional back-projection algorithm and the in aqua vivo method. The verification results of the in aqua vivo method were statistically analyzed for 751 lung cancer patients treated with IMRT and 50 lung cancer patients treated with VMAT. Results: The improvements by applying the in aqua vivo approach are considerable. The percentage of {gamma} values {<=}1 increased on average from 66.2% to 93.1% and from 43.6% to 97.5% for the IMRT and VMAT cases, respectively. The corresponding mean {gamma} value decreased from 0.99 to 0.43 for the IMRT cases and from 1.71 to 0.40 for the VMAT cases, which is similar to the accepted clinical values for the verification of IMRT treatments of prostate, rectum, and head-and-neck cancers. The deviation between the reconstructed and planned dose at the isocenter diminished on average from 5.3% to 0.5% for the VMAT patients and was almost the same, within 1%, for the IMRT cases. The in aqua vivo verification results for IMRT and VMAT treatments of a large group of patients had a mean {gamma} of approximately 0.5, a percentage of {gamma} values {<=}1 larger than 89%, and a difference of the isocenter dose value less than 1%. Conclusions: With the in aqua vivo approach for the verification of lung cancer treatments (IMRT and VMAT), we can achieve results with the same accuracy as obtained during in vivo EPID dosimetry of sites without large inhomogeneities.

  12. 3D ultrafast ultrasound imaging in vivo

    NASA Astrophysics Data System (ADS)

    Provost, Jean; Papadacci, Clement; Esteban Arango, Juan; Imbault, Marion; Fink, Mathias; Gennisson, Jean-Luc; Tanter, Mickael; Pernot, Mathieu

    2014-10-01

    Very high frame rate ultrasound imaging has recently allowed for the extension of the applications of echography to new fields of study such as the functional imaging of the brain, cardiac electrophysiology, and the quantitative imaging of the intrinsic mechanical properties of tumors, to name a few, non-invasively and in real time. In this study, we present the first implementation of Ultrafast Ultrasound Imaging in 3D based on the use of either diverging or plane waves emanating from a sparse virtual array located behind the probe. It achieves high contrast and resolution while maintaining imaging rates of thousands of volumes per second. A customized portable ultrasound system was developed to sample 1024 independent channels and to drive a 32? × ?32 matrix-array probe. Its ability to track in 3D transient phenomena occurring in the millisecond range within a single ultrafast acquisition was demonstrated for 3D Shear-Wave Imaging, 3D Ultrafast Doppler Imaging, and, finally, 3D Ultrafast combined Tissue and Flow Doppler Imaging. The propagation of shear waves was tracked in a phantom and used to characterize its stiffness. 3D Ultrafast Doppler was used to obtain 3D maps of Pulsed Doppler, Color Doppler, and Power Doppler quantities in a single acquisition and revealed, at thousands of volumes per second, the complex 3D flow patterns occurring in the ventricles of the human heart during an entire cardiac cycle, as well as the 3D in vivo interaction of blood flow and wall motion during the pulse wave in the carotid at the bifurcation. This study demonstrates the potential of 3D Ultrafast Ultrasound Imaging for the 3D mapping of stiffness, tissue motion, and flow in humans in vivo and promises new clinical applications of ultrasound with reduced intra—and inter-observer variability.

  13. Near-infrared Molecular Probes for In Vivo Imaging

    PubMed Central

    Zhang, Xuan; Bloch, Sharon; Akers, Walter; Achilefu, Samuel

    2012-01-01

    Cellular and tissue imaging in the near-infrared (NIR) wavelengths between 700 and 900 nm is advantageous for in vivo because of the low absorption of biological molecules in this region. This Unit presents protocols for small animal imaging using planar and fluorescence lifetime imaging techniques. Included is an overview of NIR fluorescence imaging of cells and small animals using NIR organic fluorophores, nanoparticles, and multimodal imaging probes. The development, advantages, and application of NIR fluorescent probes that have been used for in vivo imaging are also summarized. The use of NIR agents in conjunction with visible dyes and considerations in selecting imaging agents are discussed. We conclude with practical considerations for the use of these dyes in cell and small animal imaging applications. PMID:22470154

  14. IN VIVO STAINING OF ASTROCYTES Specific in vivo staining of astrocytes in the whole brain

    E-print Network

    Boyer, Edmond

    become a reference for in vivo staining of the whole astrocytes population in animal modelsIN VIVO STAINING OF ASTROCYTES 1 Specific in vivo staining of astrocytes in the whole brain after or electroencephalographic recordings in vivo. The high contrast of the staining facilitates the image processing and allows

  15. Multiplexed aberration measurement for deep tissue imaging in vivo.

    PubMed

    Wang, Chen; Liu, Rui; Milkie, Daniel E; Sun, Wenzhi; Tan, Zhongchao; Kerlin, Aaron; Chen, Tsai-Wen; Kim, Douglas S; Ji, Na

    2014-10-01

    We describe an adaptive optics method that modulates the intensity or phase of light rays at multiple pupil segments in parallel to determine the sample-induced aberration. Applicable to fluorescent protein-labeled structures of arbitrary complexity, it allowed us to obtain diffraction-limited resolution in various samples in vivo. For the strongly scattering mouse brain, a single aberration correction improved structural and functional imaging of fine neuronal processes over a large imaging volume. PMID:25128976

  16. Positron emitters for {ital in vivo} plant studies

    SciTech Connect

    Fares, Y.; Goeschl, J.D.; Magnuson, C.E.; McKinney, C.J.; Musser, R.L.; Strain, B.R. [Phytotron and Botany Department, Duke University, Durham, North Carolina 27706 (United States)

    1997-02-01

    The use of short-lived positron emitter isotopes in studying the dynamics of biological systems provides an indepth understanding of the regulating functions of the system, that is otherwise unattainable. When we coupled such studies with tracer kinetics models, and a system approach of data analysis, {ital in vivo} simultaneous processes and their interactions are understood. The techniques applied, results of their applications and system analysis of data are reported. {copyright} {ital 1997 American Institute of Physics.}

  17. Multiplexed aberration measurement for deep tissue imaging in vivo

    PubMed Central

    Wang, Chen; Liu, Rui; Milkie, Daniel E.; Sun, Wenzhi; Tan, Zhongchao; Kerlin, Aaron; Chen, Tsai-Wen; Kim, Douglas S.; Ji, Na

    2014-01-01

    We describe a multiplexed aberration measurement method that modulates the intensity or phase of light rays at multiple pupil segments in parallel to determine their phase gradients. Applicable to fluorescent-protein-labeled structures of arbitrary complexity, it allows us to obtain diffraction-limited resolution in various samples in vivo. For the strongly scattering mouse brain, a single aberration correction improves structural and functional imaging of fine neuronal processes over a large imaging volume. PMID:25128976

  18. In vivo imaging with oligonucleotides for diagnosis and drug development

    PubMed Central

    Tavitian, B

    2003-01-01

    Molecular imaging, the science that combines non-invasive in vivo imaging and molecular biology, has begun to use labelled oligonucleotides as radiotracers. Antisense oligonucleotides target gene expression at the RNA level, while aptamer oligonucleotides are designed to hit proteins of interest. Oligonucleotides for imaging cover a large range of applications, from the invention of new contrast agents for diagnosis to exquisite research tools for the development of new drugs. PMID:12746268

  19. Determination of Optimal Rhodamine Flurophore for In Vivo Optical Imaging

    PubMed Central

    Longmire, Michelle; Ogawa, Mikako; Hama, Yukihiro; Kosaka, Nobuyuki; Regino, Celeste A.S.; Choyke, Peter L.; Kobayashi, Hisataka

    2009-01-01

    Optical imaging has the potential to improve the efficacy of surgical and endoscopic approaches to cancer treatment; however, the optimal type of fluorescent probe has not yet been established. It is well known that rhodamine-core-derived fluorophores offer a combination of desirable properties such as good photostability, high extinction coefficient, and high fluorescence quantum yield. However, despite the ubiquitous use of rhodamine fluorophores for in vivo optical imaging, it remains to be determined, however, if unique chemical properties among individual rhodamine core family members affect fluorophore parameters critical to in vivo optical imaging applications. These parameters include: preserved fluorescence intensity in low pH environments, similar to that of the endolysosome; efficient fluorescence signal despite conformational changes to targeting proteins as may occur in harsh subcellular environments; persistence of fluorescence after cellular internalization; and sufficient signal-to-background ratios to permit the identification of fluorophore-targeted tumors. In the present study, we conjugated 4 common rhodamine-core based fluorescent dyes to a clinically feasible and quickly internalizing D-galactose receptor targeting reagent, galactosamine serum albumin (GmSA), and conducted a series of in vitro and in vivo experiments using a metastatic ovarian cancer mouse model to determine if differences exist among rhodamine fluorophores and if so, which rhodamine core possesses optimal characteristics for in vivo imaging applications. Herein, we demonstrate that the rhodamine-fluorophore, TAMRA, is the most robust of the 4 common rhodamine fluorophores for in vivo optical imaging of ovarian cancer metastases to the peritoneum. PMID:18610943

  20. In Vivo and Ex Vivo Transcutaneous Glucose Detection Using Surface-Enhanced Raman Spectroscopy

    NASA Astrophysics Data System (ADS)

    Ma, Ke

    Diabetes mellitus is widely acknowledged as a large and growing health concern. The lack of practical methods for continuously monitoring glucose levels causes significant difficulties in successful diabetes management. Extensive validation work has been carried out using surface-enhanced Raman spectroscopy (SERS) for in vivo glucose sensing. This dissertation details progress made towards a Raman-based glucose sensor for in vivo, transcutaneous glucose detection. The first presented study combines spatially offset Raman spectroscopy (SORS) with SERS (SESORS) to explore the possibility of in vivo, transcutaneous glucose sensing. A SERS-based glucose sensor was implanted subcutaneously in Sprague-Dawley rats. SERS spectra were acquired transcutaneously and analyzed using partial least-squares (PLS). Highly accurate and consistent results were obtained, especially in the hypoglycemic range. Additionally, the sensor demonstrated functionality at least17 days after implantation. A subsequent study further extends the application of SESORS to the possibility of in vivo detection of glucose in brain through skull. Specifically, SERS nanoantennas were buried in an ovine tissue behind a bone with 8 mm thickness and detected by using SESORS. In addition, quantitative detection through bones by using SESORS was also demonstrated. A device that could measure glucose continuously as well as noninvasively would be of great use to patients with diabetes. The inherent limitation of the SESORS approach may prevent this technique from becoming a noninvasive method. Therefore, the prospect of using normal Raman spectroscopy for glucose detection was re-examined. Quantitative detection of glucose and lactate in the clinically relevant range was demonstrated by using normal Raman spectroscopy with low power and short acquisition time. Finally, a nonlinear calibration method called least-squares support vector machine regression (LS-SVR) was investigated for analyzing spectroscopic data sets of glucose detection. Comparison studies were demonstrated between LS-SVR and PLS. LS-SVR demonstrated significant improvements in accuracy over PLS for glucose detection, especially when a global calibration model was required. The improvements imparted by LS-SVR open up the possibility of developing an accurate prediction algorithm for Raman-based glucose sensing applicable to a large human population. Overall, these studies show the high promise held by the Raman-based sensor for the challenge of optimal glycemic control.

  1. Ultra-performance liquid chromatography tandem mass spectrometry method for the determination of AZ66, a sigma receptor ligand, in rat plasma and its application to in vivo pharmacokinetics

    PubMed Central

    Jamalapuram, Seshulatha; Vuppala, Pradeep Kumar; Abdelazeem, Ahmed H.; McCurdy, Christopher R.; Avery, Bonnie A.

    2014-01-01

    Methamphetamine abuse continues as a major problem in the USA owing to its powerful psychological addictive properties. AZ66, 3-[4-(4-cyclohexylpiperazine-1-yl)pentyl]-6-fluorobenzo[d]thiazole-2(3H)-one, an optimized sigma receptor ligand, is a promising therapeutic agent against methamphetamine. To study the in vivo pharmacokinetics of this novel sigma receptor ligand in rats, a sensitive ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS) method was developed in rat plasma and validated. The developed method requires a small volume of plasma (100 ?L) and a simple liquid–liquid extraction. The chromatographic separations were achieved in 3.3 min using an Acquity UPLC BEH Shield RP18 column. The mass spectrophotometric detection was carried out using a Waters Micromass Quattro MicroTM triple-quadrupole system. Multiple reaction monitoring was used for the quantitation with transitions m/z 406?m/z 181 for AZ66 and m/z 448?m/z 285 for aripiprazole. The method was validated over a concentration range of 1–3500 ng/mL and the lower limit of quantitation was determined to be 1 ng/mL. Validation of the assay demonstrated that the developed UPLC/MS/MS method was sensitive, accurate and selective for the determination of AZ66 in rat plasma. The present method has been successfully applied to an i.v. pharmacokinetic study in Sprague–Dawley rats. PMID:23558564

  2. Development of a method to quantify clindamycin in vitreous humor of rabbits' eyes by UPLC-MS/MS: application to a comparative pharmacokinetic study and in vivo ocular biocompatibility evaluation.

    PubMed

    Fernandes-Cunha, Gabriella M; Gouvea, Dayana Rubio; Fulgêncio, Gustavo de Oliveira; Rezende, Cíntia M F; da Silva, Gisele Rodrigues; Bretas, Juliana M; Fialho, Sílvia Ligório; Lopes, Norberto Peporine; Silva-Cunha, Armando

    2015-01-01

    Ocular toxoplasmosis may result in uveitis in the posterior segment of the eye, leading to severe visual complications. Clindamycin-loaded poly(lactide-co-glycolide) (PLGA) implants could be applied to treat the ocular toxoplasmosis. In this study, the pharmacokinetic profiles of the drug administrated by PLGA implants and by intravitreal injections in rabbits' eyes were evaluated. The implant released the drug for 6 weeks while the drug administrated by intravitreal injections remained in the vitreous cavity for 2 weeks. Compared to the injected drug, the implants containing clindamycin had higher values of area under the curve (AUC) (39.2 vs 716.7 ng week mL(-1)) and maximum vitreous concentration (Cmax) (8.7 vs 13.83 ng mL(-1)). The implants prolonged the delivery of clindamycin and increased the contact of the drug with the eyes' tissues. Moreover, the in vivo ocular biocompatibility of the clindamycin-loaded PLGA implants was evaluated regarding to the clinical examination of the eyes and the measurement of the intraocular pressure (IOP) during 6 weeks. The implantable devices caused no ocular inflammatory process and induced the increase of the IOP in the fourth week of the study. The IOP augmentation could be related to the maximum concentration of clindamycin released from the implants. In conclusion, the PLGA implants based on clindamycin may be a therapeutic alternative to treat ocular toxoplasmosis. PMID:25459934

  3. The Transfer of Property in the Conflict of Laws: Choice of Law Rules in Inter Vivos Transfers of Property

    Microsoft Academic Search

    Janeen M. Carruthers

    This book provides a detailed and up-to-date exposition of English and Scottish rules of choice of law in inter vivos transfers of property. It traces the development of the lex situs rule, and its application to inter vivos dealings with immovable property, tangible movable property (including the special case of cultural property), and intangible movable property (including indirectly held securities).

  4. In vivo imaging of sulfotransferases

    DOEpatents

    Barrio, Jorge R; Kepe, Vladimir; Small, Gary W; Satyamurthy, Nagichettiar

    2013-02-12

    Radiolabeled tracers for sulfotransferases (SULTs), their synthesis, and their use are provided. Included are substituted phenols, naphthols, coumarins, and flavones radiolabeled with .sup.18F, .sup.123I, .sup.124I, .sup.125I, or .sup.11C. Also provided are in vivo techniques for using these and other tracers as analytical and diagnostic tools to study sulfotransferase distribution and activity, in health and disease, and to evaluate therapeutic interventions.

  5. In vivo fluorescence lifetime tomography

    NASA Astrophysics Data System (ADS)

    Nothdurft, Ralph E.; Patwardhan, Sachin V.; Akers, Walter; Ye, Yunpeng; Achilefu, Samuel; Culver, Joseph P.

    2009-03-01

    Local molecular and physiological processes can be imaged in vivo through perturbations in the fluorescence lifetime (FLT) of optical imaging agents. In addition to providing functional information, FLT methods can quantify specific molecular events and multiplex diagnostic and prognostic information. We have developed a fluorescence lifetime diffuse optical tomography (DOT) system for in vivo preclinical imaging. Data is captured using a time-resolved intensified charge coupled device (ICCD) system to measure fluorescence excitation and emission in the time domain. Data is then converted to the frequency domain, and we simultaneously reconstruct images of yield and lifetime using an extension to the normalized Born approach. By using differential phase measurements, we demonstrate DOT imaging of short lifetimes (from 350 ps) with high precision (+/-5 ps). Furthermore, this system retains the efficiency, speed, and flexibility of transmission geometry DOT. We demonstrate feasibility of FLT-DOT through a progressive series of experiments. Lifetime range and repeatability are first measured in phantoms. Imaging of subcutaneous implants then verifies the FLT-DOT approach in vivo in the presence of inhomogeneous optical properties. Use in a common research scenario is ultimately demonstrated by imaging accumulation of a targeted near-infrared (NIR) fluorescent-labeled peptide probe (cypate-RGD) in a mouse with a subcutaneous tumor.

  6. Optical Oxygen Micro- and Nanosensors for Plant Applications

    PubMed Central

    Ast, Cindy; Schmälzlin, Elmar; Löhmannsröben, Hans-Gerd; van Dongen, Joost T.

    2012-01-01

    Pioneered by Clark's microelectrode more than half a century ago, there has been substantial interest in developing new, miniaturized optical methods to detect molecular oxygen inside cells. While extensively used for animal tissue measurements, applications of intracellular optical oxygen biosensors are still scarce in plant science. A critical aspect is the strong autofluorescence of the green plant tissue that interferes with optical signals of commonly used oxygen probes. A recently developed dual-frequency phase modulation technique can overcome this limitation, offering new perspectives for plant research. This review gives an overview on the latest optical sensing techniques and methods based on phosphorescence quenching in diverse tissues and discusses the potential pitfalls for applications in plants. The most promising oxygen sensitive probes are reviewed plus different oxygen sensing structures ranging from micro-optodes to soluble nanoparticles. Moreover, the applicability of using heterologously expressed oxygen binding proteins and fluorescent proteins to determine changes in the cellular oxygen concentration are discussed as potential non-invasive cellular oxygen reporters. PMID:22969334

  7. Optical Oxygen Sensors for Applications in Microfluidic Cell Culture

    PubMed Central

    Grist, Samantha M.; Chrostowski, Lukas; Cheung, Karen C.

    2010-01-01

    The presence and concentration of oxygen in biological systems has a large impact on the behavior and viability of many types of cells, including the differentiation of stem cells or the growth of tumor cells. As a result, the integration of oxygen sensors within cell culture environments presents a powerful tool for quantifying the effects of oxygen concentrations on cell behavior, cell viability, and drug effectiveness. Because microfluidic cell culture environments are a promising alternative to traditional cell culture platforms, there is recent interest in integrating oxygen-sensing mechanisms with microfluidics for cell culture applications. Optical, luminescence-based oxygen sensors, in particular, show great promise in their ability to be integrated with microfluidics and cell culture systems. These sensors can be highly sensitive and do not consume oxygen or generate toxic byproducts in their sensing process. This paper presents a review of previously proposed optical oxygen sensor types, materials and formats most applicable to microfluidic cell culture, and analyzes their suitability for this and other in vitro applications. PMID:22163408

  8. In vivo heterogeneous tomographic bioluminescence imaging via a higher-order approximation forward model

    NASA Astrophysics Data System (ADS)

    Liu, Kai; Tian, Jie, Sr.; Qin, Chenghu; Yang, Xin; Zhu, Shouping; Han, Dong; Wu, Ping; Dai, Xiaoqian

    2011-03-01

    In vivo bioluminescence imaging (BLI) has played a more and more important role in biomedical research of small animals. Tomographic bioluminescence imaging (TBI) further translates the BLI optical information into three-dimensional bioluminescent source distribution, which could greatly facilitate applications in related studies. Although the diffusion approximation (DA) is one of the most widely-used forward models, higher-order approximations are still needed for in vivo small animal imaging. In this work, as a relatively accurate and higher-order approximation theory, a simplified spherical harmonics approximation (SPN) is applied for heterogeneous tomographic bioluminescence imaging in vivo. Furthermore, coupled with the SPN, a generalized graph cuts optimization approach is utilized, making BLT reconstructions fast and suit for the whole body of small animals. Heterogeneous in vivo experimental reconstructions via the higher-order approximation model demonstrate higher tomographic imaging quality, which is shown the capability for practical biomedical tomographic imaging applications.

