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1

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

PubMed Central

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.

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

2009-01-01

2

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

PubMed

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

Eteshola, Edward; Pandian, Ramasamy P; Lee, Stephen C; Kuppusamy, Periannan

2009-04-01

3

Lifetime-based photoacoustic oxygen sensing in vivo  

PubMed Central

Abstract. The determination of oxygen levels in blood and other tissues in vivo is critical for ensuring proper body functioning, for monitoring the status of many diseases, such as cancer, and for predicting the efficacy of therapy. Here we demonstrate, for the first time, a lifetime-based photoacoustic technique for the measurement of oxygen in vivo, using an oxygen sensitive dye, enabling real time quantification of blood oxygenation. The results from the main artery in the rat tail indicated that the lifetime of the dye, quantified by the photoacoustic technique, showed a linear relationship with the blood oxygenation levels in the targeted artery.

Ray, Aniruddha; Rajian, Justin Rajesh; Lee, Yong-Eun Koo; Wang, Xueding; Kopelman, Raoul

2012-01-01

4

Polycythemia and oxygen sensing.  

PubMed

Polycythemias can be differentiated based on the responsiveness of erythroid progenitors to circulating cytokines. Primary polycythemias are characterized by an augmented response due to acquired somatic or inherited germ-line mutations that are expressed within hematopoietic progenitors causing increased proliferation or decreased apoptosis and resulting in accumulation of red blood cells. In terms of oxygen requirements, primary polycythemias can be viewed as the production of hemoglobin fully dissociated from the tissue oxygen needs and from the oxygen sensing pathway. Polycythemia vera (PV) is the most common primary polycythemia. PV bone marrow progenitors cells can form erythroid colonies in the absence of exogenous erythropoietin in vitro. These endogenous erythroid colonies (EEC) are useful in differentiating PV and secondary polycythemias. They also can differentiate PV where this feature is independent of Epo signalling from primary familial and congenital polycythemia. In this autosomal dominant primary polycythemia, at variance with PV, EEC formation is abolished by anti-Epo and anti-Epo receptor neutralising antibodies. Mutations of the EPOR have been described and resulted in nine cases in truncated EPORs lacking the cytoplasmic carboxy-terminal of the receptor which possesses a negative growth regulatory domain. However, recent data suggest that different mutations may cause PFCP in most cases. Secondary polycythemia can be viewed as either physiological response to satisfy the oxygen needs of the tissues, resulting for instance from high affinity hemoglobins or BPG mutase deficiency, or as the result of germ-line or somatic mutations disturbing the oxygen sensing pathway or its target: Epo. Chuvash polycythemia is a frequently symptomatic disorder with an autosomal recessive inheritance and inappropriately high Epo levels. The erythroid progenitors are hypersensitive to Epo linking this condition to both primary and secondary polycythemia. A germline missense mutation at nucleotide 598 in both alleles of the von Hippel-Lindau gene results in increased hypoxia inducible factor-1 (HIF-1) expression in normoxic conditions. HIF-1 controls the expression of many genes including Epo. Identifying causal defects in other situations like post-renal transplant erythrocytosis and cases of autosomal dominant polycythemia with high Epo levels will help further understanding of the regulation of erythropoiesis. PMID:15217714

Maran, Jey; Prchal, Josef

2004-06-01

5

Oxygen sensing in the body  

Microsoft Academic Search

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

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

2006-01-01

6

Oxygen sensing and hypoxic selection for tumors  

US Patent & Trademark Office Database

The present invention is directed to oxygen sensing. The present invention provides methods for correcting (or providing for) oxygen sensing in cells. Preferably, the methods involve supplying the SDHD gene or cybS protein, or any of the other components of the mitochondrial complex II cytochrome b oxygen sensing complex in an amount which restores oxygen sensing to affected cells, and thus facilitates normoxic conditions. More specifically, the present invention relates to mutations in the SDHD gene in human cancers and their use in the diagnosis and prognosis of mammalian cancer.

Baysal; Bora E. (Pittsburgh, PA); Ferrell; Robert E. (Pittsburgh, PA); Devlin; Bernie J. (Pittsburgh, PA); Willett-Brozick; Joan E. (Pittsburgh, PA)

2002-10-22

7

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

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.

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

2010-01-01

8

A miniature inexpensive, oxygen sensing element  

SciTech Connect

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.

Arenz, R.W.

1991-10-07

9

Oxygen Sensing: Getting Pumped by Sterols  

NSDL National Science Digital Library

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.

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

2005-06-21

10

Quality assessment of packaged foods by optical oxygen sensing  

NASA Astrophysics Data System (ADS)

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.

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

2005-11-01

11

Oxygen sensing in neuroendocrine cells and other cell types: Pheochromocytoma (PC12) cells as an experimental model  

Microsoft Academic Search

A steady supply of oxygen is an absolute requirement for mammalian cells to maintain normal cellular functions. To answer\\u000a the challenge that oxygen deprivation represents, mammals have evolved specialized cell types that can sense changes in oxygen\\u000a tension and alter gene expression to enhance oxygen delivery to hypoxic areas. These oxygen-sensing cells are rare and difficult\\u000a to study in vivo.

Zachary Spicer; David E. Millhorn

2003-01-01

12

Multifunctional mesoporous nanocomposites with magnetic, optical, and sensing features: synthesis, characterization, and their oxygen-sensing performance.  

PubMed

In this paper, the fabrication, characterization, and application in oxygen sensing are reported for a novel multifunctional nanomaterial of [Ru(bpy)(2)phen-MMS] (bpy, 2,2'-bipyridyl; phen, phenathrolin) which was simply prepared by covalently grafting the ruthenium(II) polypyridyl compounds into the channels of magnetic mesoporous silica nanocomposites (MMS). Scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, N(2) adsorption-desorption, a superconducting quantum interference device, UV-vis spectroscopy, and photoluminescence spectra were used to characterize the samples. The well-designed multifunctional nanocomposites show superparamagnetic behavior and ordered mesoporous characteristics and exhibit a strong red-orange metal-to-ligand charge transfer emission. In addition, the obtained nanocomposites give high performance in oxygen sensing with high sensitivity (I(0)/I(100) = 5.2), good Stern-Volmer characteristics (R(2) = 0.9995), and short response/recovery times (t? = 6 s and t? = 12 s). The magnetic, mesoporous, luminescent, and oxygen-sensing properties of this multifunctional nanostructure make it hold great promise as a novel multifunctional oxygen-sensing system for chemical/biosensor. PMID:23286606

Wang, Yanyan; Li, Bin; Zhang, Liming; Song, Hang

2013-01-11

13

Development of polymeric nanoprobes with improved lifetime dynamic range and stability for intracellular oxygen sensing.  

PubMed

A class of core-shell nanoparticles possessing a layer of biocompatible shell and hydrophobic core with embedded oxygen-sensitive platinum-porphyrin (PtTFPP) dyes is developed via a radical-initiated microemulsion co-polymerization strategy. The influences of host matrices and the PtTFPP incorporation manner on the photophysical properties and the oxygen-sensing performance of the nanoparticles are investigated. Self-loading capability with cells and intracellular-oxygen-sensing ability of the as-prepared nanoparticle probes in the range 0%-20% oxygen concentration are confirmed. Polymeric nanoparticles with optimized formats are characterized by their relatively small diameter (<50 nm), core-shell structures with biocompatible shells, covalent-attachment-imparted leak-free construction, improved lifetime dynamic range (up to 44 ?s), excellent storage stability and photostability, and facile cell uptake. The nanoparticles' small sensor diameter and core-shell structure with biocompatible shell make them suitable for intracellular detection applications. For intracellular detection applications, the leak-free feature of the as-prepared nanoparticle sensor effectively minimizes potential chemical interferences and cytotoxicity. As a salient feature, improved lifetime dynamic range of the sensor is expected to enable precise oxygen detection and control in specific practical applications in stem-cell biology and medical research. Such a feature-packed nanoparticle oxygen sensor may find applications in precise oxygen-level mapping of living cells and tissue. PMID:23519925

Liu, Heng; Yang, Hui; Hao, Xian; Xu, Haijiao; Lv, Yi; Xiao, Debao; Wang, Hongda; Tian, Zhiyuan

2013-03-21

14

Two-photon oxygen sensing with quantum dot-porphyrin conjugates.  

PubMed

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

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

2013-08-26

15

The Role of Redox Changes in Oxygen Sensing  

PubMed Central

The specialized oxygen-sensing tissues include the carotid body and arterial smooth muscle cells in the pulmonary artery (PA) and ductus arteriosus (DA). We discuss the evidence that changes in oxygen tension are sensed through changes in redox status. “Redox” changes imply the giving or accepting of electrons. This might occur through the direct tunneling of electrons from mitochondria or redox couples to an effector protein (eg. ion channel). Alternatively, the electron might be transferred through reactive oxygen species from mitochondria or an NADPH oxidase isoform. The PA's response to hypoxia and DA's response to normoxia result from reduction or oxidation, respectively. These opposing redox stimuli lead to K+ channel inhibition, membrane depolarization and an increase in cytosolic calcium and/or calcium sensitization that causes contraction. In the neuroendocrine cells (the type 1 cell of the carotid body, neuroepithelial body and adrenomedullary cells), the response is secretion. We examine the roles played by superoxide anion, hydrogen peroxide and the anti-oxidant enzymes in the signaling of oxygen tensions.

Weir, E. Kenneth; Archer, Stephen L.

2010-01-01

16

Development of oxygen sensing in the gills of zebrafish.  

PubMed

Previous studies have described the morphology, innervation and O(2)-chemoreceptive properties of neuroepithelial cells (NECs) of the zebrafish gill filaments. The present work describes the ontogenesis of these cells, and the formation of functional O(2)-sensing pathways in developing zebrafish. Confocal immunofluorescence was performed on whole-mount gill preparations using antibodies against serotonin (5-HT) and a zebrafish-derived neuronal marker (zn-12) to identify the appearance and innervation of gill NECs during larval stages. NECs were first expressed in gill filament primordia of larvae at 5 days postfertilization (d.p.f.) and were fully innervated by 7 d.p.f. In vivo ventilation frequency analysis revealed that a behavioural response to hypoxia (11.2+/-2.8 min(-1)) developed in embryos as early as 2 d.p.f., and a significant increase (P<0.05) in the ventilatory response to hypoxia (200.8+/-23.0 min(-1)) coincided with innervation of NECs of the filaments. In addition, exogenous application of quinidine, a blocker of O(2)-sensitive background K(+) channels in NECs, induced hyperventilation in adults in a dose-dependent manner and revealed the development of a quinidine-sensitive ventilatory response in 7 d.p.f. larvae. This study shows that NEC innervation in the gill filaments may account for the development of a functional O(2)-sensing pathway and the hyperventilatory response to hypoxia in zebrafish larvae. At earlier stages, however, O(2)-sensing must occur through another pathway. The possibility that a new type of 5-HT-positive NEC of the gill arches may account for this earlier hypoxic response is discussed. PMID:15802677

Jonz, Michael G; Nurse, Colin A

2005-04-01

17

In Vivo Applications of Fiberoptic Chemical Sensors  

Microsoft Academic Search

\\u000a As stated at the beginning of this volume, the term “biosensor” refers to sensors that use biomolecules in the molecular recognition\\u000a or transduction processes. Although there have been many proposals to use fiberoptic biosensors in vivo, almost all the work\\u000a to date has been in vitro. In the more general class of fiberoptic chemical sensors, in vivo applications have progressed

Amos Gottlieb; Skip Divers; Henry K. Hui

18

Models for oxygen sensing in yeast: implications for oxygen-regulated gene expression in higher eucaryotes  

Microsoft Academic Search

Adaptation to changes in oxygen tension in cells, tissues, and organisms depends on changes in the level of expression of a large and diverse set of proteins. It is likely that most cells and tissues possess an oxygen sensing apparatus and signal transduction pathways for regulating expression of oxygen-responsive genes. Although progress has been made in understanding the transcriptional machinery

Robert O Poyton

1999-01-01

19

Cellular mechanisms of oxygen sensing at the carotid body: heme proteins and ion channels  

Microsoft Academic Search

The purpose of this article is to highlight some recent concepts on oxygen sensing mechanisms at the carotid body chemoreceptors. Most available evidence suggests that glomus (type I) cells are the initial site of transduction and they release transmitters in response to hypoxia, which in turn depolarize the nearby afferent nerve ending, leading to an increase in sensory discharge. Two

Nanduri R Prabhakar; Jeffrey L Overholt

2000-01-01

20

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

PubMed Central

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.

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

2013-01-01

21

Fiber Optic Oxygen Sensing using Metal-Halide Cluster Luminescence  

Microsoft Academic Search

The quantitative detection of oxygen is important for industrial, automotive and medical applications. Although a number of fiber sensors have been developed based on oxygen quenching of the fluorescence from organometallic compounds, we report on a fiber probe that utilizes the luminescence from hexanuclear molybdenum chloride clusters(R. N. Ghosh et al, Appl. Phys. Lett. 75, 2885 (1999)). The advantages of

Ruby N. Ghosh

2000-01-01

22

Quality assessment of packaged foods by optical oxygen sensing  

Microsoft Academic Search

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

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

2005-01-01

23

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

NSDL National Science Digital Library

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.

Cormac Taylor (UCD Conway Institute); Jennifer C McElwain (UCD School of Biology and Environmental Sciences)

2010-10-01

24

Phosphorescent Platinum(II) and Palladium(II) Complexes with Azatetrabenzoporphyrins--New Red Laser Diode-Compatible Indicators for Optical Oxygen Sensing  

PubMed Central

A new class of oxygen indicators is described. Platinum(II) and palladium(II) complexes of azatetrabenzoporphyrins occupy an intermediate position between tetrabenzoporphyrins and phthalocyanines and combine features of both. The new dyes are excitable in the red part of the spectrum and possess strong room-temperature NIR phosphorescence. Other features include excellent spectral compatibility with the red laser diodes and 632.8 nm line of He?Ne laser, excellent photostability, and significantly shorter decay times than for the respective meso-tetraphenyltetrabenzoporphyrins. Applicability of the complexes for optical oxygen sensing is demonstrated.

2010-01-01

25

Recombinant PAS-heme domains of oxygen sensing proteins: High level production and physical characterization  

Microsoft Academic Search

Details of a high-level recombinant production method for the heme-PAS domains of heme oxygen sensing proteins from Sinorhizobium meliloti (Sm) (formerly Rhizobium meliloti, Rm), Bradyrhizobium japonicum (Bj), and Escherichia coli (Ec) are described. Using a newly proposed, concise, and unambiguous naming system (also described here) these proteins are: SmFixLH128–264, BjFixLH140–270, and EcDosH1–147. In addition, high-level production of BjFixL140–505, the soluble

Christine Suquet; Marina Savenkova; James D. Satterlee

2005-01-01

26

Synthesis, structure and oxygen-sensing properties of iridium(III)-containing coordination polymers with different cations.  

PubMed

Four iridium(III)-containing coordination polymers 1-4 using Ir(ppy)(2)(H(2)dcbpy)PF(6) (L-H(2), ppy = 2-phenylpyridine, H(2)dcbpy = 4,4'-dicarboxy-2,2'-bipyridine) as the bridging ligand, [ZnL(2)]·3DMF·5H(2)O (1), [CdL(2)(H(2)O)(2)]·3DMF·6H(2)O (2), [CoL(2)(H(2)O)(2)]·2DMF·8H(2)O (3) and [NiL(2)(H(2)O)(2)]·3DMF·6H(2)O (4), have been synthesized and structurally characterized. The emissions from 1-4 are ascribed to a metal-to-ligand charge transfer transition (MLCT). The absolute emission quantum yields for 1-4 in single crystals were measured in air to be 0.274, 0.193, 0.001 and 0.002, respectively. The noteworthy oxygen-sensing properties of 1-4 as well as L-H(2) in a single crystal were also evaluated. The Stern-Volmer quenching constant, K(SV) values, of 1-4 and L-H(2) can be deduced to be 0.834, 2.820, 1.328, 1.111 and 2.476, respectively. The results show promising K(SV) values (e.g.2) that are competitive or even larger than those of many known Ir-complexes. Moreover, the short response time (e.g. compound 2) and recovery times toward oxygen of 1-4 have been measured in their single crystal forms. The reversibility experiments for 1-4 were carried out for seven repeated cycles. As a result, >75% recovery of intensity for 1 and 2 on each cycle demonstrates a high degree of reproducibility during the sensing process. It should be noted that iridium(III)-containing coordination polymers with high emission intensity and notable oxygen sensing properties are obscure, especially in the single crystal form. This, in combination with its fine reversibility, leads to success in single crystal oxygen recognition based on photoluminescence imaging. The detection limit could be 0.50% for gaseous oxygen. Moreover, the temperature effect of compound 2 in a single crystal upon application as an oxygen sensor was expected. PMID:22222947

Ho, Mei-Lin; Chen, Yi-An; Chen, Tsai-Chen; Chang, Pei-Jen; Yu, Yi-Ping; Cheng, Kum-Yi; Shih, Chien-Hung; Lee, Gene-Hsiang; Sheu, Hwo-Shuenn

2012-01-05

27

Synthesis and characterization of a new trifunctional magnetic photoluminescent oxygen-sensing nanomaterial  

NASA Astrophysics Data System (ADS)

Magnetic Fe2O3 nanoparticles coated with SiO2 chemically doped with a Ru(II) complex were prepared using a simple solution based method. Field-emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) showed that the Fe2O3 nanoparticles with a mean diameter of ~115 nm were successfully coated with Ru(II) complex-chemically doped SiO2 shell with a thickness of ~30 nm. The obtained nanocomposite material showed a strong magnetic response to a varying magnetic field, exhibited the bright red triplet metal-to-ligand charge transfer (3MLCT) emission, and its photoluminescent intensity was sensitive to oxygen concentration. Compared with the Ru(II) complex in silica gels, the Ru(II) complex in the magnetic-optical-oxygen-sensing nanocomposite demonstrated improved thermodynamic stability of emissions. These nanocomposites are also nontoxic and easily conjugated with biomolecules. Their magnetic, photoluminescent and oxygen-sensing properties make them promising candidates for cell separation, biomarkers and optical oxygen sensors, which can measure the O2 concentration in biological bodies.

Liu, Lina; Li, Bin; Ying, Jun; Wu, Xiudong; Zhao, Haifeng; Ren, Xinguang; Zhu, Dongxia; Su, Zhongmin

2008-12-01

28

The human carotid body transcriptome with focus on oxygen sensing and inflammation - a comparative analysis  

PubMed Central

The carotid body (CB) is the key oxygen sensing organ. While the expression of CB specific genes is relatively well studied in animals, corresponding data for the human CB are missing. In this study we used five surgically removed human CBs to characterize the CB transcriptome with microarray and PCR analyses, and compared the results with mice data. In silico approaches demonstrated a unique gene expression profile of the human and mouse CB transcriptomes and an unexpected upregulation of both human and mouse CB genes involved in the inflammatory response compared to brain and adrenal gland data. Human CBs express most of the genes previously proposed to be involved in oxygen sensing and signalling based on animal studies, including NOX2, AMPK, CSE and oxygen sensitive K+ channels. In the TASK subfamily of K+ channels, TASK-1 is expressed in human CBs, while TASK-3 and TASK-5 are absent, although we demonstrated both TASK-1 and TASK-3 in one of the mouse reference strains. Maxi-K was expressed exclusively as the spliced variant ZERO in the human CB. In summary, the human CB transcriptome shares important features with the mouse CB, but also differs significantly in the expression of a number of CB chemosensory genes. This study provides key information for future functional investigations on the human carotid body.

Mkrtchian, Souren; Kahlin, Jessica; Ebberyd, Anette; Gonzalez, Constancio; Sanchez, Diego; Balbir, Alexander; Kostuk, Eric W; Shirahata, Machiko; Fagerlund, Malin Jonsson; Eriksson, Lars I

2012-01-01

29

Mitochondrial cytochrome redox states and respiration in acute pulmonary oxygen sensing.  

PubMed

Hypoxic pulmonary vasoconstriction (HPV) is an essential mechanism to optimise lung gas exchange. We aimed to decipher the proposed oxygen sensing mechanism of mitochondria in HPV. Cytochrome redox state was assessed by remission spectrophotometry in intact lungs and isolated pulmonary artery smooth muscle cells (PASMC). Mitochondrial respiration was quantified by high-resolution respirometry. Alterations were compared with HPV and hypoxia-induced functional and molecular readouts on the cellular level. Aortic and renal arterial smooth muscle cells (ASMC and RASMC, respectively) served as controls. The hypoxia-induced decrease of mitochondrial respiration paralleled HPV in isolated lungs. In PASMC, reduction of respiration and mitochondrial cytochrome c and aa3 (complex IV), but not of cytochrome b (complex III) matched an increase in matrix superoxide levels as well as mitochondrial membrane hyperpolarisation with subsequent cytosolic calcium increase. In contrast to PASMC, RASMC displayed a lower decrease in respiration and no rise in superoxide, membrane potential or intracellular calcium. Pharmacological inhibition of mitochondria revealed analogous kinetics of cytochrome redox state and strength of HPV. Our data suggest inhibition of complex IV as an essential step in mitochondrial oxygen sensing of HPV. Concomitantly, increased superoxide release from complex III and mitochondrial membrane hyperpolarisation may initiate the cytosolic calcium increase underlying HPV. PMID:20516051

Sommer, N; Pak, O; Schörner, S; Derfuss, T; Krug, A; Gnaiger, E; Ghofrani, H A; Schermuly, R T; Huckstorf, C; Seeger, W; Grimminger, F; Weissmann, N

2010-06-01

30

Low oxygen sensing and balancing in plant seeds: a role for nitric oxide  

PubMed Central

Storage product accumulation in seeds of major crop species is limited by their low internal oxygen concentration. Adjustment of energy and storage metabolism to oxygen deficiency (hypoxia) in seeds is highly relevant for agriculture and biotechnology. However, the mechanisms of low-oxygen sensing and balancing remain a mystery. Here, it is shown that normal hypoxia in seeds of soybean (Glycine max) and pea (Pisum sativum) triggers a nitrite-dependent increase in endogenous nitric oxide (NO) concentrations. NO, in turn, reduces the oxygen consumption of seeds, generating a localized decrease in both ATP availability and biosynthetic activity. Increasing oxygen availability reduces endogenous NO concentrations, thereby abolishing mitochondrial and metabolic inhibition. This auto-regulatory and reversible oxygen balancing, via NO, avoids seed anoxia and suggests a key role for NO in regulating storage activity. This hypothesis is reinforced by changes in energy status (ATP:ADP ratio), steady-state metabolite concentrations and biosynthetic fluxes under NO treatment. The proposed mechanism of low-oxygen sensing and balancing in plants offers the prospect of a new field of study in crop biotechnology. New Phytologist (2007) 176: 813–823

Borisjuk, Ljudmilla; Macherel, David; Benamar, Abdelilah; Wobus, Ulrich; Rolletschek, Hardy

2007-01-01

31

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

SciTech Connect

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.