  9. Wide-field in vivo background free imaging by selective magnetic modulation of nanodiamond fluorescence

    PubMed Central

    Sarkar, Susanta K.; Bumb, Ambika; Wu, Xufeng; Sochacki, Kem A.; Kellman, Peter; Brechbiel, Martin W.; Neuman, Keir C.

    2014-01-01

    The sensitivity and resolution of fluorescence-based imaging in vivo is often limited by autofluorescence and other background noise. To overcome these limitations, we have developed a wide-field background-free imaging technique based on magnetic modulation of fluorescent nanodiamond emission. Fluorescent nanodiamonds are bright, photo-stable, biocompatible nanoparticles that are promising probes for a wide range of in vitro and in vivo imaging applications. Our readily applied background-free imaging technique improves the signal-to-background ratio for in vivo imaging up to 100-fold. This technique has the potential to significantly improve and extend fluorescent nanodiamond imaging capabilities on diverse fluorescence imaging platforms. PMID:24761300

  10. In vivo and in vitro Skin Permeation of Butyl Methoxydibenzoylmethane from Lipospheres

    Microsoft Academic Search

    V. Iannuccelli; G. Coppi; S. Sergi; M. Mezzena; S. Scalia

    2008-01-01

    Lipid microparticles (lipospheres) loaded with butyl methoxydibenzoylmethane (BMDBM), a widely used UV-A sunscreen agent, were prepared by melt technique and evaluated for skin permeation both in vivo, by tape stripping method, and in vitro, by a flow-through diffusion chamber. Following in vivo human skin application of an O\\/W emulsion containing 2% of BMDBM loaded in lipospheres, 15% of the applied

  11. Iodine-131 radiolabeling of poly ethylene glycol-coated gold nanorods for in vivo imaging.

    PubMed

    Eskandari, Najmeh; Yavari, Kamal; Outokesh, Mohammad; Sadjadi, Sodeh; Ahmadi, Seyed Javad

    2013-01-01

    Gold nanorods (GNRs) can be used in various biomedical applications; however, very little is known about their in vivo tissue distribution by radiolabeling. Here, we have developed a rapid and simple method with high yield and without disturbing their optical properties for radiolabeling of gold rods with iodine-131 in order to track in vivo tissue uptake of GNRs after systemic administration by biodistribution analysis and ?-imaging. Following intravenous injection into rat, PEGylated GNRs have much longer blood circulation times. PMID:24285135

  12. Spatial light modulator based active wide-field illumination for ex vivo and in vivo quantitative NIR FRET imaging

    PubMed Central

    Zhao, Lingling; Abe, Ken; Rajoria, Shilpi; Pian, Qi; Barroso, Margarida; Intes, Xavier

    2014-01-01

    Fluorescence lifetime imaging is playing an increasing role in drug development by providing a sensitive method to monitor drug delivery and receptor-ligand interactions. However, the wide dynamic range of fluorescence intensity emitted by ex vivo and in vivo samples presents challenges in retrieving information over the whole subject accurately and quantitatively. To overcome this challenge, we developed an active wide-field illumination (AWFI) strategy based on a spatial light modulator that acquires optimal fluorescence signals by enhancing the dynamic range, signal to noise ratio, and estimation of lifetime-based parameters. We demonstrate the ability of AWFI to estimate Förster resonance energy transfer (FRET) donor fraction from dissected organs with high accuracy (standard deviation <6%) over the whole field of view, in contrast with the homogenous wide-field illumination. We further report its successful application to quantitative FRET imaging in a live mouse. AWFI allows improved detection of weak signals and enhanced quantitative accuracy in ex vivo and in vivo molecular fluorescence quantitative imaging. The technique allows for robust quantitative estimation of the bio-distribution of molecular probes and lifetime-based parameters over an extended imaging field exhibiting a large range of fluorescence intensities and at a high acquisition speed (less than 1 min). PMID:24688826

  13. In vivo pool-based shRNA screens to identify modulators of disease progression in hematopoietic malignancies

    E-print Network

    Meacham, Corbin Elizabeth

    2012-01-01

    shRNA screens have been very effective in identifying novel cancer genes in mammalian cells, but they have primarily been limited to in vitro applications in tumor cell lines. Whereas in vivo retroviral mutagenesis screens ...

  14. Simultaneous two-voxel localized 1H-observed 13C-edited spectroscopy for in vivo MRS on rat brain at 9.4 T: Application to the investigation of excitotoxic lesions

    NASA Astrophysics Data System (ADS)

    Doan, Bich-Thuy; Autret, Gwennhael; Mispelter, Joël; Méric, Philippe; Même, William; Montécot-Dubourg, Céline; Corrèze, Jean-Loup; Szeremeta, Frédéric; Gillet, Brigitte; Beloeil, Jean-Claude

    2009-05-01

    13C spectroscopy combined with the injection of 13C-labeled substrates is a powerful method for the study of brain metabolism in vivo. Since highly localized measurements are required in a heterogeneous organ such as the brain, it is of interest to augment the sensitivity of 13C spectroscopy by proton acquisition. Furthermore, as focal cerebral lesions are often encountered in animal models of disorders in which the two brain hemispheres are compared, we wished to develop a bi-voxel localized sequence for the simultaneous bilateral investigation of rat brain metabolism, with no need for external additional references. Two sequences were developed at 9.4 T: a bi-voxel 1H-( 13C) STEAM-POCE (Proton Observed Carbon Edited) sequence and a bi-voxel 1H-( 13C) PRESS-POCE adiabatically decoupled sequence with Hadamard encoding. Hadamard encoding allows both voxels to be recorded simultaneously, with the same acquisition time as that required for a single voxel. The method was validated in a biological investigation into the neuronal damage and the effect on the Tri Carboxylic Acid cycle in localized excitotoxic lesions. Following an excitotoxic quinolinate-induced localized lesion in the rat cortex and the infusion of U- 13C glucose, two 1H-( 13C) spectra of distinct (4 × 4 × 4 mm 3) voxels, one centred on the injured hemisphere and the other on the contralateral hemisphere, were recorded simultaneously. Two 1H bi-voxel spectra were also recorded and showed a significant decrease in N-acetyl aspartate, and an accumulation of lactate in the ipsilateral hemisphere. The 1H-( 13C) spectra could be recorded dynamically as a function of time, and showed a fall in the glutamate/glutamine ratio and the presence of a stable glutamine pool, with a permanent increase of lactate in the ipsilateral hemisphere. This bi-voxel 1H-( 13C) method can be used to investigate simultaneously both brain hemispheres, and to perform dynamic studies. We report here the neuronal damage and the effect on the Tri Carboxylic Acid cycle in localized excitotoxic lesions.

  15. Interstitial leukocyte migration in vivo

    PubMed Central

    Lam, Pui-ying; Huttenlocher, Anna

    2013-01-01

    Rapid leukocyte motility is essential for immunity and host defense. There has been progress in understanding the molecular signals that regulate leukocyte motility both in vitro and in vivo. However, a gap remains in understanding how complex signals are prioritized to result in directed migration, which is critical for both adaptive and innate immune function. Here we focus on interstitial migration and how external cues are translated into intracellular signaling pathways that regulate leukocyte polarity, directional sensing and motility in three-dimensional spaces. PMID:23797028

  16. Biophotonics techniques for structural and functional imaging, in vivo

    PubMed Central

    Ardeshirpour, Yasaman; Gandjbakhche, Amir H.; Najafizadeh, Laleh

    2014-01-01

    In vivo optical imaging is being conducted in a variety of medical applications, including optical breast cancer imaging, functional brain imaging, endoscopy, exercise medicine, and monitoring the photodynamic therapy and progress of neoadjuvant chemotherapy. In the past three decades, in vivo diffuse optical breast cancer imaging has shown promising results in cancer detection, and monitoring the progress of neoadjuvant chemotherapy. The use of near infrared spectroscopy for functional brain imaging has been growing rapidly. In fluorescence imaging, the difference between autofluorescence of cancer lesions compared to normal tissues were used in endoscopy to distinguish malignant lesions from normal tissue or inflammation and in determining the boarders of cancer lesions in surgery. Recent advances in drugs targeting specific tumor receptors, such as AntiBodies (MAB), has created a new demand for developing non-invasive in vivo imaging techniques for detection of cancer biomarkers, and for monitoring their down regulations during therapy. Targeted treatments, combined with new imaging techniques, are expected to potentially result in new imaging and treatment paradigms in cancer therapy. Similar approaches can potentially be applied for the characterization of other disease-related biomarkers. In this chapter, we provide a review of diffuse optical and fluorescence imaging techniques with their application in functional brain imaging and cancer diagnosis. PMID:22433452

  17. Simultaneous in vivo positron emission tomography and magnetic resonance imaging

    PubMed Central

    Catana, Ciprian; Procissi, Daniel; Wu, Yibao; Judenhofer, Martin S.; Qi, Jinyi; Pichler, Bernd J.; Jacobs, Russell E.; Cherry, Simon R.

    2008-01-01

    Positron emission tomography (PET) and magnetic resonance imaging (MRI) are widely used in vivo imaging technologies with both clinical and biomedical research applications. The strengths of MRI include high-resolution, high-contrast morphologic imaging of soft tissues; the ability to image physiologic parameters such as diffusion and changes in oxygenation level resulting from neuronal stimulation; and the measurement of metabolites using chemical shift imaging. PET images the distribution of biologically targeted radiotracers with high sensitivity, but images generally lack anatomic context and are of lower spatial resolution. Integration of these technologies permits the acquisition of temporally correlated data showing the distribution of PET radiotracers and MRI contrast agents or MR-detectable metabolites, with registration to the underlying anatomy. An MRI-compatible PET scanner has been built for biomedical research applications that allows data from both modalities to be acquired simultaneously. Experiments demonstrate no effect of the MRI system on the spatial resolution of the PET system and <10% reduction in the fraction of radioactive decay events detected by the PET scanner inside the MRI. The signal-to-noise ratio and uniformity of the MR images, with the exception of one particular pulse sequence, were little affected by the presence of the PET scanner. In vivo simultaneous PET and MRI studies were performed in mice. Proof-of-principle in vivo MR spectroscopy and functional MRI experiments were also demonstrated with the combined scanner. PMID:18319342

  18. Biophotonics techniques for structural and functional imaging, in vivo.

    PubMed

    Ardeshirpour, Yasaman; Gandjbakhche, Amir H; Najafizadeh, Laleh

    2012-01-01

    In vivo optical imaging is being conducted in a variety of medical applications, including optical breast cancer imaging, functional brain imaging, endoscopy, exercise medicine, and monitoring the photodynamic therapy and progress of neoadjuvant chemotherapy. In the past three decades, in vivo diffuse optical breast cancer imaging has shown promising results in cancer detection, and monitoring the progress of neoadjuvant chemotherapy. The use of near infrared spectroscopy for functional brain imaging has been growing rapidly. In fluorescence imaging, the difference between autofluorescence of cancer lesions compared to normal tissues were used in endoscopy to distinguish malignant lesions from normal tissue or inflammation and in determining the boarders of cancer lesions in surgery. Recent advances in drugs targeting specific tumor receptors, such as AntiBodies (MAB), has created a new demand for developing non-invasive in vivo imaging techniques for detection of cancer biomarkers, and for monitoring their down regulations during therapy. Targeted treatments, combined with new imaging techniques, are expected to potentially result in new imaging and treatment paradigms in cancer therapy. Similar approaches can potentially be applied for the characterization of other disease-related biomarkers. In this chapter, we provide a review of diffuse optical and fluorescence imaging techniques with their application in functional brain imaging and cancer diagnosis. PMID:22433452

  19. In vivo imaging with persistent luminescence silicate-based nanoparticles

    NASA Astrophysics Data System (ADS)

    Maldiney, Thomas; Viana, Bruno; Bessière, Aurélie; Gourier, Didier; Bessodes, Michel; Scherman, Daniel; Richard, Cyrille

    2013-08-01

    We have recently introduced the use of persistent luminescence nanoparticles, or long-lasting phosphors, for in vivo bioimaging applications in living animals. Red long-lasting phosphors possess an emission located in the tissue transparency window, between 600 and 900 nm, allowing their use for in vivo imaging. Thanks to their optical properties, such nanoparticles can be excited before their systemic injection, allowing highly sensitive detection without any autofluorescence. This article reviews our recent work describing the influence of crystal size and surface state on the biodistribution of persistent luminescence nanoparticles after intravenous injection in living animal. Moreover, additional results from this collaboration show that a proper understanding of the fundamental mechanism associated with persistent luminescence in these silicate-based structures constitutes a major help to improve the optical properties of PLNP, resulting in a longer observation of the nanoprobes in vivo. Such promising results offer interesting perspectives for the development of persistent luminescence nanophosphors intended for long-term applications in living animals.

  20. against Tumor Antigen In Vivo

    E-print Network

    The priming of an immune response against a major histocompatibility complex class I-restricted antigen expressed by nonhematopoietic cells involves the transfer of that antigen to a host bone marrow-derived antigen presenting cell (APC) for presentation to CD8 + T lymphocytes. Dendritic cells (DC), as bone marrow-derived APC, are first candidates for presentation of tumorassociated antigens (TAA). The aim of this study was to see whether DC are able to prime in vivo antigen-specific cytotoxic T lymphocytes after exposure to a soluble protein antigen in vitro. Lacking a well-defined murine TAA, we took advantage of 13-galactosidase (13-gal)transduced tumor cell lines as a model in which [3-gal operationally functions as TAA. For in vivo priming both a DC line, transduced or not transduced with the gene coding for murine GM-CSF, and fresh bone marrow-derived DC (bm-DC), loaded in vitro with soluble ~-gal, were used. Priming with either granulocyte macrophage colony-stimulating factor-transduced DC line or fresh bm-DC but not with untransduced DC line generated CTL able to lyse 13-gal-transfected target cells. Furthermore, GM-CSF was necessary for the DC line to efficiently present soluble [3-gal as an H-2Ld-restricted peptide to a [3-gal--specific CTL clone. Data also show that a long-lasting immunity against tumor challenge can be induced using

  1. In vivo studies of opiate receptors

    Microsoft Academic Search

    J. James Frost; Robert F. Dannals; Timothy Duelfer; H. Donald Burns; Hayden T. Ravert; B. Langstroem; V. Balasubramanian; Henry N. Wagner

    1984-01-01

    To study opiate receptors noninvasively in vivo using positron emission tomography, techniques for preferentially labeling opiate receptors in vivo can be used. The rate at which receptor-bound ligand clears from the brain in vivo can be predicted by measuring the equilibrium dissociation constant (KD) at 37 degrees C in the presence of 100 mM sodium chloride and 100 microM guanyl-5'-imidodiphosphate,

  2. Nanoparticle PEBBLE sensors in live cells and in vivo

    PubMed Central

    Smith, Ron

    2009-01-01

    Nanoparticle sensors have been developed for imaging and dynamic monitoring, in live cells and in vivo, of the molecular or ionic components, constructs, forces and dynamics, all in real time, during biological/chemical/physical processes. With their biocompatible small size and inert matrix, nanoparticle sensors have been successfully applied for non-invasive real-time measurements of analytes and fields in cells and rodents, with spatial, temporal, physical and chemical resolution. This review describes the diverse designs of nanoparticle sensors for ions and small molecules, physical fields and biological features, as well as the characterization, properties, and applications of these nanosensors to in vitro and in vivo measurements. Their floating as well as localization ability in biological media is captured by the acronym PEBBLE: photonic explorer for bioanalysis with biologically localized embedding. PMID:20098636

  3. In vivo recordings of brain activity using organic transistors

    PubMed Central

    Khodagholy, Dion; Doublet, Thomas; Quilichini, Pascale; Gurfinkel, Moshe; Leleux, Pierre; Ghestem, Antoine; Ismailova, Esma; Hervé, Thierry; Sanaur, Sébastien; Bernard, Christophe; Malliaras, George G.

    2013-01-01

    In vivo electrophysiological recordings of neuronal circuits are necessary for diagnostic purposes and for brain-machine interfaces. Organic electronic devices constitute a promising candidate because of their mechanical flexibility and biocompatibility. Here we demonstrate the engineering of an organic electrochemical transistor embedded in an ultrathin organic film designed to record electrophysiological signals on the surface of the brain. The device, tested in vivo on epileptiform discharges, displayed superior signal-to-noise ratio due to local amplification compared with surface electrodes. The organic transistor was able to record on the surface low-amplitude brain activities, which were poorly resolved with surface electrodes. This study introduces a new class of biocompatible, highly flexible devices for recording brain activity with superior signal-to-noise ratio that hold great promise for medical applications. PMID:23481383

  4. In vivo photoacoustic imaging of osteosarcoma on animal model

    NASA Astrophysics Data System (ADS)

    Yu, Menglei; Ye, Fei; Hu, Jun

    2011-01-01

    Osteosarcoma is the commonest primary malignant tumor of bone, and the second highest cause of cancer-related death in the paediatric age group. Although there are several methods for osteosarcoma detection, e.g. X-ray, CT, MRI and bone scan, they are not satisfied methods because they can hardly detect osteosarcoma in early stage. Photoacoustic imaging (PAI) is an emerging hybrid imaging modality that is noninvasive, nonionizing, with high sensitivity, satisfactory imaging depth and good temporal and spatial resolution. In order to explore this new method to detect osteosarcoma, we established SD rat models with osteosarcoma and utilized PAI to reconstruct the osteosarcoma image in vivo. This is the first time detecting osteosarcoma in vivo using PAI, and the results suggested that PAI has potential clinical application for detecting osteosarcoma in the early stage.

  5. Manganese ferrite-based nanoparticles induce ex vivo, but not in vivo, cardiovascular effects

    PubMed Central

    Nunes, Allancer DC; Ramalho, Laylla S; Souza, Álvaro PS; Mendes, Elizabeth P; Colugnati, Diego B; Zufelato, Nícholas; Sousa, Marcelo H; Bakuzis, Andris F; Castro, Carlos H

    2014-01-01

    Magnetic nanoparticles (MNPs) have been used for various biomedical applications. Importantly, manganese ferrite-based nanoparticles have useful magnetic resonance imaging characteristics and potential for hyperthermia treatment, but their effects in the cardiovascular system are poorly reported. Thus, the objectives of this study were to determine the cardiovascular effects of three different types of manganese ferrite-based magnetic nanoparticles: citrate-coated (CiMNPs); tripolyphosphate-coated (PhMNPs); and bare magnetic nanoparticles (BaMNPs). The samples were characterized by vibrating sample magnetometer, X-ray diffraction, dynamic light scattering, and transmission electron microscopy. The direct effects of the MNPs on cardiac contractility were evaluated in isolated perfused rat hearts. The CiMNPs, but not PhMNPs and BaMNPs, induced a transient decrease in the left ventricular end-systolic pressure. The PhMNPs and BaMNPs, but not CiMNPs, induced an increase in left ventricular end-diastolic pressure, which resulted in a decrease in a left ventricular end developed pressure. Indeed, PhMNPs and BaMNPs also caused a decrease in the maximal rate of left ventricular pressure rise (+dP/dt) and maximal rate of left ventricular pressure decline (?dP/dt). The three MNPs studied induced an increase in the perfusion pressure of isolated hearts. BaMNPs, but not PhMNPs or CiMNPs, induced a slight vasorelaxant effect in the isolated aortic rings. None of the MNPs were able to change heart rate or arterial blood pressure in conscious rats. In summary, although the MNPs were able to induce effects ex vivo, no significant changes were observed in vivo. Thus, given the proper dosages, these MNPs should be considered for possible therapeutic applications. PMID:25031535

  6. Reprogramming in vivo produces teratomas and iPS cells with totipotency features.

    PubMed

    Abad, María; Mosteiro, Lluc; Pantoja, Cristina; Cañamero, Marta; Rayon, Teresa; Ors, Inmaculada; Graña, Osvaldo; Megías, Diego; Domínguez, Orlando; Martínez, Dolores; Manzanares, Miguel; Ortega, Sagrario; Serrano, Manuel

    2013-10-17

    Reprogramming of adult cells to generate induced pluripotent stem cells (iPS cells) has opened new therapeutic opportunities; however, little is known about the possibility of in vivo reprogramming within tissues. Here we show that transitory induction of the four factors Oct4, Sox2, Klf4 and c-Myc in mice results in teratomas emerging from multiple organs, implying that full reprogramming can occur in vivo. Analyses of the stomach, intestine, pancreas and kidney reveal groups of dedifferentiated cells that express the pluripotency marker NANOG, indicative of in situ reprogramming. By bone marrow transplantation, we demonstrate that haematopoietic cells can also be reprogrammed in vivo. Notably, reprogrammable mice present circulating iPS cells in the blood and, at the transcriptome level, these in vivo generated iPS cells are closer to embryonic stem cells (ES cells) than standard in vitro generated iPS cells. Moreover, in vivo iPS cells efficiently contribute to the trophectoderm lineage, suggesting that they achieve a more plastic or primitive state than ES cells. Finally, intraperitoneal injection of in vivo iPS cells generates embryo-like structures that express embryonic and extraembryonic markers. We conclude that reprogramming in vivo is feasible and confers totipotency features absent in standard iPS or ES cells. These discoveries could be relevant for future applications of reprogramming in regenerative medicine. PMID:24025773

  7. Quercetin in w\\/o microemulsion: In vitro and in vivo skin penetration and efficacy against UVB-induced skin damages evaluated in vivo

    Microsoft Academic Search

    Fabiana T. M. C. Vicentini; Thaís R. M. Simi; José O. Del Ciampo; Nilce O. Wolga; Dimitrius L. Pitol; Mamie M. Iyomasa; M. Vitória L. B. Bentley; Maria J. V. Fonseca

    2008-01-01

    The present study evaluated the potential of a w\\/o microemulsion as a topical carrier system for delivery of the antioxidant quercetin. Topical and transdermal delivery of quercetin were evaluated in vitro using porcine ear skin mounted on a Franz diffusion cell and in vivo on hairless-skin mice. Skin irritation by topical application of the microemulsion containing quercetin, and the protective

  8. In vivo dosimetry in brachytherapy

    SciTech Connect

    Tanderup, Kari [Department of Oncology, Aarhus University Hospital, Aarhus 8000 (Denmark); Department of Clinical Medicine, Aarhus University, Aarhus 8000 (Denmark); Beddar, Sam [Department of Radiation Oncology, University of Texas MD Anderson Cancer Center, Houston, Texas 77030 (United States); Andersen, Claus E.; Kertzscher, Gustavo [Center of Nuclear Technologies, Technical University of Denmark, Roskilde 4000 (Denmark); Cygler, Joanna E. [Department of Physics, Ottawa Hospital Cancer Centre, Ottawa, Ontario K1H 8L6 (Canada)

    2013-07-15

    In vivo dosimetry (IVD) has been used in brachytherapy (BT) for decades with a number of different detectors and measurement technologies. However, IVD in BT has been subject to certain difficulties and complexities, in particular due to challenges of the high-gradient BT dose distribution and the large range of dose and dose rate. Due to these challenges, the sensitivity and specificity toward error detection has been limited, and IVD has mainly been restricted to detection of gross errors. Given these factors, routine use of IVD is currently limited in many departments. Although the impact of potential errors may be detrimental since treatments are typically administered in large fractions and with high-gradient-dose-distributions, BT is usually delivered without independent verification of the treatment delivery. This Vision 20/20 paper encourages improvements within BT safety by developments of IVD into an effective method of independent treatment verification.

  9. In vivo label-free photoacoustic microscopy of cell nuclei by excitation of DNA and RNA

    PubMed Central

    Yao, Da-Kang; Maslov, Konstantin; Shung, Kirk K.; Zhou, Qifa; Wang, Lihong V.

    2010-01-01

    Imaging of cell nuclei plays a critical role in cancer diagnosis and prognosis. In order to image noninvasively cell nuclei in vivo without staining, we developed ultraviolet photoacoustic microscopy (UV-PAM), in which 266-nm-wavelength ultraviolet light excites unlabeled DNA and RNA in cell nuclei to produce photoacoustic waves. We applied UV-PAM to ex vivo imaging of cell nuclei in a mouse lip and a mouse small intestine, and to in vivo imaging of the cell nuclei in the mouse skin. The UV-PAM images of unstained cell nuclei match the optical micrographs of the histologically stained cell nuclei. Given intrinsic optical contrast and high spatial resolution, in vivo label-free UV-PAM has potential for unique biological and clinical application. PMID:21165116

  10. Imaging axonal transport of mitochondria in vivo

    E-print Network

    Cai, Long

    Imaging axonal transport of mitochondria in vivo Thomas Misgeld1,2,5, Martin Kerschensteiner1,3,5, Florence M Bareyre1,3, Robert W Burgess4 & Jeff W Lichtman1 Neuronal mitochondria regulate synaptic and sustained changes in anterograde and retrograde transport. In vivo imaging of mitochondria will be a useful

  11. In Vivo Models of Biofilm Infection

    Microsoft Academic Search

    Kendra P. Rumbaugh; Nancy L. Carty

    \\u000a The in vivo biofilm models that have been described use a variety of animals ranging from goats to chinchillas, and mimic\\u000a diseases including dental caries, endocarditis, pneumonia, keratitis, and otitis media. Representatives of these in vivo models\\u000a are listed in Table 16.1.

  12. Luminescent probes for optical in vivo imaging

    Microsoft Academic Search

    Isabelle Texier; Veronique Josserand; Elisabeth Garanger; Jesus Razkin; Zhaohui Jin; Pascal Dumy; Marie Favrot; Didier Boturyn; Jean-Luc Coll

    2005-01-01

    Going along with instrumental development for small animal fluorescence in vivo imaging, we are developing molecular fluorescent probes, especially for tumor targeting. Several criteria have to be taken into account for the optimization of the luminescent label. It should be adapted to the in vivo imaging optical conditions : red-shifted absorption and emission, limited overlap between absorption and emission for

  13. In vivo imaging of Drosophila melanogaster

    E-print Network

    Perrimon, Norbert

    embryos or small-animal extremities. For whole-body visualization of those targets, a new imaging approachIn vivo imaging of Drosophila melanogaster pupae with mesoscopic fluorescence tomography Claudio, advances in optical micro- scopy for in vivo imaging have focused on physical methods to reject scattered

  14. Pilot in vivo toxicological investigation of boron nitride nanotubes.

    PubMed

    Ciofani, Gianni; Danti, Serena; Genchi, Giada Graziana; D'Alessandro, Delfo; Pellequer, Jean-Luc; Odorico, Michaël; Mattoli, Virgilio; Giorgi, Mario

    2012-01-01

    Boron nitride nanotubes (BNNTs) have attracted huge attention in many different research fields thanks to their outstanding chemical and physical properties. During recent years, our group has pioneered the use of BNNTs for biomedical applications, first of all assessing their in vitro cytocompatibility on many different cell lines. At this point, in vivo investigations are necessary before proceeding toward realistic developments of the proposed applications. In this communication, we report a pilot toxicological study of BNNTs in rabbits. Animals were injected with a 1 mg/kg BNNT solution and blood tests were performed up to 72 hours after injection. The analyses aimed at evaluating any acute alteration of hematic parameters that could represent evidence of functional impairment in blood, liver, and kidneys. Even if preliminary, the data are highly promising, as they showed no adverse effects on all the evaluated parameters, and therefore suggest the possibility of the realistic application of BNNTs in the biomedical field. PMID:22275819

  15. Pilot in vivo toxicological investigation of boron nitride nanotubes

    PubMed Central

    Ciofani, Gianni; Danti, Serena; Genchi, Giada Graziana; D’Alessandro, Delfo; Pellequer, Jean-Luc; Odorico, Michaël; Mattoli, Virgilio; Giorgi, Mario

    2012-01-01

    Boron nitride nanotubes (BNNTs) have attracted huge attention in many different research fields thanks to their outstanding chemical and physical properties. During recent years, our group has pioneered the use of BNNTs for biomedical applications, first of all assessing their in vitro cytocompatibility on many different cell lines. At this point, in vivo investigations are necessary before proceeding toward realistic developments of the proposed applications. In this communication, we report a pilot toxicological study of BNNTs in rabbits. Animals were injected with a 1 mg/kg BNNT solution and blood tests were performed up to 72 hours after injection. The analyses aimed at evaluating any acute alteration of hematic parameters that could represent evidence of functional impairment in blood, liver, and kidneys. Even if preliminary, the data are highly promising, as they showed no adverse effects on all the evaluated parameters, and therefore suggest the possibility of the realistic application of BNNTs in the biomedical field. PMID:22275819

  16. Oxygen sensing glucose biosensors based on alginate nano-micro systems

    NASA Astrophysics Data System (ADS)

    Chaudhari, Rashmi; Joshi, Abhijeet; Srivastava, Rohit

    2014-04-01

    Clinically glucose monitoring in diabetes management is done by point-measurement. However, an accurate, continuous glucose monitoring, and minimally invasive method is desirable. The research aims at developing fluorescence-mediated glucose detecting biosensors based on near-infrared radiation (NIR) oxygen sensitive dyes. Biosensors based on Glucose oxidase (GOx)-Rudpp loaded alginate microspheres (GRAM) and GOx-Platinum-octaethylporphyrin (PtOEP)-PLAalginate microsphere system (GPAM) were developed using air-driven atomization and characterized using optical microscopy, CLSM, fluorescence spectro-photometry etc. Biosensing studies were performed by exposing standard solutions of glucose. Uniform sized GRAM and GPAM with size 50+/-10?m were formed using atomization. CLSM imaging of biosensors suggests that Rudpp and PtOEP nanoparticles are uniformly distributed in alginate microspheres. The GRAM and GPAM showed a good regression constant of 0.974 and of 0.9648 over a range of 0-10 mM of glucose with a high sensitivity of 3.349%/mM (625 nm) and 2.38%/mM (645 nm) at 10 mM of glucose for GRAM and GPAM biosensor. GRAM and GPAM biosensors show great potential in development of an accurate and minimally invasive glucose biosensor. NIR dye based assays can aid sensitive, minimally-invasive and interference-free detection of glucose in diabetic patients.

  17. Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D. J. Osborn

    2003-09-30

    Spectroscopy of Mo{sub 6}Cl{sub 12} immobilized in a sol-gel matrix and heated to 200 C has been performed. Oxygen quenching of the luminescence was observed. Aging Mo{sub 6}Cl{sub 12} to temperatures above 250 C converts the canary yellow Mo{sub 6}Cl{sub 12} to a non-luminescent gray solid. Preliminary experiments point to oxidation of the clusters as the likely cause of thermally induced changes in the physical and optical properties of the clusters.

  18. Oxygen-sensing scaffolds for 3-dimensional cell and tissue culture.

    PubMed

    Jenkins, James; Dmitriev, Ruslan I; Morten, Karl; McDermott, Kieran W; Papkovsky, Dmitri B

    2015-04-01

    Porous membrane scaffolds are widely used materials for three-dimensional cell cultures and tissue models. Additional functional modification of such scaffolds can significantly extend their use and operational performance. Here we describe hybrid microporous polystyrene-based scaffolds impregnated with a phosphorescent O2-sensitive dye PtTFPP, optimized for live cell fluorescence microscopy and imaging of O2 distribution in cultured cells. Modified scaffolds possess high brightness, convenient spectral characteristics (534nm excitation, 650nm emission), stable and robust response to pO2 in phosphorescence intensity and lifetime imaging modes (>twofold response over 21/0% O2), such as confocal PLIM. They are suitable for prolonged use under standard culturing conditions without affecting cell viability, and for multi-parametric imaging analysis of cultured cells and tissue samples. We tested the O2 scaffolds with cultured cancer cells (HCT116), multicellular aggregates (PC12) and rat brain slices and showed that they can inform on tissue oxygenation at different depths and cell densities, changes in respiration activity, viability and responses to drug treatment. Using this method multiplexed with staining of dead cells (CellTox Green) and active mitochondria (TMRM), we demonstrated that decreased O2 (20-24?M) in scaffold corresponds to highest expression of tyrosine hydroxylase in PC12 cells. Such hypoxia is also beneficial for action of hypoxia-specific anti-cancer drug tirapazamine (TPZ). Thus, O2 scaffolds allow for better control of conditions in 3D tissue cultures, and are useful for a broad range of biomaterials and physiological studies. PMID:25653216

  19. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III

    2003-05-01

    More Mo{sub 6}Cl{sub 12} has been synthesized. We have found that previous ambiguous x-ray powder diffraction results are due to disruption of long-range order in the crystals during the heating stage of the synthesis. The quartz cell heaters have been redesigned and are able to heat the substrate. Initial films have been fabricated and are currently under investigation to determine optimal deposition conditions.

  20. A cell viability assay based on monitoring respiration by optical oxygen sensing.

    PubMed

    O'Riordan, T C; Buckley, D; Ogurtsov, V; O'Connor, R; Papkovsky, D B

    2000-02-15

    A cell viability assay based on monitoring of the metabolic activity of living cells via their consumption of dissolved oxygen has been developed. It uses a microwell plate format and disposable phosphorescent sensor inserts incorporated into each sample. The wells are subsequently sealed from ambient oxygen using a layer of mineral oil, and periodically scanned from underneath with a simple fiber-optic phosphorescent phase detector. Thus, dissolved oxygen levels and time profiles of cell respiration can be determined noninvasively and compared to each other. The system was tested by monitoring the viability of the fission yeast Schizosaccharomyces pombe. In comparison with the conventional cell densitometry assay, the optical oxygen sensor method could reliably monitor lower numbers of cells (10(4)-10(5) vs 10(6)-10(7) cells/ml for densitometry), and accurately determine culture viability within 1 h. The assay was then applied to determine the viability of samples treated with toxic agents such as azide and in response to expression of a physiological inducer of cell death, the Bcl-2 family member Bak. The results obtained confirm that measurement of cell respiration by this assay can serve as a predictable, reliable, and fast method for high-throughput determination of cell viability and growth. PMID:10660466

  1. FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS

    SciTech Connect

    Gregory L. Baker; Ruby N. Ghosh; D.J. Osborn III

    2003-01-01

    In this report, initial results pertaining to the synthesis molybdenum clusters and characterization using absorption, optical microscopy, and x-ray powder diffraction are discussed. The synthesis was performed according to literature [1], but results from x-ray powder diffraction indicate that the synthesis did not give the desired compound. The absorption and optical microscopy indicate that the compound synthesized has properties similar to the desired Mo{sub 6}Cl{sub 12} clusters [2,3], so it is unclear as of yet what happened. The sample cell for performing high temperature spectroscopy on thin films of the molybdenum clusters at elevated temperature in a controlled gas environment was designed and an initial prototype was built.

  2. Sol–gel immobilized room-temperature phosphorescent metal-chelate as luminescent oxygen sensing material

    Microsoft Academic Search

    J. M Costa-Fernández; M. E Diaz-Garc??a; A Sanz-Medel

    1998-01-01

    The chelate formed by 8-hydroxy-7-iodo-5-quinolinesulfonic acid (ferron) with aluminium exhibits strong room-temperature phosphorescence (RTP) when retained on a solid support. In a previous paper we have found that sol–gel technology is a very useful approach for developing RTP optical sensors as a new way to immobilize lumiphors. Sol–gel active phases proved to exhibit a high physical rigidity that enhanced relative

  3. Dissolved Oxygen Sensing in a Flow Stream using Molybdenum Chloride Optical Indicators

    Microsoft Academic Search

    Reza Loloee; Per A. Askeland; Ruby N. Ghosh

    2007-01-01

    Dissolved oxygen concentration is considered the most important water quality variable in fish culture. Reliable and continuous (24\\/7) oxygen monitoring of dissolved oxygen (DO) in the 1-11 mg\\/L range would be of great benefit to the aquaculture industry. We briefly review selected DO sensors from both the Clark and optical sensor categories as well a comparing their differences, both advantages

  4. A Cell Viability Assay Based on Monitoring Respiration by Optical Oxygen Sensing

    Microsoft Academic Search

    Tomás C. O'Riordan; Deirdre Buckley; Vladimir Ogurtsov; Rosemary O'Connor; Dmitri B. Papkovsky

    2000-01-01

    A cell viability assay based on monitoring of the metabolic activity of living cells via their consumption of dissolved oxygen has been developed. It uses a microwell plate format and disposable phosphorescent sensor inserts incorporated into each sample. The wells are subsequently sealed from ambient oxygen using a layer of mineral oil, and periodically scanned from underneath with a simple

  5. Novel luminescent Ir(III) dyes for developing highly sensitive oxygen sensing films

    Microsoft Academic Search

    M. Marin-Suarezdel Toro; J. F. Fernandez-Sanchez; E. Baranoff; M. Graetzel; A. Fernandez-Gutierrez

    2010-01-01

    New sensing films have been developed for the detection of molecular oxygen. These films are based on luminescent Ir(III) dyes incorporated either into polystyrene (with and without plasticizer) or metal oxide, nanostructured material. The preparation and characterization of each film have been investigated in detail. Due to their high sensitivity for low oxygen concentration, the parameters pO2(S=1\\/2) and ?I1% have

  6. Primary endosymbiosis and the evolution of light and oxygen sensing in photosynthetic eukaryotes

    PubMed Central

    Rockwell, Nathan C.; Lagarias, J. Clark; Bhattacharya, Debashish

    2015-01-01

    The origin of the photosynthetic organelle in eukaryotes, the plastid, changed forever the evolutionary trajectory of life on our planet. Plastids are highly specialized compartments derived from a putative single cyanobacterial primary endosymbiosis that occurred in the common ancestor of the supergroup Archaeplastida that comprises the Viridiplantae (green algae and plants), red algae, and glaucophyte algae. These lineages include critical primary producers of freshwater and terrestrial ecosystems, progenitors of which provided plastids through secondary endosymbiosis to other algae such as diatoms and dinoflagellates that are critical to marine ecosystems. Despite its broad importance and the success of algal and plant lineages, the phagotrophic origin of the plastid imposed an interesting challenge on the predatory eukaryotic ancestor of the Archaeplastida. By engulfing an oxygenic photosynthetic cell, the host lineage imposed an oxidative stress upon itself in the presence of light. Adaptations to meet this challenge were thus likely to have occurred early on during the transition from a predatory phagotroph to an obligate phototroph (or mixotroph). Modern algae have recently been shown to employ linear tetrapyrroles (bilins) to respond to oxidative stress under high light. Here we explore the early events in plastid evolution and the possible ancient roles of bilins in responding to light and oxygen.

  7. The effect of high light intensities on luminescence lifetime based oxygen sensing.

    PubMed

    Larndorfer, Christoph; Borisov, Sergey M; Lehner, Philipp; Klimant, Ingo

    2014-12-21

    This study highlights possible errors in luminescence lifetime measurements when using bright optical oxygen sensors with high excitation light intensities. An analysis of the sensor with a mathematical model shows that high light intensities will cause a depopulation of the ground state of the luminophore, which results in a non-linear behaviour of the luminescence emission light with respect to the excitation light. The effect of this non-linear behaviour on different lifetime determination methods, including phase-fluorometry, is investigated and in good agreement with the output of the model. Furthermore, the consequences of increasingly high light intensities on phase fluorometric lifetime measurements are illustrated for different oxygen sensors based on benzoporphyrin indicators. For the specific case of PdTPTBPF-based sensors an error as high as 50% is possible under high light conditions (0.25 mol m(-2) s(-1) ? 50 mW mm(-2)). A threshold of applied excitation light intensity is derived, thus enabling the point at which errors become significant to be estimated. Strategies to further avoid such errors are presented. The model also predicts a similar depopulation of the ground state of the quencher; however, the effect of this process was not seen in lab measurements. Possible explanations for this deviation are discussed. PMID:25364788

  8. Global Oxygen Sensing and Visualization in Water using Luminescent Probe on Anodized Aluminum

    NASA Astrophysics Data System (ADS)

    Ozaki, Tatsuya; Ishikawa, Hitoshi; Iijima, Yoshimi; Sakaue, Hirotaka

    2008-11-01

    The extension of pressure-sensitive paint (PSP) technique as a wind tunnel technology to a global oxygen visualization and detection in water is presented. The topic includes the development of anodized-aluminum pressure-sensitive paint (AA-PSP) as a global oxygen sensor in water as well as its calibration and demonstration. Based on the luminophore study, platinum porphyrin is selected as a luminophore, because it is not dissolved in water. It is found that the luminescent increase is over 20 percent after 8 days immersed in water. Even though the signal increases after water immersion, its oxygen sensitivity is the same, which is 0.4. This AA-PSP is used to visualize oxygen rich water (20 mg/l) impinged in less oxygen water (3 mg/l). Even though the difference of water is only the amount of oxygen, we can visualize the water jet with its mixing process using a fast frame rate camera at the frame rate of 100 Hz. In the final version, we will include the oxygen map combined with the visualization result.