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

2012-11-21

32

Novel optical oxygen sensing material: metalloporphyrin dispersed in fluorinated poly(aryl ether ketone) films  

Microsoft Academic Search

A series of new fluorine-containing poly(aryl ether ketone)s (8F-PEKEK(Ar); Ar: 2-2-bis(4-hydroxyphenyl)-1,1,1,3,3,3-hexafluoropropane (6FBA), 2,2-bis(4-hydroxyphenyl)propane (BA), 2-(4-hydroxyphenyl)-2-(3-hydroxyphenyl)propane (3,4?-BA) or 9,9?-bis(4-hydroxyphenyl)fluorine (HF)) are synthesized and applied to the matrix of optical oxygen sensing using phosphorescence quenching of metalloporphyrins, platinum and palladium octaethylporphyrin, (PtOEP and PdOEP) by oxygen. The phosphorescence intensity of PtOEP and PdOEP in 8F-PEKEK(Ar) films decreased with increase of oxygen concentration.

Yutaka Amao; Yumi Tabuchi; Yuhiko Yamashita; Kunio Kimura

2002-01-01

33

Emerging novel functions of the oxygen-sensing prolyl hydroxylase domain enzymes.  

PubMed

Oxygen-sensing prolyl hydroxylase domain enzymes (PHDs) target hypoxia-inducible factor (HIF)-? subunits for proteasomal degradation in normoxia through hydroxylation. Recently, novel mechanisms of PHD activation and function have been unveiled. Interestingly, PHD3 can unexpectedly amplify HIF signaling through hydroxylation of the glycolytic enzyme pyruvate kinase (PK) muscle isoform 2 (PKM2). Recent studies have also yielded insight into HIF-independent PHD functions, including the control of ?-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor trafficking in synaptic transmission and the activation of transient receptor potential cation channel member A1 (TRPA1) ion channels by oxygen levels in sensory nerves. Finally, PHD activation has been shown to involve the iron chaperoning function of poly(rC) binding protein (PCBP)1 and the (R)-enantiomer of 2-hydroxyglutarate (2-HG). The intersection of these regulatory pathways and interactions highlight the complexity of PHD regulation and function. PMID:23200187

Wong, Brian W; Kuchnio, Anna; Bruning, Ulrike; Carmeliet, Peter

2012-11-28

34

Oxygen sensing, hypoxia-inducible factor-1 and the regulation of mammalian gene expression.  

PubMed

A great many aspects of the anatomy and physiology of large animals are constrained by the need to match oxygen supply to cellular metabolism and appear likely to involve the regulation of gene expression by oxygen. Some insight into possible underlying mechanisms has been provided by studies of erythropoietin, a haemopoietic growth factor which stimulates red cell production in response to hypoxia. Studies of hypoxia-inducible cis-acting sequences from the erythropoietin gene have led to the recognition of a widespread transcriptional response to hypoxia based on the activation of a DNA-binding complex termed hypoxia-inducible factor-1 (HIF-1). Perturbation of the transcriptional response by particular transition metal ions, iron chelators and certain redox-active agents have suggested a specific oxygen sensing mechanism, perhaps involving a haem protein in a flavoprotein/cytochrome system. In addition to erythropoietin, HIF-1-responsive genes include examples with functions in cellular energy metabolism, iron metabolism, catecholamine metabolism, vasomotor control and angiogenesis, suggesting an important role in the coordination of oxygen supply and cellular metabolism. In support of this, we have demonstrated an important role for HIF-1 in tumour angiogenesis. HIF-1 itself consists of a heterodimer of two basic-helix-loop-helix proteins of the PAS family, termed HIF-1alpha and HIF-1beta, although other closely related members of this family may also contribute to the response to hypoxia. We have fused domains of HIF-1 genes to heterologous transcription factors to assay for regulatory function. These experiments have defined several domains in HIF-1alpha which can independently confer the hypoxia-inducible property, and they suggest a mechanism of HIF-1 activation in which post-translational activation/derepression of HIF-1alpha is amplified by changes in HIF-1alpha abundance most probably arising from suppression of proteolytic breakdown. Pursuit of the mechanism(s) underlying these processes should ultimately lead to better definition of the oxygen-sensing process. PMID:9510527

Ratcliffe, P J; O'Rourke, J F; Maxwell, P H; Pugh, C W

1998-04-01

35

Thiosulfate: a readily accessible source of hydrogen sulfide in oxygen sensing.  

PubMed

H2S derived from organic thiol metabolism has been proposed serve as an oxygen sensor in a variety of systems because of its susceptibility to oxidation and its ability to mimic hypoxic responses in numerous oxygen-sensing tissues. Thiosulfate, an intermediate in oxidative H2S metabolism can alternatively be reduced and regenerate H2S. We propose that this contributes to the H2S-mediated oxygen-sensing mechanism. H2S formation from thiosulfate in buffers and in a variety of mammalian tissues and in lamprey dorsal aorta was examined in real time using a polarographic H2S sensor. Inferences of intracellular H2S production were made by examining hypoxic pulmonary vasoconstriction (HPV) in bovine pulmonary arteries under conditions in which increased H2S production would be expected and in mouse and rat aortas, where reducing conditions should mediate vasorelaxation. In Krebs-Henseleit (mammalian) and Cortland (lamprey) buffers, H2S was generated from thiosulfate in the presence of the exogenous reducing agent, DTT, or the endogenous reductant dihydrolipoic acid (DHLA). Both the magnitude and rate of H2S production were greatly increased by these reductants in the presence of tissue, with the most notable effects occurring in the liver. H2S production was only observed when tissues were hypoxic; exposure to room air, or injecting oxygen inhibited H2S production and resulted in net H2S consumption. Both DTT and DHLA augmented HPV, and DHLA dose-dependently relaxed precontracted mouse and rat aortas. These results indicate that thiosulfate can contribute to H2S signaling under hypoxic conditions and that this is not only a ready source of H2S production but also serves as a means of recycling sulfur and thereby conserving biologically relevant thiols. PMID:23804280

Olson, Kenneth R; Deleon, Eric R; Gao, Yan; Hurley, Kevin; Sadauskas, Victor; Batz, Catherine; Stoy, Gilbrian F

2013-06-26

36

Laser diode based oxygen sensing: A comparison of VCSEL and DFB laser diodes emitting in the 762 nm region  

Microsoft Academic Search

The performance of VCSEL and DFB laser diodes for spectroscopic based high sensitivity oxygen sensing is compared. Detectivities of < 20 ppm m using the DFB laser diode and < 7 × 103 ppm m using the VCSEL were determined utilising wavelength modulation spectroscopy and harmonic detection. We assess factors influencing the relative performance of these devices, including spectral resolution,

V. Weldon; J. O'Gorman; J. J. Pérez-Camacho; D. McDonald; J. Hegarty; J. C. Connolly; N. A. Morris; R. U. Martinelli; J. H. Abeles

1997-01-01

37

A phosphorescent copper(I) complex: Synthesis, characterization, photophysical property, and oxygen-sensing behavior  

NASA Astrophysics Data System (ADS)

In this paper, we report the synthesis, crystal structure, photophysical properties, and electronic nature of a phosphorescent Cu(I) complex of [Cu(Phen-Np)(POP)]BF 4, where Phen-Np and POP stand for 2-(naphthalen-1-yl)-1H-imidazo[4,5-f][1,10]phenanthroline and bis(2-(diphenylphosphanyl)phenyl) ether, respectively. [Cu(Phen-Np)(POP)]BF 4 renders a yellow phosphorescence peaking at 545 nm, with a long excited state lifetime of 4.69 ?s. Density functional calculation reveals that the emission comes from a triplet metal-to-ligand-charge-transfer excited state. We electrospun composite nanofibers of [Cu(Phen-Np)(POP)]BF 4 and polystyrene (PS), hoping to explore the possibility of using the composite nanofibers as an oxygen sensing material. The finally obtained samples with average diameter of ˜300 nm exhibit a maximum sensitivity of 7.2 towards molecular oxygen with short response time of 7 s due to the large surface-area-to-volume ratio of nanofibrous membranes. No photobleaching is detected in these samples.

Wen, Caihong; Tao, Guoquan; Xu, Xinhua; Feng, Xiaoqing; Luo, Rongcheng

2011-09-01

38

Thickness Dependency of Thin Film Samaria Doped Ceria for Oxygen Sensing  

SciTech Connect

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.

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

2010-11-23

39

Oxygen sensing characteristics of limiting current-type sensors with microstructural and structural variations in diffusion barrier  

Microsoft Academic Search

Several compositional mixtures of Al2O3 (mean diameter = 1.0 ?m)-YSZ (yttria stabilized zirconia, mean diameter = 0.05 ?m) porous layers were used to study the oxygen sensing characteristics and mechanisms of the limiting current-type sensors with various microstructures of the diffusion barriers. The pore size, the porosity, the limiting current, and the degree of the normal diffusion increased with Al2O3

Jong-Heun Lee; Hoin Kim; Byung Ki Kim

1996-01-01

40

Synthesis, photophysical and oxygen-sensing properties of a novel Eu3+ complex incorporated in mesoporous MCM-41  

NASA Astrophysics Data System (ADS)

A novel Eu3+ complex of Eu(DPIQ)(TTA)3 (DPIQ=10H-dipyrido [f,h] indolo [3,2-b] quinoxaline, TTA=2-thenoyltrifluoroacetonate) was synthesized and encapsulated in the mesoporous MCM-41, hoping to explore an oxygen-sensing system based on the long-lived Eu3+ emitter. The Eu(DPIQ)(TTA)3/MCM-41 composites were characterized by infrared spectra (IR), ultraviolet-visible (UV-vis) absorption spectra, small-angle X-ray diffraction (SAXRD), luminescence intensity quenching upon various oxygen concentrations, and fluorescence decay analysis. The results indicated that the composites exhibited the characteristic emission of the Eu3+ ion and the fluorescence intensity of 5D0-7F2 obviously decreased with increasing oxygen concentrations. The oxygen sensing properties of the composites with different loading levels of Eu(DPIQ)(TTA)3 complex were investigated. A sensitivity of 3.04, a short response time of 7 s, and good linearity were obtained for the composites with a loading level of 20 mg/g. These results are the best reported values for optical oxygen-sensing materials based on Eu3+ complexes so far.

Zuo, Qinghui; Li, Bin; Zhang, Liming; Wang, Yinghui; Liu, Yanhong; Zhang, Jun; Chen, Ying; Guo, Lifan

2010-07-01

41

Enzyme substrate recognition in oxygen sensing: how the HIF trap snaps.  

PubMed

The transcriptional activator HIF (hypoxia-inducible factor) is a focal point of biomedical research because many situations in physiology and in pathology coincide with hypoxia. The effects of HIF activation may be a facet of normal growth, as in embryonic development, they may counterbalance a disease, as seen in the stimulation of erythropoiesis in anaemia, and they may be part of the pathological processes, as exemplified by tumour angiogenesis. The oxygen-sensitive alpha-subunits of HIF are primarily regulated by the enzymatic hydroxylation that induces rapid proteasomal degradation. The HIFalpha hydroxylases belong to a superfamily of dioxygenases that require the co-substrates oxygen and 2-oxoglutarate as well as the cofactors Fe2+ and ascorbate. The regulation of enzyme turnover by the concentration of the cosubstrate oxygen constitutes the interface between tissue oxygen level and the activity of HIF. The HIFalpha prolyl hydroxylases, termed PHDs/EGLNs (prolyl hydroxylase domain proteins/EGL nine homologues), bind to a conserved Leu-Xaa-Xaa-Leu-Ala-Pro motif present in all substrates identified so far. This recognition motif is present twice in HIF1alpha, which gives rise to a NODD [N-terminal ODD (oxygen-dependent degradation domain)] containing Pro402 of HIF1alpha and a CODD (C-terminal ODD) where Pro564 is hydroxylated. PHD1/EGLN2 and PHD2/EGLN1 hydroxylate both ODDs with higher activity towards CODD, whereas PHD3/EGLN3 is specific for CODD. The reason for this behaviour has been unclear. In this issue of the Biochemical Journal, Villar and colleagues demonstrate that distinct PHD/EGLN domains, that are remote from the catalytic site, function in substrate discrimination. This elegant study improves our understanding of the interaction of the oxygen-sensing PHDs/EGLNs with their substrates, which include, but are not limited to, the HIFalpha proteins. PMID:17990984

Metzen, Eric

2007-12-01

42

Aptamers against extracellular targets for in vivo applications  

Microsoft Academic Search

Oligonucleotides are multifunctional molecules which can interfere with gene expression by different mechanism such as antisense, RNA interference, ribozymes, etc. For most in vivo diagnostic and therapeutic applications, oligonucleotides need to be delivered to the intracellular compartment of a specific organ, a difficult task which limits considerably their use. However, aptamer oligonucleotides which target extracellular markers obviate this problem. Aptamers

C. Pestourie; B. Tavitian; F. Duconge

2005-01-01

43

Theory and in Vivo Application of Electroporative Gene Delivery  

Microsoft Academic Search

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

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

2000-01-01

44

In Vivo Application of Photocleavable Protein Interaction Reporter Technology  

PubMed Central

Summary In vivo protein structures and protein-protein interactions are critical to the function of proteins in biological systems. As a complementary approach to traditional protein interaction identification methods, cross-linking strategies are beginning to provide additional data on protein and protein complex topological features. Previously, photocleavable protein interaction reporter (pcPIR) technology was demonstrated by cross-linking pure proteins and protein complexes and the use of ultraviolet light to cleave or release cross-linked peptides to enable identification. In the present report, the pcPIR strategy is applied to E. coli cells and in vivo protein interactions and topologies are measured. More than 1600 labeled peptides from E. coli were identified, indicating many protein sites react with pcPIR in vivo. From those labeled sites, 53 in vivo inter-cross-linked peptide pairs were identified and manually validated. Approximately half of the interactions have been reported using other techniques, although detailed structures exist for very few. Three proteins or protein complexes with detailed crystallography structures are compared to the cross-linking results obtained from in vivo application of pcPIR technology.

Yang, Li; Zheng, Chunxiang; Weisbrod, Chad R.; Tang, Xiaoting; Munske, Gerhard R.; Hoopmann, Michael R.; Eng, Jimmy K.; Bruce, James E.

2012-01-01

45

Extremely high oxygen sensing of individual ZnSnO3 nanowires arising from grain boundary barrier modulation  

NASA Astrophysics Data System (ADS)

Extremely high oxygen sensing is realized from individual ZnSnO3 nanowires with abundant grain boundaries. The current across one single ZnSnO3 nanowire increases by about six orders of magnitude, from 1.20×10-7 to 3.78×10-1 ?A, as the oxygen pressure decreases from 3.7×104 to 1.0×10-4 Pa. Such a drastic sensing is ascribed to grain boundary barrier modulation. This interpretation is confirmed by the sensing experiments under UV illumination. The results demonstrate a promising approach to realize miniaturized and highly sensitive oxygen sensors.

Xue, X. Y.; Feng, P.; Wang, Y. G.; Wang, T. H.

2007-07-01

46

Biodegradable luminescent porous silicon nanoparticles for in vivo applications  

PubMed Central

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.

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

2011-01-01

47

Applications of nuclear technologies for in-vivo elemental analysis  

SciTech Connect

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.

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

1982-01-01

48

In vivo Coherent Raman Imaging for Neuroscience Applications  

NASA Astrophysics Data System (ADS)

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.

Cote, Daniel

2010-08-01

49

[Clinical application of bone regeneration by in vivo tissue engineering].  

PubMed

With regard to the graft materials, cortical bone block, vascularized bone flap and particulate cancellous bone and marrow (PCBM) have been used for the reconstruction of maxillofacial skeleton. Needless to say, the aim of cortical bone block and vascularized bone flap transfer is the transplantation of bony tissue of its own. On the other hand, the main objective of PCBM grafting is the transplant of osteogenic stem cells derived from uncommitted marrow mesenchymal cells. After PCBM grafting, active new bone formation occurs from osteogenic stem cells followed by bone remodeling and replacement of host bone. This process means that PCBM grafting is the method of bone regeneration that is based on in vivo tissue engineering. In this paper, clinical application of PCBM grafting for the reconstruction of maxillofacial skeleton is introduced by showing the repair of maxillary bony defect of cleft lip and palate patients, alveolar ridge augmentation and the reconstruction of large mandibular segmental defects. PMID:19043190

Iino, Mitsuyoshi; Mori, Yoshiyuki; Chikazu, Daichi; Saijyo, Hideto; Ohkubo, Kazumi; Takato, Tsuyoshi

2008-12-01

50

Engineered Biocompatible Nanoparticles for in Vivo Imaging Applications  

PubMed Central

Iron?platinum alloy nanoparticles (FePt NPs) are extremely promising candidates for the next generation of contrast agents for magnetic resonance (MR) diagnostic imaging and MR-guided interventions, including hyperthermic ablation of solid cancers. FePt has high Curie temperature, saturation magnetic moment, magneto-crystalline anisotropy, and chemical stability. We describe the synthesis and characterization of a family of biocompatible FePt NPs suitable for biomedical applications, showing and discussing that FePt NPs can exhibit low cytotoxicity. The importance of engineering the interface of strongly magnetic NPs using a coating allowing free aqueous permeation is demonstrated to be an essential parameter in the design of new generations of diagnostic and therapeutic MRI contrast agents. We report effective cell internalization of FePt NPs and demonstrate that they can be used for cellular imaging and in vivo MRI applications. This opens the way for several future applications of FePt NPs, including regenerative medicine and stem cell therapy in addition to enhanced MR diagnostic imaging.

2010-01-01

51

Study on a phosphorescent copper(I) complex and its oxygen-sensing performances upon polystyrene and MCM-41 matrixes  

NASA Astrophysics Data System (ADS)

In this paper, we synthesize a new ligand of 1-ethyl-2-(naphthalen-1-yl)-1H-imidazo[4,5-f][1,10]phenanthroline (Phen-Np-Et) and its corresponding Cu(I) complex of [Cu(Phen-Np-Et)(POP)]BF4, where POP is bis(2-(diphenylphosphanyl)phenyl) ether. The single-crystal structure, electronic nature and photophysical property of [Cu(Phen-Np-Et)(POP)]BF4 are discussed in detail. It is found that the yellow emission from [Cu(Phen-Np-Et)(POP)]BF4 owns a long excited state lifetime of 287 ?s under pure N2 atmosphere. Theoretical calculation on [Cu(Phen-Np-Et)(POP)]+ suggests that the emission comes from a triplet metal-to-ligand-charge-transfer excited state. Then, [Cu(Phen-Np-Et)(POP)]BF4 are doped into two matrixes of polystyrene and MCM-41 to investigate the oxygen-sensing performance. Finally, sensitivity maxima of 9.6 and 3.6 are achieved by the composite nanofibers of [Cu(Phen-Np-Et)(POP)]BF4/polystyrene and the [Cu(Phen-Np-Et)(POP)]BF4/MCM-41, respectively. Both samples are highly sensitive toward molecular oxygen, owing to the large surface-area-to-volume ratios of nanofibrous membranes and MCM-41 matrix.

Xu, Xiao-yong; Xiao, Han-ning; Xu, Yi-ming; Zhang, Ming-jun

52

Study on a phosphorescent copper(I) complex and its oxygen-sensing performances upon polystyrene and MCM-41 matrixes.  

PubMed

In this paper, we synthesize a new ligand of 1-ethyl-2-(naphthalen-1-yl)-1H-imidazo[4,5-f][1,10]phenanthroline (Phen-Np-Et) and its corresponding Cu(I) complex of [Cu(Phen-Np-Et)(POP)]BF(4), where POP is bis(2-(diphenylphosphanyl)phenyl) ether. The single-crystal structure, electronic nature and photophysical property of [Cu(Phen-Np-Et)(POP)]BF(4) are discussed in detail. It is found that the yellow emission from [Cu(Phen-Np-Et)(POP)]BF(4) owns a long excited state lifetime of 287 ?s under pure N(2) atmosphere. Theoretical calculation on [Cu(Phen-Np-Et)(POP)](+) suggests that the emission comes from a triplet metal-to-ligand-charge-transfer excited state. Then, [Cu(Phen-Np-Et)(POP)]BF(4) are doped into two matrixes of polystyrene and MCM-41 to investigate the oxygen-sensing performance. Finally, sensitivity maxima of 9.6 and 3.6 are achieved by the composite nanofibers of [Cu(Phen-Np-Et)(POP)]BF(4)/polystyrene and the [Cu(Phen-Np-Et)(POP)]BF(4)/MCM-41, respectively. Both samples are highly sensitive toward molecular oxygen, owing to the large surface-area-to-volume ratios of nanofibrous membranes and MCM-41 matrix. PMID:22580147

Xu, Xiao-yong; Xiao, Han-ning; Xu, Yi-ming; Zhang, Ming-jun

2012-04-23

53

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

PubMed

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

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

2013-04-22

54

FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS  

SciTech Connect

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. Alkali salts of Mo{sub 6}Cl{sub 12} were synthesized and heated to 280 C for one hour in air. Optical measurements of the thermally treated material confirm the potential of the salts as lumophores in high temperature fiber optic sensors. In addition sol-gel films containing Mo{sub 6}Cl{sub 12} were dip coated on quartz substrates and heated at 200 C for one hour. Conditions were developed for successfully immobilizing monomeric complexes that are compatible with sol-gel processing.

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

2004-07-01

55

Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants  

SciTech Connect

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.

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

2006-09-30

56

Novel luminescent Ir(III) dyes for developing highly sensitive oxygen sensing films.  