  9. Development of an integrated microfluidic platform for oxygen sensing and delivery

    E-print Network

    Vollmer, Adam P

    2005-01-01

    Treatment for end stage lung disease has failed to benefit from advances in medical technology that have produced new treatments for cardiovascular disease, certain cancers, and other major illnesses in recent years. As a ...

  10. Chromatin and oxygen sensing in the context of JmjC histone demethylases

    PubMed Central

    Shmakova, Alena; Batie, Michael; Druker, Jimena; Rocha, Sonia

    2014-01-01

    Responding appropriately to changes in oxygen availability is essential for multicellular organism survival. Molecularly, cells have evolved intricate gene expression programmes to handle this stressful condition. Although it is appreciated that gene expression is co-ordinated by changes in transcription and translation in hypoxia, much less is known about how chromatin changes allow for transcription to take place. The missing link between co-ordinating chromatin structure and the hypoxia-induced transcriptional programme could be in the form of a class of dioxygenases called JmjC (Jumonji C) enzymes, the majority of which are histone demethylases. In the present review, we will focus on the function of JmjC histone demethylases, and how these could act as oxygen sensors for chromatin in hypoxia. The current knowledge concerning the role of JmjC histone demethylases in the process of organism development and human disease will also be reviewed. PMID:25145438

  11. Green luminescent iridium(III) complex immobilized in fluoropolymer film as optical oxygen-sensing material

    Microsoft Academic Search

    Yutaka Amao; Yuichi Ishikawa; Ichiro Okura

    2001-01-01

    An optical oxygen sensor based on the luminescence intensity changes of tris(2-phenylpyridine anion) iridium(III) complex ([Ir(ppy)3]) immobilized in fluoropolymer, poly(styrene-co-2,2,2-trifluoroethyl methacrylate) (poly(styrene-co-TFEM)) film was developed. The luminescence intensity of [Ir(ppy)3] in poly(styrene-co-TFEM) film decreased with increase of oxygen concentration. The ratio I0\\/I100 (a sensitivity of the film, where I0 and I100 represent the detected luminescence intensities from a film exposed

  12. Absolute calibration in vivo measurement systems

    SciTech Connect

    Kruchten, D.A.; Hickman, D.P.

    1991-02-01

    Lawrence Livermore National Laboratory (LLNL) is currently investigating a new method for obtaining absolute calibration factors for radiation measurement systems used to measure internally deposited radionuclides in vivo. Absolute calibration of in vivo measurement systems will eliminate the need to generate a series of human surrogate structures (i.e., phantoms) for calibrating in vivo measurement systems. The absolute calibration of in vivo measurement systems utilizes magnetic resonance imaging (MRI) to define physiological structure, size, and composition. The MRI image provides a digitized representation of the physiological structure, which allows for any mathematical distribution of radionuclides within the body. Using Monte Carlo transport codes, the emission spectrum from the body is predicted. The in vivo measurement equipment is calibrated using the Monte Carlo code and adjusting for the intrinsic properties of the detection system. The calibration factors are verified using measurements of existing phantoms and previously obtained measurements of human volunteers. 8 refs.

  13. Clinical application of BASING and spectral/spatial water and lipid suppression pulses for prostate cancer staging and localization by in vivo 3D 1H magnetic resonance spectroscopic imaging.

    PubMed

    Males, R G; Vigneron, D B; Star-Lack, J; Falbo, S C; Nelson, S J; Hricak, H; Kurhanewicz, J

    2000-01-01

    In previous in situ point-resolved spectroscopy (PRESS) three-dimensional (3D) 1H magnetic resonance (MR) spectroscopic imaging studies, it has been demonstrated that the ratio of prostatic metabolites can noninvasively discriminate prostate cancer from surrounding normal tissue. However, in these studies, conventional chemical shift selective suppression (CHESS) and short-time inversion recovery (STIR) techniques often resulted in inadequate water and lipid suppression. To improve suppression and spatial coverage, the newly developed T1 insensitive dual band selective inversion with gradient dephasing (BASING) Bandstop Filter and dual phase-compensating spectral/spatial spin-echo pulses have been implemented in a clinical setting. In phantom studies, no change in metabolic profiles was observed with application of either BASING or spectral/spatial pulses. In a study of 17 prostate cancer patients, the use of either BASING or spectral/spatial pulses allowed for suppression of water (BASING 99.80 +/- 0.14% and spectral/spatial 99.73 +/- 0.47%) and lipid (BASING 98.56 +/- 1.03% and spectral/spatial 98.44 +/- 1.90%) without a significant difference in the prostatic metabolite ratios. Spectral/spatial suppression has the added advantage of reducing the chemical shift dependence of the PRESS volume, but optimal performance requires high-speed gradients with negligible eddy current effects. BASING suppression is less reliant on accurate pulse and gradient timings and can be implemented easily with no loss in performance on clinical MR scanners with conventional gradients. PMID:10642727

  14. Lessons learned from vivo-morpholinos: How to avoid vivo-morpholino toxicity

    PubMed Central

    Ferguson, David P.; Dangott, Lawrence J.; Lightfoot, J. Timothy

    2014-01-01

    Vivo-morpholinos are a promising tool for gene silencing. These oligonucleotide analogs transiently silence genes by blocking either translation or pre-mRNA splicing. Little to no toxicity has been reported for vivo-morpholino treatment. However, in a recent study conducted in our lab, treatment of mice with vivo-morpholinos resulted in high mortality rates. We hypothesized that the deaths were the result of oligonucleotide hybridization, causing an increased cationic charge associated with the dendrimer delivery moiety of the vivo-morpholino. The cationic charge increased blood clot formation in whole blood treated with vivo-morpholinos, suggesting that clotting could have caused cardiac arrest in the deceased mice. Therefore, we investigate the mechanism by which some vivo-morpholinos increase mortality rates and propose techniques to alleviate vivo-morpholino toxicity. PMID:24806225

  15. DNA nanotubes as intracellular delivery vehicles in vivo.

    PubMed

    Sellner, Sabine; Kocabey, Samet; Nekolla, Katharina; Krombach, Fritz; Liedl, Tim; Rehberg, Markus

    2015-06-01

    DNA-based nanoconstructs possess great potential for biomedical applications. However, the in vivo behavior of such constructs at the microscopic tissue/cell level as well as their inflammatory potential is largely unknown. Unmethylated CpG sequences of DNA are recognized by Toll-like receptor 9 (TLR9), and thus initiate an innate immune response. In this study, we investigated the use of DNA-based nanotubes as carrier systems for CpG delivery and their effect on immune cells in vivo and in real time. DNA nanotubes were microinjected into skeletal muscle of anesthetized mice. Using in vivo microscopy, we observed that the DNA tubes were internalized within minutes by tissue-resident macrophages and localized in their endosomes. Only microinjection of CpG-decorated DNA nanotubes but not of plain DNA nanotubes or CpG oligonucleotides induced a significant recruitment of leukocytes into the muscle tissue as well as activation of the NF-?B pathway in surrounding cells. These results suggest that DNA nanotubes are promising delivery vehicles to target tissue macrophages, whereupon the immunogenic potential depends on the decoration with CpG oligonucleotides. PMID:25890742

  16. In-vivo Reflectance Measurements from Human Skin

    NASA Astrophysics Data System (ADS)

    Delgado, J. A.; Cornejo, A.; Cunill, M.; Báez, J. J.; Matos, R.; Anasagasti, L.; Santiago, C.

    2006-09-01

    We evaluate the potential of using a standard commercial spectrophotometer, specifically designed to meet the growing requirement for color control in the digital-imaging application field, to perform in-vivo diffuse reflectance measurements from adult human skin. We report and discuss diffuse reflectance spectra for three practical situations: a) reflectance versus skin type, b) reflectance from normal skin with different grade of solar exposition, c) reflectance from normal skin versus reflectance from seborrheic keratosis. Results show, that using the above spectrophotometer we can easily differentiate two sites of different solar exposition. Besides, significant differences are found in the normal skin diffuse reflectance for patients with different skin types.

  17. Functionalized Magnetic Nanoparticles as an In Vivo Delivery System

    NASA Astrophysics Data System (ADS)

    Taira, Shu; Moritake, Shinji; Hatanaka, Takahiro; Ichiyanagi, Yuko; Setou, Mitsutoshi

    We developed extremely small functionalized magnetic nanoparticles (MNPs) for use as an in vivo delivery system for pharmaceuticals and biomolecules. We functionalized the MNPs (d = 3 nm) by silanization of amino groups on the particles with (3-aminopropyl)triethoxysilane for subsequent cross-linking with pharmaceuticals and biomolecules. The MNPs were successfully introduced into living cells without any further modification, such as the use of cationic residues, to enhance endocytic internalization. The particles could be incorporated into the subcutaneous tissue of a mouse’s ear through the skin of the ear and could be localized by application of an external magnetic field.

  18. Viscous optical clearing agent for in vivo optical imaging

    NASA Astrophysics Data System (ADS)

    Deng, Zijian; Jing, Lijia; Wu, Ning; lv, Pengyu; Jiang, Xiaoyun; Ren, Qiushi; Li, Changhui

    2014-07-01

    By allowing more photons to reach deeper tissue, the optical clearing agent (OCA) has gained increasing attention in various optical imaging modalities. However, commonly used OCAs have high fluidity, limiting their applications in in vivo studies with oblique, uneven, or moving surfaces. In this work, we reported an OCA with high viscosity. We measured the properties of this viscous OCA, and tested its successful performances in the imaging of a living animal's skin with two optical imaging modalities: photoacoustic microscopy and optical coherence tomography. Our results demonstrated that the viscous OCA has a great potential in the study of different turbid tissues using various optical imaging modalities.

  19. Viscous optical clearing agent for in vivo optical imaging.

    PubMed

    Deng, Zijian; Jing, Lijia; Wu, Ning; Lv, Pengyu; Jiang, Xiaoyun; Ren, Qiushi; Li, Changhui

    2014-01-01

    By allowing more photons to reach deeper tissue, the optical clearing agent (OCA) has gained increasing attention in various optical imaging modalities. However, commonly used OCAs have high fluidity, limiting their applications in in vivo studies with oblique, uneven, or moving surfaces. In this work, we reported an OCA with high viscosity. We measured the properties of this viscous OCA, and tested its successful performances in the imaging of a living animal’s skin with two optical imaging modalities: photoacoustic microscopy and optical coherence tomography. Our results demonstrated that the viscous OCA has a great potential in the study of different turbid tissues using various optical imaging modalities. PMID:25069008

  20. In Vivo Methods for the Assessment of Topical Drug Bioavailability

    PubMed Central

    Herkenne, Christophe; Alberti, Ingo; Naik, Aarti; Kalia, Yogeshvar N.; Mathy, François-Xavier; Préat, Véronique

    2007-01-01

    This paper reviews some current methods for the in vivo assessment of local cutaneous bioavailability in humans after topical drug application. After an introduction discussing the importance of local drug bioavailability assessment and the limitations of model-based predictions, the focus turns to the relevance of experimental studies. The available techniques are then reviewed in detail, with particular emphasis on the tape stripping and microdialysis methodologies. Other less developed techniques, including the skin biopsy, suction blister, follicle removal and confocal Raman spectroscopy techniques are also described. PMID:17985216

  1. Fructation In Vivo: Detrimental and Protective Effects of Fructose

    PubMed Central

    Semchyshyn, H. M.

    2013-01-01

    There is compelling evidence that long-term intake of excessive fructose can have deleterious side effects in different experimental models. However, the role of fructose in vivo remains controversial, since acute temporary application of fructose is found to protect yeast as well as animal tissues against exogenous oxidative stress. This review suggests the involvement of reactive carbonyl and oxygen species in both the cytotoxic and defensive effects of fructose. Potential mechanisms of the generation of reactive species by fructose in the nonenzymatic reactions, their implication in the detrimental and protective effects of fructose are discussed. PMID:23984346

  2. Technical development of a new semispherical radiofrequency bipolar device (RONJA): ex vivo and in vivo studies.

    PubMed

    Vavra, Petr; Penhaker, Marek; Grepl, Jan; Jurcikova, Jana; Palecek, Jiri; Crha, Michal; Nowakova, Jana; Hasal, Martin; Skrobankova, Martina; Ostruszka, Petr; Ihnat, Peter; Delongova, Patricie; Salounova, Dana; Habib, Nagy A; Zonca, Pavel

    2014-01-01

    The aim of this study is to inform about the development of a new semispherical surgical instrument for the bipolar multielectrode radiofrequency liver ablation. Present tools are universal; however they have several disadvantages such as ablation of healthy tissue, numerous needle punctures, and, therefore, longer operating procedure. Our newly designed and tested semispherical surgical tool can solve some of these disadvantages. By conducting an in vivo study on a set of 12 pigs, randomly divided into two groups, we have compared efficiency of the newly developed instrument with the commonly used device. Statistical analysis showed that there were no significant differences between the groups. On average, the tested instrument RONJA had shorter ablation time in both liver lobes and reduced the total operating time. The depth of the thermal alteration was on average 4 mm larger using the newly tested instrument. The new radiofrequency method described in this study could be used in open liver surgery for the treatment of small liver malignancies (up to 2 cm) in a single application with the aim of saving healthy liver parenchyma. Further experimental studies are needed to confirm these results before clinical application of the method in the treatment of human liver malignancies. PMID:24812620

  3. Technical Development of a New Semispherical Radiofrequency Bipolar Device (RONJA): Ex Vivo and In Vivo Studies

    PubMed Central

    Vavra, Petr; Jurcikova, Jana; Crha, Michal; Skrobankova, Martina; Ostruszka, Petr; Ihnat, Peter; Delongova, Patricie; Salounova, Dana; Habib, Nagy A.; Zonca, Pavel

    2014-01-01

    The aim of this study is to inform about the development of a new semispherical surgical instrument for the bipolar multielectrode radiofrequency liver ablation. Present tools are universal; however they have several disadvantages such as ablation of healthy tissue, numerous needle punctures, and, therefore, longer operating procedure. Our newly designed and tested semispherical surgical tool can solve some of these disadvantages. By conducting an in vivo study on a set of 12 pigs, randomly divided into two groups, we have compared efficiency of the newly developed instrument with the commonly used device. Statistical analysis showed that there were no significant differences between the groups. On average, the tested instrument RONJA had shorter ablation time in both liver lobes and reduced the total operating time. The depth of the thermal alteration was on average 4?mm larger using the newly tested instrument. The new radiofrequency method described in this study could be used in open liver surgery for the treatment of small liver malignancies (up to 2?cm) in a single application with the aim of saving healthy liver parenchyma. Further experimental studies are needed to confirm these results before clinical application of the method in the treatment of human liver malignancies. PMID:24812620

  4. Use of an optical clearing agent during noninvasive laser coagulation of the canine vas deferens, ex vivo and in vivo

    NASA Astrophysics Data System (ADS)

    Cilip, Christopher M.; Ross, Ashley E.; Jarow, Jonathan P.; Fried, Nathaniel M.

    2010-02-01

    Development of a noninvasive vasectomy technique may eliminate male fear of complications and result in a more popular procedure. This study explores application of an optical clearing agent (OCA) to the scrotal skin to reduce both the laser power necessary for successful noninvasive laser vasectomy and the probability of scrotal skin burns. A mixture of DMSO/glycerol was noninvasively delivered into the scrotal skin using a Madajet. Near-infrared laser radiation with a range of average powers (7.0-11.7 W) was delivered in conjunction with a range of cryogen spray cooling rates (0.20-0.33 Hz) to the skin surface in a canine model, ex vivo and in vivo. Burst pressure (BP) measurements were conducted to quantify the strength of vas closure. A 30-min application of the OCA improved skin transparency by 26 +/- 5 %, reducing the average power necessary for successful noninvasive laser vasectomy from 9.2 W without OCA (BP = 291 +/- 31 mmHg) to 7.0 W with OCA (BP = 292 +/- 19 mmHg). Control studies without OCA at 7.0 W failed to coagulate the vas with burst pressures (82 +/- 28 mmHg) significantly below typical ejaculation pressures (136 +/- 29 mmHg). Application of an optical clearing agent reduced the laser power necessary for successful noninvasive thermal coagulation of the vas by approximately 25%. This technique may result in the use of a less expensive laser system and eliminate the formation of scrotal skin burns during the procedure.

  5. Expanding room for tetrazine ligations in the in vivo chemistry toolbox.

    PubMed

    Se?kut?, Jolita; Devaraj, Neal K

    2013-10-01

    There is tremendous interest in developing and refining methods to predictably perform chemical reactions within the framework of living systems. Here we review recent advances in applying tetrazine cycloadditions to live cell and in vivo chemistry. We highlight new syntheses of the tetrazine and dienophile precursors useful for in vivo studies. We briefly overview the use of this reaction in combination with unnatural amino acid technology and discuss applications involving the imaging of glycans on live cells. An emerging area is the use of tetrazine ligations for the development of in vivo imaging probes such as those used for positron emission tomography. We summarize recent applications involving tetrazine cycloadditions performed in live mice for pretargeted imaging of cancer cell biomarkers. PMID:24021760

  6. Expanding Room for Tetrazine Ligations in the in vivo Chemistry Toolbox

    PubMed Central

    Še kut, Jolita; Devaraj, Neal K.

    2014-01-01

    There is tremendous interest in developing and refining methods to predictably perform chemical reactions within the framework of living systems. Here we review recent advances in applying tetrazine cycloadditions to live cell and in vivo chemistry. We highlight new syntheses of the tetrazine and dienophile precursors useful for in vivo studies. We briefly overview the use of this reaction in combination with unnatural amino acid technology and discuss applications involving the imaging of glycans on live cells. An emerging area is the use of tetrazine ligations for the development of in vivo imaging probes such as those used for positron emission tomography. We summarize recent applications involving tetrazine cycloadditions performed in live mice for pretargeted imaging of cancer cell biomarkers. PMID:24021760

  7. In vivo generator for radioimmunotherapy

    DOEpatents

    Mausner, Leonard F. (Stony Brook, NY); Srivastava, Suresh G. (Setauket, NY); Straub, Rita F. (Brookhaven, NY)

    1988-01-01

    The present invention involves labeling monoclonal antibodies with intermediate half-life radionuclides which decay to much shorter half-life daughters with desirable high energy beta emissions. Since the daughter will be in equilibrium with the parent, it can exert an in-situ tumoricidal effect over a prolonged period in a localized fashion, essentially as an "in-vivo generator". This approach circumvents the inverse relationship between half-life and beta decay energy. Compartmental modeling was used to determine the relative distribution of dose from both parent and daughter nuclei in target and non-target tissues. Actual antibody biodistribution data have been used to fit realistic rate constants for a model containing tumor, blood, and non-tumor compartments. These rate constants were then used in a variety of simulations for two generator systems, Ba-128/Cs-128 (t.sub.1/2 =2.4d/3.6m) and Pd-112/Ag-112 (t.sub.1/2 =0.9d/192m). The results show that higher tumor/background dose ratios may be achievable by virtue of the rapid excretion of a chemically different daughter during the uptake and clearance phases. This modeling also quantitatively demonstrates the favorable impact on activity distribution of a faster monoclonal antibody tumor uptake, especially when the antibody is labeled with a radionuclide with a comparable half-life.

  8. In vivo facilitated diffusion model

    E-print Network

    Max Bauer; Ralf Metzler

    2013-01-23

    Under dilute in vitro conditions transcription factors rapidly locate their target sequence on DNA by using the facilitated diffusion mechanism. However, whether this strategy of alternating between three-dimensional bulk diffusion and one-dimensional sliding along the DNA contour is still beneficial in the crowded interior of cells is highly disputed. Here we use a simple model for the bacterial genome inside the cell and present a semi-analytical model for the in vivo target search of transcription factors within the facilitated diffusion framework. Without having to resort to extensive simulations we determine the mean search time of a lac repressor in a living E. coli cell by including parameters deduced from experimental measurements. The results agree very well with experimental findings, and thus the facilitated diffusion picture emerges as a quantitative approach to gene regulation in living bacteria cells. Furthermore we see that the search time is not very sensitive to the parameters characterizing the DNA configuration and that the cell seems to operate very close to optimal conditions for target localization. Local searches as implied by the colocalization mechanism are only found to mildly accelerate the mean search time within our model.

  9. Novel peptides functionally targeting in vivo human lung cancer discovered by in vivo peptide displayed phage screening.