PubMed

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 p(O2) (S=1/2) and DeltaI(1%) have been also evaluated in order to establish the most sensitive membrane for controlling concentrations between 0 and 10% and low oxygen concentrations (lower than 1%), respectively. The results show that the use of nanostructured material increased the sensitivity of the film; the most sensitive membrane for controlling O(2) between 0 and 10% is based on N1001 immobilized in AP200/19 (k(sv)=2848+/-101 bar(-1) and p(O2) (S=1/2)=0.0006), and the complex N969 incorporated into AP200/19 seems to be the most suitable for applications in oxygen trace sensing (DeltaI(1%)=93.13+/-0.13%). PMID:20602945

Toro, M Marin-Suarezdel; Fernandez-Sanchez, J F; Baranoff, E; Nazeeruddin, Md K; Graetzel, M; Fernandez-Gutierrez, A

2010-05-16

57

FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS  

SciTech Connect

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.

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

2005-01-01

58

FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS  

SciTech Connect

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.

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

2005-07-01

59

FIBER OPTICAL MICRO-DETECTORS FOR OXYGEN SENSING IN POWER PLANTS  

SciTech Connect

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.

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

2005-04-01

60

Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants  

SciTech Connect

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.

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

2006-01-01

61

Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants  

SciTech Connect

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.

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

2005-10-01

62

Application of compressed sensing to in vivo 3D 19F CSI  

NASA Astrophysics Data System (ADS)

The application of CS to 3D 19F CSI enables in vivo applications in significantly reduced measurement times while preserving the relevant spectroscopic and biological information.. Display OmittedResearch highlightsApplication of CS to 3D 19F CSI. CS algorithm without regularization parameters and strict data consistency. Detailed investigation on the effect of different noise levels and acceleration factors. Reduced in vivo measurement time.

Kampf, T.; Fischer, A.; Basse-Lüsebrink, T. C.; Ladewig, G.; Breuer, F.; Stoll, G.; Jakob, P. M.; Bauer, W. R.

2010-12-01

63

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

Code of Federal Regulations, 2010 CFR

... Tissue culture media for human ex vivo tissue and cell culture processing applications... Tissue culture media for human ex vivo tissue and cell culture processing applications...maintenance of tissues and cells of human origin. The solutions...

2010-04-01

64

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

Code of Federal Regulations, 2010 CFR

... Tissue culture media for human ex vivo tissue and cell culture processing applications... Tissue culture media for human ex vivo tissue and cell culture processing applications...maintenance of tissues and cells of human origin. The solutions...

2009-04-01

65

In vivo magnetic resonance spectroscopy: basic methodology and clinical applications  

Microsoft Academic Search

The clinical use of in vivo magnetic resonance spectroscopy (MRS) has been limited for a long time, mainly due to its low\\u000a sensitivity. However, with the advent of clinical MR systems with higher magnetic field strengths such as 3 Tesla, the development\\u000a of better coils, and the design of optimized radio-frequency pulses, sensitivity has been considerably improved. Therefore,\\u000a in vivo

Marinette van der Graaf

2010-01-01

66

Polycythemia and oxygen sensing  

Microsoft Academic Search

Polycythemias can be differentiated based on the responsiveness of erythroid progenitors to circulating cytokines. Primary polycythemias are characterized by an augmented response due to acquired somatic or inherited germ-line mutations that are expressed within hematopoietic progenitors causing increased proliferation or decreased apoptosis and resulting in accumulation of red blood cells. In terms of oxygen requirements, primary polycythemias can be viewed

Jey Maran; Josef Prchal

2004-01-01

67

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

PubMed Central

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.

Yigit, Mehmet V; Medarova, Zdravka

2012-01-01

68

Hematopoietic Stem Cells: Transcriptional Regulation, Ex Vivo Expansion and Clinical Application  

PubMed Central

Maintenance of ex vivo hematopoietic stem cells (HSC) pool and its differentiated progeny is regulated by complex network of transcriptional factors, cell cycle proteins, extracellular matrix, and their microenvironment through an orchestrated fashion. Strides have been made to understand the mechanisms regulating in vivo quiescence and proliferation of HSCs to develop strategies for ex vivo expansion. Ex vivo expansion of HSCs is important to procure sufficient number of stem cells and as easily available source for HSC transplants for patients suffering from hematological disorders and malignancies. Our lab has established a nanofiber-based ex vivo expansion strategy for HSCs, while preserving their stem cell characteristics. Ex vivo expanded cells were also found biologically functional in various disease models. However, the therapeutic potential of expanded stem cells at clinical level still needs to be verified. This review outlines transcriptional factors that regulate development of HSCs and their commitment, genes that regulate cell cycle status, studies that attempt to develop an effective and efficient protocol for ex vivo expansion of HSCs and application of HSC in various non-malignant and malignant disorders. Overall the goal of the current review is to deliver an understanding of factors that are critical in resolving the challenges that limit the expansion of HSCs in vivo and ex vivo.

Aggarwal, R.; Lu, J.; Pompili, V.J.; Das, H.

2012-01-01

69

A New Crosslinkable Oxygen Sensor Covalently Bonded into Poly(2-hydroxyethyl methacrylate)-CO-Polyacrylamide Thin Film for Dissolved Oxygen Sensing  

PubMed Central

A new oxygen sensor, compound 2, was synthesized through a chemical modification of a popularly used oxygen sensor of platinum(II)-5,10,15,20-tetrakis-(2,3,4,5,6-pentafluorophenyl)-porphyrin (PtTFPP). The new sensor compound 2 possesses four crosslinkable methacrylate functional moieties, enabling it to be polymerized and crosslinked with other monomers for polymer sensing film (also called membrane) preparation. Using this characteristic, compound 2 was covalently bonded to hydrophilic poly(2-hydroxyethyl methacrylate)-co-polyacrylamide (referred to as PHEMA to simplify) and hydrophobic polystyrene (PS) films. To better understand the advantages and disadvantages of chemical crosslinking approaches and the influence of polymer matrices on sensing performance, PtTFPP was physically incorporated into the same PHEMA and PS matrices to compare. Response to dissolved oxygen (DO), leaching of the sensor molecules from their matrices, photostability of the sensors, and response time to DO changes were studied. It was concluded that the chemical crosslinking of the sensor compound 2 in polymer matrices: (i) alleviated the leaching problem of sensor molecules which usually occurred in the physically doped sensing systems and (ii) significantly improved sensors’ photostability. The PHEMA matrix was demonstrated to be more suitable for oxygen sensing than PS, because for the same sensor molecule, the oxygen sensitivity in PHEMA film was higher than that in PS and response time to DO change in the PHEMA film was faster than that in PS. It was the first time oxygen sensing films were successfully prepared using biocompatible hydrophilic PHEMA as a matrix, which does not allow leaching of the sensor molecules from the polymer matrix, has a faster response to DO changes than that of PS, and does not present cytotoxicity to human lung adenocarcinoma epithelial cells (A549). It is expected that the new sensor compound 2 and its similar compounds with chemically crosslinking characteristics can be widely applied to generate many interesting oxygen sensing materials for studying biological phenomena.

Tian, Yanqing; Shumway, Bradley R.; Meldrum, Deirdre R.

2010-01-01

70

In Vivo Application of Optogenetics for Neural Circuit Analysis  

PubMed Central

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.

2012-01-01

71

In-vivo neutron activation analysis: principles and clinical applications  

SciTech Connect

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.

Cohn, S.H.

1982-01-01

72

Clinical applications of in vivo neutron-activation analysis  

SciTech Connect

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.

Cohn, S.H.

1982-01-01

73

Towards in vivo application of RNA interference - new toys, old problems  

PubMed Central

RNA interference (RNAi) is the sequence-specific degradation of mRNA by short double-stranded RNA molecules. The technology, introduced only 5 years ago, has stimulated many fantasies regarding the future of functional gene analysis and gene therapy. Given its ease of application, its high efficiency and remarkable specificity, RNAi holds great promise for broad in vitro and in vivo application in all areas of biomedicine. Despite its potential, the major obstacle to the use of RNAi (as for all previous gene silencing approaches) is the need for efficient and sustained delivery of small interfering RNA into primary mammalian cells, and specific targeting of particular cell types in vivo.

Rutz, Sascha; Scheffold, Alexander

2004-01-01

74

Versatility of biodegradable biopolymers: degradability and an in vivo application  

Microsoft Academic Search

Biodegradable materials have various important applications in the biomedical field. There are basically two groups of polyesters which have significant importance in this field. These are polylactides and polyhydroxybutyrates. Both groups degrade via hydrolysis with the rates of degradation depending on medium properties such as pH, temperature, solvent and presence of biocatalysts, as well as on chemical compositions. In order

V. Has?rc?; K. Lewandrowski; J. D. Gresser; D. L. Wise; D. J. Trantolo

2001-01-01

75

Optogenetic tools for in vivo applications in neonatal mice  

NASA Astrophysics Data System (ADS)

Spontaneous neural activities exist early in development and their spatiotemporal patterns play important roles in the development of sensory maps such as maps of retinotopy in the visual system. We summarized different optogenetic tools, including transgenic mouse lines, viral-mediated transfection and electroporation methods to enable the expression of light-gated channelrhodopsin (ChR2) in retinal ganglion cells (RGCs) before the onset of vision. Patch-clamp and extracellular recording experiments verified that activities of ChR2-expressing cells were precisely manipulated by the patterns of optical stimuli. In chronic stimulation experiments, light-emitting diodes controlled the activity patterns of ChR2-expressing RGCs in vivo. Changes in the retinotopic map in Superior Colliculus (SC) were examined by quantifying the relative sizes of fluorescently labeled target zones. Our results revealed that various optogenetic and optical tools can manipulate retinal activities with precise temporal patterns. These techniques can be readily used in studying the development of the central nervous system of neonatal rodents.

Zhang, Yue; Qin, Nan; Diao, Yupu; Guan, Yangtai; Fan, Lu; Crair, Michael C.; Zhang, Jiayi

2012-10-01

76

Histotripsy for Pediatric Cardiac Applications: In Vivo Neonatal Pig Model  

NASA Astrophysics Data System (ADS)

This study investigated the in vivo feasibility of using histotripsy to non-invasively create a flow channel between the ventricles by generating a perforation of the ventricular septum, clinically referred to as a ventricular septum defect (VSD). The overall goal is to develop a non-invasive procedure to aid in the treatment of neonatal patients with complex congenital heart diseases such as Hypoplastic Left Heart Syndrome (HLHS). Histotripsy is a therapeutic ultrasound technique that produces mechanical fractionation of soft tissue through controlled cavitation. The study was conducted in a live and intact neonatal pig model. The ventricular septum in the neonatal pig heart was treated with histotripsy delivered by a spherically focused 1 MHz transducer positioned outside the chest wall. Histotripsy treatment was applied using 5-cycle ultrasound pulses at 1 kHz pulse repetition frequency with 12-18 MPa peak negative pressure. The treatment was guided and monitored with ultrasound imaging. In all nine subjects treated, a bubble cloud was generated on the ventricular septum using histotripsy, and visualized with ultrasound imaging. Within 20 seconds to 4 minutes following the initiation of a bubble cloud, a VSD was created in all nine pigs and confirmed by the detection of blood flow through the ventricular septum with color Doppler ultrasound. Gross morphology and histology on all hearts showed a demarcated perforation in the ventricular septum. This study shows that a VSD can be created in an intact neonatal animal using extracorporeal histotripsy under real-time ultrasound guidance.

Miller, Ryan M.; Owens, Gabe; Ensing, Gregory; Ludomirsky, Achiau; Cain, Charles; Xu, Zhen

2010-03-01

77

Biocompatible PEGylated gold nanorods as colored contrast agents for targeted in vivo cancer applications  

NASA Astrophysics Data System (ADS)

In this contribution, we report the use of a PEGylated gold nanorods formulation as a colored dye for tumor labeling in vivo. We have demonstrated that the nanorod-targeted tumor site can be easily differentiated from the background tissues by the 'naked eye' without the need of sophisticated imaging instruments. In addition to tumor labeling, we have also performed in vivo toxicity and biodistribution studies of PEGylated gold nanorods in vivo by using BALB/c mice as the model. In vivo toxicity studies indicated no mortality or adverse effects or weight changes in BALB/c mice treated with PEGylated gold nanorods. This finding will provide useful guidelines in the future development of diagnostic probes for cancer diagnosis, optically guided tumor surgery, and lymph node mapping applications.

Kopwitthaya, Atcha; Yong, Ken-Tye; Hu, Rui; Roy, Indrajit; Ding, Hong; Vathy, Lisa A.; Bergey, Earl J.; Prasad, Paras N.

2010-08-01

78

Application of POLARIC™ fluorophores in an in vivo tumor model.  

PubMed

Fluorescent and luminescent tools are commonly used to study the dynamics of cancer progression and metastases in real?time. Fluorophores have become essential tools to study biological events. However, few can sustain fluorescence long enough during long-term studies. In the present study, we focused on a series of new amphiphilic fluorophores known as POLARIC™, which emit strong fluorescence in lipid bilayers and can be readily modified using the Suzuki-Miyaura cross?coupling reaction. Appropriate chemical modifications of substituent groups can improve target-site specificity, reduce cytotoxicity and prolong emission. Therefore, in contrast to conventional fluorescent probes, these fluorophores show promise for long-term monitoring of biological processes. In the present study, we conducted long-term observations of tumor growth and metastasis using a POLARIC derivative as a novel fluorescent probe. For this purpose, we studied the metastatic melanoma cell line A375-SM, which proliferates at a high rate. We compared the characteristics of the POLARIC probe with the commercially available fluorescent dye PKH26 and fluorescent protein mRFP1. A375-SM cells were labeled with these fluorescent probes and orthotopically implanted into nude mice. The fluorescence emitted by POLARIC was detected more than five weeks after implantation without causing detectable harmful effects on tumor growth. By contrast, fluorescence of cells labeled with PKH26 could not be detected at this same time. Furthermore, POLARIC-, but not PKH26-labeled cells, were also detected in lung metastases. These results indicate that labeling cells with POLARIC fluorophores can significantly extend the time course of in vivo studies on tumor cell growth. PMID:23877868

Maishi, Nako; Kawamoto, Taisuke; Ohga, Noritaka; Yamada, Koji; Akiyama, Kosuke; Yamamoto, Kazuyuki; Osawa, Takahiro; Hida, Yasuhiro; Hida, Kyoko

2013-07-18

79

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

Microsoft Academic Search

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

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

2003-01-01

80

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

Microsoft Academic Search

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

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

2010-01-01

81

What does it take to improve existing fluorescent proteins for in vivo imaging applications?  

PubMed

Although fluorescent proteins are ubiquitously used as genetic tracers and imaging agents, there is significant room for improvement. This chapter discusses how new improved fluorescent proteins can be designed. It focuses on the design of far-red and infrared fluorescent proteins, since the currently-available red fluorescent proteins are not optimal for in vivo applications. PMID:22700415

Zimmer, Marc

2012-01-01

82

Application of locked nucleic acids to improve aptamer in vivo stability and targeting function  

Microsoft Academic Search

Aptamers are powerful candidates for molecular imaging applications due to a number of attractive features, including rapid blood clearance and tumor penetration. We carried out structure-activity rela- tionship (SAR) studies with the Tenascin-C binding aptamer TTA1, which is a promising candidate for application in tumor imaging with radioisotopes. The aim was to improve its in vivo stability and target binding.

Kathrin S. Schmidt; Sandra Borkowski; Jens Kurreck; Andrew W. Stephens; Rolf Bald; Maren Hecht; Matthias Friebe; Ludger Dinkelborg; Volker A. Erdmann

2004-01-01

83

In vivo confocal microscopy in dermatology: from research to clinical application.  

PubMed

Confocal laser scanning microscopy (CLSM) represents an emerging technique for the noninvasive histomorphological analysis of skin in vivo and has shown its applicability for dermatological research as well as its value as an adjunct tool in the clinical management of skin cancer patients. Herein, we aim to give an overview on the current clinical indications for CLSM in dermatology and also highlight the diverse applications of CLSM in dermatological research. PMID:23338938

Ulrich, Martina; Lange-Asschenfeldt, Susanne

2013-06-01

84

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

NASA Astrophysics Data System (ADS)

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.

Schultz, Kristin N.; Kennedy, Robert T.

2008-07-01

85

Towards a versatile platform based on magnetic nanoparticles for in vivo applications  

Microsoft Academic Search

Magnetic nanoparticles have attracted wide attention because of their usefulness as contrast agents for magnetic resonance\\u000a imaging (MRI) or colloidal mediators for cancer magnetic hyperthermia. This paper examines these in vivo applications through an understanding of the problems involved and the current and future possibilities for resolving them.\\u000a A special emphasis is made on magnetic nanoparticle requirements from a physical

E Duguet; S Vasseur; S Mornet; G Goglio; A Demourgues; J Portier; F Grasset; P Veverka; E Pollert

2006-01-01

86

In vivo molecular imaging using nanomaterials: general in vivo characteristics of nano-sized reagents and applications for cancer diagnosis.  

PubMed

Nanoparticles present a new collection of contrast agents for the field of in vivo molecular imaging. This review focuses on promising molecular imaging probes for optical and magnetic resonance imaging based on four representative nanomaterial(s) platforms: quantum dots, upconversion phosphors, superparamagnetic iron oxides, and dendrimer-based agents. Quantum dots are extremely efficient fluorescent nanoparticles with size-tunable emission properties, enabling high sensitivity and greater depth penetration. Their heavy metal composition and long retention in the body, however, pose concerns for clinical translational applications. Upconversion phosphors generate excellent signal-to-background contrast because they emit light with higher energy than the excitation photons and autofluorescence signals. For MRI, iron oxide particles also generate excellent signal and have been used in liver imaging and for cell tracking studies. As they are metabolized through endogenous iron salvage pathways, they have already been introduced as clinical contrast agents. Lastly, dendrimers, a 'soft' nanoparticle, can be used as a structural basis for the attachment of small molecule imaging agents and/or targeting groups. This array of nanoparticles should offer insights into the uses and potentials of nanoparticles for the molecular imaging. PMID:20455640

Rosenblum, Lauren T; Kosaka, Nobuyuki; Mitsunaga, Makoto; Choyke, Peter L; Kobayashi, Hisataka

2010-05-10

87

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

NASA Astrophysics Data System (ADS)

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.

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

2010-08-01

88

The in vivo transport of elastic vesicles into human skin: effects of occlusion, volume and duration of application  

Microsoft Academic Search

In the present study, several aspects of elastic vesicle transport into human skin were investigated in vivo. Surfactant-based elastic vesicles were applied onto human skin in vivo and subsequently a series of tape-strippings were performed, which were visualised by freeze fracture electron microscopy. Factors of investigation for non-occlusive treatment were the duration of application and the volume of application. In

P. Loan Honeywell-Nguyen; H. W. Wouter Groenink; Anko M. de Graaff; Joke A. Bouwstra

2003-01-01

89

Oxygen sensing in yeast: Evidence for the involvement of the respiratory chain in regulating the transcription of a subset of hypoxic genes  

PubMed Central

Oxygen availability affects the transcription of a number of genes in nearly all organisms. Although the molecular mechanisms for sensing oxygen are not precisely known, heme is thought to play a pivotal role. Here, we address the possibility that oxygen sensing in yeast, as in mammals, involves a redox-sensitive hemoprotein. We have found that carbon monoxide (CO) completely blocks the anoxia-induced expression of two hypoxic genes, OLE1 and CYC7, partially blocks the induction of a third gene, COX5b, and has no effect on the expression of other hypoxic or aerobic genes. In addition, transition metals (Co and Ni) induce the expression of OLE1 and CYC7 in a concentration-dependent manner under aerobic conditions. These findings suggest that the redox state of an oxygen-binding hemoprotein is involved in controlling the expression of at least two hypoxic yeast genes. By using mutants deficient in each of the two major yeast CO-binding hemoproteins (cytochrome c oxidase and flavohemoglobin), respiratory inhibitors, and cob1 and ?0 mutants, we have found that the respiratory chain is involved in the anoxic induction of these two genes and that cytochrome c oxidase is likely the hemoprotein “sensor.” Our findings also indicate that there are at least two classes of hypoxic genes in yeast (CO sensitive and CO insensitive) and imply that multiple pathways/mechanisms are involved in modulating the expression of hypoxic yeast genes.

Kwast, Kurt E.; Burke, Patricia V.; Staahl, Brett T.; Poyton, Robert O.

1999-01-01

90

Fabricated Micro-Nano Devices for In vivo and In vitro Biomedical Applications.  

PubMed

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. WIREs Nanomed Nanobiotechnol 2013, 5:544-568. doi: 10.1002/wnan.1227 Conflict of interest: The authors have declared no conflicts of interest for this article. For further resources related to this article, please visit the WIREs website. PMID:23894041

Barkam, Swetha; Saraf, Shashank; Seal, Sudipta

2013-07-26

91

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

SciTech Connect

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)

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

1981-01-01

92

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

PubMed

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 imaging enables 3D mapping of the distribution of a given free radical while spectral-spa-tial EPR imaging enables mapping of the spectral information at each spatial position, and, from the observed line width, the localized tissue oxygenation can be determined. A variety of spatial, and spectral-spatial EPR imaging applications have been performed. These techniques, along with the use of biocompatible paramagnetic probes including particulate suspensions and soluble nitroxide radicals, enable spatial imaging of the redox state and oxygenation in a variety of biomedical applications. With spectral-spatial EPR imaging, oxygenation was mapped within the gastrointestinal (GI) tract of living mice, enabling measurement of the oxygen gradient from the proximal to the distal GI tract. Using spatial EPR imaging, the distribution and metabolism of nitroxide radicals within the major organs of the body of living mice was visualized and anatomically co-registered by proton MRI enabling in vivo mapping of the redox state and radical clearance. EPR imaging techniques have also been applied to non-invasively measure the distribution and metabolism of topically applied nitroxide redox probes in humans, providing information regarding the penetration of the label through the skin and measurement of its redox clearance. Thus, EPR spectroscopy and imaging has provided important information in a variety of applications ranging from small animal models of disease to topical measurement of redox state in humans. PMID:12162454

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

93

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

Microsoft Academic Search

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

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

2009-01-01

94

AirSR, a [2Fe-2S] Cluster-Containing Two-Component System, Mediates Global Oxygen Sensing and Redox Signaling in Staphylococcus aureus  

PubMed Central

Oxygen sensing and redox signaling significantly affect bacterial physiology and host-pathogen interaction. Here we show that a Staphylococcus aureus two-component system, AirSR (anaerobic iron-sulfur cluster-containing redox sensor regulator, formerly YhcSR), responds to oxidation signals (O2, H2O2, NO, etc) by using a redox-active [2Fe-2S] cluster in the sensor kinase AirS. Mutagenesis studies demonstrate that the [2Fe-2S] cluster is essential for the kinase activity of AirS. We have also discovered that a homolog of IscS (SA1450) in S. aureus is active as a cysteine desulfurase, which enables the in vitro reconstitution of the [2Fe-2S] cluster in AirS. Phosphorylation assays show that the oxidized AirS with a [2Fe-2S]2+ cluster is the fully active form of the kinase but not the apo-AirS nor the reduced AirS possessing a [2Fe-2S]+ cluster. Over-oxidation by prolonged exposure to O2 or contact with H2O2 or NO led to inactivation of AirS. Transcriptome analysis revealed that mutation of airR impacts the expression of ~355 genes under anaerobic conditions. Moreover, the mutant strain displayed increased resistance toward H2O2, vancomycin, norfloxacin, and ciprofloxacin under anaerobic conditions. Together, our results show that S. aureus AirSR is a redox-dependent global regulatory system that plays important roles in gene regulation using a redox active Fe-S cluster under O2-limited conditions.

Sun, Fei; Ji, Quanjiang; Jones, Marcus B.; Deng, Xin; Liang, Haihua; Frank, Bryan; Telser, Joshua; Peterson, Scott N.; Bae, Taeok; He, Chuan

2011-01-01

95

An Automated Method to Quantify Microglia Morphology and Application to Monitor Activation State Longitudinally In Vivo  

PubMed Central

Microglia are specialized immune cells of the brain. Upon insult, microglia initiate a cascade of cellular responses including a characteristic change in cell morphology. To study the dynamics of microglia immune response in situ, we developed an automated image analysis method that enables the quantitative assessment of microglia activation state within tissue based solely on cell morphology. Per cell morphometric analysis of fluorescently labeled microglia is achieved through local iterative threshold segmentation, which reduces errors caused by signal-to-noise variation across large volumes. We demonstrate, utilizing systemic application of lipopolysaccharide as a model of immune challenge, that several morphological parameters, including cell perimeter length, cell roundness and soma size, quantitatively distinguish resting versus activated populations of microglia within tissue comparable to traditional immunohistochemistry methods. Furthermore, we provide proof-of-concept data that monitoring soma size enables the longitudinal assessment of microglia activation in the mouse neocortex imaged via 2-photon in vivo microscopy. The ability to quantify microglia activation automatically by shape alone allows unbiased and rapid analysis of both fixed and in vivo central nervous system tissue.

Kozlowski, Cleopatra; Weimer, Robby M.

2012-01-01

96

In vivo optical investigation of short term skin water contact and moisturizer application using NIR spectroscopy.  

PubMed

Nowadays, a number of noninvasive methods and instruments are available to inspect the biophysical properties and effects of various applicants on human skin, providing quantitative measurements and more details regarding the interactions between skin and various products. Such methods include Near Infrared Spectroscopy (NIRS), a technique which over the years, has gained quite a reputation in being able to accurately determine moisture levels and water contents due to its sensitivity to hydrogen bonding. This paper reports preliminary results of an in vivo study carried out on the skin of a small number of human participants, investigating the optical response of human skin after direct short-term contact with water followed by application of a moisturizer, using a highly advanced spectrophotometer in the region of 900-2100nm, and equipped with a reflectance fibre optic probe. Results obtained here certainly raise some questions regarding the optical characteristics of different skin types and the influence of frequent moisturizer use, as well as the varying response between different water bands in the NIR region. Future work will focus on gaining more knowledge about these, in order to further improve optical skin measurements, and hopefully support the design and development of a portable and/or miniaturized optical device that could provide reliable, accurate and fast skin hydration readings in real time. PMID:24110207

Qassem, M; Kyriacou, P A

2013-07-01

97

Application of the laser capture microdissection technique for molecular definition of skeletal cell differentiation in vivo.  

PubMed

Laser capture microdissection (LCM) method allows selection of individual or clustered cells from intact tissues. This technology enables one to pick cells from tissues that are difficult to study individually, sort the anatomical complexity of these tissues, and make the cells available for molecular analyses. Following the cells' extraction, the nucleic acids and proteins can be isolated and used for multiple applications that provide an opportunity to uncover the molecular control of cellular fate in the natural microenvironment. Utilization of LCM for the molecular analysis of cells from skeletal tissues will enable one to study differential patterns of gene expression in the native intact skeletal tissue with reliable interpretation of function for known genes as well as to discover novel genes. Variability between samples may be caused either by differences in the tissue samples (different areas isolated from the same section) or some variances in sample handling. LCM is a multi-task technology that combines histology, microscopy work, and dedicated molecular biology. The LCM application will provide results that will pave the way toward high throughput profiling of tissue-specific gene expression using Gene Chip arrays. Detailed description of in vivo molecular pathways will make it possible to elaborate on control systems to apply for the repair of genetic or metabolic diseases of skeletal tissues. PMID:18463821

Benayahu, Dafna; Socher, Rina; Shur, Irena

2008-01-01

98

Application of gold quenching of luminescence to improve oxygen sensing using a ruthenium (4,7-diphenyl-1,10-phenanthroline) 3Cl 2:TEOS thin film  

Microsoft Academic Search

A new method for enhancement of luminescence from sol–gel based sensors is demonstrated, utilising the quenching effect and filtering effect of a gold film. Ruthenium (4,7-diphenyl-1,10-phenanthroline)3Cl2 is encapsulated in a trimethoxysilane (TEOS) xerogel and spin-coated to an approximate thickness of less than 100nm. Using off axis illumination of the xerogel film, luminescence was collected using a photodiode below the plane

Philip J. R. Roche; Maurice C.-K. Cheung; Ka Yi Yung; Andrew G. Kirk; Vamsy P. Chodavarpu; Frank V. Bright

2010-01-01

99

In vitro and in vivo characterization of novel biomaterial for transdermal application.  

PubMed

Polymers have become an indispensable part in the design of a conventional as well as novel drug delivery system. Gum Copal (GC), a novel biomaterial obtained from Agathis species, is evaluated in the present study for its potential application as a matrix former in transdermal drug delivery systems. GC was initially characterized for various physicochemical properties and then mechanical characterization of the Plasticized films of GC was investigated. Verapamil hydrochloride (VH), owing to its pharmacokinetic properties, was selected as the model drug for the present work. Matrix type transdermal films of VH with GC, alone and in combination with polyvinyl pyrrolidone (PVP K-30), were developed and evaluated for various physicochemical properties. In-vitro drug release study was carried out using paddle over disk method and in-vitro skin permeation study was performed using human cadaver skin. On the basis of physicochemical properties, in-vitro drug release study and permeation performance, formulation F5 containing GC: PVP K-30 (60:40) was selected as an optimized formulation for in vivo study. Animal studies were carried out using Dawley rats and the data obtained from the plasma drug analysis showed that peak drug concentration of about 244.94 ± 1.25 ng/mL was achieved in 6 h after the application of the patch and plasma drug concentration was maintained till 24 h. Skin irritancy test results proved the suitability of the biomaterial for transdermal application. The drug polymer interaction studies carried out using UV, FTIR and TLC analysis indicated that drug and polymer were compatible. Due to reasonably good mechanical properties, low water vapor transmission and sustained release capability, GC seems to be a promising film former for transdermal drug delivery. PMID:21696352

Mundada, Atish S; Avari, Jasmine G

2011-09-01

100

In vivo bio-safety evaluations and diagnostic/therapeutic applications of chemically designed mesoporous silica nanoparticles.  

PubMed

The remarkable progress of nanotechnology and its application in biomedicine have greatly expanded the ranges and types of biomaterials from traditional organic material-based nanoparticles (NPs) to inorganic biomaterials or organic/inorganic hybrid nanocomposites due to the unprecedented advantages of the engineered inorganic material-based NPs. Colloidal mesoporous silica NPs (MSNs), one of the most representative and well-established inorganic materials, have been promoted into biology and medicine, and shifted from extensive in vitro research towards preliminary in vivo assays in small-animal disease models. In this comprehensive review, the recent progresses in chemical design and engineering of MSNs-based biomaterials for in vivo biomedical applications has been detailed and overviewed. Due to the intrinsic structural characteristics of elaborately designed MSNs such as large surface area, high pore volume and easy chemical functionalization, they have been extensively investigated for therapeutic, diagnostic and theranostic (concurrent diagnosis and therapy) purposes, especially in oncology. Systematic in vivo bio-safety evaluations of MSNs have revealed the evidences that the in vivo bio-behaviors of MSNs are strongly related to their preparation prodecures, particle sizes, geometries, surface chemistries, dosing parameters and even administration routes. In vivo pharmacokinetics and pharmacodynamics further demonstrated the effectiveness of MSNs as the passively and/or actively targeted drug delivery systems (DDSs) for cancer chemotherapy. Especially, the advance of nano-synthetic chemistry enables the production of composite MSNs for advanced in vivo therapeutic purposes such as gene delivery, stimuli-responsive drug release, photothermal therapy, photodynamic therapy, ultrasound therapy, or anti-bacteria in tissue engineering, or as the contrast agents for biological and diagnostic imaging. Additionally, the critical issues and potential challenges related to the chemical design/synthesis of MSNs-based "magic bullet" by advanced nano-synthetic chemistry and in vivo evaluation have been discussed to highlight the issues scientists face in promoting the translation of MSNs-based DDSs into clinical trials. PMID:23681931

Chen, Yu; Chen, Hangrong; Shi, Jianlin

2013-05-17

101

Studies on dental erosion: An in vivo-in vitro model of endogenous dental erosion-its application to testing protection b flouride gel application  

Microsoft Academic Search

Background: The objective in this study was to develop an in vivo-in vitro model of endogenous erosion, with a view to exploring the potential for some degree of its control by the use of topical fluoride gel application to teeth. Methods: Six volunteers each wore a small clasp retained palatal acrylic appliance to which six sterilized enamel tiles were bonded.

L. Jones; D. Lekkas; D. Hunt; J. McIntyre; W. Rafir

2002-01-01

102

Multiphoton fluorescence recovery after photobleaching: Advancements for novel in vivo applications  

NASA Astrophysics Data System (ADS)

Multiphoton fluorescence recovery after photobleaching (MP-FRAP) is a laser microscopy technique used to probe the transport properties of macromolecules in biological systems. MP-FRAP utilizes two-photon fluorescence and photobleaching to produce a three-dimensionally resolved diffusion coefficient for an ensemble of molecules in the region of the two-photon focal volume. This thesis describes two fundamental improvements to the MP-FRAP technique, which are vital steps to enable MP-FRAP to be applied to the complex in vivo environment. In Chapter 1, we lay the groundwork for our discussion of these advancements by introducing the MP-FRAP technique and the physics upon which it is based. We begin with a description of fluorescence and diffusion and discuss their importance in biomedical research. Next, we describe how two-photon fluorescence and photobleaching are applied to a diffusing system to measure the diffusion coefficient via fluorescence recovery after photobleaching (FRAP). Then, we take the reader through the evolution of FRAP, which leads to the application of two- photon fluorescence and photobleaching to produce MP-FRAP. Along the way, we highlight applications and advancements of the FRAP techniques, and introduce fluorescence correlation spectroscopy, a popular complement to FRAP. In Chapter 2, we collect the experimental methods for the studies presented in Chapters 3 and 4. We begin with an in-depth discussion of our work to build and troubleshoot our MP-FRAP apparatus, followed by a detailed description of our data analysis protocol. Next, we delve into the specific methods for producing computer generated data and fits, as well as in vitro and in vivo experimental data, for our work in Chap. 3 on improving MP-FRAP to measure diffusion in the presence of convective flow. We end with a description of the Monte Carlo algorithm we developed for our work in Chap. 4 to model diffusion and multiphoton fluorescence recovery after photobleaching in the presence of reflective boundaries of various geometries. In Chapter 3, we develop an improved analytical model of multiphoton fluorescence recovery after photobleaching that includes the effects of convective flows within a system. We use computer generated data and fits to explore the effect of convective flow on the shape and speed of fluorescence recovery, and to estimate the range of diffusion coefficients and flow speeds over which this new "diffusion-convection" model yields accurate diffusion coefficients (as compared to the diffusion-only model). We then demonstrate the validity of the diffusion-convection model through in vitro experimentation in systems with known diffusion coefficients and known flow speeds, and show that the diffusion-convection model enables accurate determination of the diffusion coefficient via MP-FRAP, even when significant flows are present. We conclude by demonstrating the effectiveness of the diffusion-convection model in vivo by measuring the diffusion coefficient and flow speed within tumor vessels of 4T1 murine mammary adenocarcinomas implanted in the dorsal skinfold chamber. In Chapter 4, we present our work that allows MP-FRAP to be performed accurately near reflective boundaries of various geometries. Using Monte Carlo techniques, we first generate an initial distribution of bleached molecules, then simulate their diffusion away from the initial distribution, thereby producing fluorescence vs. time recovery curves in the region of the initial bleached distribution. These curves are then fit to the standard analytical MP-FRAP model to produce a diffusion coefficient. By introducing solid barriers into the model in the region of the initial bleached distribution, we learn how the presence of harriers of different geometries affects the measurement of diffusion via MP-FRAP. Finally, we supply ranges of barrier positions for each geometry within which MP-FR AP can confidently be employed to measure accurate diffusion coefficients.

Sullivan, Kelley Diane

103

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

PubMed Central

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.

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

2010-01-01

104

A miniaturized pressure sensor with inherent biofouling protection designed for in vivo applications.  

PubMed

The design, fabrication, and measurement results for a diaphragm-based single crystal silicon sensor element of size 820 ?m × 820 ?m × 500 ?m are presented. The sensor element is designed for in vivo applications with respect to size and measurement range. Moreover, it is optimized for longtime operation in the human body through a built-in protection preventing biofouling on the piezoresistors. The sensitivity is about 20 mV/V for a change from 500 to 1500 mbar absolute pressure. This result is comparable to conventional sized micromachined pressure sensors. The output signal is not found to be influenced by exposure to 60 °C for three hours, a normal temperature load for a typical sterilization process for medical devices (Ethylene Oxide Sterilization). The hysteresis is low; < 0.25% of full scale output signal. The sensor element withstands an overload pressure of 3000 mbar absolute pressure. Observed decrease in the output signal with temperatures and observed nonlinearity can easily be handled by traditional electronic compensation techniques. PMID:22254697

Clausen, I; Moe, S T; Tvedt, L G W; Vogl, A; Wang, D T

2011-01-01

105

Application of tetra-isopalmitoyl ascorbic acid in cosmetic formulations: stability studies and in vivo efficacy.  

PubMed

Liposoluble vitamin C derivatives, such as tetra-isopalmitoyl ascorbic acid (IPAA), are often used in dermocosmetic products due to their higher stability than vitamin C free form as well as its proposed effects in skin; however, there are no studies analyzing IPAA stability or its in vivo effects when present in dermocosmetic formulations. Thus, this study aimed to evaluate chemical stability and pre-clinical and clinical efficacy of dermocosmetic formulations containing IPAA in skin hydration and microrelief. Chemical stability of the formulations added with 1% IPAA was evaluated by heat stress during 35 days by HPLC. For pre-clinical evaluation, experimental formulations were topically applied on hairless skin mice during 5 days and animal skins were analyzed by non-invasive biophysic techniques (water content of stratum corneum, TEWL, viscoelasticity, and microrelief) and by histopathological studies. For clinical efficacy tests, the formulations were topically applied to the forearm and face of human volunteers, and 3h and 15 days after applications, the skins were evaluated by the same non-invasive techniques mentioned before. Results showed that formulations containing IPAA had medium stability and had pronounced moisturizing effects on stratum corneum and on viable epidermis. These formulations also improved skin microrelief especially in relation to skin smoothness and roughness. PMID:22974986

Maia Campos, Patrícia M B G; Gianeti, Mirela D; Camargo, Flávio B; Gaspar, Lorena R

2012-09-05

106

Radiolabelling of engineered nanoparticles for in vitro and in vivo tracing applications using cyclotron accelerators.  

PubMed

We present in this article an outline of some cyclotron-based irradiation techniques that can be used to directly radiolabel industrially manufactured nanoparticles, as well as two techniques for synthesis of labelled nanoparticles using cyclotron-generated radioactive precursor materials. These radiolabelled nanoparticles are suitable for a range of different in vitro and in vivo tracing studies of relevance to the field of nanotoxicology. A basic overview is given of the relevant physics of nuclear reactions regarding both ion-beam and neutron production of radioisotopes. The various issues that determine the practicality and usefulness of the different methods are discussed, including radioisotope yield, nuclear reaction kinetics, radiation and thermal damage, and radiolabel stability. Experimental details are presented regarding several techniques applied in our laboratories, including direct light-ion activation of dry nanoparticle samples, neutron activation of nanoparticles and suspensions using an ion-beam driven activator, spark-ignition generation of nanoparticle aerosols using activated electrode materials, and radiochemical synthesis of nanoparticles using cyclotron-produced isotopes. The application of these techniques is illustrated through short descriptions of some selected results thus far achieved. It is shown that these cyclotron-based methods offer a very useful range of options for nanoparticle radiolabelling despite some experimental difficulties associated with their application. For direct nanoparticle radiolabelling, if care is taken in choosing the experimental conditions applied, useful activity levels can be achieved in a wide range of nanoparticle types, without causing substantial thermal or radiation damage to the nanoparticle structure. Nanoparticle synthesis using radioactive precursors presents a different set of issues and offers a complementary and equally valid approach when laboratory generation of the nanoparticles is acceptable for the proposed studies, and where an appropriate radiolabel can be incorporated into the nanoparticles during synthesis. PMID:21479952

Gibson, N; Holzwarth, U; Abbas, K; Simonelli, F; Kozempel, J; Cydzik, I; Cotogno, G; Bulgheroni, A; Gilliland, D; Ponti, J; Franchini, F; Marmorato, P; Stamm, H; Kreyling, W; Wenk, A; Semmler-Behnke, M; Buono, S; Maciocco, L; Burgio, N

2011-04-11

107

Application of in vivo ESR spectroscopy to measurement of cerebrovascular ROS generation in stroke  

Microsoft Academic Search

This study used an in vivo ESR spectroscopy\\/spin probe technique to measure directly the generation of reactive oxygen species (ROS) in the brain after cerebral ischemia-reperfusion. Transient middle cerebral artery occlusion (MCAO) was induced in rats by inserting a nylon thread into the internal carotid artery for 1 h. The in vivo generation of ROS and its location in the

Mayumi Yamato; Toru Egashira; Hideo Utsumi

2003-01-01

108

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

PubMed Central

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.

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

109

Biosensors based on inorganic nanoparticles with biomimetic properties: Biomedical applications and in vivo cytotoxicity measurements  

NASA Astrophysics Data System (ADS)

The rapid progress of nanotechnology and advanced nanomaterials production offer significant opportunities for designing powerful biosensing devices with enhanced performances. This thesis introduces ceria (CeO 2) nanoparticles and its congeners as a new class of materials with huge potential in bioanalytical and biosensing applications. Unique redox, catalytic and oxygen storage/release properties of ceria nanoparticles, originating from their dual oxidation state are used to design biomedical sensors with high sensitivity and low oxygen dependency. This thesis describes a new approach for fabrication of implantable microbiosensors designed for monitoring neurological activity in physiological conditions. Understanding the mechanisms involved in neurological signaling and functioning is of great physiological importance. In this respect, the development of effective methods that allow accurate detection and quantification of biological analytes (i.e. L-glutamate and glucose) associated with neurological processes is of paramount importance. The performance of most analytical techniques currently used to monitor L-glutamate and glucose is suboptimal and only a limited number of approaches address the problem of operation in oxygen-restricted conditions, such as ischemic brain injury. Over the past couple of years, enzyme based biosensors have been used to investigate processes related to L-glutamate release/uptake and the glucose cycle within the brain. However, most of these sensors, based on oxidoreductase enzymes, do not work in conditions of limited oxygen availability. This thesis presents the development of a novel sensing technology for the detection of L-glutamate and glucose in conditions of oxygen deprivation. This technology provides real-time assessment of the concentrations of these analytes with high sensitivity, wide linear range, and low oxygen dependence. The fabrication, characterization and optimization of enzyme microbiosensors are discussed. This work introduces a new generic approach of improving the sensitivity of oxidase-based enzymatic assays and indicates that ceria and its mixture with other metal oxide nanoparticles could be used to minimize the problems associated with variations of the oxygen. These materials have great potential in bioanalytical and biotechnological applications and offer great opportunities for development of implantable sensing devices for in vivo and in vitro monitoring of analytes of clinical relevance. Additionally, this thesis evaluates the toxicity of different metal and metal oxide nanoparticles by using zebrafish embryos as a toxicological target. Because of their similarities with other vertebrates, rapid development and low cost, zebrafish embryos are ideal animal models for probing toxicological effects of engineered nanomaterials. Among the nanomaterials tested, nickel nanoparticles were characterized by high toxicity and induced delayed development and morphological malformations, while metal oxides nanoparticles (i.e. ceria nanoparticles) had no toxic effects.

Ispas, Cristina R.

110

A Microwave Applicator for in Vivo Rapid Inactivation of Enzymes in the Central Nervous System.  

National Technical Information Service (NTIS)

The paper describes modifications of microwave techniques for in vivo rapid inactivation of brain enzymes. These modified techniques offer greater rapidity and homigeneity of inactivation. The microwave-treated brain remains suitable for regional dissecti...

R. H. Lenox O. P. Gandhi J. L. Meyerhoff H. Mark Grove

1974-01-01

111

Application of Electrical Stimulation for Functional Tissue Engineering In Vitro and In Vivo.  

National Technical Information Service (NTIS)

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 stimulati...

G. V. Novakovic H. Park L. Freed M. Radisic R. Langer

2004-01-01

112

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

Microsoft Academic Search

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

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

1995-01-01

113

Synthesis and properties of new phosphorescent red light-excitable platinum(II) and palladium(II) complexes with Schiff bases for oxygen sensing and triplet-triplet annihilation-based upconversion.  