    PubMed

    Lee, Kyoung Jin; Lee, Jae Hee; Chung, Hye Kyung; Choi, Jinhyang; Park, Jaesook; Park, Seok Soon; Ju, Eun Jin; Park, Jin; Shin, Seol Hwa; Park, Hye Ji; Ko, Eun Jung; Suh, Nayoung; Kim, InKi; Hwang, Jung Jin; Song, Si Yeol; Jeong, Seong-Yun; Choi, Eun Kyung

    2015-02-01

    Discovery of the cancer-specific peptidic ligands have been emphasized for active targeting drug delivery system and non-invasive imaging. For the discovery of useful and applicable peptidic ligands, in vivo peptide-displayed phage screening has been performed in this study using a xenograft mouse model as a mimic microenvironment to tumor. To seek human lung cancer-specific peptides, M13 phage library displaying 2.9 × 10(9) random peptides was intravenously injected into mouse model bearing A549-derived xenograft tumor through the tail vein. Then the phages emerged from a course of four rounds of biopanning in the xenograft tumor tissue. Novel peptides were categorized into four groups according to a sequence-homology phylogenicity, and in vivo tumor-targeting capacity of these peptides was validated by whole body imaging with Cy5.5-labeled phages in various cancer types. The result revealed that novel peptides accumulated only in adenocarcinoma lung cancer cell-derived xenograft tissue. For further confirmation of the specific targeting ability, in vitro cell-binding assay and immunohistochemistry in vivo tumor tissue were performed with a selected peptide. The peptide was found to bind intensely to lung cancer cells both in vitro and in vivo, which was efficiently compromised with unlabeled phages in an in vitro competition assay. In conclusion, the peptides specifically targeting human lung cancer were discovered in this study, which is warranted to provide substantive feasibilities for drug delivery and imaging in terms of a novel targeted therapeutics and diagnostics. PMID:25366491

  10. Gravitational physiology of human immune cells: a review of in vivo, ex vivo and in vitro studies

    NASA Technical Reports Server (NTRS)

    Cogoli, A.

    1996-01-01

    The study of the function of immune cells in microgravity has been studied for more than 20 years in several laboratories. It is clear today that the immune system is depressed in more than 50% of the astronauts during and after space flight and that the activation of T lymphocytes by mitogens in vitro changes dramatically. This article gives an overview of the gravitational studies conducted by our laboratory in Spacelab, in MIR station, in sounding rockets and on the ground in the clinostat and the centrifuge. Three experimental approaches are followed in our work: (i) Ex vivo studies are performed with blood samples drawn from astronauts; (ii) in vivo studies are based on the application of seven antigens to the skin of the astronauts; (iii) in vitro studies are carried out with immune cells purified from the blood of healthy donors (not astronauts). The data from our in vivo and ex vivo studies are in agreement with those of other laboratories and show that the immunological function is depressed in the majority of astronauts as a consequence of the stress of space flight rather than by a direct influence of gravity on the cell. Immune depression may become a critical hazard on long duration flights on space stations or to other planets. In vitro experiments show that cultures of free-floating lymphocytes and monocytes undergo a dramatic depression of activation by the mitogen concanavalin A, while activation is more than doubled when the cells are attached to microcarrier beads. Such effects may be attributed to both direct and indirect effects of gravitational unloading on basic biological mechanisms of the cell. While the in vitro data are very important to clarify certain aspects of the biological mechanism of T cells activation, they are not descriptive of the changes of the immunological function of the astronauts.

  11. Deltex1 antagonizes HIF-1? and sustains the stability of regulatory T cells in vivo

    PubMed Central

    Hsiao, Huey-Wen; Hsu, Tzu-Sheng; Liu, Wen-Hsien; Hsieh, Wan-Chen; Chou, Ting-Fang; Wu, Yu-Jung; Jiang, Si-Tse; Lai, Ming-Zong

    2015-01-01

    Application of regulatory T cells (Tregs) in transplantation, autoimmunity and allergy has been extensively explored, but how Foxp3 and Treg stability is regulated in vivo is incompletely understood. Here, we identify a requirement for Deltex1 (DTX1), a contributor to T-cell anergy and Foxp3 protein level maintenance in vivo. Dtx1?/? Tregs are as effective as WT Tregs in the inhibition of CD4+CD25? T-cell activation in vitro. However, the suppressive ability of Dtx1?/? Tregs is greatly impaired in vivo. We find that Foxp3 expression is diminished when Dtx1?/? Tregs are co-transferred with effector T cells in vivo. DTX1 promotes the degradation of HIF-1?. Knockout of HIF-1? restores the Foxp3 stability and rescues the defective suppressive activity in Dtx1?/? Treg cells in vivo. Our results suggest that DTX1 exerts another level of control on Treg stability in vivo by sustaining the expression of Foxp3 protein in Tregs. PMID:25695215

  12. Comparative Investigations on the Retrieval Capabilities of Various Baskets and Graspers in Four Ex Vivo Models

    Microsoft Academic Search

    Taras Ptashnyk; Armando Cueva-Martinez; Maurice Stephan Michel; Peter Alken; Kai Uwe Köhrmann

    2002-01-01

    Objectives: The increasing application of ureterorenoscopy for the treatment of urolithiasis has produced a myriad of different help-tools for stone retrieval. In this study, we compared the retrieval capabilities of different baskets and graspers in ex vivo models and attempted to find the most appropriate tool for stone extraction considering the location and size of stone, number of stones and

  13. Intracellular and Computational Characterization of the Intracortical Inhibitory Control of Synchronized Thalamic Inputs In Vivo

    E-print Network

    Destexhe, Alain

    inputs in vivo. J. Neuro- On the other hand, topical application of penicillin on physiol. 78: 335 to the hypothesisdles or following thalamic stimulation; 2) reversed IPSPs with that, in the penicillin epilepsy model the penicillin studies would be that during spindles, thalamic on reconstructed pyramidal cells constrained

  14. Time dependence of enamel fluoride acquisition from APF gels II. In vivo study

    Microsoft Academic Search

    Stephen H. Y. Wei; Eilly W. S. Lau

    1988-01-01

    In vivo enamel fluoride (F) uptake as a function of time after a single application of a new topical APF gel, Minute- Gel TM (gel A) , was compared to a control APF gel, Nupro ® (gel B). The retained F in enamel after different post-treatment intervals also was assessed. Forty orthodontic patients aged 10-16 with four premolars scheduled for

  15. Anticancer activity of peach and plum extracts against human breast cancer in vitro and in vivo

    E-print Network

    Noratto Dongo, Giuliana Doris

    2009-05-15

    and autochthonous tumors, which may reduce the predictive power of xenografts to clinical tumors.87, 88 Furthermore, it has been reported that about one- third of the compounds with an in vivo activity tested by xenograft models, had correlation with activity... .............................................11 Xenograft model ...............................................................................12 Application of xenograft models in breast cancer research ..............13 Xenograft models and natural compounds...

  16. The Effect of Age on Hepatic Gene Transfer with Self-Inactivating Lentiviral Vectors in Vivo

    E-print Network

    Ford, James

    of Pediatrics and Program in Gene Therapy, Department of Genetics, Stanford University, Stanford, California INTRODUCTION Lentiviral vectors (LVs) have promising potential for gene therapy applications to correctThe Effect of Age on Hepatic Gene Transfer with Self-Inactivating Lentiviral Vectors in Vivo Frank

  17. Adventitious shoot formation on leaf cuttings in vivo, a tool in horticulture

    Microsoft Academic Search

    J. B. M. Custers

    1986-01-01

    Adventitious shoot formation implies the regeneration or development of shoots from fully differentiated tissue. Its application has, after the rise of in vitro culture, assumed large proportions. Then the question arose whether in vivo adventitious shoot formation could not be applied more widely in commercial horticulture. To answer this question investigations were made on the regeneration of leaf cuttings and

  18. Peri-implant care with the CO 2 laser: In vitro and in vivo results

    Microsoft Academic Search

    Herbert Deppe; Hans-Henning Horch; Helmut Greim; Thomas Brill; Stefan Wagenpfeil; Karl Donath

    2005-01-01

    Background: Numerous applications for dental lasers have been proposed for both clinical use and experimental purposes. A new indication might be the sterilization of exposed implant surfaces in order to rehabilitate ailing implants. The purposes of this study were to assess CO2 laser parameters for the decontamination process in vitro and to evaluate the method in vivo.Methods: In vitro, temperature

  19. Ginkgo biloba extract EGb ® 761 increases endothelial nitric oxide production in vitro and in vivo

    Microsoft Academic Search

    A. Koltermann; A. Hartkorn; E. Koch; R. Fürst; A. M. Vollmar; S. Zahler

    2007-01-01

    .  Beneficial effects of Ginkgo biloba on peripheral arterial occlusive disease have been repeatedly shown in clinical trials, especially after use of EGb 761, a standardized special extract. Since the underlying mechanisms are widely unknown, we aimed to elucidate the molecular\\u000a basis on which EGb 761 protects against endothelial dysfunction in vitro and in vivo. Application of therapeutically feasible doses of

  20. In Vivo Gene Therapy of Hemophilia B: Sustained Partial Correction in Factor IX-Deficient Dogs

    Microsoft Academic Search

    Mark A. Kay; Steven Rothenberg; Charles N. Landen; Dwight A. Bellinger; Frances Leland; Carol Toman; Milton Finegold; Arthur R. Thompson; M. S. Read; Kenneth M. Brinkhous; Savio L. C. Woo

    1993-01-01

    The liver represents a model organ for gene therapy. A method has been developed for hepatic gene transfer in vivo by the direct infusion of recombinant retroviral vectors into the portal vasculature, which results in the persistent expression of exogenous genes. To determine if these technologies are applicable for the treatment of hemophilia B patients, preclinical efficacy studies were done

  1. Using a priori knowledge to classify in vivo images of the lung

    E-print Network

    Paris-Sud XI, Université de

    Using a priori knowledge to classify in vivo images of the lung Chesner D´esir1 , Caroline perspectives for this very challenging medical application. 1 Introduction The lungs are the essential, that includes the trachea, bronchi, and bronchioles, and (ii) the gas-exchange region, or lung parenchyma, made

  2. Engineering intracellular active transport systems as in vivo biomolecular tools.

    SciTech Connect

    Bachand, George David; Carroll-Portillo, Amanda

    2006-11-01

    Active transport systems provide essential functions in terms of cell physiology and metastasis. These systems, however, are also co-opted by invading viruses, enabling directed transport of the virus to and from the cell's nucleus (i.e., the site of virus replication). Based on this concept, fundamentally new approaches for interrogating and manipulating the inner workings of living cells may be achievable by co-opting Nature's active transport systems as an in vivo biomolecular tool. The overall goal of this project was to investigate the ability to engineer kinesin-based transport systems for in vivo applications, specifically the collection of effector proteins (e.g., transcriptional regulators) within single cells. In the first part of this project, a chimeric fusion protein consisting of kinesin and a single chain variable fragment (scFv) of an antibody was successfully produced through a recombinant expression system. The kinesin-scFv retained both catalytic and antigenic functionality, enabling selective capture and transport of target antigens. The incorporation of a rabbit IgG-specific scFv into the kinesin established a generalized system for functionalizing kinesin with a wide range of target-selective antibodies raised in rabbits. The second objective was to develop methods of isolating the intact microtubule network from live cells as a platform for evaluating kinesin-based transport within the cytoskeletal architecture of a cell. Successful isolation of intact microtubule networks from two distinct cell types was demonstrated using glutaraldehyde and methanol fixation methods. This work provides a platform for inferring the ability of kinesin-scFv to function in vivo, and may also serve as a three-dimensional scaffold for evaluating and exploiting kinesin-based transport for nanotechnological applications. Overall, the technology developed in this project represents a first-step in engineering active transport system for in vivo applications. Further development could potentially enable selective capture of intracellular antigens, targeted delivery of therapeutic agents, or disruption of the transport systems and consequently the infection and pathogenesis cycle of biothreat agents.

  3. Entomopathogenic nematode production and application technology

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Production and application technology is critical for the success of entomopathogenic nematodes (EPNs) in biological control. Production approaches include in vivo, and in vitro methods (solid or liquid fermentation). For laboratory use and small scale field experiments, in vivo production of EPNs...

  4. In Vivo Imaging of Tissue Physiological Function

    Cancer.gov

    The National Cancer Institute's Radiation Biology Branch is seeking statements of capability or interest from parties interested in collaborative research to further develop, evaluate, or commercialize methods for in vivo imaging.

  5. Development of the in vivo flow cytometer

    E-print Network

    Novak, John P. (John Peter), 1957-

    2004-01-01

    An in vivo flow cytometer has been developed that allows the real-time detection and quantification of circulating cells containing fluorescent proteins or labeled with fluorochrome molecules in live animals, without the ...

  6. High-throughput in vivo vertebrate screening

    E-print Network

    Pardo-Martin, Carlos

    We demonstrate a high-throughput platform for cellular-resolution in vivo chemical and genetic screens on zebrafish larvae. The system automatically loads zebrafish from reservoirs or multiwell plates, and positions and ...

  7. Neuronal activity regulates extracellular tau in vivo

    PubMed Central

    Yamada, Kaoru; Holth, Jerrah K.; Liao, Fan; Stewart, Floy R.; Mahan, Thomas E.; Jiang, Hong; Cirrito, John R.; Patel, Tirth K.; Hochgräfe, Katja; Mandelkow, Eva-Maria

    2014-01-01

    Tau is primarily a cytoplasmic protein that stabilizes microtubules. However, it is also found in the extracellular space of the brain at appreciable concentrations. Although its presence there may be relevant to the intercellular spread of tau pathology, the cellular mechanisms regulating tau release into the extracellular space are not well understood. To test this in the context of neuronal networks in vivo, we used in vivo microdialysis. Increasing neuronal activity rapidly increased the steady-state levels of extracellular tau in vivo. Importantly, presynaptic glutamate release is sufficient to drive tau release. Although tau release occurred within hours in response to neuronal activity, the elimination rate of tau from the extracellular compartment and the brain is slow (half-life of ?11 d). The in vivo results provide one mechanism underlying neuronal tau release and may link trans-synaptic spread of tau pathology with synaptic activity itself. PMID:24534188

  8. The potential of photoacoustic microscopy as a tool to characterize the in vivo degradation of surgical sutures

    PubMed Central

    Aguirre, Juan; Morales-Dalmau, Jordi; Funk, Lutz; Jara, Francesc; Turon, Pau; Durduran, Turgut

    2014-01-01

    The ex vivo and in vivo imaging, and quantitative characterization of the degradation of surgical sutures (?500 ?m diameter) up to ?1cm depth is demonstrated using a custom dark-field photo-acoustic microscope (PAM). A practical algorithm is developed to accurately measure the suture diameter during the degradation process. The results from tissue simulating phantoms and mice are compared to ex vivo measurements with an optical microscope demonstrating that PAM has a great deal of potential to characterize the degradation process of surgical sutures. The implications of this work for industrial applications are discussed. PMID:25136508

  9. Genetic targeting of chemical indicators in vivo.

    PubMed

    Yang, Guoying; de Castro Reis, Fernanda; Sundukova, Mayya; Pimpinella, Sofia; Asaro, Antonino; Castaldi, Laura; Batti, Laura; Bilbao, Daniel; Reymond, Luc; Johnsson, Kai; Heppenstall, Paul A

    2015-02-01

    Fluorescent protein reporters have become the mainstay for tracing cellular circuitry in vivo but are limited in their versatility. Here we generated Cre-dependent reporter mice expressing the Snap-tag to target synthetic indicators to cells. Snap-tag labeling worked efficiently and selectively in vivo, allowing for both the manipulation of behavior and monitoring of cellular fluorescence from the same reporter. PMID:25486061

  10. Technologies of directed protein evolution in vivo

    Microsoft Academic Search

    Artem Blagodatski; Vladimir L. Katanaev

    2011-01-01

    Directed evolution of proteins for improved or modified functionality is an important branch of modern biotechnology. It has\\u000a traditionally been performed using various in vitro methods, but more recently, methods of in vivo artificial evolution come\\u000a into play. In this review, we discuss and compare prokaryotic and eukaryotic-based systems of directed protein evolution in\\u000a vivo, highlighting their benefits and current

  11. Quantum Dots for Live Cell and In Vivo Imaging

    PubMed Central

    Walling, Maureen A; Novak, Jennifer A; Shepard, Jason R. E

    2009-01-01

    In the past few decades, technology has made immeasurable strides to enable visualization, identification, and quantitation in biological systems. Many of these technological advancements are occurring on the nanometer scale, where multiple scientific disciplines are combining to create new materials with enhanced properties. The integration of inorganic synthetic methods with a size reduction to the nano-scale has lead to the creation of a new class of optical reporters, called quantum dots. These semiconductor quantum dot nanocrystals have emerged as an alternative to organic dyes and fluorescent proteins, and are brighter and more stable against photobleaching than standard fluorescent indicators. Quantum dots have tunable optical properties that have proved useful in a wide range of applications from multiplexed analysis such as DNA detection and cell sorting and tracking, to most recently demonstrating promise for in vivo imaging and diagnostics. This review provides an in-depth discussion of past, present, and future trends in quantum dot use with an emphasis on in vivo imaging and its related applications. PMID:19333416

  12. Blood flow and blood velocity measurement in vivo by electromagnetic induction

    Microsoft Academic Search

    D. G. Wyatt

    1984-01-01

    The application of electromagnetic induction to the measurement of blood flow and blood velocityin vivo is reviewed. The electronics requirements are discussed and the associated problems are surveyed. An account is given of\\u000a Bevir's virtual-current theory and its application to various flow and velocity measuring devices. The devices at present\\u000a known to be in use or under development are reviewed

  13. Triplet photosensitizers: from molecular design to applications.

    PubMed

    Zhao, Jianzhang; Wu, Wanhua; Sun, Jifu; Guo, Song

    2013-06-21

    Triplet photosensitizers (PSs) are compounds that can be efficiently excited to the triplet excited state which subsequently act as catalysts in photochemical reactions. The name is originally derived from compounds that were used to transfer the triplet energy to other compounds that have only a small intrinsic triplet state yield. Triplet PSs are not only used for triplet energy transfer, but also for photocatalytic organic reactions, photodynamic therapy (PDT), photoinduced hydrogen production from water and triplet-triplet annihilation (TTA) upconversion. A good PS should exhibit strong absorption of the excitation light, a high yield of intersystem crossing (ISC) for efficient production of the triplet state, and a long triplet lifetime to allow for the reaction with a reactant molecule. Most transition metal complexes show efficient ISC, but small molar absorption coefficients in the visible spectral region and short-lived triplet excited states, which make them unsuitable as triplet PSs. One obstacle to the development of new triplet PSs is the difficulty in predicting the ISC of chromophores, especially of organic compounds without any heavy atoms. This review article summarizes some molecular design rationales for triplet PSs, based on the molecular structural factors that facilitate ISC. The design of transition metal complexes with large molar absorption coefficients in the visible spectral region and long-lived triplet excited states is presented. A new method of using a spin converter to construct heavy atom-free organic triplet PSs is discussed, with which ISC becomes predictable, C60 being an example. To enhance the performance of triplet PSs, energy funneling based triplet PSs are proposed, which show broadband absorption in the visible region. Applications of triplet PSs in photocatalytic organic reactions, hydrogen production, triplet-triplet annihilation upconversion and luminescent oxygen sensing are briefly introduced. PMID:23450221

  14. Ex Vivo and In Vivo Models for Endoscopic Submucosal Dissection Training

    PubMed Central

    Gonzalez, Nicolas; Arnau, Maria Rosa

    2012-01-01

    Endoscopic submucosal dissection is a technically challenging but highly effective technique for the treatment of well selected early neoplasms in the digestive tract. Although it is frequently performed in East Asian countries, the Western world has not adopted this technique yet, probably due in part to the difficulty to learn it. Ex vivo and in vivo animal models are invaluable tools to overcome at least the beginning of the learning curve, although the initial step is the acquisition of basic knowledge about early diagnosis of neoplasias, and observing real procedures in expert centers. The practical issues, advantages, and disadvantages of the ex vivo and in vivo models are discussed. PMID:23251881

  15. Oxygen sensitivity and biocompatibility of an implantable paramagnetic probe for repeated measur e ments of tissue oxygenation

    Microsoft Academic Search

    Guruguhan Meenakshisundaram; Edward Eteshola; Ramasamy P. Pandian; Anna Bratasz; Karuppaiyah Selvendiran; Stephen C. Lee; Murali C. Krishna; Harold M. Swartz; Periannan Kuppusamy

    2009-01-01

    The use of oxygen-sensing water-insoluble paramagnetic probes, such as lithium octa-n-butoxynaphthalocyanine (LiNc-BuO), enables repeated measurements of pO2 from the same location in tissue by electron paramagnetic resonance (EPR) spectroscopy. In order to facilitate direct in vivo application, and hence eventual clinical applicability, of LiNc-BuO, we encapsulated LiNc-BuO microcrystals in polydimethylsiloxane\\u000a (PDMS), an oxygen-permeable and bioinert polymer, and developed an implantable

  16. Fiber optic confocal microscope: In vivo precancer detection

    NASA Astrophysics Data System (ADS)

    Carlson, Kristen Dawn

    Cancer is a significant public health problem worldwide. Many cancers originate as precancerous lesions in the epithelium which, when removed in sufficient time, can prevent progression to cancer. However, current detection techniques are typically time-consuming and expensive, limiting their acceptance and accessibility. Optical techniques, such as confocal microscopy, have significant potential to provide clinicians with real-time, high-resolution images of cells and tissue without tissue removal. These images of cell morphology and tissue architecture can be used to characterize tissue and determine the presence or extent of precancer and cancer. This dissertation explores the instrumentation and application of fiber optic reflectance confocal microscopy for in vivo precancer detection. The first part of the dissertation presents in vivo imaging of suspicious lesions in the human uterine cervix and oral mucosa using a fiber bundle based confocal microscope with a complex glass miniature objective lens. Images are analyzed quantitatively and qualitatively to determine the potential of this technology in vivo. An analysis of nuclear density from images of 30 cervical epithelium sites shows differentiation between normal and precancerous sites. Similarly, images from 20 oral mucosa sites demonstrate changes in nuclear density and tissue architecture indicative of progression of precancer and cancer. In addition to this multi-fiber confocal microscope used with a glass objective lens for the clinical studies, imaging of tissue samples has been accomplished with the same confocal system using an injection molded plastic miniature objective lens demonstrating comparable optical quality for a significantly less expensive optical component. Finally, a benchtop prototype of a single fiber confocal microscope using a gimbaled two-axis MEMS scanner has been designed and constructed. Imaging of a resolution target and cellular samples demonstrates sufficient resolution and field of view for cellular imaging. The results from the imaging studies presented here indicate that in vivo confocal microscopy has the potential to improve early precancer detection in epithelial tissue. Advances in imaging technology will continue to reduce the cost of imaging systems and improve the imaging capability, leading to an inexpensive, real-time, minimally-invasive tool for in vivo imaging.