PubMed

New Pt(II) and Pd(II) complexes with donor-acceptor Schiff bases are conveniently prepared in only two steps. The complexes efficiently absorb in the red part of the spectrum (? > 10(5) M(-1) cm(-1)) and show moderate to strong room-temperature phosphorescence in the near-infrared (NIR) region. Particularly, Pt(II) complexes possess phosphorescence quantum yields (?) of ~10%, but the emission of the respective Pd(II) complexes is less efficient (? ? 1%-2%). The complexes exhibit solvatochromic behavior, in which the absorption and emission spectra shift bathochromically in polar solvents. The Pt(II) complexes are embedded in polystyrene to produce oxygen-sensing materials. The Pd(II) and Pt(II) complexes are demonstrated to be efficient sensitizers in triplet-triplet annihilation-based upconversion systems. PMID:23231719

Borisov, Sergey M; Saf, Robert; Fischer, Roland; Klimant, Ingo

2012-12-11

114

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...

115

Advanced in vivo applications of blue light photoreceptors as alternative fluorescent proteins.  

PubMed

The ultimate ambition in cell biology, microbiology and biomedicine is to unravel complex physiological and pathophysiological processes within living organisms. To conquer this challenge, fluorescent proteins (FPs) are used as versatile in vivo reporters and biosensors to study gene regulation as well as the synthesis, localization and function of proteins in living cells. The most widely used FPs are the green fluorescent protein (GFP) and its derivatives and relatives. Their use as in vivo reporter proteins, however, is sometimes restricted by different environmental and cellular factors. Consequently, a whole range of alternative, cofactor-dependent reporter proteins have been developed recently. In this perspective, we summarize the advantages and limitations of the novel class of cyan-green fluorescent flavoproteins in comparison to members of the GFP family and discuss some correlated consequences for the use of FPs as in vivo reporters. PMID:23660639

Drepper, Thomas; Gensch, Thomas; Pohl, Martina

2013-07-01

116

Application of the front detection photopiroelectric configuration to the study of in vivo human skin  

NASA Astrophysics Data System (ADS)

We report a novel method for measurements in vivo of the penetration of topically applied substances by inverse photopyroelectric configuration. This configuration was used to obtain the thermal effusivity, as a function of time, of in vivo human skin with ointments. This thermal magnitude was employed to characterize the penetration on the anterior-face of the volunteers forearm. This thermal effusivity was fitted with an exponential function in order to obtain a parameter (characteristic time) for the penetration. The substances used were a sunscreen and Vick Vaporub ointment. We found that the sunscreen have a characteristic time bigger that the Vick Vaporub ointment. The feasibility of skin hydration studies are discussed.

Gutierrez-Juarez, G.; Pichardo-Molina, J. L.; Rocha-Osornio, L. N.; Huerta-Franco, R.; Ivanov, R.; Huerta-Franco, B.; Cordova-Fraga, T.; Vargas-Luna, M.

2005-06-01

117

Application of vesicles to rat skin in vivo: a confocal laser scanning microscopy study  

Microsoft Academic Search

A major problem in (trans)dermal drug delivery is the low penetration rate of most substances through the barrier of the skin, the stratum corneum. One of the methods to increase the penetration rate across the skin is encapsulation of a (model) drug in lipid vesicles. In this study fluorescently labelled liposomes were applied on rat skin, in vivo. Bilayer labelled

Marly E. M. J. van Kuijk-Meuwissen; Luc Mougin; Hans E. Junginger; Joke A. Bouwstra

1998-01-01

118

Polylactide nanoparticles containing stably incorporated cyanine dyes for in vitro and in vivo imaging applications  

Microsoft Academic Search

Stably incorporating fluorescent molecules to polymeric nanoparticles (NPs) or micelles can facilitate the prolonged tracking of these drug-delivery vehicles in vitro and in vivo. However, incorporation of fluorescent molecules, usually charged and thereby water-soluble, through the encapsulation strategy to hydrophobic polymer matrices is challenging. The encapsu- lated fluorescent agents are also subject to rapid release when the polymeric NPs are

Rong Tong; Virginia J. Coyle; Li Tang; Anne M. Barger; Timothy M. Fan; Jianjun Cheng

2010-01-01

119

In vivo molecular imaging using nanomaterials: General in vivo characteristics of nano-sized reagents and applications for cancer diagnosis (Review)  

PubMed Central

Nanoparticles present a new collection of contrast agents for the field of in vivo molecular imaging. This review focuses on promising molecular imaging probes for optical and magnetic resonance imaging based on four representative nanomaterial(s) platforms: quantum dots, upconversion phosphors, superparamagnetic iron oxides, and dendrimer-based agents. Quantum dots are extremely efficient fluorescent nanoparticles with size-tunable emission properties, enabling high sensitivity and greater depth penetration. Their heavy metal composition and long retention in the body, however, pose concerns for clinical translational applications. Upconversion phosphors generate excellent signal-to-background contrast because they emit light with higher energy than the excitation photons and autofluorescence signals. For MRI, iron oxide particles also generate excellent signal and have been used in liver imaging and for cell tracking studies. As they are metabolized through endogenous iron salvage pathways, they have already been introduced as clinical contrast agents. Lastly, dendrimers, a ‘soft’ nanoparticle, can be used as a structural basis for the attachment of small molecule imaging agents and/or targeting groups. This array of nanoparticles should offer insights into the uses and potentials of nanoparticles for the molecular imaging.

ROSENBLUM, LAUREN T.; KOSAKA, NOBUYUKI; MITSUNAGA, MAKOTO; CHOYKE, PETER L.; KOBAYASHI, HISATAKA

2012-01-01

120

In Vivo Performance of a Liposomal Vascular Contrast Agent for CT and MR-Based Image Guidance Applications  

Microsoft Academic Search

Purpose  This study evaluated the in vivo performance of a liposome formulation that co-encapsulates iohexol and gadoteridol as a multimodal contrast agent for computed\\u000a tomography (CT) and magnetic resonance (MR)-based image guidance applications.\\u000a \\u000a \\u000a \\u000a Materials and Methods  The pharmacokinetics and biodistribution studies were conducted in Balb-C mice using high performance liquid chromatography\\u000a (HPLC) and inductively coupled plasma atomic emission spectrometry (ICP-AES) to detect

Jinzi Zheng; Jubo Liu; Mike Dunne; David A. Jaffray; Christine Allen

2007-01-01

121

[In vitro determination of the pressure-diameter relationship and velocity profiles by ultrasonic technics. In vivo application].  

PubMed

A good knowledge of arterial flow mechanics and of the phenomena associated with fluid-boundary interactions is necessary for the determination of some fundamental parameters such as velocity, pressure and pressure-diameter relationship during a cardiac cycle. Ultrasonic techniques were developed on a test bench and directly applied to animals without major modification. On such a test bench allowing a good simulation of physiological type flows, velocity field and pressure-diameter relationship were determined. In vivo application of these techniques allowed a systematic analysis of velocity profiles in the rabbit abdominal aorta and a precise approach of rheological properties of the vascular wall. PMID:3332291

Rieu, R; Friggi, A; Farahifar, D; Cassot, F

1987-01-01

122

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

NASA Astrophysics Data System (ADS)

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.

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

2004-10-01

123

A novel in vivo inducible expression system in Edwardsiella tarda for potential application in bacterial polyvalence vaccine.  

PubMed

Recombinant bacterial vector vaccine is an attractive vaccination strategy to induce the immune response to a carried protective antigen, and the main concern of bacterial vector vaccine is to establish a stable antigen expression system in vector bacteria. Edwardsiella tarda is an important facultative intracellular pathogen of both animals and humans, and its attenuated derivates are excellent bacterial vectors for use in recombinant vaccine design. In this study, we design an in vivo inducible expression system in E. tarda and establish potential recombinant E. tarda vector vaccines. With wild type strain E. tarda EIB202 as a vector, 53 different bacteria-originated promoters were examined for iron-responsive transcription in vitro, and the promoters P(dps) and P(yncE) showed high transcription activity. The transcription profiles in vivo of two promoters were further assayed, and P(dps) revealed an enhanced in vivo inducible transcription in macrophage, larvae and adult zebra fish. The gapA34 gene, encoding the protective antigen GAPDH from the fish pathogen Aeromonas hydrophila LSA34, was introduced into the P(dps)-based protein expression system, and transformed into attenuated E. tarda strains. The resultant recombinant vector vaccine WED/pUTDgap was evaluated in turbot (Scophtalmus maximus). Over 60% of the vaccinated fish survived under the challenge with A. hydrophila LSA34 and E. tarda EIB202, suggesting that the P(dps)-based antigen delivery system had great potential in bacterial vector vaccine application. PMID:21964456

Mu, Wei; Guan, Lingyu; Yan, Yijian; Liu, Qin; Zhang, Yuanxing

2011-09-25

124

Tracking stem cells for cardiovascular applications in vivo: focus on imaging techniques  

PubMed Central

Despite rapid translation of stem cell therapy into clinical practice, the treatment of cardiovascular disease using embryonic stem cells, adult stem and progenitor cells or induced pluripotent stem cells has not yielded satisfactory results to date. Noninvasive stem cell imaging techniques could provide greater insight into not only the therapeutic benefit, but also the fundamental mechanisms underlying stem cell fate, migration, survival and engraftment in vivo. This information could also assist in the appropriate choice of stem cell type(s), delivery routes and dosing regimes in clinical cardiovascular stem cell trials. Multiple imaging modalities, such as MRI, PET, SPECT and CT, have emerged, offering the ability to localize, monitor and track stem cells in vivo. This article discusses stem cell labeling approaches and highlights the latest cardiac stem cell imaging techniques that may help clinicians, research scientists or other healthcare professionals select the best cellular therapeutics for cardiovascular disease management.

Fu, Yingli; Azene, Nicole; Xu, Yi; Kraitchman, Dara L

2011-01-01

125

The application of PEDRI to the study of free radicals in vivo  

NASA Astrophysics Data System (ADS)

Proton-electron double-resonance imaging (PEDRI) has considerable value for study of the distribution and elimination pathways of nitroxide free radicals (NFRs). This has been illustrated by its use in studies of kidney function in the living rat in which the NFR proxyl carboxylic acid (PCA) has been employed as a `tracer'. The technique, at its present stage of development, can demonstrate location of PCA in enough detail to observe the passage through kidney cortex and medulla differentially, and to see the NFR within the major abdominal blood vessels. These studies are helping towards an understanding of the metabolic fate of PCA, as well as providing information about kidney performance after challenge with a nephrotoxin. In addition, nitric oxide complexes, formed in vivo by providing rats with a nitrite-rich diet, have been observed ex vivo using PEDRI and field-cycled DNP.

Foster, M. A.; Seimenis, I.; Lurie, D. J.

1998-07-01

126

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

NASA Astrophysics Data System (ADS)

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.

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

1995-03-01

127

Miniature Uncooled Infrared Sensitive Detectors for in Vivo Biomedical Imaging Applications  

SciTech Connect

Broadband infrared (OR) radiation detectors have been developed using miniature, inexpensive, mass produced microcantilevers capable of detecting temperature differences as small as lea(-6) K. Microcantilevers made out of semiconductor materials can be used either as uncurled photon or thermal detectors. Mounted on a probe mm in diameter a number of microcantilevers can be accommodated in the working channel of existing endoscopes for in vivo proximity focus measurements inside the human body.

Datskos, P. G.; Demos, S. G.; Rajic, S.

1998-06-01

128

Continuous glucose sensor using novel genetically engineered binding polypeptides towards in vivo applications  

Microsoft Academic Search

As the importance of blood-glucose control for both diabetic and non-diabetic patients continues to increase, there is a need for more advanced glucose-sensing technologies. In particular, an in vivo glucose sensor is needed that exhibits high accuracy when operating in a continuous manner for a relatively long period of time (3–5 days). Development of such sensors has been hampered, as

Jonathan Siegrist; Troy Kazarian; Charles Ensor; Smita Joel; Marc Madou; Ping Wang; Sylvia Daunert

2010-01-01

129

Elastography: elasticity imaging using ultrasound with application to muscle and breast in vivo.  

PubMed

Changes in tissue elasticity are generally correlated with its pathological state. In many cases, despite the difference in elasticity, the small size of a lesion or its location deep in the body preclude its detection by palpation. In general, such a lesion may or may not possess echogenic properties that would make it ultrasonically detectable. Elastography is an ultrasonic method for imaging the elasticity of compliant tissues. The method estimates the local longitudinal strain of tissue elements by ultrasonically assessing the one dimensional local displacements. This information can be combined with first order theoretical estimates of the local stress to yield a quantitative measure of the local elastic properties of tissue. The elasticity information is displayed in the form of a gray scale image called an elastogram. An experimental system for elastography in phantoms based on a single element transducer has been described previously [1]. Here we introduce a new elastography system based on a linear array transducer that is suitable for in vivo scanning. We describe tissue mimicking phantom experiments and preliminary in vivo breast and muscle elastograms confirming the feasibility of performing elastography in vivo. An elastogram of a breast containing an 8 mm palpable cancer nodule clearly shows the lesion. Elastograms and their corresponding sonograms show some similarities and differences in the depiction of tissue structures. PMID:8346612

Céspedes, I; Ophir, J; Ponnekanti, H; Maklad, N

1993-04-01

130

Measuring T2 In Vivo With J-Difference Editing: Application to GABA at 3 Tesla  

PubMed Central

Purpose To develop an experimental approach for determining in vivo transverse relaxation rates (T2) of metabolites that are detected by spectral editing without using simulations, and to demonstrate this approach to measure the T2 of ?-aminobutyric acid (GABA). Materials and Methods The proposed method first determines the TE-dependence of the edited signals using measurements in a pure phantom solution (10 mM ?-aminobutyric acid; GABA); the phantom T2 is also determined. Once the editing echo time (TE) -modulation pattern is known, it can then be used to determine T2 in vivo. The method was applied to measure GABA T2 in the occipital lobe of five healthy adult subjects at 3T, using a J-difference editing method. Unwanted macromolecular contributions to the GABA signal were also measured. Results The in vivo T2 of edited GABA signal was 88 ± 12 ms; this preliminary result is somewhat shorter than other metabolite T2 values in the literature at this field strength. Conclusion Spectral editing methods are now widely used to detect low concentration metabolites, such as GABA, but to date no edited acquisition methods have been proposed for the measurement of transverse relaxation times (T2). The method described has been successfully applied to measuring the T2 of GABA.

Edden, Richard A.E.; Intrapiromkul, Jarunee; Zhu, He; Cheng, Ying; Barker, Peter B.

2012-01-01

131

Biodegradation and in vivo biocompatibility of a degradable, polar/hydrophobic/ionic polyurethane for tissue engineering applications.  

PubMed

A degradable, polar/hydrophobic/ionic polyurethane (D-PHI) scaffold was optimized in in vitro studies to yield mechanical properties appropriate to replicate vascular graft tissue while eliciting a more wound-healing phenotype macrophage when compared to established materials. The objectives of this study were to characterize the biodegradation (in vitro and in vivo) and assess the in vivo biocompatibility of D-PHI, comparing it to a well-established, commercially-available scaffold biomaterial, polylactic glycolic acid (PLGA), recognized as being degradable, non-cytotoxic, and showing good biocompatibility. PLGA and D-PHI were formed into 6 mm diameter disk-shaped scaffolds (2 mm thick) of similar porosity (?82%) and implanted subcutaneously in rats. Both PLGA and D-PHI scaffolds were well-tolerated at the 7 d time point in vivo. In vitro D-PHI scaffolds degraded slowly (only 12 wt% in PBS in vitro after 120 d at 37 °C). In vivo, D-PHI scaffolds degraded at a more controlled rate (7 wt% loss over the acute 7 d implant phase and subsequently a linear profile of degradation leading to a 21 wt% mass loss by 100 d (chronic period)) than PLGA scaffolds which showed an initial more rapid degradation (14 wt% over 7 d), followed by minimal change between 7 and 30 d, and then a very rapid breakdown of the scaffold over the next 60 d. Histological examination of D-PHI scaffolds showed tissue ingrowth into the pores increased with time whereas PLGA scaffolds excluded cells/tissue from its porous structure as it degraded. The results of this study suggest that D-PHI has promising qualities for use as an elastomeric scaffold material for soft TE applications yielding well integrated tissue within the scaffold and a controlled rate of degradation stabilizing the form and shape of the implant. PMID:21641638

McBane, Joanne E; Sharifpoor, Soroor; Cai, Kuihua; Labow, Rosalind S; Santerre, J Paul

2011-06-08

132

A biocompatible in vivo ligation reaction and its application for noninvasive bioluminescent imaging of protease activity in living mice.  

PubMed

The discovery of biocompatible reactions had a tremendous impact on chemical biology, allowing the study of numerous biological processes directly in complex systems. However, despite the fact that multiple biocompatible reactions have been developed in the past decade, very few work well in living mice. Here we report that D-cysteine and 2-cyanobenzothiazoles can selectively react with each other in vivo to generate a luciferin substrate for firefly luciferase. The success of this "split luciferin" ligation reaction has important implications for both in vivo imaging and biocompatible labeling strategies. First, the production of a luciferin substrate can be visualized in a live mouse by bioluminescence imaging (BLI) and furthermore allows interrogation of targeted tissues using a "caged" luciferin approach. We therefore applied this reaction to the real-time noninvasive imaging of apoptosis associated with caspase 3/7. Caspase-dependent release of free D-cysteine from the caspase 3/7 peptide substrate Asp-Glu-Val-Asp-D-Cys (DEVD-(D-Cys)) allowed selective reaction with 6-amino-2-cyanobenzothiazole (NH(2)-CBT) in vivo to form 6-amino-D-luciferin with subsequent light emission from luciferase. Importantly, this strategy was found to be superior to the commercially available DEVD-aminoluciferin substrate for imaging of caspase 3/7 activity. Moreover, the split luciferin approach enables the modular construction of bioluminogenic sensors, where either or both reaction partners could be caged to report on multiple biological events. Lastly, the luciferin ligation reaction is 3 orders of magnitude faster than Staudinger ligation, suggesting further applications for both bioluminescence and specific molecular targeting in vivo. PMID:23463944

Godinat, Aurélien; Park, Hyo Min; Miller, Stephen C; Cheng, Ke; Hanahan, Douglas; Sanman, Laura E; Bogyo, Matthew; Yu, Allen; Nikitin, Gennady F; Stahl, Andreas; Dubikovskaya, Elena A

2013-03-29

133

Ring-shaped light illumination ultrasound-modulated optical tomography and its application for sentinel lymph node mapping ex vivo  

NASA Astrophysics Data System (ADS)

We have succeeded in implementing ring-shaped light illumination ultrasound-modulated optical tomography (UOT) in both transmission and reflection modes. These systems used intense acoustic bursts and a charge-coupled device camera-based speckle contrast detection method. By mounting an ultrasound transducer into an optical condenser, we can combine the illuminating light component with the ultrasound transducer. Thus, the UOT system is more clinically applicable than previous orthogonal mode systems. Furthermore, we have successfully imaged an ex vivo methyleneblue-dyed sentinel lymph node (SLN) embedded deeper than 12 mm, the mean depth of human sentinel lymph nodes, in chicken breast tissue. These UOT systems offer several advantages: noninvasiveness, nonionizing radiation, portability, cost-effectiveness, and possibility of combination with ultrasound pulse-echo imaging and photoacoustic imaging. One potential application of the UOT systems is mapping SLNs in axillary staging for breast cancer patients.

Kim, Chulhong; Song, Kwang Hyun; Maslov, Konstantin; Wang, Lihong V.

2009-02-01

134

Oxygen sensing by the prolyl-4-hydroxylase PHD2 within the nuclear compartment and the influence of compartmentalisation on HIF-1 signalling.  

PubMed

Hypoxia-inducible factors (HIFs) regulate more than 200 genes involved in cellular adaptation to reduced oxygen availability. HIFs are heterodimeric transcription factors that consist of one of three HIF-? subunits and a HIF-? subunit. Under normoxic conditions the HIF-? subunit is hydroxylated by members of a family of prolyl-4-hydroxylase domain (PHD) proteins, PHD1, PHD2 and PHD3, resulting in recognition by von-Hippel-Lindau protein, ubiquitylation and proteasomal degradation. It has been suggested that PHD2 is the key regulator of HIF-1? stability in vivo. Previous studies on the intracellular distribution of PHD2 have provided evidence for a predominant cytoplasmic localisation but also nuclear activity of PHD2. Here, we investigated functional nuclear transport signals in PHD2 and identified amino acids 196-205 as having a crucial role in nuclear import, whereas amino acids 6-20 are important for nuclear export. Fluorescence resonance energy transfer (FRET) showed that an interaction between PHD2 and HIF-1? occurs in both the nuclear and cytoplasmic compartments. However, a PHD2 mutant that is restricted to the cytoplasm does not interact with HIF-1? and shows less prolyl hydroxylase activity for its target HIF-1? than wild-type PHD2 located in the nucleus. Here, we present a new model by which PHD2-mediated hydroxylation of HIF-1? predominantly occurs in the cell nucleus and is dependent on very dynamic subcellular trafficking of PHD2. PMID:22946054

Pientka, Friederike Katharina; Hu, Jun; Schindler, Susann Gaby; Brix, Britta; Thiel, Anika; Jöhren, Olaf; Fandrey, Joachim; Berchner-Pfannschmidt, Utta; Depping, Reinhard

2012-09-03

135

Anti-Bladder-Tumor Effect of Baicalein from Scutellaria baicalensis Georgi and Its Application In Vivo  

PubMed Central

Some phytochemicals with the characteristics of cytotoxicity and/or antimetastasis have generated intense interest among the anticancer studies. In this study, a natural flavonoid baicalein was evaluated in bladder cancer in vitro and in vivo. Baicalein inhibits 5637 cell proliferation. It arrests cells in G1 phase at 100??M and in S phase below 75??M. The protein expression of cyclin B1 and cyclin D1 is reduced by baicalein. Baicalein-induced p-ERK plays a minor role in cyclin B1 reduction. Baicalein-inhibited p65NF-?B results in reduction of cell growth. Baicalein-induced pGSK(ser9) has a little effect in increasing cyclin B1/D1 expression instead. The translation inhibitor cycloheximide blocks baicalein-reduced cyclin B1, suggesting that the reduction is caused by protein synthesis inhibition. On the other hand, neither cycloheximide nor proteasome inhibitor MG132 completely blocks baicalein-reduced cyclin D1, suggesting that baicalein reduces cyclin D1 through protein synthesis inhibition and proteasomal degradation activation. In addition, baicalein also inhibits cell invasion by inhibiting MMP-2 and MMP-9 mRNA expression and activity. In mouse orthotopic bladder tumor model, baicalein slightly reduces tumor size but with some hepatic toxicity. In summary, these results demonstrate the anti-bladder-tumor properties of the natural compound baicalein which shows a slight anti-bladder-tumor effect in vivo.

Wu, Jin-Yi; Li, Yi-Zhen; Chang, Yi-Sheng; Lai, Yi-Chien; Laio, Yu-Han; Wu, Jiann-Der; Liu, Yi-Wen

2013-01-01

136

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

SciTech Connect

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.

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

1989-10-01

137

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

PubMed

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

Acharya, Amitabha

2013-06-01

138

In vitro and in vivo evaluation of an alumina-zirconia composite for arthroplasty applications.  

PubMed

In order to improve the reliability and the mechanical properties of orthopaedic hip prosthesis, new ceramic composites starting with nanosized powders of alumina and zirconia have been recently developed. The aim of the present study was to investigate the biological tolerance of one of these sintered ceramics and of its alumina and zirconia constitutive nanosized powders with both in vitro and in vivo approaches. At first, osteoblasts and fibroblasts were cultured either upon sintered ceramic discs with polished or rough surfaces or in the presence of the corresponding alumina or zirconia powders at various concentrations. Thereafter, we chronically injected these powders in the knee articulation of rats. In vitro, the materials showed no deleterious effect on cell proliferation, extra-cellular matrix production (human type I collagen and fibronectin) or on cell morphology. In vivo, the histological examination showed only a very moderate and non-specific granulomatous response of the synovial membrane but no major inflammation as clinically described with metals or polyethylene wear debris. Besides its improved physical properties, this recently developed alumina-zirconia composite showed satisfactory biocompatibility. PMID:20053439

Roualdes, Olivier; Duclos, Marie-Eve; Gutknecht, Dan; Frappart, Lucien; Chevalier, Jérôme; Hartmann, Daniel J

2010-01-06

139

Application of In Vivo Induced Antigen Technology (IVIAT) to Bacillus anthracis  

PubMed Central

In vivo induced antigen technology (IVIAT) is an immuno-screening technique that identifies bacterial antigens expressed during infection and not during standard in vitro culturing conditions. We applied IVIAT to Bacillus anthracis and identified PagA, seven members of a N-acetylmuramoyl-L-alanine amidase autolysin family, three P60 family lipoproteins, two transporters, spore cortex lytic protein SleB, a penicillin binding protein, a putative prophage holin, respiratory nitrate reductase NarG, and three proteins of unknown function. Using quantitative real-time PCR comparing RNA isolated from in vitro cultured B. anthracis to RNA isolated from BALB/c mice infected with virulent Ames strain B. anthracis, we confirmed induced expression in vivo for a subset of B. anthracis genes identified by IVIAT, including L-alanine amidases BA3767, BA4073, and amiA (pXO2-42); the bacteriophage holin gene BA4074; and pagA (pXO1-110). The exogenous addition of two purified putative autolysins identified by IVIAT, N-acetylmuramoyl-L-alanine amidases BA0485 and BA2446, to vegetative B. anthracis cell suspensions induced a species-specific change in bacterial morphology and reduction in viable bacterial cells. Many of the proteins identified in our screen are predicted to affect peptidoglycan re-modeling, and our results support significant cell wall structural remodeling activity during B. anthracis infection. Identification of L-alanine amidases with B. anthracis specificity may suggest new potential therapeutic targets.

Rollins, Sean M.; Peppercorn, Amanda; Young, John S.; Drysdale, Melissa; Baresch, Andrea; Bikowski, Margaret V.; Ashford, David A.; Quinn, Conrad P.; Handfield, Martin; Hillman, Jeffrey D.; Lyons, C. Rick; Koehler, Theresa M.; Calderwood, Stephen B.; Ryan, Edward T.

2008-01-01

140

Anti-Bladder-Tumor Effect of Baicalein from Scutellaria baicalensis Georgi and Its Application In Vivo.  

PubMed

Some phytochemicals with the characteristics of cytotoxicity and/or antimetastasis have generated intense interest among the anticancer studies. In this study, a natural flavonoid baicalein was evaluated in bladder cancer in vitro and in vivo. Baicalein inhibits 5637 cell proliferation. It arrests cells in G1 phase at 100? ? M and in S phase below 75? ? M. The protein expression of cyclin B1 and cyclin D1 is reduced by baicalein. Baicalein-induced p-ERK plays a minor role in cyclin B1 reduction. Baicalein-inhibited p65NF- ? B results in reduction of cell growth. Baicalein-induced pGSK(ser9) has a little effect in increasing cyclin B1/D1 expression instead. The translation inhibitor cycloheximide blocks baicalein-reduced cyclin B1, suggesting that the reduction is caused by protein synthesis inhibition. On the other hand, neither cycloheximide nor proteasome inhibitor MG132 completely blocks baicalein-reduced cyclin D1, suggesting that baicalein reduces cyclin D1 through protein synthesis inhibition and proteasomal degradation activation. In addition, baicalein also inhibits cell invasion by inhibiting MMP-2 and MMP-9 mRNA expression and activity. In mouse orthotopic bladder tumor model, baicalein slightly reduces tumor size but with some hepatic toxicity. In summary, these results demonstrate the anti-bladder-tumor properties of the natural compound baicalein which shows a slight anti-bladder-tumor effect in vivo. PMID:23573134

Wu, Jin-Yi; Tsai, Kun-Wei; Li, Yi-Zhen; Chang, Yi-Sheng; Lai, Yi-Chien; Laio, Yu-Han; Wu, Jiann-Der; Liu, Yi-Wen

2013-03-17

141

Synthesis, Cellular Delivery and In vivo Application of Dendrimer-based pH Sensors.  

PubMed

The development of fluorescent indicators represented a revolution for life sciences. Genetically encoded and synthetic fluorophores with sensing abilities allowed the visualization of biologically relevant species with high spatial and temporal resolution. Synthetic dyes are of particular interest thanks to their high tunability and the wide range of measureable analytes. However, these molecules suffer several limitations related to small molecule behavior (poor solubility, difficulties in targeting, often no ratiometric imaging allowed). In this work we introduce the development of dendrimer-based sensors and present a procedure for pH measurement in vitro, in living cells and in vivo. We choose dendrimers as ideal platform for our sensors for their many desirable properties (monodispersity, tunable properties, multivalency) that made them a widely used scaffold for several biomedical devices. The conjugation of fluorescent pH indicators to the dendrimer scaffold led to an enhancement of their sensing performances. In particular dendrimers exhibit reduced cell leakage, improved intracellular targeting and allow ratiometric measurements. These novel sensors were successfully employed to measure pH in living HeLa cells and in vivo in mouse brain. PMID:24056638

Albertazzi, Lorenzo; Storti, Barbara; Brondi, Marco; Sulis Sato, Sebastian; Michele Ratto, Gian; Signore, Giovanni; Beltram, Fabio

2013-09-10

142

Manufacture of IRDye800CW-coupled Fe3O4 nanoparticles and their applications in cell labeling and in vivo imaging  

Microsoft Academic Search

BACKGROUND: In recent years, near-infrared fluorescence (NIRF)-labeled iron nanoparticles have been synthesized and applied in a number of applications, including the labeling of human cells for monitoring the engraftment process, imaging tumors, sensoring the in vivo molecular environment surrounding nanoparticles and tracing their in vivo biodistribution. These studies demonstrate that NIRF-labeled iron nanoparticles provide an efficient probe for cell labeling.

Yong Hou; Yingxun Liu; Zhongping Chen; Ning Gu; Jinke Wang

2010-01-01

143

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

NASA Astrophysics Data System (ADS)

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.

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

2011-08-01

144

A polymer-based neural microimplant for optogenetic applications: design and first in vivo study.  

PubMed

In optogenetics, neurons are genetically modified to become sensitive to light and thus, they can be stimulated or inhibited by light of certain wavelengths. In this work, we describe the fabrication of a polymer-based shaft electrode as a tool for optogenetics. This device can conduct light as well as fluids to a target brain region and record electrical neural signals from the same part of the tissue simultaneously. It is intended to facilitate optogenetic in vivo experiments with those novel multimodal neural probes or polymer optrodes. We used microsystems technology to integrate an SU-8 based waveguide and fluidic channel into a polyimide-based electrode shaft to allow simultaneous optical stimulation, fluid delivery, and electrophysiological recording in awake behaving animals. In a first acute proof-of-concept experiment in genetically modified mice, our device recorded single unit activity that was modulated by laser light transmitted into the tissue via the integrated waveguide. PMID:23306183

Rubehn, Birthe; Wolff, Steffen B E; Tovote, Philip; Lüthi, Andreas; Stieglitz, Thomas

2013-02-21

145

A new in vivo Raman probe for enhanced applicability to the body.  

PubMed

This paper describes a new in vivo Raman probe that allows investigation of areas of the body that are otherwise difficult to access. It is coupled to a previously described commercially available in vivo Raman spectrometer that samples the skin through an optical flat. In the work presented here, the laser light emerges from a smaller pen-shaped probe. It thus works on the same principles as the original spectrometer, while its relative performance in terms of signal-to-noise ratio of the spectra and obtained spatial resolution is only slightly diminished. It allows the window to be placed against the subject in more curved and recessed areas of subject's body and also for them to be more comfortable while the measurements take place. Results from three areas of the body that have previously been very difficult to study are described, the mouth, axilla, and scalp. Results from the scalp and axilla strata cornea (SC) show significant differences from the "normal" SC of the volar forearm. For instance, the scalp is observed to have lower amounts of natural moisturizing factors (NMF) compared to the volar forearm within the same subjects. Also for both the axilla and scalp the lipids show a change in order as compared to the lipids in the volar forearm and also differences from each other. The potential significance of these observations is discussed. Further, we show how we can probe the mouth, in this case observing the presence of the astringent tea polyphenol epigallocatechin gallate within the oral mucosa. PMID:22800645

Pudney, Paul D A; Bonnist, Eleanor Y M; Caspers, Peter J; Gorce, Jean-Philippe; Marriot, Chris; Puppels, Gerwin J; Singleton, Scott; van der Wolf, Martin J G

2012-07-13

146

Development and application of in vivo molecular traps reveals that dynein light chain occupancy differentially affects dynein-mediated processes  

PubMed Central

The ability to rapidly and specifically regulate protein activity combined with in vivo functional assays and/or imaging can provide unique insight into underlying molecular processes. Here we describe the application of chemically induced dimerization of FKBP to create nearly instantaneous high-affinity bivalent ligands capable of sequestering cellular targets from their endogenous partners. We demonstrate the specificity and efficacy of these inducible, dimeric “traps” for the dynein light chains LC8 (Dynll1) and TcTex1 (Dynlt1). Both light chains can simultaneously bind at adjacent sites of dynein intermediate chain at the base of the dynein motor complex, yet their specific function with respect to the dynein motor or other interacting proteins has been difficult to dissect. Using these traps in cultured mammalian cells, we observed that induction of dimerization of either the LC8 or TcTex1 trap rapidly disrupted early endosomal and lysosomal organization. Dimerization of either trap also disrupted Golgi organization, but at a substantially slower rate. Using either trap, the time course for disruption of each organelle was similar, suggesting a common regulatory mechanism. However, despite the essential role of dynein in cell division, neither trap had a discernable effect on mitotic progression. Taken together, these studies suggest that LC occupancy of the dynein motor complex directly affects some, but not all, dynein-mediated processes. Although the described traps offer a method for rapid inhibition of dynein function, the design principle can be extended to other molecular complexes for in vivo studies.

Varma, Dileep; Dawn, Amrita; Ghosh-Roy, Anindya; Weil, Sarah J.; Ori-McKenney, Kassandra M.; Zhao, Yanqiu; Keen, James; Vallee, Richard B.; Williams, John C.

2010-01-01

147

Application of LCModel for quality control and quantitative in vivo 1H MR spectroscopy by short echo time STEAM sequence.  

PubMed

The linear combination of model spectra (LCModel) calculation of a parameter for long-term quality control, kT, was introduced, representing the ratio of the temporal and nominal intensities of CH3 groups of lactate and acetate in a quality control phantom. This procedure is a part of the quality assurance of the scanner using fully automatic measurement and calculation of kT parameters, and utilizing Shewhart regulation control charts for continuous evaluation of the magnetic resonance (MR) scanner setting. The application of the kT parameter for the correction of in vivo data increases the precision of molar concentration determination by about 4%. This was tested by the quantitative in vivo MR determination of the molar concentrations of 13 prominent metabolites (N-acetylaspartate (NAA), N-acetylaspartylglutamate, creatine and phosphocreatine (Cr), choline-containing compounds (Cho), myo-inositol, scyllo-inositol, gamma-aminobutyric acid, glutamine, glutamate, glucose, lactate, alanine, taurine) in the white matter and hippocampus of the brain in groups of volunteers, using a short echo time stimulated echo acquisition mode sequence (echo time = 10 ms) and the LCModel technique. The repeatability of the measurement of prominent metabolites such as NAA, Cr and Cho was found to be around 10% (relative standard deviation, n = 6); precision in a group of volunteers (n = 20 and 28, respectively) was in the range of approximately 13-20%. For other metabolites, which are measured with a lower signal-to-noise ratio, the precision can be much lower. PMID:10697221

Hájek, M; Burian, M; Dezortová, M

2000-02-01

148

Application of the moving-actuator type pump as a ventricular assist device: in vitro and in vivo studies.  

PubMed

A moving actuator type pump has been developed as a multifunctional Korean artificial heart (AnyHeart). The pump consists of a moving actuator as an energy converter, right and left sacs, polymer (or mechanical) valves, and a rigid polyurethane housing. The actuator containing a brushless DC motor moves back and forth on an epicyclical gear train to produce a pendular motion, which compresses both sacs alternately. Of its versatile functions of ventricular assist device and total artificial heart use, we have evaluated the system performance as a single or biventricular assist device through in vitro and in vivo experiments. Pump performance and anatomical feasibility were tested using various animals of different sizes. In the case of single ventricular assist device (VAD) use, one of the sacs remained empty and a mini-compliance chamber was attached to either an outflow or inflow port of the unused sac. The in vitro and in vivo studies show acceptable performance and pump behavior. Further extensive study is required to proceed to human application. PMID:12117296

Lee, H S; Rho, Y R; Park, C Y; Hwang, C M; Kim, W G; Sun, K; Choi, M J; Lee, K K; Cheong, J T; Shim, E B; Min, B G

2002-06-01

149

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

NASA Astrophysics Data System (ADS)

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.

Gali, Adam; Zólyomi, Viktor; Somogyi, Bálint

2013-03-01

150

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)

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. .

Kehayias, J. J.

2001-07-01

151

Human Application of Ex-Vivo Expanded Umbilical Cord-Derived Mesenchymal Stem Cells: Enhance Hematopoiesis after Cord Blood Transplantation.  

PubMed

Delayed hematopoietic reconstitution after cord blood (CB) transplantation (CBT) needs to be overcome. Bone marrow-derived mesenchymal stem cells (BMMSCs) have been found to enhance engraftment after hematopoietic stem cell transplantation. However, getting BMMSCs involves an invasive procedure. In this study, umbilical cord-derived mesenchymal stem cells (UCMSCs) were isolated from Wharton's jelly and cryopreserved in the UCMSC bank. Compared with BMMSCs, we found that UCMSCs had superior proliferative potential. We found that NOD/SCID mice cotransplanted with CB and UCMSCs demonstrated the significant human CD45+ cells engraftment than those transplanted with CB alone. Then, twenty patients with high-risk leukemia prospectively randomized to receive cotransplantation of CB and ex-vivo expanded banked UCMSCs or to receive CBT alone. No serious adverse events were observed in the patients receiving UCMSCs infusion. The time to neutrophil engraftment and platelet engraftment was significantly shorter in the 8 patients receiving cotransplantation than that in the 12 patients receiving CBT alone (p = 0.003 and p = 0.004, respectively). According, application of ex-vivo expanded banked UCMSCs in humans appears to be feasible and safe. UCMSCs can enhance engraftment after CBT, but further studies are warranted. PMID:23452720

Wu, Kang-Hsi; Tsai, Chris; Wu, Han-Ping; Sieber, Martin; Peng, Ching-Tien; Chao, Yu-Hua

2013-02-26

152

Thermally cross-linked superparamagnetic iron oxide nanoparticles: synthesis and application as a dual imaging probe for cancer in vivo.  

PubMed

We report the fabrication and characterization of thermally cross-linked superparamagnetic iron oxide nanoparticles (TCL-SPION) and their application to the dual imaging of cancer in vivo. Unlike dextran-coated cross-linked iron oxide nanoparticles, which are prepared by a chemical cross-linking method, TCL-SPION are prepared by a simple, thermal cross-linking method using a Si-OH-containing copolymer. The copolymer, poly(3-(trimethoxysilyl)propyl methacrylate-r-PEG methyl ether methacrylate-r-N-acryloxysuccinimide), was synthesized by radical polymerization and used as a coating material for as-synthesized magnetite (Fe3O4) SPION. The polymer-coated SPION was further heated at 80 degrees C to induce cross-linking between the -Si(OH)3 groups in the polymer chains, which finally generated TCL-SPION bearing a carboxyl group as a surface functional group. The particle size, surface charge, presence of polymer-coating layers, and the extent of thermal cross-linking were characterized and confirmed by various measurements, including dynamic light scattering, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. The carboxyl TCL-SPION was converted to amine-modified TCL-SPION and then finally to Cy5.5 dye-conjugated TCL-SPION for use in dual (magnetic resonance/optical) in vivo cancer imaging. When the Cy5.5 TCL-SPION was administered to Lewis lung carcinoma tumor allograft mice by intravenous injection, the tumor was unambiguously detected in T2-weighted magnetic resonance images as a 68% signal drop as well as in optical fluorescence images within 4 h, indicating a high level of accumulation of the nanomagnets within the tumor site. In addition, ex vivo fluorescence images of the harvested tumor and other major organs further confirmed the highest accumulation of the Cy5.5 TCL-SPION within the tumor. It is noteworthy that, despite the fact that TCL-SPION does not bear any targeting ligands on its surface, it was highly effective for tumor detection in vivo by dual imaging. PMID:17892287

Lee, Haerim; Yu, Mi Kyung; Park, Sangjin; Moon, Sungmin; Min, Jung Jun; Jeong, Yong Yeon; Kang, Hae-Won; Jon, Sangyong

2007-09-25

153

In Vivo Imaging of Changes in Tumor Oxygenation During Growth and After Treatment  

PubMed Central

A novel procedure for in vivo imaging of the oxygen partial pressure (pO2) in implanted tumors is reported. The procedure uses electron paramagnetic resonance imaging (EPRI) of oxygen-sensing nanoprobes embedded in the tumor cells. Unlike existing methods of pO2 quantification, wherein the probes are physically inserted at the time of measurement, the new approach uses cells that are preinternalized (labeled) with the oxygen-sensing probes, which become permanently embedded in the developed tumor. Radiation-induced fibrosarcoma (RIF-1) cells, internalized with nanoprobes of lithium octa-n-butoxy-naphthalocyanine (LiNc-BuO), were allowed to grow as a solid tumor. In vivo imaging of the growing tumor showed a heterogeneous distribution of the spin probe, as well as oxygenation in the tumor volume. The pO2 images obtained after the tumors were exposed to a single dose of 30-Gy X-radiation showed marked redistribution as well as an overall increase in tissue oxygenation, with a maximum increase 6 hr after irradiation. However, larger tumors with a poorly perfused core showed no significant changes in oxygenation. In summary, the use of in vivo EPR technology with embedded oxygen-sensitive nanoprobes enabled noninvasive visualization of dynamic changes in the intracellular pO2 of growing and irradiated tumors.

Bratasz, Anna; Pandian, Ramasamy P.; Deng, Yuanmu; Petryakov, Sergey; Grecula, John C.; Gupta, Nilendu; Kuppusamy, Periannan

2008-01-01

154

Dendrimer-Based Fluorescent Indicators: In Vitro and In Vivo Applications  

Microsoft Academic Search

BackgroundThe development of fluorescent proteins and synthetic molecules whose fluorescence properties are controlled by the environment makes it possible to monitor physiological and pathological events in living systems with minimal perturbation. A large number of small organic dyes are available and routinely used to measure biologically relevant parameters. Unfortunately their application is hindered by a number of limitations stemming from

Lorenzo Albertazzi; Marco Brondi; Giovanni M. Pavan; Sebastian Sulis Sato; Giovanni Signore; Barbara Storti; Gian Michele Ratto; Fabio Beltram

2011-01-01

155

TOPICAL APPLICATION OF BLEACHING PHENOLS; IN VIVO STUDIES AND MECHANISM OF ACTION RELEVANT TO MELANOMA TREATMENT  

PubMed Central

Skin depigmentation represents a well established treatment for extensive vitiligo and may likewise be suited to prevent tumor recurrences and as a prophylactic treatment of familial melanoma, as common bleaching agents are cytotoxic to melanocytes. Effective melanoma prevention requires a bleaching agent-induced loss of exposed melanocytes supported by an immune response to distant pigment cells. Studies on human explant cultures treated with depigmenting agents, 4-tertiary butyl phenol (4-TBP) or monobenzyl ether of hydroquinone (MBEH) revealed a significant increase in the migration of Langerhans cells towards the dermis only upon MBEH treatment thus suggesting selective elicitation of an immune response. To assess the depigmenting potential of bleaching agents in vivo, 4-TBP and MBEH were topically applied to C57BL/6 wild type as well as k14-SCF transgenic, epidermally pigmented mice. MBEH induced significant skin depigmentation in both strains, not observed upon 4-TBP treatment. Cytokine expression patterns in MBEH treated skin support activation of a Th1 mediated immune response corresponding to an influx of T cells and macrophages. Importantly, despite insensitivity of tumor cells to MBEH induced cytotoxicity, significantly retarded tumor growth was observed in B16 challenged k14-SCF mice pretreated with MBEH, likely due to an abundance of cytotoxic T cells accompanied by an increased expression of Th1 and Th17 cytokines. These data support the use of MBEH as a prophylactic treatment for melanoma.