  17. In Vivo Imaging of Stepwise Vessel Occlusion in Cerebral Photothrombosis of Mice by 19F MRI

    PubMed Central

    Kleinschnitz, Christoph; Kampf, Thomas; Jakob, Peter M.; Stoll, Guido

    2011-01-01

    Background 19F magnetic resonance imaging (MRI) was recently introduced as a promising technique for in vivo cell tracking. In the present study we compared 19F MRI with iron-enhanced MRI in mice with photothrombosis (PT) at 7 Tesla. PT represents a model of focal cerebral ischemia exhibiting acute vessel occlusion and delayed neuroinflammation. Methods/Principal Findings Perfluorocarbons (PFC) or superparamagnetic iron oxide particles (SPIO) were injected intravenously at different time points after photothrombotic infarction. While administration of PFC directly after PT induction led to a strong 19F signal throughout the entire lesion, two hours delayed application resulted in a rim-like 19F signal at the outer edge of the lesion. These findings closely resembled the distribution of signal loss on T2-weighted MRI seen after SPIO injection reflecting intravascular accumulation of iron particles trapped in vessel thrombi as confirmed histologically. By sequential administration of two chemically shifted PFC compounds 0 and 2 hours after illumination the different spatial distribution of the 19F markers (infarct core/rim) could be visualized in the same animal. When PFC were applied at day 6 the fluorine marker was only detected after long acquisition times ex vivo. SPIO-enhanced MRI showed slight signal loss in vivo which was much more prominent ex vivo indicative for neuroinflammation at this late lesion stage. Conclusion Our study shows that vessel occlusion can be followed in vivo by 19F and SPIO-enhanced high-field MRI while in vivo imaging of neuroinflammation remains challenging. The timing of contrast agent application was the major determinant of the underlying processes depicted by both imaging techniques. Importantly, sequential application of different PFC compounds allowed depiction of ongoing vessel occlusion from the core to the margin of the ischemic lesions in a single MRI measurement. PMID:22194810

  18. In Vivo Delivery of Nucleic Acids via Glycopolymer Vehicles Affords Therapeutic Infarct Size Reduction In Vivo

    PubMed Central

    Tranter, Michael; Liu, Yemin; He, Suiwen; Gulick, James; Ren, Xiaoping; Robbins, Jeffrey; Jones, W. Keith; Reineke, Theresa M.

    2012-01-01

    Using a new class of nontoxic and degradable glycopolymer-based vehicles termed poly(glycoamidoamine)s, we demonstrate virus-like delivery efficacy of oligodeoxynucleotide (ODN) decoys to cardiomyoblasts (H9c2), primary cardiomyocytes, and the mouse heart. These glycopolymers bind and compact ODN decoys into nanoparticle complexes that are internalized by the cell membrane and mediate nuclear uptake of DNA in 90+% of cultured primary cardiomyocytes and 87% of the mouse myocardium. Experimental results reveal that decoys delivered via these glycopolymers block the activation of the transcription factor NF-?B, a major contributor to ischemia/reperfusion injury. Decoy complexes formed with glycopolymer T4 significantly blocked downstream gene expression of Cox-2 and limited myocardial infarction in vivo, phenocopying a transgenic mouse model. These promising delivery vehicles may facilitate high-throughput genetic approaches in animal models. Additionally, the low toxicity, biodegradation, and outstanding delivery efficacy suggest that these nanomedicines may be clinically applicable for gene regulatory therapy. PMID:22186793

  19. Combined Raman spectroscopy and autofluoresence imaging method for in vivo skin tumor diagnosis

    NASA Astrophysics Data System (ADS)

    Zakharov, V. P.; Bratchenko, I. A.; Myakinin, O. O.; Artemyev, D. N.; Khristoforova, Y. A.; Kozlov, S. V.; Moryatov, A. A.

    2014-09-01

    The fluorescence and Raman spectroscopy (RS) combined method of in vivo detection of malignant human skin cancer was demonstrated. The fluorescence analysis was used for detection of abnormalities during fast scanning of large tissue areas. In suspected cases of malignancy the Raman spectrum analysis of biological tissue was performed to determine the type of neoplasm. A special RS phase method was proposed for in vivo identification of skin tumor. Quadratic Discriminant Analysis was used for tumor type classification on phase planes. It was shown that the application of phase method provides a diagnosis of malignant melanoma with a sensitivity of 89% and a specificity of 87%.

  20. An in vivo study of turbidity suppression by optical phase conjugation (TSOPC) on rabbit ear

    PubMed Central

    Cui, Meng; McDowell, Emily J.; Yang, Changhuei

    2010-01-01

    We present a holography-based in vivo optical phase conjugation experiment performed on a living rabbit ear. The motion of live tissues caused the phase conjugate signal to decay with a consistent decay time of less than two seconds. We monitor the signal decay time variation after the ear is excised to postulate different mechanisms that cause the signal decay. The experimental findings address the minimum speed limit of a broad range of optical time reversal experiments for in vivo applications on tissues. PMID:20173817

  1. In vivo biodistribution of amino-functionalized ceria nanoparticles in rats using positron emission tomography.

    PubMed

    Rojas, Santiago; Gispert, Juan Domingo; Abad, Sergio; Buaki-Sogo, Mireia; Victor, Victor M; Garcia, Hermenegildo; Herance, Jose Raúl

    2012-12-01

    A variety of nanoparticles have been proposed for several biomedical applications. To gauge the therapeutic potential of these nanoparticles, in vivo biodistribution is essential and mandatory. In the present study, ceria nanoparticles (5 nm average particle size) were labeled with (18)F to study their in vivo biodistribution in rats by positron emission tomography (PET). The (18)F isotope was anchored by reaction of N-succinimidyl 4-[(18)F]fluorobenzoate ((18)F-SFB) with a modified nanoparticle surface obtained by silylation with 3-aminopropylsilyl. Radiolabeled ceria nanoparticles accumulated mainly in lungs, spleen, and liver. Metabolic products of the radiolabeled nanoparticulate material were excreted into the urinary tract. PMID:23140442

  2. Specific in vivo labeling with GFP retroviruses, lentiviruses, and adenoviruses for imaging

    NASA Astrophysics Data System (ADS)

    Hoffman, Robert M.; Kishimoto, Hiroyuki; Fujiwara, Toshiyoshi

    2008-02-01

    Fluorescent proteins have revolutionized the field of imaging. Our laboratory pioneered in vivo imaging with fluorescent proteins. Fluorescent proteins have enabled imaging at the subcellular level in mice. We review here the use of different vectors carrying fluorescent proteins to selectively label normal and tumor tissue in vivo. We show that a GFP retrovirus and telomerase-driven GFP adenovirus can selectively label tumors in mice. We also show that a GFP lentivirus can selectively label the liver in mice. The practical application of these results are discussed.

  3. Extradiscal ultrasound thermal therapy (ExDUSTT): evaluation in ex vivo and in vivo spine models (Invited Paper)

    NASA Astrophysics Data System (ADS)

    Diederich, Chris J.; Kinsey, Adam; Nau, William H.; Shu, Richard; Lotz, Jeffrey C.

    2005-04-01

    The application of heat to intervertebral discs is being clinically investigated for the treatment of discogenic back pain. The purpose of this study was to develop and test the feasibility of small ultrasound applicators that can be endoscopically placed adjacent to the disc, and deliver heating energy into the disc without puncturing the annular wall. Prototype devices were fabricated using curvilinear transducers (2.5-3.5 mm wide x 10 mm long, 5.4 - 6.5 MHz) that produce a narrow penetrating beam extending along the length of the ultrasound element. The transducer was affixed to either a flexible or rigid delivery catheter, and enclosed within an asymmetric coupling balloon with water-cooling flow. Bench measurements demonstrated 35-60% acoustic efficiencies, high-power output capabilities, and lightly focused beam patterns. The heating characteristics of these devices were evaluated with ex vivo and in vivo experiments within lumbar and cervical spine segments from sheep models and human cadaveric spine. The applicators were positioned adjacent to the annular wall of the surgically exposed discs. Ultrasound energy was focused directly into the disc to avoid heating the vertebral bodies. Multi-point thermocouple probes were placed throughout the disc to characterize the resultant temperature distributions. These studies demonstrated that ultrasound energy from these applicators penetrated the annular wall of the disc, and produced thermal coagulative temperatures of >60-65°C as far as 10 mm into the tissue. This study also showed that lower power levels and temperatures delivered for 10 minutes can generate a cytotoxic thermal dose of t43°C >240 min penetrating 5-10 mm from the annular wall.

  4. In Vivo Mitochondrial Oxygen Tension Measured by a Delayed Fluorescence Lifetime Technique

    PubMed Central

    Mik, Egbert G.; Johannes, Tanja; Zuurbier, Coert J.; Heinen, Andre; Houben-Weerts, Judith H. P. M.; Balestra, Gianmarco M.; Stap, Jan; Beek, Johan F.; Ince, Can

    2008-01-01

    Mitochondrial oxygen tension (mitoPO2) is a key parameter for cellular function, which is considered to be affected under various pathophysiological circumstances. Although many techniques for assessing in vivo oxygenation are available, no technique for measuring mitoPO2 in vivo exists. Here we report in vivo measurement of mitoPO2 and the recovery of mitoPO2 histograms in rat liver by a novel optical technique under normal and pathological circumstances. The technique is based on oxygen-dependent quenching of the delayed fluorescence lifetime of protoporphyrin IX. Application of 5-aminolevulinic acid enhanced mitochondrial protoporphyrin IX levels and induced oxygen-dependent delayed fluorescence in various tissues, without affecting mitochondrial respiration. Using fluorescence microscopy, we demonstrate in isolated hepatocytes that the signal is of mitochondrial origin. The delayed fluorescence lifetime was calibrated in isolated hepatocytes and isolated perfused livers. Ultimately, the technique was applied to measure mitoPO2 in rat liver in vivo. The results demonstrate mitoPO2 values of ?30–40 mmHg. mitoPO2 was highly sensitive to small changes in inspired oxygen concentration around atmospheric oxygen level. Ischemia-reperfusion interventions showed altered mitoPO2 distribution, which flattened overall compared to baseline conditions. The reported technology is scalable from microscopic to macroscopic applications, and its reliance on an endogenous compound greatly enhances its potential field of applications. PMID:18641065

  5. Melanopsin Is Highly Resistant to Light and Chemical Bleaching in Vivo*S

    E-print Network

    Palczewski, Krzysztof

    Melanopsin Is Highly Resistant to Light and Chemical Bleaching in Vivo*S Received for publication understood. Results: In situ melanopsin was partially bleached by high-intensity UV light or hydroxylamine and could be restored by cis-retinal only. Conclusion: Melanopsin resistance to light and UV bleaching

  6. In vivo continuous directed evolution.

    PubMed

    Badran, Ahmed H; Liu, David R

    2015-02-01

    The development and application of methods for the laboratory evolution of biomolecules has rapidly progressed over the last few decades. Advancements in continuous microbe culturing and selection design have facilitated the development of new technologies that enable the continuous directed evolution of proteins and nucleic acids. These technologies have the potential to support the extremely rapid evolution of biomolecules with tailor-made functional properties. Continuous evolution methods must support all of the key steps of laboratory evolution - translation of genes into gene products, selection or screening, replication of genes encoding the most fit gene products, and mutation of surviving genes - in a self-sustaining manner that requires little or no researcher intervention. Continuous laboratory evolution has been historically used to study problems including antibiotic resistance, organismal adaptation, phylogenetic reconstruction, and host-pathogen interactions, with more recent applications focusing on the rapid generation of proteins and nucleic acids with useful, tailor-made properties. The advent of increasingly general methods for continuous directed evolution should enable researchers to address increasingly complex questions and to access biomolecules with more novel or even unprecedented properties. PMID:25461718

  7. In vivo studies of opiate receptors

    SciTech Connect

    Frost, J.J.; Dannals, R.F.; Duelfer, T.; Burns, H.D.; Ravert, H.T.; Langstroem, B.; Balasubramanian, V.; Wagner, H.N. Jr.

    1984-01-01

    To study opiate receptors noninvasively in vivo using positron emission tomography, techniques for preferentially labeling opiate receptors in vivo can be used. The rate at which receptor-bound ligand clears from the brain in vivo can be predicted by measuring the equilibrium dissociation constant (KD) at 37 degrees C in the presence of 100 mM sodium chloride and 100 microM guanyl-5'-imidodiphosphate, the drug distribution coefficient, and the molecular weight. A suitable ligand for labeling opiate receptors in vivo is diprenorphine, which binds to mu, delta, and kappa receptors with approximately equal affinity in vitro. However, in vivo diprenorphine may bind predominantly to one opiate receptor subtype, possibly the mu receptor. To predict the affinity for binding to the opiate receptor, a Hansch correlation was determined between the 50% inhibitory concentration for a series of halogen-substituted fentanyl analogs and electronic, lipophilic, and steric parameters. Radiochemical methods for the synthesis of carbon-11-labeled diprenorphine and lofentanil are presented.

  8. Combining pharmacology and whole-cell patch recording from CNS neurons, in vivo

    PubMed Central

    Rose, Gary J.; Alluri, Rishi K.; Vasquez-Opazo, Gustavo A.; Odom, Stephen E.; Graham, Jalina A.; Leary, Christopher J.

    2013-01-01

    Whole-cell patch neurophysiology and pharmacological manipulations have provided unprecedented insight into the functions of central neurons, but their combined use has been largely restricted to in vitro preparations. We describe a method for performing whole-cell patch recording and focal application of pharmacological agents in vivo. A key feature of this technique involves iontophoresis of glutamate to establish proximity of drug and recording pipettes. We show data from iontophoresis of glutamate during extracellular and whole-cell recordings made in vivo from auditory neurons in the midbrain of the leopard frog, Rana pipiens, and the effects of blocking GABAA receptors while making a whole-cell recording. This methodology should accelerate our understanding of the roles of particular neurotransmitter systems in normal and pathological conditions, and facilitate investigation of the in vivo effects of drugs and the mechanisms underlying computations. PMID:23261772

  9. In vitro and in vivo antioxidant activities of polysaccharide purified from aloe vera (Aloe barbadensis) gel.

    PubMed

    Kang, Min-Cheol; Kim, Seo Young; Kim, Yoon Taek; Kim, Eun-A; Lee, Seung-Hong; Ko, Seok-Chun; Wijesinghe, W A J P; Samarakoon, Kalpa W; Kim, Young-Sun; Cho, Jin Hun; Jang, Hyeang-Su; Jeon, You-Jin

    2014-01-01

    The in vitro and in vivo antioxidant potentials of a polysaccharide isolated from aloe vera gel were investigated. Enzymatic extracts were prepared from aloe vera gel by using ten digestive enzymes including five carbohydrases and five proteases. Among them, the highest yield was obtained with the Viscozyme extract and the same extract showed the best radical scavenging activity. An active polysaccharide was purified from the Viscozyme extract using ethanol-added separation and anion exchange chromatography. Purified aloe vera polysaccharide (APS) strongly scavenged radicals including DPPH, hydroxyl and alkyl radicals. In addition, APS showed a protective effect against AAPH-induced oxidative stress and cell death in Vero cells as well as in the in vivo zebrafish model. In this study, it is proved that both the in vitro and in vivo antioxidant potentials of APS could be further utilized in relevant industrial applications. PMID:24274519

  10. In vivo photoacoustic flow cytometry for monitoring of circulating single cancer cells and contrast agents

    NASA Astrophysics Data System (ADS)

    Zharov, Vladimir P.; Galanzha, Ekaterina I.; Shashkov, Evgeny V.; Khlebtsov, Nicolai G.; Tuchin, Valery V.

    2006-12-01

    A new photoacoustic flow cytometry was developed for real-time detection of circulating cells, nanoparticles, and contrast agents in vivo. Its capability, integrated with photothermal and optical clearing methods, was demonstrated using a near-infrared tunable laser to characterize the in vivo kinetics of Indocyanine Green alone and single cancer cells labeled with gold nanorods and Indocyanine Green in the vasculature of the mouse ear. In vivo applications are discussed, including selective nanophotothermolysis of metastatic squamous cells, label-free detection of melanoma cells, study of pharmokinetics, and immune response to apoptotic and necrotic cells, with potential translation to humans. The threshold sensitivity is estimated as one cancer cell in the background of 107 normal blood cells.

  11. In vivo photoacoustic flow cytometry for monitoring of circulating single cancer cells and contrast agents.

    PubMed

    Zharov, Vladimir P; Galanzha, Ekaterina I; Shashkov, Evgeny V; Khlebtsov, Nicolai G; Tuchin, Valery V

    2006-12-15

    A new photoacoustic flow cytometry was developed for real-time detection of circulating cells, nanoparticles, and contrast agents in vivo. Its capability, integrated with photothermal and optical clearing methods, was demonstrated using a near-infrared tunable laser to characterize the in vivo kinetics of Indocyanine Green alone and single cancer cells labeled with gold nanorods and Indocyanine Green in the vasculature of the mouse ear. In vivo applications are discussed, including selective nanophotothermolysis of metastatic squamous cells, label-free detection of melanoma cells, study of pharmokinetics, and immune response to apoptotic and necrotic cells, with potential translation to humans. The threshold sensitivity is estimated as one cancer cell in the background of 10(7) normal blood cells. PMID:17130924

  12. Real time monitoring of superoxide dynamics in vivo through fluorescent proteins using a sensitive fiber probe

    NASA Astrophysics Data System (ADS)

    Chang, Yu-Chung; Ken, Chuian-Fu; Hsu, Che-Wei; Liu, Ya-Ging

    2014-03-01

    Superoxide anion is the primary oxygen free radical generated in mitochondria that causes intracellular oxidative stress. The lack of a method to directly monitor superoxide concentration in vivo in real time has severely hindered our understanding on its pathophysiology. We made transgenic zebrafish to specifically express fluorescent proteins, which are recently developed as reversible superoxide-specific indicators, in the liver. A fiber-optic fluorescent probe was used to noninvasively monitor superoxide generation in the liver in real time. The fish were placed in microfluidic channels for manipulation and reagents administration. Several superoxide-inducing and scavenging reagents were administrated onto the fish to investigate their effects on superoxide anion balancing. The biochemical dynamics of superoxide due to the application reagents were revealed in the transient behaviors of fluorescence time courses. With the ability to monitor superoxide dynamics in vivo in real time, this method can be used as an in vivo pharmaceutical screening platform.