Hariharan, Vidhya; Toole, Timothy; Klarquist, Jared; Longley, Jack B; Mosenson, Jeffrey; Le Poole1, I. Caroline

2012-01-01

156

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

PubMed

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

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

157

Imaging the Function of P-Glycoprotein With Radiotracers: Pharmacokinetics and In Vivo Applications  

PubMed Central

P-glycoprotein (P-gp), an efflux transporter, controls the pharmacokinetics of various compounds under physiological conditions. P-gp-mediated drug efflux has been suggested as playing a role in various disorders, including multidrug-resistant cancer and medication-refractory epilepsy. However, P-gp inhibition has had, to date, little or no clinically significant effect in multidrug-resistant cancer. To enhance our understanding of its in vivo function under pathophysiological conditions, substrates of P-gp have been radiolabeled and imaged using single-photon emission computed tomography (SPECT) and positron emission tomography (PET). To accurately quantify P-gp function, a radiolabeled P-gp substrate should be selective for P-gp, produce a large signal after P-gp blockade, and generate few radiometabolites that enter the target tissue. Furthermore, quantification of P-gp function via imaging requires pharmacological inhibition of P-gp, which requires knowledge of P-gp density at the target site. By meeting these criteria, imaging can elucidate the function of P-gp in various disorders and improve the efficacy of treatments.

Kannan, P; John, C; Zoghbi, SS; Halldin, C; Gottesman, MM; Innis, RB; Hall, MD

2009-01-01

158

In vivo biochemistry: applications for small molecule biosensors in plant biology.  

PubMed

Revolutionary new technologies, namely in the areas of DNA sequencing and molecular imaging, continue to impact new discoveries in plant science and beyond. For decades we have been able to determine properties of enzymes, receptors and transporters in vitro or in heterologous systems, and more recently been able to analyze their regulation at the transcriptional level, to use GFP reporters for obtaining insights into cellular and subcellular localization, and tp measure ion and metabolite levels with unprecedented precision using mass spectrometry. However, we lack key information on the location and dynamics of the substrates of enzymes, receptors and transporters, and on the regulation of these proteins in their cellular environment. Such information can now be obtained by transitioning from in vitro to in vivo biochemistry using biosensors. Genetically encoded fluorescent protein-based sensors for ion and metabolite dynamics provide highly resolved spatial and temporal information, and are complemented by sensors for pH, redox, voltage, and tension. They serve as powerful tools for identifying missing processes (e.g., glucose transport across ER membranes), components (e.g., SWEET sugar transporters for cellular sugar efflux), and signaling networks (e.g., from systematic screening of mutants that affect sugar transport or cytosolic and vacuolar pH). Combined with the knowledge of properties of enzymes and transporters and their interactions with the regulatory machinery, biosensors promise to be key diagnostic tools for systems and synthetic biology. PMID:23587939

Jones, Alexander M; Grossmann, Guido; Danielson, Jonas Åh; Sosso, Davide; Chen, Li-Qing; Ho, Cheng-Hsun; Frommer, Wolf B

2013-04-12

159

Formulation of meloxicam gel for topical application: In vitro and in vivo evaluation.  

PubMed

Skin delivery of NSAIDs offers several advantages over the oral route associated with potential side effects. In the present investigation, topical gel of meloxicam (MLX) was formulated using N-methyl pyrrolidone (NMP) as a solubilizer and Carbopol Ultrez 10® as a gelling polymer. MLX gel was evaluated with respect to different physicochemical parameters such as pH, viscosity and spreadability. Irritation potential of MLX gel was studied on rabbits. Permeation of MLX gel was studied using freshly excised rat skin as a membrane. Anti-inflammatory activity of MLX gel was studied in rats and compared with the commercial formulation of piroxicam (Pirox® gel, 0.5% m/m). Accelerated stability studies were carried out for MLX gel for 6 months according to ICH guidelines. MLX gel was devoid of any skin irritation in rabbits. After 12 h, cumulative permeation of MLX through excised rat skin was 3.0 ± 1.2 mg cm-2 with the corresponding flux value of 0.24 ± 0.09 mg cm-2 h-1. MLX gel exhibited significantly higher anti-inflammatory activity in rats compared to Pirox® gel. Physicochemically stable and non-irritant MLX gel was formulated which could deliver significant amounts of active substance across the skin in vitro and in vivo to elicit the anti-inflammatory activity. PMID:21134852

Bachhav, Yogeshwar G; Patravale, Vandana B

2010-06-01

160

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

Microsoft Academic Search

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.

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

2008-01-01

161

In vivo sustained dermal delivery and pharmacokinetics of interferon alpha in biphasic vesicles after topical application.  

PubMed

Biphasic vesicles, a novel nanostructured lipid-based delivery system show potential for topical application of interferon alpha (IFN ?) for the treatment of human papillomavirus (HPV) infections (anogenital warts). Dermal delivery of IFN ? encapsulated in biphasic vesicles (BPV-IFN ?), applied topically to the skin, was characterized in a guinea pig model. BPV-IFN ? (1g, 2 MIU/g) was topically applied either as a single or multiple treatments on the skin of guinea pigs. As a comparison with currently used regimens, IFN ? solution was administered intravenously or intradermally. Skin and serum samples were collected over 96 h, IFN ? levels were determined by an antiviral assay, and half-life (t?/?) and elimination (k) rates were calculated. Topical BPV-IFN ? treatment resulted in maximum skin levels (about 100,000 U/100 cm(2)) of IFN ? within 6h and maintained for 72-96 h. Clearance from the skin after intradermal injections was initially fast (t?/? 0.62 h, k 1.1179 h(-1)), followed by a slower steady decrease after 6h. After intravenous and intradermal administration, IFN ? was rapidly cleared from the serum, t?/? 0.75 h, k 0.9271 h(-1) and t?/? 1.28 h, k 0.5421 h(-1), respectively, whereas after topical application, IFN ? levels remained below 100 U/mL. Topical application of BPV- IFN ? resulted in sustained delivery of biologically active IFN ? locally into skin with minimal systemic exposure. PMID:23500117

King, Martin; Kumar, Praveen; Michel, Deborah; Batta, Ravinderjit; Foldvari, Marianna

2013-03-14

162

In vivo response to starch-based scaffolds designed for bone tissue engineering applications.  

PubMed

Our purpose was to evaluate the in vivo endosseous response to three starch-based scaffolds implanted in rats (n = 54). We implanted the three scaffold groups; a 50/50 (wt %) blend of corn starch and ethylene-vinyl alcohol (SEVA-C), the same composition coated with a biomimetic calcium phosphate (Ca-P) layer (SEVA-C/CaP), and a 50/50 (wt %) blend of corn starch and cellulose acetate (SCA), all produced by extrusion with blowing agents, into distal femurs proximal to the epiphyseal plate, for 1, 3, or 6 weeks. Our results showed that at 1 week considerable reparative bone formed around all scaffold groups, although the bone was separated from the scaffold by an intervening soft tissue interfacial zone that comprised two distinct compartments: the surface of the scaffold was occupied by multinucleate giant cells and the compartment between these cells and the surrounding bone was occupied by a streaming fibrous-like tissue. The extracellular matrix of the latter was continuous with the extracellular bone matrix itself, labeled positively for osteocalcin and appeared mineralized by back-scattered electron imaging. All three scaffolds showed a similar tissue response, with the soft tissue interface diminishing with time. No bone contact was observed with SEVA-C at any time point, only transitory bone contact was observed with SEVA-C/CaP at 3 weeks, but SCA exhibited direct bone contact at 6 weeks where 56.23 +/- 6.46% of the scaffold surface was occupied by bone. We conclude that all materials exhibited a favorable bony response and that the rapidly forming initial "connective tissue" seen around all scaffolds was a very early form of bone formation. PMID:17109411

Salgado, A J; Coutinho, O P; Reis, R L; Davies, J E

2007-03-15

163

Polylactide nanoparticles containing stably incorporated cyanine dyes for in vitro and in vivo imaging applications.  

PubMed

Stably incorporating fluorescent molecules to polymeric nanoparticles (NPs) or micelles can facilitate the prolonged tracking of these drug-delivery vehicles in vitro and in vivo. However, incorporation of fluorescent molecules, usually charged and thereby water-soluble, through the encapsulation strategy to hydrophobic polymer matrices is challenging. The encapsulated fluorescent agents are also subject to rapid release when the polymeric NPs are exposed to biological media. To address this issue, we developed Cy5-conjugated polylactide (Cy5-PLA) NPs through Cy5/(BDI)ZnN(TMS)2 [(BDI) = 2-((2,6-diisopropylphenyl)amido)-4-((2,6-diisopropylphenyl)-imino)-2-pentene]-mediated ring-opening polymerization of lactide (LA) followed by nanoprecipitation. This process allows for covalent conjugation of Cy5 to PLA with quantitative incorporation efficiency and formulation of Cy5-PLA NPs with controlled particles size (approximately 100 nm). As much as 80% of Cy5 was still present in the Cy5-PLA NPs after theses NPs were incubated in PBS at 37 degrees C for 12 days. Cy5-PLA NPs were conjugated to the A10 RNA aptamer that binds to the prostate-specific membrane antigen (PSMA). The resulting Cy5-PLA/aptamer NPs were found to only bind to and get internalized by LNCaP and canine prostate adenocarcinoma cells (PSMA-positive), but not to PC3 cells (PSMA-negative). The Cy5-PLA NPs were administered to balb/c mice intravenously and found to have excellent signals with low-background fluorescence in various organs. PMID:20146347

Tong, Rong; Coyle, Virginia J; Tang, Li; Barger, Anne M; Fan, Timothy M; Cheng, Jianjun

2010-09-01

164

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 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...

2013-04-01

165

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

PubMed Central

The in vivo activities of amoxicillin and amoxicillin-clavulanate against 17 strains of Streptococcus pneumoniae 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 infection with 105 to 106 CFU, groups of mice were treated with 7 mg of amoxicillin per kg of body weight alone or combined with clavulanate (ratio, 4:1) every 8 h for 1 and 4 days. There was an excellent correlation between the MIC of amoxicillin (0.03 to 5.6 mg/liter) and (i) the change in log10 CFU/thigh at 24 h and (ii) survival after 4 days of therapy. Organisms for which MICs were 2 mg/liter or less were killed at 1.4 to 4.2 and 1.6 to 4.1 log10 CFU/thigh at 24 h by amoxicillin and amoxicillin-clavulanate, respectively. The four strains for which MICs were >4 mg/liter grew 0.2 to 2.6 and 0.6 to 2.3 logs at 24 h despite therapy with amoxicillin and amoxicillin-clavulanate, respectively. Infection was uniformly fatal by 72 h in untreated mice. Amoxicillin therapy resulted in no mortality with organisms for which MICs were 1 mg/liter or less, 20 to 40% mortality with organisms for which MICs were 2 mg/liter, and 80 to 100% mortality with organisms for which MICs were 4.0–5.6 mg/liter. Lower and higher doses (0.5, 2, and 20 mg/kg) of amoxicillin were studied against organisms for which MICs were near the breakpoint. These studies demonstrate that a reduction of 1 log10 or greater in CFU/thigh at 24 h is consistently observed when amoxicillin levels exceed the MIC for 25 to 30% of the dosing interval. These studies would support amoxicillin (and amoxicillin-clavulanate) MIC breakpoints of 1 mg/liter for susceptible, 2 mg/liter for intermediate, and 4 mg/liter for resistant strains of S. pneumoniae.

Andes, D.; Craig, W. A.

1998-01-01

166

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

PubMed

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 degrees independent active sectors per tube. A flexible delivery catheter facilitated water cooling (100 ml min(-1)) within an expandable urethral balloon (35 mm long x 10 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 approximately 90 degrees-100 degrees acoustic output patterns from each 120 degrees 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 degrees C maximum temperature, t43 = 240 min thermal dose contours, and TTC tissue sections) that extended radially from the applicator toward the border of the prostate (approximately15 mm) during a short power application (approximately 8-16 W per active sector, 8-15 min), with approximately 200 degrees or 360 degrees 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 approximately 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. PMID:18561684

Kinsey, Adam M; Diederich, Chris J; Rieke, Viola; Nau, William H; Pauly, Kim Butts; Bouley, Donna; Sommer, Graham

2008-05-01

167

In vitro and in vivo characterization of novel biodegradable polymers for application as drug-eluting stent coatings.  

PubMed

We have used a series of in vitro and in vivo tests to assess the suitability of two new degradable polymers for application as coatings for drug-eluting stents. The first is a family of urethane-linked multi-block copolymers (MBCP) that comprise blocks of lactide, glycolide, epsilon-caprolactone and/or poly(ethylene glycol) chain-extended with 1,4-butanediisocyanate (SynBiosys polymers). The second is a family of maltodextrin (MD) modified with fatty acid sidechains to yield a hydrophobic polymer (Eureka() SOLO polymers). We coated stainless-steel stents with two representative urethane-linked MBCPs and one hydrophobic MD polymer alone or in combination with the anti-restenotic drug sirolimus. Urethane-linked MBCPs formed uniform coatings on the stent substrates, withstood crimping and expansion on balloon catheters, completely released sirolimus from the coating within 30 days, and degraded within 30-60 days in PBS. The hydrophobic MD polymer formed uniform coatings, exhibited somewhat slower release of sirolimus (approx. 85% within 30 days), degraded within 60 days in PBS when sirolimus was incorporated in the coating, but showed very slow degradation in the absence of drug. We implanted stents coated with urethane-linked MBCPs or hydrophobic MD polymers in a porcine coronary artery model and used histological analysis at 28- and 90-day end-points to assess the biological response to the materials. Measures of stenosis and inflammation for urethane-linked MBCP and hydrophobic MD polymer coatings were not statistically different from bare metal controls at 28 and 90 days, suggesting that the polymers show good vascular biocompatibility. Endothelialization was nearly complete at 28 days and complete at 90 days for all formulations. Urethane-linked MBCP polymer-only and drug-eluting coatings and hydrophobic MD drug-eluting coatings were nearly completely degraded within 90 days in vivo whereas roughly half of hydrophobic MD polymer-only coatings remained after 90 days. Taken together, our in vitro and in vivo results suggest that SynBiosys urethane-linked MBCP and Eureka SOLO hydrophobic MD polymer families possess the physical and chemical properties and vascular biocompatibility necessary for further investigation for use in the next generation of drug-eluting stents. PMID:20233507

Lockwood, Nathan A; Hergenrother, Robert W; Patrick, Laura M; Stucke, Sean M; Steendam, Rob; Pacheco, Erica; Virmani, Renu; Kolodgie, Frank D; Hubbard, Brad

2010-01-01

168

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

NASA Astrophysics Data System (ADS)

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.

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

169

Cellular oxygen sensing in health and disease  

Microsoft Academic Search

To avoid localised problems resulting from excess or inadequate oxygen, all cells and tissues have the ability to sense and\\u000a respond to changes in oxygen levels. Despite their rich blood supply, the kidneys have unique properties with respect to oxygen\\u000a that enable them to act as specialised organs, sensing oxygen delivery as well as rendering them prone to hypoxic injury.

David R. Mole; Peter J. Ratcliffe

2008-01-01

170

TRANSCRIPTION: Oxygen Sensing Gets a Second Wind  

NSDL National Science Digital Library

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.

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

2002-02-01

171

Regulating cellular oxygen sensing by hydroxylation  

Microsoft Academic Search

Oxygen homeostasis under conditions of limited O2 supply requires hypoxia-dependent gene regulation. The transcription factor complex hypoxia-inducible factor 1 (HIF-1) has been recognized as the master regulator that mediates the adaptational genetic response to ensure restoration of energy supply. This review will focus on the recent advances in understanding the hypoxia-induced cellular response with particular respect to cellular O2 sensing

Joachim Fandrey; Thomas A. Gorr; Max Gassmann

2006-01-01

172

Antioxidative peptides: enzymatic production, in vitro and in vivo antioxidant activity and potential applications of milk-derived antioxidative peptides.  

PubMed

The beneficial effects of food-derived antioxidants in health promotion and disease prevention are being increasingly recognized. Recently, there has been a particular focus on milk-derived peptides; as a source of antioxidants, these peptides are inactive within the sequence of the parent protein but can be released during enzyme hydrolysis. Once released, the peptides have been shown to possess radical scavenging, metal ion chelation properties and the ability to inhibit lipid peroxidation. A variety of methods have been used to evaluate in vitro antioxidant activity, however, there is no standardised methodology, which hinders comparison of data. This review provides an overview on the generation of antioxidative peptides from milk proteins, the proposed mechanisms of protein/peptide induced antioxidant activity, in vitro measurement of antioxidant activity, in vivo evaluation of plasma antioxidant capacity and the bioavailability of antioxidative peptides. The understanding gained from other food proteins is referred to where specific data on milk-derived peptides are limited. The potential applications and health benefits of antioxidant peptides are discussed with a particular focus on the aging population. The regulatory requirements for peptide-based antioxidant functional foods are also considered. PMID:22968663

Power, O; Jakeman, P; FitzGerald, R J

2012-09-12

173

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

NASA Astrophysics Data System (ADS)

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.

Leamy, Patrick J.

174

The effect of intrastriatal application of directly and indirectly acting dopamine agonists and antagonists on the in vivo release of acetylcholine measured by brain microdialysis  

Microsoft Academic Search

The effect of intrastriatal application of D-1, D-2 and indirect dopaminergic drugs on the release of striatal acetylcholine as a function of the post-implantation intervals was studied using in vivo microdialysis. The dopamine, D-2 agonists LY 171555 and (-)N0437 inhibited the release of striatal acetylcholine to 40% of control values 16–24 h after implantation of the dialysis cannula. When LY

P. De Boer; G. Damsma; Q. Schram; J. C. Stoof; J. Zaagsma; B. H. C. Westerink

1992-01-01

175

Manufacture of IRDye800CW-coupled Fe3O4 nanoparticles and their applications in cell labeling and in vivo imaging  

PubMed Central

Background In recent years, near-infrared fluorescence (NIRF)-labeled iron nanoparticles have been synthesized and applied in a number of applications, including the labeling of human cells for monitoring the engraftment process, imaging tumors, sensoring the in vivo molecular environment surrounding nanoparticles and tracing their in vivo biodistribution. These studies demonstrate that NIRF-labeled iron nanoparticles provide an efficient probe for cell labeling. Furthermore, the in vivo imaging studies show excellent performance of the NIR fluorophores. However, there is a limited selection of NIRF-labeled iron nanoparticles with an optimal wavelength for imaging around 800 nm, where tissue autofluorescence is minimal. Therefore, it is necessary to develop additional alternative NIRF-labeled iron nanoparticles for application in this area. Results This study manufactured 12-nm DMSA-coated Fe3O4 nanoparticles labeled with a near-infrared fluorophore, IRDye800CW (excitation/emission, 774/789 nm), to investigate their applicability in cell labeling and in vivo imaging. The mouse macrophage RAW264.7 was labeled with IRDye800CW-labeled Fe3O4 nanoparticles at concentrations of 20, 30, 40, 50, 60, 80 and 100 ?g/ml for 24 h. The results revealed that the cells were efficiently labeled by the nanoparticles, without any significant effect on cell viability. The nanoparticles were injected into the mouse via the tail vein, at dosages of 2 or 5 mg/kg body weight, and the mouse was discontinuously imaged for 24 h. The results demonstrated that the nanoparticles gradually accumulated in liver and kidney regions following injection, reaching maximum concentrations at 6 h post-injection, following which they were gradually removed from these regions. After tracing the nanoparticles throughout the body it was revealed that they mainly distributed in three organs, the liver, spleen and kidney. Real-time live-body imaging effectively reported the dynamic process of the biodistribution and clearance of the nanoparticles in vivo. Conclusion IRDye800CW-labeled Fe3O4 nanoparticles provide an effective probe for cell-labeling and in vivo imaging.

2010-01-01

176

Epidural application of spinal instrumentation particulate wear debris: a comprehensive evaluation of neurotoxicity using an in vivo animal model.  