  13. In Vivo Gene Therapy of Hemophilia B: Sustained Partial Correction in Factor IX-Deficient Dogs

    NASA Astrophysics Data System (ADS)

    Kay, Mark A.; Rothenberg, Steven; Landen, Charles N.; Bellinger, Dwight A.; Leland, Frances; Toman, Carol; Finegold, Milton; Thompson, Arthur R.; Read, M. S.; Brinkhous, Kenneth M.; Woo, Savio L. C.

    1993-10-01

    The liver represents a model organ for gene therapy. A method has been developed for hepatic gene transfer in vivo by the direct infusion of recombinant retroviral vectors into the portal vasculature, which results in the persistent expression of exogenous genes. To determine if these technologies are applicable for the treatment of hemophilia B patients, preclinical efficacy studies were done in a hemophilia B dog model. When the canine factor IX complementary DNA was transduced directly into the hepatocytes of affected dogs in vivo, the animals constitutively expressed low levels of canine factor IX for more than 5 months. Persistent expression of the clotting. factor resulted in reductions of whole blood clotting and partial thromboplastin times of the treated animals. Thus, long-term treatment of hemophilia B patients may be feasible by direct hepatic gene therapy in vivo.

  14. In vivo induced antigenic determinants of Actinobacillus actinomycetemcomitans.

    PubMed

    Cao, Sam Linsen; Progulske-Fox, Ann; Hillman, Jeffrey D; Handfield, Martin

    2004-08-01

    Actinobacillus actinomycetemcomitans is a Gram-negative capnophilic rod and the etiological agent of localized aggressive periodontitis. The genome-wide survey of A. actinomycetemcomitans using in vivo induced antigen technology (IVIAT) has previously resulted in the discovery of antigenic determinants expressed specifically in diseased patients. The present study evaluated the potential of these antigens as putative disease markers, and investigating their contribution to the pathogenesis of the microorganism. Sera from patients had a significantly greater antibody titer than sera from healthy controls against six antigens, which supports the in vivo expression of these antigens, and suggests their usefulness as disease markers. A. actinomycetemcomitans invasion of epithelium-derived HeLa cells resulted in the induction of all three genes tested, as evidenced by real-time PCR. Isogenic mutants of these three genes were constructed and the adhesion and intracellular survival of the mutants was assayed in a competition assay with the wild-type strain. A significant defect in the intracellular survival of two of these mutant strains (orf1402 and orf859) was found. This defect could not be attributed to an adhesion defect. In contrast, a mutation in vapA, a homologue of a novel putative transcriptional regulator, out-competed the wild-type strain in the same assay. The virulent phenotype was restored for a mutant strain in orf859 upon complementation. This data provided new insight into the pathogenic personality of A. actinomycetemcomitans in vivo and supported the use of HeLa cells as a valid in vitro host-pathogen interactions model for that microorganism. IVIAT is applicable to most pathogens and will undoubtedly lead to the discovery of novel therapies, antibiotics and diagnostic tools. PMID:15268943

  15. T cells enhance gold nanoparticle delivery to tumors in vivo

    PubMed Central

    2011-01-01

    Gold nanoparticle-mediated photothermal therapy (PTT) has shown great potential for the treatment of cancer in mouse studies and is now being evaluated in clinical trials. For this therapy, gold nanoparticles (AuNPs) are injected intravenously and are allowed to accumulate within the tumor via the enhanced permeability and retention (EPR) effect. The tumor is then irradiated with a near infrared laser, whose energy is absorbed by the AuNPs and translated into heat. While reliance on the EPR effect for tumor targeting has proven adequate for vascularized tumors in small animal models, the efficiency and specificity of tumor delivery in vivo, particularly in tumors with poor blood supply, has proven challenging. In this study, we examine whether human T cells can be used as cellular delivery vehicles for AuNP transport into tumors. We first demonstrate that T cells can be efficiently loaded with 45 nm gold colloid nanoparticles without affecting viability or function (e.g. migration and cytokine production). Using a human tumor xenograft mouse model, we next demonstrate that AuNP-loaded T cells retain their capacity to migrate to tumor sites in vivo. In addition, the efficiency of AuNP delivery to tumors in vivo is increased by more than four-fold compared to injection of free PEGylated AuNPs and the use of the T cell delivery system also dramatically alters the overall nanoparticle biodistribution. Thus, the use of T cell chaperones for AuNP delivery could enhance the efficacy of nanoparticle-based therapies and imaging applications by increasing AuNP tumor accumulation. PMID:21711861

  16. In Vivo Ultrasonic Detection of Polyurea Crosslinked Silica Aerogel Implants

    PubMed Central

    Sabri, Firouzeh; Sebelik, Merry E.; Meacham, Ryan; Boughter, John D.; Challis, Mitchell J.; Leventis, Nicholas

    2013-01-01

    Background Polyurea crosslinked silica aerogels are highly porous, lightweight, and mechanically strong materials with great potential for in vivo applications. Recent in vivo and in vitro studies have demonstrated the biocompatibility of this type of aerogel. The highly porous nature of aerogels allows for exceptional thermal, electric, and acoustic insulating capabilities that can be taken advantage of for non-invasive external imaging techniques. Sound-based detection of implants is a low cost, non-invasive, portable, and rapid technique that is routinely used and readily available in major clinics and hospitals. Methodology In this study the first in vivo ultrasound response of polyurea crosslinked silica aerogel implants was investigated by means of a GE Medical Systems LogiQe diagnostic ultrasound machine with a linear array probe. Aerogel samples were inserted subcutaneously and sub-muscularly in a) fresh animal model and b) cadaveric human model for analysis. For comparison, samples of polydimethylsiloxane (PDMS) were also imaged under similar conditions as the aerogel samples. Conclusion/significance Polyurea crosslinked silica aerogel (X-Si aerogel) implants were easily identified when inserted in either of the regions in both fresh animal model and cadaveric model. The implant dimensions inferred from the images matched the actual size of the implants and no apparent damage was sustained by the X-Si aerogel implants as a result of the ultrasonic imaging process. The aerogel implants demonstrated hyperechoic behavior and significant posterior shadowing. Results obtained were compared with images acquired from the PDMS implants inserted at the same location. PMID:23799093

  17. T cells enhance gold nanoparticle delivery to tumors in vivo

    NASA Astrophysics Data System (ADS)

    Kennedy, Laura C.; Bear, Adham S.; Young, Joseph K.; Lewinski, Nastassja A.; Kim, Jean; Foster, Aaron E.; Drezek, Rebekah A.

    2011-12-01

    Gold nanoparticle-mediated photothermal therapy (PTT) has shown great potential for the treatment of cancer in mouse studies and is now being evaluated in clinical trials. For this therapy, gold nanoparticles (AuNPs) are injected intravenously and are allowed to accumulate within the tumor via the enhanced permeability and retention (EPR) effect. The tumor is then irradiated with a near infrared laser, whose energy is absorbed by the AuNPs and translated into heat. While reliance on the EPR effect for tumor targeting has proven adequate for vascularized tumors in small animal models, the efficiency and specificity of tumor delivery in vivo, particularly in tumors with poor blood supply, has proven challenging. In this study, we examine whether human T cells can be used as cellular delivery vehicles for AuNP transport into tumors. We first demonstrate that T cells can be efficiently loaded with 45 nm gold colloid nanoparticles without affecting viability or function (e.g. migration and cytokine production). Using a human tumor xenograft mouse model, we next demonstrate that AuNP-loaded T cells retain their capacity to migrate to tumor sites in vivo. In addition, the efficiency of AuNP delivery to tumors in vivo is increased by more than four-fold compared to injection of free PEGylated AuNPs and the use of the T cell delivery system also dramatically alters the overall nanoparticle biodistribution. Thus, the use of T cell chaperones for AuNP delivery could enhance the efficacy of nanoparticle-based therapies and imaging applications by increasing AuNP tumor accumulation.

  18. In vivo imaging of rat cortical bone porosity by synchrotron phase contrast micro computed tomography

    NASA Astrophysics Data System (ADS)

    Pratt, I. V.; Belev, G.; Zhu, N.; Chapman, L. D.; Cooper, D. M. L.

    2015-01-01

    Cortical bone is a dynamic tissue which undergoes adaptive and pathological changes throughout life. Direct longitudinal tracking of this remodeling process holds great promise for improving our understanding of bone development, maintenance and senescence. The application of in vivo micro-computed tomography (micro-CT) has enabled longitudinal tracking of trabecular bone microarchitecture with commercially available scanners generally operating in the 10–20?µm voxel range with absorbed doses reported between 0.5 and 1?Gy. Imaging of cortical bone microarchitecture (porosity) requires higher resolution and thus in vivo imaging of these structures has not been achieved due to excessive radiation dose. In this study we tested the hypothesis that synchrotron propagation phase contrast micro-CT can enable in vivo imaging of cortical porosity in rats at doses comparable to those currently employed for trabecular bone imaging. Synchrotron imaging experiments were conducted at the Canadian Light Source using the bending magnet beamline of the BioMedical Imaging and Therapy (BMIT) facility. Protocol optimization (propagation distance, projection number) was conducted ex vivo on rat (Sprague-Dawley) forelimbs with dose determined by ion chamber and lithium fluoride crystal thermoluminescent dosimeters. Comparative ex vivo imaging was performed using laboratory in vivo scanning systems, identifying a range of doses between 1.2–3.6?Gy for common protocols. A final in vivo synchrotron protocol involving a 2.5?Gy dose was implemented with live rats. The resulting images demonstrated improved delineation of cortical porosity through the improved edge enhancement effect of phase contrast, opening the door to novel experimental studies involving the longitudinal tracking of remodeling.

  19. In vivo imaging of rat cortical bone porosity by synchrotron phase contrast micro computed tomography.

    PubMed

    Pratt, I V; Belev, G; Zhu, N; Chapman, L D; Cooper, D M L

    2015-01-01

    Cortical bone is a dynamic tissue which undergoes adaptive and pathological changes throughout life. Direct longitudinal tracking of this remodeling process holds great promise for improving our understanding of bone development, maintenance and senescence. The application of in vivo micro-computed tomography (micro-CT) has enabled longitudinal tracking of trabecular bone microarchitecture with commercially available scanners generally operating in the 10-20?µm voxel range with absorbed doses reported between 0.5 and 1?Gy. Imaging of cortical bone microarchitecture (porosity) requires higher resolution and thus in vivo imaging of these structures has not been achieved due to excessive radiation dose. In this study we tested the hypothesis that synchrotron propagation phase contrast micro-CT can enable in vivo imaging of cortical porosity in rats at doses comparable to those currently employed for trabecular bone imaging. Synchrotron imaging experiments were conducted at the Canadian Light Source using the bending magnet beamline of the BioMedical Imaging and Therapy (BMIT) facility. Protocol optimization (propagation distance, projection number) was conducted ex vivo on rat (Sprague-Dawley) forelimbs with dose determined by ion chamber and lithium fluoride crystal thermoluminescent dosimeters. Comparative ex vivo imaging was performed using laboratory in vivo scanning systems, identifying a range of doses between 1.2-3.6?Gy for common protocols. A final in vivo synchrotron protocol involving a 2.5?Gy dose was implemented with live rats. The resulting images demonstrated improved delineation of cortical porosity through the improved edge enhancement effect of phase contrast, opening the door to novel experimental studies involving the longitudinal tracking of remodeling. PMID:25489926

  20. Biofilm formation by vaginal Lactobacillus in vivo.

    PubMed

    Ventolini, G; Mitchell, E; Salazar, M

    2015-05-01

    Biofilm formation by nonpathogenic bacteria is responsible for their stable maintenance in vivo ecosystems as it promotes long-term permanence on the host's vaginal mucosa. Biofilm formation by Lactobacilli has been reported in vitro but not in vivo. We hypothesize the presence of biofilm formation in vivo could be also documented by microscope photographs (MP) of wet mounts obtained from uninfected vaginal samples satisfying rigorous scientific identification criteria. We analyzed 400 MP from our database, and we were able to determine that 12 MP from 6 different patients contained clues of the formation of biofilm by Lactobacilli. The most probable lactobacillus involved is presumed to be Lactobacillus jensenii. The documentation of biofilm formation by vaginal Lactobacilli at fresh wet mount preparation is significant and has several important clinical preventive and therapeutic implications. PMID:25725906

  1. In vivo magnetic resonance spectroscopy of liver tumors and metastases

    PubMed Central

    ter Voert, EGW; Heijmen, L; van Laarhoven, HWM; Heerschap, A

    2011-01-01

    Primary liver cancer is the fifth most common malignancy in men and the eighth in women worldwide. The liver is also the second most common site for metastatic spread of cancer. To assist in the diagnosis of these liver lesions non-invasive advanced imaging techniques are desirable. Magnetic resonance (MR) is commonly used to identify anatomical lesions, but it is a very versatile technique and also can provide specific information on tumor pathophysiology and metabolism, in particular with the application of MR spectroscopy (MRS). This may include data on the type, grade and stage of tumors, and thus assist in further management of the disease. The purpose of this review is to summarize and discuss the available literature on proton, phosphorus and carbon-13-MRS as performed on primary liver tumors and metastases, with human applications as the main perspective. Upcoming MRS approaches with potential applications to liver tumors are also included. Since knowledge of some technical background is indispensable to understand the results, a basic introduction of MRS and some technical issues of MRS as applied to tumors and metastases in the liver are described as well. In vivo MR spectroscopy of tumors in a metabolically active organ such as the liver has been demonstrated to provide important information on tumor metabolism, but it also is challenging as compared to applications on some other tissues, in particular in humans, mostly because of its abdominal location where movement may be a disturbing factor. PMID:22215937

  2. In vivo high-resolution structural imaging of large arteries in small rodents using two-photon laser scanning microscopy

    NASA Astrophysics Data System (ADS)

    Megens, Remco T. A.; Reitsma, Sietze; Prinzen, Lenneke; Oude Egbrink, Mirjam G. A.; Engels, Wim; Leenders, Peter J. A.; Brunenberg, Ellen J. L.; Reesink, Koen D.; Janssen, Ben J. A.; Ter Haar Romeny, Bart M.; Slaaf, Dick W.; van Zandvoort, Marc A. M. J.

    2010-01-01

    In vivo (molecular) imaging of the vessel wall of large arteries at subcellular resolution is crucial for unraveling vascular pathophysiology. We previously showed the applicability of two-photon laser scanning microscopy (TPLSM) in mounted arteries ex vivo. However, in vivo TPLSM has thus far suffered from in-frame and between-frame motion artifacts due to arterial movement with cardiac and respiratory activity. Now, motion artifacts are suppressed by accelerated image acquisition triggered on cardiac and respiratory activity. In vivo TPLSM is performed on rat renal and mouse carotid arteries, both surgically exposed and labeled fluorescently (cell nuclei, elastin, and collagen). The use of short acquisition times consistently limit in-frame motion artifacts. Additionally, triggered imaging reduces between-frame artifacts. Indeed, structures in the vessel wall (cell nuclei, elastic laminae) can be imaged at subcellular resolution. In mechanically damaged carotid arteries, even the subendothelial collagen sheet (~1 ?m) is visualized using collagen-targeted quantum dots. We demonstrate stable in vivo imaging of large arteries at subcellular resolution using TPLSM triggered on cardiac and respiratory cycles. This creates great opportunities for studying (diseased) arteries in vivo or immediate validation of in vivo molecular imaging techniques such as magnetic resonance imaging (MRI), ultrasound, and positron emission tomography (PET).

  3. Tracking the Relative In Vivo Pharmacokinetics of Nanoparticles with PARACEST MRI

    PubMed Central

    Ali, M. Meser; Yoo, Byunghee; Pagel, Mark D.

    2014-01-01

    A noninvasive assay that tracks the relative in vivo pharmacokinetics of two nanoparticles may accelerate the development of nanoparticles for biomedical applications, and may provide a method to select personalized nanomedicines for individual patients. To develop an in vivo competitive assay, two MRI contrast agents that could be selectively detected through paramagnetic chemical exchange saturation transfer (PARACEST) were conjugated to a second generation and fifth generation polyamidoamine (PAMAM) dendrimer. The CEST effects of each agent was calibrated relative to concentration. The effects of T1 relaxivities of these dendritic PARACEST magnetic resonance imaging (MRI) contrast agents were found to be negligible relative to their CEST effects with respect to changes in image contrast, which facilitated the measurement of the ratios of their chemical exchange lifetimes. Injection of both contrast agents into a mouse model of mammary carcinoma resulted in a temporal increase in the CEST effect from each agent in the flank tumor. Although the in vivo CEST effects could not be used to determine the absolute concentrations of each agent within the tumor, the ratio of the in vivo CEST effects was used to measure the ratio of the concentrations of the agents. This result demonstrated that the relative in vivo pharmacokinetics of two nanoparticles may be evaluated using PARACEST MRI. PMID:19298054

  4. Iodine-125 radiolabeling of silver nanoparticles for in vivo SPECT imaging

    PubMed Central

    Chrastina, Adrian; Schnitzer, Jan E

    2010-01-01

    Silver nanoparticles are increasingly finding applications in medicine; however, little is known about their in vivo tissue distribution. Here, we have developed a rapid method for radiolabeling of silver nanoparticles with iodine-125 in order to track in vivo tissue uptake of silver nanoparticles after systemic administration by biodistribution analysis and single-photon emission computerized tomography (SPECT) imaging. Poly(N-vinyl-2 -pyrrolidone)-capped silver nanoparticles with an average size of 12 nm were labeled by chemisorption of iodine-125 with a > 80% yield of radiolabeling efficiency. Radiolabeled silver nanoparticles were intravenously injected in Balb/c mice, and the in vivo distribution pattern of these nanoparticles was evaluated by noninvasive whole-body SPECT imaging, which revealed uptake of the nanoparticles in the liver and spleen. Biodistribution analysis confirmed predominant accumulation of the silver nanoparticles in the spleen (41.5%ID/g) and liver (24.5%ID/g) at 24 h. Extensive uptake in the tissues of the reticuloendothelial system suggests that further investigation of silver nanoparticle interaction with hepatic and splenic tissues at the cellular level is critical for evaluation of the in vivo effects and potential toxicity of silver nanoparticles. This method enables rapid iodine-125 radiolabeling of silver nanoparticles with a specific activity sufficient for in vivo imaging and biodistribution analysis. PMID:20856841

  5. In vivo detection of magnetic labeled oxidized multi-walled carbon nanotubes by magnetic resonance imaging

    NASA Astrophysics Data System (ADS)

    Li, Ruibin; Wu, Ren’an; Zhao, Liang; Qin, Hongqiang; Wu, Jianlin; Zhang, Jingwen; Bao, Ruyi; Zou, Hanfa

    2014-12-01

    Functionalized carbon nanotubes (f-CNTs) have been widely used in bio-medicine as drug carriers, bio-sensors, imaging agents and tissue engineering additives, which demands better understanding of their in vivo behavior because of the increasing exposure potential to humans. However, there are limited studies to investigate the in vivo biodistribution and elimination of f-CNTs. In this study, superparamagnetic iron oxides (SPIOs) were used to label oxidized multiwalled carbon nanotubes (o-MWCNTs) for in vivo distribution study of o-MWCNTs by magnetic resonance imaging (MRI). SPIO labeled o-MWCNTs ((SPIO)o-MWCNTs) were prepared by a hydrothermal reaction process, and characterized by TEM, XRD and magnetometer. (SPIO)o-MWCNTs exhibited superparamagnetic property, excellent biocompatibility and stability. The intravenously injected (SPIO)o-MWCNTs were observed in liver, kidney and spleen, while the subcutaneously injected (SPIO)o-MWCNTs could be only detected in sub mucosa. Most of the intravenously injected (SPIO)o-MWCNTs could be eliminated from liver, spleen, kidney and sub mucosa on 4 d post injection (P.I.). However, the residual o-MWCNTs could induce 30–40% MRI signal-to-noise ratio changes in these tissues even on 30 d P.I. This in vivo biodistribution and elimination information of o-MWCNTs will greatly facilitate the application of f-CNT based nanoproducts in biomedicine. In addition, the magnetic labeling method provides an approach to investigate the in vivo biodistribution and clearance of other nanomaterials.

  6. Use of in vivo magnetic resonance spectroscopy for studying metabolic diseases

    PubMed Central

    Hwang, Jong-Hee; Choi, Cheol Soo

    2015-01-01

    Owing to the worldwide obesity epidemic and the sedentary lifestyle in industrialized countries, the number of people with metabolic diseases is explosively increasing. Magnetic resonance spectroscopy (MRS), which is fundamentally similar to magnetic resonance imaging, can detect metabolic changes in vivo noninvasively. With its noninvasive nature, 1H, 13C and 31P MRS are being actively utilized in clinical and biomedical metabolic studies to detect lipids and important metabolites without ionizing radiation. 1H MRS can quantify lipid content in liver and muscle and can detect other metabolites, such as 2-hydroxyglutarate, in vivo. Of interest, many studies have indicated that hepatic and intramyocellular lipid content is inversely correlated with insulin sensitivity in humans. Thus, lipid content can be utilized as an in vivo biomarker for detecting early insulin resistance. Employing 13C MRS, hepatic glycogen synthesis and breakdown can be directly detected, whereas 31P MRS provides in vivo adenosine triphosphate (ATP) synthesis rates by saturation transfer methods in addition to ATP content. These in vivo data can be very difficult to assess by other methods and offer a critical piece of metabolic information. To aid the reader in understanding these new methods, fundamentals of MRS are described in this review in addition to promising future applications of MRS and its limitations. PMID:25656949

  7. In vivo molecular and morphological imaging by real time confocal mini-microscopy

    NASA Astrophysics Data System (ADS)

    Goetz, Martin; Gregor, Sebastian; Fottner, Christian; Garcia-Lazaro, Jose; Schirrmacher, Esther; Kempski, Oliver; Bartenstein, Peter; Weber, Mathias; Biesterfeld, Stefan; Galle, Peter R.; Neurath, Markus F.; Kiesslich, Ralf

    2006-02-01

    We evaluated a newly developed miniaturized confocal laser microscopy probe for real-time in vivo molecular and morphological imaging of normal, inflammatory, and malignant tissue in rodents. In the rigid mini-microscopy probe (diameter 7 mm), a single line laser delivers an excitation wavelength of 488 nm. Optical slice thickness is 7 ?m, lateral resolution 0.7 ?m. The range of the z-axis is 0 - 250 ?m below the tissue surface. Organ systems were examined in vivo in rodent models of human diseases. FITC-labeled Lycopersion esculentum lectin was injected or selected cell populations stained for molecular targeting. Morphological imaging was performed using fluorescein sodium, FITC-labeled dextran, and/or acriflavine hydrochloride. Cellular and subcellular details could be readily visualised in vivo at high resolution. Tissue characteristics of different organs were rendered at real time. Selective blood cell staining allowed observation of blood flow and cell migration. Inflammatory diseases such as hepatitis were diagnosed, and tumors were characterized under microscopic control in vivo. Confocal mini-microscopy allows real time in vivo molecular and morphological histologic imaging at high resolution of normal and diseased tissue. Since confocal microscopy is applicable to humans, this technology will have a high impact on different faculties in medicine.

  8. In Vivo Cellular Imaging for Translational Medical Research

    PubMed Central

    Arbab, Ali S; Janic, Branislava; Haller, Jodi; Pawelczyk, Edyta; Liu, Wei; Frank, Joseph A

    2009-01-01

    Personalized treatment using stem, modified or genetically engineered, cells is becoming a reality in the field of medicine, in which allogenic or autologous cells can be used for treatment and possibly for early diagnosis of diseases. Hematopoietic, stromal and organ specific stem cells are under evaluation for cell-based therapies for cardiac, neurological, autoimmune and other disorders. Cytotoxic or genetically altered T-cells are under clinical trial for the treatment of hematopoietic or other malignant diseases. Before using stem cells in clinical trials, translational research in experimental animal models are essential, with a critical emphasis on developing noninvasive methods for tracking the temporal and spatial homing of these cells to target tissues. Moreover, it is necessary to determine the transplanted cell’s engraftment efficiency and functional capability. Various in vivo imaging modalities are in use to track the movement and incorporation of administered cells. Tagging cells with reporter genes, fluorescent dyes or different contrast agents transforms them into cellular probes or imaging agents. Recent reports have shown that magnetically labeled cells can be used as cellular magnetic resonance imaging (MRI) probes, demonstrating the cell trafficking to target tissues. In this review, we will discuss the methods to transform cells into probes for in vivo imaging, along with their advantages and disadvantages as well as the future clinical applicability of cellular imaging method and corresponding imaging modality. PMID:19768136

  9. In vivo transcostal histotripsy therapy without aberration correction

    NASA Astrophysics Data System (ADS)

    Kim, Y.; Vlaisavljevich, E.; Owens, G. E.; Allen, S. P.; Cain, C. A.; Xu, Z.

    2014-06-01

    This study investigates the in vivo therapeutic capabilities of transcostal histotripsy without using aberration correction mechanisms and its thermal impact on overlying tissues. Non-invasive liver treatments were conducted in eight pigs, with four lesions generated through transcostal windows with full ribcage obstruction and four lesions created through transabdominal windows without rib coverage. Treatments were performed by a 750 kHz focused transducer using 5 cycle pulses at 200 Hz PRF, with estimated in situ peak negative pressures of 13-17 MPa. Temperatures on overlying tissues including the ribs were measured with needle thermocouples inserted superficially beneath the skin. Treatments of approximately 40 min were applied, allowing overlying tissue temperatures to reach saturation. Lesions yielded statistically comparable ablation volumes of 3.6 ± 1.7 cm3 and 4.5 ± 2.0 cm3 in transcostal and transabdominal treatments, respectively. The average temperature increase observed in transcostal treatments was 3.9 ± 2.1 °C, while transabdominal treatments showed an increase of 1.7 ± 1.3 °C. No damage was seen on the ribcage or other overlying tissues. These results indicate that histotripsy can achieve effective treatment through the ribcage in vivo without requiring correction mechanisms, while inducing no substantial thermal effects or damage to overlying tissues. Such capabilities could benefit several non-invasive therapy applications involving transcostal treatment windows.

  10. Photoacoustic tomography of pathological tissue in ex vivo mouse hearts

    NASA Astrophysics Data System (ADS)

    Holotta, Markus; Grossauer, Harald; Kremser, Christian; Torbica, Pavle; Völkl, Jakob; Degenhart, Gerald; Esterhammer, Regina; Nuster, Robert; Paltauf, Günther; Jaschke, Werner

    2010-02-01

    In the present study, we evaluate the applicability of ex-vivo photoacoustic imaging (PAI) in organs of small animals. We used photoacoustic tomography (PAT) to visualize infarcted areas within mouse hearts and compared it to other imaging techniques (MRI and ?CT). In order to induce ischemia an in-vivo ligation of the Ramus interventricularis anterior (RIVA, left anterior descending, LAD) was performed on nine wild type C41 mice. After varying survival periods the mice were sacrificed. The hearts were excised and immediately transferred into a formaldehyde solution for conservation. Various wavelengths in the visible and near infrared region (500 nm - 1000 nm) had been tested to find the best representation of the ischemic regions. Samples were illuminated with nanosecond laser pulses delivered by an Nd:YAG pumped optical parametric oscillator. Ultrasound detection was achieved by an optical Mach-Zehnder interferometer working as an integrating line detector. For acoustic coupling the samples were located inside a water tank. The voxel data are computed from the measurement data by a Fourier-domain based reconstruction algorithm, followed by a sequence of inverse Radon transforms. Results clearly show the capability of PAI to detect pathological tissue and the possibility to produce three-dimensional images with resolutions well below 100 ?m. Different wavelengths allow the representation of structure inside an organ or on the surface even without contrast enhancing tracers.

  11. In vivo immobilization of D-hydantoinase in Escherichia coli.

    PubMed

    Chen, Shan-Yu; Chien, Yi-Wen; Chao, Yun-Peng

    2014-07-01

    D-P-Hydroxyphenylglycine (D-HPG) is a precursor required for the synthesis of semi-synthetic antibiotics. This unnatural amino acid can be produced by a transformation reaction mediated by D-hydantoinase (D-HDT) and d-amidohydrolase. In this study, a method was developed to integrate production and immobilization of recombinant D-HDT in vivo. This was approached by first fusion of the gene encoding D-HDT with phaP (encoding phasin) of Ralstonia eutropha H16. The fusion gene was then expressed in the Escherichia coli strain that carried a heterologous synthetic pathway for polyhydroxyalkanoate (PHA). As a result, d-HDT was found to associate with isolated PHA granules. Further characterization illustrated that D-HDT immobilized on PHA exhibited the maximum activity at pH 9 and 60°C and had a half-life of 95 h at 40°C. Moreover, PHA-bound d-HDT could be reused for 8 times with the conversion yield exceeding 90%. Overall, it illustrates the feasibility of this approach to facilitate in vivo immobilization of enzymes in heterologous E. coli strain, which may open a new avenue of enzyme application in industry. PMID:24508023

  12. Vitamin E Reverses Multidrug Resistance In Vitro and In Vivo

    PubMed Central

    Tang, Jingling; Fu, Qiang; Wang, Yongjun; Racette, Kelly; Wang, Dun; Liu, Feng

    2013-01-01

    Multidrug resistance (MDR) is a major obstacle to successful and effective chemotherapeutic treatments of cancers. This study explored the reversal effects of vitamin E on MDR tumor cells in vitro and in vivo, elucidating the potential mechanism of this reversal. VE at a concentration of 50 ?M exhibited a significant reversal of the MDR effect (compared to only PTX in DMSO, p < 0.05) in two human MDR cell lines (H460/taxR and KB-8-5). The MDR cell xenograft model was established to investigate the effect of VE on reversing MDR in vivo. Mice intravenously injected with Taxol (10 mg/kg) with VE (500 mg/kg, IP) showed an ability to overcome the MDR. VE and its derivatives can significantly increase intracellular accumulation of rhodamine 123 and doxorubicin (P-gp substrate), but not alter the levels of P-gp expression. These treatments also did not decrease the levels of intracellular ATP, but were still able to inhibit the verapamil-induced ATPase activity of P-gp. The new application of VE as an MDR sensitizer will be attractive due to the safety of this treatment. PMID:23624302

  13. Dexpanthenol modulates gene expression in skin wound healing in vivo.

    PubMed

    Heise, R; Skazik, C; Marquardt, Y; Czaja, K; Sebastian, K; Kurschat, P; Gan, L; Denecke, B; Ekanayake-Bohlig, S; Wilhelm, K-P; Merk, H F; Baron, J M

    2012-01-01

    Topical application of dexpanthenol is widely used in clinical practice for the improvement of wound healing. Previous in vitro experiments identified a stimulatory effect of pantothenate on migration, proliferation and gene regulation in cultured human dermal fibroblasts. To correlate these in vitro findings with the more complex in vivo situation of wound healing, a clinical trial was performed in which the dexpanthenol-induced gene expression profile in punch biopsies of previously injured and dexpanthenol-treated skin in comparison to placebo-treated skin was analyzed at the molecular level by Affymetrix® GeneChip analysis. Upregulation of IL-6, IL-1?, CYP1B1, CXCL1, CCL18 and KAP 4-2 gene expression and downregulation of psorasin mRNA and protein expression were identified in samples treated topically with dexpanthenol. This in vivo study might provide new insight into the molecular mechanisms responsible for the effect of dexpanthenol in wound healing and shows strong correlations to previous in vitro data using cultured dermal fibroblasts. PMID:22759998

  14. In vivo bone aluminum measurements in patients with renal disease

    SciTech Connect

    Ellis, K.J.; Kelleher, S.P.

    1986-01-01

    Contamination of the dialysis solution with trace amounts of aluminum and long-term use of aluminum-based phosphate binders have led to increased body burden of aluminum in patients with end-stage renal disease. A significant clinical problem associated with aluminum-overload is the early diagnosis of aluminum-induced dialysis dementia and osteomalacic osteodystrophy. There are few, if any, blood or urine indices that provide an early monitor of this bone disease, especially in the asymptomatic patient. Although a bone biopsy is usually the basis for the final clinical diagnosis, this procedure is not recommended for routine monitoring of patients. The present technique demonstrates the direct in vivo measurement of bone aluminum levels in patients with renal failure. The interference normally present from activation of bone phosphorus is eliminated by using a thermal/epithermal neutron beam. For the clinical management of the patients, the Al/Ca ratio for the hand may be more useful than an absolute measurement of the total body or skeletal aluminum burden. The relationship between the increased serum Al levels following disferrioxamine infusion and the direct in vivo measurement of bone aluminum using the Al/Ca ratio are currently under investigation. The neutron activation procedure presented in this pilot study is a promising new technique with an immediate clinical application. 5 refs., 3 figs., 1 tab.

  15. Metabolic Flux and Compartmentation Analysis in the Brain In vivo

    PubMed Central

    Lanz, Bernard; Gruetter, Rolf; Duarte, João M. N.

    2013-01-01

    Through significant developments and progresses in the last two decades, in vivo localized nuclear magnetic resonance spectroscopy (MRS) became a method of choice to probe brain metabolic pathways in a non-invasive way. Beside the measurement of the total concentration of more than 20 metabolites, 1H MRS can be used to quantify the dynamics of substrate transport across the blood-brain barrier by varying the plasma substrate level. On the other hand, 13C MRS with the infusion of 13C-enriched substrates enables the characterization of brain oxidative metabolism and neurotransmission by incorporation of 13C in the different carbon positions of amino acid neurotransmitters. The quantitative determination of the biochemical reactions involved in these processes requires the use of appropriate metabolic models, whose level of details is strongly related to the amount of data accessible with in vivo MRS. In the present work, we present the different steps involved in the elaboration of a mathematical model of a given brain metabolic process and its application to the experimental data in order to extract quantitative brain metabolic rates. We review the recent advances in the localized measurement of brain glucose transport and compartmentalized brain energy metabolism, and how these reveal mechanistic details on glial support to glutamatergic and GABAergic neurons. PMID:24194729

  16. In Vivo Nanotoxicity Testing using the Zebrafish Embryo Assay

    PubMed Central

    Rizzo, Larissa Y.; Golombek, Susanne K.; Mertens, Marianne E.; Pan, Yu; Laaf, Dominic; Broda, Janine; Jayapaul, Jabadurai; Möckel, Diana; Subr, Vladimir; Hennink, Wim E.; Storm, Gert; Simon, Ulrich; Jahnen-Dechent, Willi; Kiessling, Fabian; Lammers, Twan

    2013-01-01

    Nanoparticles are increasingly used for biomedical purposes. Many different diagnostic and therapeutic applications are envisioned for nanoparticles, but there are often also serious concerns regarding their safety. Given the fact that numerous new nanomaterials are being developed every day, and that not much is known about the long-term toxicological impact of exposure to nanoparticles, there is an urgent need to establish efficient methods for nanotoxicity testing. The zebrafish (Danio rerio) embryo assay has recently emerged as an interesting ‘intermediate’ method for in vivo nanotoxicity screening, enabling (semi-) high-throughput analyses in a system significantly more complex than cultured cells, but at the same time also less ‘invasive’ and less expensive than large-scale biocompatibility studies in mice or rats. The zebrafish embryo assay is relatively well-established in the environmental sciences, but it has not yet gained wide notice in the nanomedicine field. Using prototypic polymeric drug carriers, gold-based nanodiagnostics and nanotherapeutics, and iron oxide-based nanodiagnostics, we here show that toxicity testing using zebrafish embryos is easy, efficient and informative, and faithfully reflects, yet significantly extends, cell-based toxicity testing. We therefore expect that the zebrafish embryo assay will become a popular future tool for in vivo nanotoxicity screening. PMID:24179674

  17. Biodegradable optode-based nanosensors for in vivo monitoring

    PubMed Central

    Balaconis, Mary K.; Clark, Heather A.

    2012-01-01

    Optode-based fluorescent nanosensors are being developed for monitoring important diseased states such as hyponatremia and diabetes. However, traditional optode-based sensors are composed of nonbiodegradable polymers such as polyvinyl chloride (PVC) raising toxicity concerns for long-term in vivo use. Here, we report the development of the first biodegradable optode-based nanosensors that maintain sensing characteristics identical to traditional optode sensors. The polymer matrix of these sensors is composed of polycaprolactone (PCL) and a citric acid ester plasticizer. The PCL-based nanosensors yielded a dynamic and reversible response to sodium, were tuned to respond to extracellular sodium concentrations, and had a lifetime of at least 14 days at physiological temperature. When in the presence of lipase, the nanosensors degraded within 4 hours at lipase concentrations found in the liver but were present after 3 days at lipase concentrations found in serum. This development of biodegradable nanosensors is not only necessary for future in vivo applications, but it has also created a new sensor platform that can be extended to other sensing mechanisms such as for small molecules or enzymes. PMID:22725692

  18. Photoacoustic tomography of ex vivo mouse hearts with myocardial infarction

    NASA Astrophysics Data System (ADS)

    Holotta, Markus; Grossauer, Harald; Kremser, Christian; Torbica, Pavle; Völkl, Jakob; Degenhart, Gerald; Esterhammer, Regina; Nuster, Robert; Paltauf, Günther; Jaschke, Werner

    2011-03-01

    In the present study, we evaluated the applicability of ex vivo photoacoustic imaging (PAI) on small animal organs. We used photoacoustic tomography (PAT) to visualize infarcted areas within murine hearts and compared these data to other imaging techniques [magnetic resonance imaging (MRI), micro-computed tomography] and histological slices. In order to induce ischemia, an in vivo ligation of the left anterior descending artery was performed on nine wild-type mice. After varying survival periods, the hearts were excised and fixed in formaldehyde. Samples were illuminated with nanosecond laser pulses delivered by a Nd:YAG pumped optical parametric oscillator. Ultrasound detection was achieved using a Mach-Zehnder interferometer (MZI) working as an integrating line detector. The voxel data were computed using a Fourier-domain based reconstruction algorithm, followed by inverse Radon transforms. The results clearly showed the capability of PAI to visualize myocardial infarction and to produce three-dimensional images with a spatial resolution of approximately 120 ?m. Regions of affected muscle tissue in PAI corresponded well with the results of MRI and histology. Photoacoustic tomography utilizing a MZI for ultrasound detection allows for imaging of small tissue samples. Due to its high spatial resolution, good soft tissue contrast and comparatively low cost, PAT offers great potentials for imaging.

  19. Quantifying in vivo somatic mutations using transgenic mouse model systems.

    PubMed

    Swiger, Roy R

    2014-01-01

    This chapter describes the use of the bacteriophage cII positive selection somatic mutational assay with the Muta™Mouse transgenic model system. The assay is similar to others involving a transgenic target, including the cII and lacI assays in the Big Blue(®) Mouse, lacZ in the MutaMouse, and the gpt delta assay. Briefly, high-molecular-weight DNA is purified from the tissue of interest and used as substrate during in vitro packaging reactions, where the ? transgenes are excised from the genome and assembled into viable phage. Phage containing the mutational targets is then adsorbed into an appropriate bacterial host, and mutations sustained in vivo are detected and quantified by either standard recombinant screening or selection assays. Mutant frequencies are reported as the ratio of mutant phage to total phage units analyzed. The ?-based transgenic mouse assays are used to study and characterize in vivo mutagenesis as well as for mutagenicity assessment of chemicals and other agents. These models permit the enumeration of mutations sustained in virtually any tissue of the mouse and are both sensitive and robust. Application of the assays is simple, not requiring resources beyond those commonly found in most academic laboratories. PMID:24623235

  20. Depleted uranium disturbs immune parameters in zebrafish, Danio rerio: an ex vivo/in vivo experiment.

    PubMed

    Gagnaire, Béatrice; Bado-Nilles, Anne; Sanchez, Wilfried

    2014-10-01

    In this study, we investigated the effects of depleted uranium (DU), the byproduct of nuclear enrichment of uranium, on several parameters related to defence system in the zebrafish, Danio rerio, using flow cytometry. Several immune cellular parameters were followed on kidney leucocytes: cell proportion, cell mortality, phagocytosis activity and associated oxidative burst and lysosomal membrane integrity (LMI). Effects of DU were tested ex vivo after 17 h of contact between DU and freshly isolated leucocytes from 0 to 500 µg DU/L. Moreover, adult zebrafish were exposed in vivo during 3 days at 20 and 250 µg DU/L. Oxidative burst results showed that DU increased reactive oxygen species (ROS) basal level and therefore reduced ROS stimulation index in both ex vivo and in vivo experiments. ROS PMA-stimulated level was also increased at 250 µg DU/L in vivo only. Furthermore, a decrease of LMI was detected after in vivo experiments. Cell mortality was also decreased at 20 µg DU/L in ex vivo experiment. However, phagocytosis activity was not modified in both ex vivo and in vivo experiments. A reduction of immune-related parameters was demonstrated in zebrafish exposed to DU. DU could therefore decrease the ability of fish to stimulate its own immune system which could, in turn, enhance the susceptibility of fish to infection. These results encourage the development and the use of innate immune analysis by flow cytometry in order to understand the effects of DU and more generally radionuclides on fish immune system and response to infectious diseases. PMID:24723161