PubMed

Object The introduction and utilization of motion-preserving implant systems for spinal reconstruction served as the impetus for this basic scientific investigation. The effect of unintended wear particulate debris resulting from micromotion at spinal implant interconnections and bearing surfaces remains a clinical concern. Using an in vivo rabbit model, the current study quantified the neural and systemic histopathological responses following epidural application of 11 different types of medical-grade particulate wear debris produced from spinal instrumentation. Methods A total of 120 New Zealand White rabbits were equally randomized into 12 groups based on implant treatment: 1) sham (control), 2) stainless steel, 3) titanium alloy, 4) cobalt chromium alloy, 5) ultra-high molecular weight polyethylene (UHMWPe), 6) ceramic, 7) polytetrafluoroethylene, 8) polycarbonate urethane, 9) silicone, 10) polyethylene terephthalate, 11) polyester, and 12) polyetheretherketone. The surgical procedure consisted of a midline posterior approach followed by resection of the L-6 spinous process and L5-6 ligamentum flavum, permitting interlaminar exposure of the dural sac. Four milligrams of the appropriate treatment material (Groups 2-12) was then implanted onto the dura in a dry, sterile format. All particles (average size range 0.1-50 ?m in diameter) were verified to be endotoxin free prior to implantation. Five animals from each treatment group were sacrificed at 3 months and 5 were sacrificed at 6 months postoperatively. Postmortem analysis included epidural cultures and histopathological assessment of local and systemic tissue samples. Immunocytochemical analysis of the spinal cord and overlying epidural fibrosis quantified the extent of proinflammatory cytokines (tumor necrosis factor-?, tumor necrosis factor-?, interleukin [IL]-1?, IL-1?, and IL-6) and activated macrophages. Results Epidural cultures were negative for nearly all cases, and there was no evidence of particulate debris or significant histopathological changes in the systemic tissues. Gross histopathological examination demonstrated increased levels of epidural fibrosis in the experimental treatment groups compared with the control group. Histopathological evaluation of the epidural fibrous tissues showed evidence of a histiocytic reaction containing phagocytized inert particles and foci of local inflammatory reactions. At 3 months, immunohistochemical examination of the spinal cord and epidural tissues demonstrated upregulation of IL-6 in the groups in which metallic and UHMWPe debris were implanted (p < 0.05), while macrophage activity levels were greatest in the stainless-steel and UHMWPe groups (p < 0.05). By 6 months, the levels of activated cytokines and macrophages in nearly all experimental cases were downregulated and not significantly different from those of the operative controls (p > 0.05). The spinal cord had no evidence of lesions or neuropathology. However, multiple treatments in the metallic groups exhibited a mild, chronic macrophage response to particulate debris, which had diffused intrathecally. Conclusions Epidural application of spinal instrumentation particulate wear debris elicits a chronic histiocytic reaction localized primarily within the epidural fibrosis. Particles have the capacity to diffuse intrathecally, eliciting a transient upregulation in macrophage/cytokine activity response within the epidural fibrosis. Overall, based on the time periods evaluated, there was no evidence of an acute neural or systemic histopathological response to the materials included in the current project. PMID:23808583

Cunningham, Bryan W; Hallab, Nadim J; Hu, Nianbin; McAfee, Paul C

2013-06-28

177

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

Microsoft Academic Search

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

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

2005-01-01

178

Application of quantitative coronary angiography in a cineless environment: In vivo assessment of a fully automated system for clinical use  

Microsoft Academic Search

The accuracy and precision of a fully automated quantitative coronary angiography (QCA) algorithm for use in a cineless environment were determined in phantom studies and in an in vivo canine preparation. Imaging studies of 118 coronary segments in six anesthetized dogs were compared with measurements of the diameters of casts of the canine coronary arteries produced in physiologic conditions. Regression

Jack T. Cusma; Laurence A. Spero; Rob J. van der Geest; Thomas M. Bashore; Kenneth G. Morris

1995-01-01

179

Application of Monte Carlo calculation for the virtual calibration of a low-energy in vivo counting system  

Microsoft Academic Search

Internal dose assessment can be derived from the measurement of retained activity in the whole body or in an organ at a given time. In radiation protection, this assessment, so-called in vivo measurement, is performed by an external measurement of the subject with germanium detectors (in most cases). Calibration of these detectors is ensured by anthropomorphic phantoms which, for technical

Noëlle Pierrat; Loïc de Carlan; Didier Cavadore; Didier Franck

2005-01-01

180

Application of Monte Carlo calculation for the virtual calibration of a low-energy in vivo counting system  

Microsoft Academic Search

Internal dose assessment can be derived from the measurement of retained activity in the whole body or in an organ at a given time. In radiation protection, this assessment, so-called in vivo measurement, is performed by an external measurement of the subject with germanium detectors (in most cases). Calibration of these detectors is ensured by anthropomorphic phantoms, which however can

Noëlle Pierrat; Loïc de Carlan; Didier Cavadore; Didier Franck

2004-01-01

181

Variations in the large surface protein gene of hepatitis B virus in vivo: a longitudinal study and application  

Microsoft Academic Search

AIM: To develop a method for longitudinal study of variations in the large surface protein gene of hepatitis B virus (HBV) in vivo and apply it to the investigation of the dynamics of HBV quasispecies. METHODS: Two HBV carriers were included in the study. Patient 1 was a 38-year-old male who received no anti-virus therapy before and dur- ing the

Nan Yu; Jin Cui; Guo-Bao Zhou; Yun-Jiao Zhang; Jing-Li Chen

182

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

Microsoft Academic Search

\\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

Vicko Gluncic

183

Application of an in vivo mutagenesis system to assess aminothiol effects on neutron-induced genotoxic damage in mouse spleenocytes  

SciTech Connect

A cloning technique has been developed to quantitate and study {ital in vivo} somatic mutations at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus in human lymphocytes. In this paper we describe a modification of this assay to quantify HGPRT mutations in mouse spleenocytes. In particular, we have investigated the effects of the aminothiol on mutagenesis induced by single doses of whole body exposures to fission-spectrum neutrons from the JANUS reactor at Argonne National Laboratory. 7 refs., 3 tabs.

Basic, I. (Zagreb Univ. (Yugoslavia). Dept. of Animal Physiology); Grdina, D.J.; Lyons, T. (Argonne National Lab., IL (USA))

1989-01-01

184

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

Microsoft Academic Search

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

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

2004-01-01

185

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

SciTech Connect

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.

Shanks, J.V.

1989-01-01

186

Application of Twyman-Green interferometer for evaluation of in vivo breakup characteristic of the human tear film  

NASA Astrophysics Data System (ADS)

The paper presents an interferometric method of assessing the in vivo stability of the precorneal tear film. To observe dynamic effects on a human cornea the Twyman-Green interferometer with television frame speed digital registration synchronized with a laser flash was used. The instrument was applied to the human cornea in vivo. The results of the experiment, both tear film distribution and its dynamics, are presented. The proposed interferometric setup can be used to evaluate the breakup characteristics of the tear film, its distribution, and to examine its dynamic changes. The breakup profiles and their cross sections calculated from the interferogram analysis are presented. The depth of recorded breakup, calculated on the basis of interferogram analysis, amounts to about 1.5 micrometers . The proposed method has the advantage of being noncontact and applies only a low-energy laser beam to the eye. This provides noninvasive viewing of human cornea in vivo and makes it possible to observe the kinetics of its tear film deterioration.

Licznerski, Tomasz J.; Kasprzak, Henryk T.; Kowalik, Waldemar

1999-01-01

187

Development of a disposable magnetically levitated centrifugal blood pump (MedTech Dispo) intended for bridge-to-bridge applications--two-week in vivo evaluation.  

PubMed

Last year, we reported in vitro pump performance, low hemolytic characteristics, and initial in vivo evaluation of a disposable, magnetically levitated centrifugal blood pump, MedTech Dispo. As the first phase of the two-stage in vivo studies, in this study we have carried out a 2-week in vivo evaluation in calves. Male Holstein calves with body weight of 62.4–92.2 kg were used. Under general anesthesia, a left heart bypass with a MedTech Dispo pump was instituted between the left atrium and the descending aorta via left thoracotomy. Blood-contacting surface of the pump was coated with a 2-methacryloyloxyethyl phosphorylcholine polymer. Post-operatively, with activated clotting time controlled at 180–220 s using heparin and bypass flow rate maintained at 50 mL/kg/min, plasma-free hemoglobin (Hb), coagulation, and major organ functions were analyzed for evaluation of biocompatibility. The animals were electively sacrificed at the completion of the 2-week study to evaluate presence of thrombus inside the pump,together with an examination of major organs. To date, we have done 13 MedTech Dispo implantations, of which three went successfully for a 2-week duration. In these three cases, the pump produced a fairly constant flow of 50 mL/Kg/min. Neurological disorders and any symptoms of thromboembolism were not seen. Levels of plasma-free Hb were maintained very low. Major organ functions remained within normal ranges. Autopsy results revealed no thrombus formation inside the pump. In the last six cases, calves suffered from severe pneumonia and they were excluded from the analysis. The MedTech Dispo pump demonstrated sufficient pump performance and biocompatibility to meet requirements for 1-week circulatory support. The second phase (2-month in vivo study) is under way to prove the safety and efficacy of MedTech Dispo for 1-month applications. PMID:20928936

Nagaoka, Eiki; Someya, Takeshi; Kitao, Takashi; Kimura, Taro; Ushiyama, Tomohiro; Hijikata, Wataru; Shinshi, Tadahiko; Arai, Hirokuni; Takatani, Setsuo

2010-09-01

188

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

PubMed

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

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

2012-12-05

189

In vivo fluence rate measurements during Foscan®-mediated photodynamic therapy of persistent and recurrent nasopharyngeal carcinomas using a dedicated light applicator  

NASA Astrophysics Data System (ADS)

The objective of this study was to evaluate the performance of a dedicated light applicator for light delivery and fluence rate monitoring during Foscan®-mediated photodynamic therapy of nasopharyngeal carcinoma in a clinical phase I/II study. We have developed a flexible silicone applicator that can be inserted through the mouth and fixed in the nasopharyngeal cavity. Three isotropic fibers, for measuring of the fluence (rate) during therapy, were located within the nasopharyngeal tumor target area and one was manually positioned to monitor structures at risk in the shielded area. A flexible black silicon patch tailored to the patient's anatomy is attached to the applicator to shield the soft palate and oral cavity from the 652-nm laser light. Fourteen patients were included in the study, resulting in 26 fluence rate measurements in the risk volume (two failures). We observed a systematic reduction in fluence rate during therapy in 20 out of 26 illuminations, which may be related to photodynamic therapy-induced increased blood content, decreased oxygenation, or reduced scattering. Our findings demonstrate that the applicator was easily inserted into the nasopharynx. The average light distribution in the target area was reasonably uniform over the length of the applicator, thus giving an acceptably homogeneous illumination throughout the cavity. Shielding of the risk area was adequate. Large interpatient variations in fluence rate stress the need for in vivo dosimetry. This enables corrections to be made for differences in optical properties and geometry resulting in comparable amounts of light available for Foscan® absorption.

van Veen, R. L. P.; Nyst, H.; Indrasari, S. R.; Yudharto, M. A.; Robinson, D. J.; Tan, I. B.; Meewis, C.; Peters, R.; Spaniol, Stefan B.; Stewart, Fiona A.; Levendag, P. C.; Sterenborg, Henricus J. C. M.

2006-07-01

190

Development and application of a quantitative method of monitoring macroscopic cavitation in smooth surface carious lesions in vivo.  

PubMed

Replica techniques have been widely used in dental research, but not previously applied to the in vivo study of carious cavities on free smooth surfaces. A replica method was developed using an epimine resin as the impression material, and a silicone elastomer as the casting material. Cavity areas were measured on the replicas using the Reflex microscope. The sources of method error were identified and quantified. The method was successfully incorporated into a conventional clinical trial and changes in cavity size with time were investigated. PMID:8319258

Neilson, A; Pitts, N B

1993-01-01

191

The application of in vivo laser confocal microscopy to the diagnosis and evaluation of meibomian gland dysfunction  

PubMed Central

Purpose To evaluate the morphological changes of the meibomian glands (MG) in patients with meibomian gland dysfunction (MGD) compared to normal subjects by in vivo confocal microscopy and to investigate the relation of these changes to the clinical ocular surface findings and tear functions. Methods Twenty MGD patients and 15 normal subjects were recruited into this prospective study. Patients and controls underwent slit lamp examinations, tear film break-up time (BUT) measurements, fluorescein and Rose-Bengal stainings, Schirmer test I without anesthesia, tear evaporation rate assessment (TEROS), tear film lipid layer interferometry (DR-1), transillumination of the lids (meibography), MG expressibility test, and in vivo laser confocal microscopy of the lids (HRTII-RCM). Results The BUT, DR-1 tear film lipid layer interferometry grades, fluorescein and Rose-Bengal staining scores, MG drop out grade in meibography, and MG expressibility grades were significantly worse in MGD patients compared to normal controls (p<0.05). The severity of both MG dropout and MG expressibility related significantly with the BUT, DR-1 grades, and TEROS (p<0.05). The mean density of acinar units of MGs as measured by HRTII-RCM was significantly lower in MGD patients (47.6±26.6/mm2) than in control subjects (101.3±33.8/mm2; p<0.05). The mean acinar unit diameter as determined by HRTII-RCM was significantly larger in MGD patients (98.2±53.3 ?m) than in controls (41.6±11.9 ?m; p<0.05). Both the density and diameter of MG acinar units related significantly with the severity of MG dropout and MG expression grades (p<0.05). Conclusions In vivo confocal microscopy can effectively demonstrate the morphological changes of the MG in patients with MGD. Glandular acinar density and acinar unit diameter seemed to be promising new parameters of in vivo confocal microscopy, which is significantly related to the clinical ocular surface and tear function findings of MGD.

Matsumoto, Yukihiro; Sato, Enrique Adan; Ibrahim, Osama M.A.; Tsubota, Kazuo

2008-01-01

192

Development of a Small D-Enantiomeric Alzheimer's Amyloid-? Binding Peptide Ligand for Future In Vivo Imaging Applications  

PubMed Central

Alzheimer’s disease (AD) is a devastating disease affecting predominantly the aging population. One of the characteristic pathological hallmarks of AD are neuritic plaques, consisting of amyloid-? peptide (A?). While there has been some advancement in diagnostic classification of AD patients according to their clinical severity, no fully reliable method for pre-symptomatic diagnosis of AD is available. To enable such early diagnosis, which will allow the initiation of treatments early in the disease progress, neuroimaging tools are under development, making use of A?-binding ligands that can visualize amyloid plaques in the living brain. Here we investigate the properties of a newly designed series of D-enantiomeric peptides which are derivatives of ACI-80, formerly called D1, which was developed to specifically bind aggregated A?1–42. We describe ACI-80 derivatives with increased stability and A? binding properties, which were characterized using surface plasmon resonance and enzyme-linked immunosorbent assays. The specific interactions of the lead compounds with amyloid plaques were validated by ex vivo immunochemistry in transgenic mouse models of AD. The novel compounds showed increased binding affinity and are promising candidates for further development into in vivo imaging compounds.

Funke, Susanne Aileen; Bartnik, Dirk; Gluck, Julian Marius; Piorkowska, Kasia; Wiesehan, Katja; Weber, Urs; Gulyas, Balazs; Halldin, Christer; Pfeifer, Andrea; Spenger, Christian; Muhs, Andreas; Willbold, Dieter

2012-01-01

193

Application of the critical angle method to refractive index measurement of human skin in vivo under partial contact.  

PubMed

We adapted the critical angle method for measuring rough surfaces under partial contact to acquire an in vivo skin refractive index (RI). Assuming that the total reflection is the simple sum of reflection from areas that are in contact and reflection from those that are not in contact, the RI can be estimated even for partial contact with a rough surface. We found that cheek skin is sufficiently soft that a sufficiently large area can be in contact and that the critical angle was detectable. The RIs of the cheeks of adult females were measured. The RI range was about 1.51 to 1.53, at a wavelength of 550 nm, without considering systematic errors. The RIs of cheeks are significantly correlated with their conductance, which corresponds to their water content. We determined the relationship between the RI and conductance within the variation of skin under normal conditions; this relationship was theoretically obtained in previous studies. In the present study, a direct in vivo measurement method was developed that enabled us to measure the RI in daily life, although this method contains errors for several reasons, including disregarding absorption. PMID:23455964

Yoshida, Kenichiro; Ohkubo, Kohji; Ojima, Nobutoshi; Iwata, Kayoko

2013-03-01

194

Application of LC-MS/MS method for the in vivo metabolite determination of oleuropein after intravenous administration to rat.  

PubMed

A highly sensitive, specific and simple LC-MS/MS method was developed to investigate in vivo bio-transformation of oleuropein in rat. Rat urine samples collected after the intravenous administrations were determined using liquid chromatography coupled to tandem mass spectrometry with electrospray ionization in the negative-ion mode. The assay procedure involves a simple liquid-liquid extraction of parent oleuropein and the metabolite from rat urine with ethyl acetate. Chromatographic separation was operated with 0.1% formic acid aqueous and methanol in gradient program at a flow rate of 0.80?mL/min on an RP-C(18) column with a total run time of 30?min. This method has been successfully applied to simultaneous determination of oleuropein and its metabolite in rat urine. Oxygenation was found to be the major metabolic pathway of the oleuropein in rat after intravenous administration. PMID:21308708

Zhou, Ting; Qian, Tianxiu; Wang, Xiaoying; Li, Xianen; Cao, Li; Gui, Shuangying

2011-02-10

195

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

NASA Astrophysics Data System (ADS)

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.

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

2012-09-01

196

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

PubMed

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. PMID:22948192

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

2012-09-05

197

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

PubMed Central

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.

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

2012-01-01

198

Application of a Physiologically Based Pharmacokinetic Model to Assess Propofol Hepatic and Renal Glucuronidation in Isolation: Utility of In Vitro and In Vivo Data  

PubMed Central

A physiologically based pharmacokinetic (PBPK) modeling approach was used to assess the prediction accuracy of propofol hepatic and extrahepatic metabolic clearance and to address previously reported underprediction of in vivo clearance based on static in vitro–in vivo extrapolation methods. The predictive capacity of propofol intrinsic clearance data (CLint) obtained in human hepatocytes and liver and kidney microsomes was assessed using the PBPK model developed in MATLAB software. Microsomal data obtained by both substrate depletion and metabolite formation methods and in the presence of 2% bovine serum albumin were considered in the analysis. Incorporation of hepatic and renal in vitro metabolic clearance in the PBPK model resulted in underprediction of propofol clearance regardless of the source of in vitro data; the predicted value did not exceed 35% of the observed clearance. Subsequently, propofol clinical data from three dose levels in intact patients and anhepatic subjects were used for the optimization of hepatic and renal CLint in a simultaneous fitting routine. Optimization process highlighted that renal glucuronidation clearance was underpredicted to a greater extent than liver clearance, requiring empirical scaling factors of 17 and 9, respectively. The use of optimized clearance parameters predicted hepatic and renal extraction ratios within 20% of the observed values, reported in an additional independent clinical study. This study highlights the complexity involved in assessing the contribution of extrahepatic clearance mechanisms and illustrates the application of PBPK modeling, in conjunction with clinical data, to assess prediction of clearance from in vitro data for each tissue individually.

Gill, Katherine L.; Gertz, Michael; Houston, J. Brian

2013-01-01

199

Application of a physiologically based pharmacokinetic model to assess propofol hepatic and renal glucuronidation in isolation: utility of in vitro and in vivo data.  

PubMed

A physiologically based pharmacokinetic (PBPK) modeling approach was used to assess the prediction accuracy of propofol hepatic and extrahepatic metabolic clearance and to address previously reported underprediction of in vivo clearance based on static in vitro-in vivo extrapolation methods. The predictive capacity of propofol intrinsic clearance data (CLint) obtained in human hepatocytes and liver and kidney microsomes was assessed using the PBPK model developed in MATLAB software. Microsomal data obtained by both substrate depletion and metabolite formation methods and in the presence of 2% bovine serum albumin were considered in the analysis. Incorporation of hepatic and renal in vitro metabolic clearance in the PBPK model resulted in underprediction of propofol clearance regardless of the source of in vitro data; the predicted value did not exceed 35% of the observed clearance. Subsequently, propofol clinical data from three dose levels in intact patients and anhepatic subjects were used for the optimization of hepatic and renal CLint in a simultaneous fitting routine. Optimization process highlighted that renal glucuronidation clearance was underpredicted to a greater extent than liver clearance, requiring empirical scaling factors of 17 and 9, respectively. The use of optimized clearance parameters predicted hepatic and renal extraction ratios within 20% of the observed values, reported in an additional independent clinical study. This study highlights the complexity involved in assessing the contribution of extrahepatic clearance mechanisms and illustrates the application of PBPK modeling, in conjunction with clinical data, to assess prediction of clearance from in vitro data for each tissue individually. PMID:23303442

Gill, Katherine L; Gertz, Michael; Houston, J Brian; Galetin, Aleksandra

2013-01-09

200

Comparison of elastic scattering spectroscopy with histology in ex vivo prostate glands: potential application for optically guided biopsy and directed treatment.  

PubMed

The false-negative rate of ultrasound-guided sextant prostate biopsy has been estimated to be as high as 35 %. A significant percentage (10-35 %) of these prostate cancers diagnosed at a second or later attempt are high grade and, therefore, potentially lethal. We discuss the feasibility for performing optically guided biopsy using elastic scattering spectroscopy (ESS) to reduce sampling errors and improve sensitivity. ESS measurements were performed on 42 prostate glands ex vivo and correlated with standard histopathological assessment. Sliced glands were examined with wavelength ranges of 330-760 nm. The ESS portable system used a new fiber-optic probe with integrated cutting tool, designed specifically for ex vivo pathology applications. ESS spectra were grouped by diagnosis from standard histopathological procedure and then classified using linear support vector machine. Preliminary data are encouraging. ESS data showed strong spectral trends correlating with the histopathological assignments. The classification results showed a sensitivity of 0.83 and specificity of 0.87 for distinguishing dysplastic prostatic tissue from benign prostatic tissue. Similar results were obtained for distinguishing dysplastic prostatic tissue from prostatitis with a sensitivity and specificity of 0.80 and 0.88, respectively. The negative predictive values obtained with ESS are better than those obtained with transrectal ultrasound (TRUS)-guided core-needle biopsy. PMID:23247663

A'amar, O M; Liou, L; Rodriguez-Diaz, E; De Las Morenas, A; Bigio, I J

2012-12-18

201

A computer-assisted rapid-scanning spectrophotometer with applications to tissues in vitro and in vivo.  

PubMed

A flexible approach is described to acquire, store, retrieve, and manipulate the large volume of data provided by a rapid-scanning spectrophotometer designed for biological preparations. Data can be displayed in spectral and kinetic formats. This spectrophotometer provides a means to simultaneously measure the concentrations of components of light scattering preparations and to record chemical reactions as they occur. Although application of this instrument for the study of reduction/oxidation shifts of mitochondrial respiratory chain components is emphasized, the design is suitable for many spectrophotometric applications. PMID:2996832

Pikarsky, S M; LaManna, J C; Sick, T J; Rosenthal, M

1985-10-01

202

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

Microsoft Academic Search

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

Patrick J. Leamy

2003-01-01

203

Modelling and testing of a piezoelectric ultrasonic micro-motor suitable for in vivo micro-robotic applications  

NASA Astrophysics Data System (ADS)

A piezoelectric ultrasonic resonant micro-motor is developed with a stator diameter of 241 ?m and an overall diameter of 400 ?m. The motor is shown to produce a start-up torque of 1.2 nN m and a peak output power of 0.25 ?W as designed, with a preload of 46.6 ?N. An increase in preload to 2264 ?N improved the performance to a start-up torque of 29 nN m and a peak output power of 9.1 ?W. The motor is five times smaller than the current smallest piezoelectric ultrasonic resonant motor produced by Kanda et al. The motor is designed to operate at approximately 771 kHz, matching the fundamental axial, second harmonic torsional and electro-mechanical resonant frequencies. This is achieved through the use of a novel design process that uses scaling theories to greatly reduce the computational time to design the device. The resultant size and performance of the motor make it the first motor design capable of meeting the requirements of a drive system in a tetherless swimming in vivo micro-robot.

Watson, B.; Friend, J.; Yeo, L.

2010-11-01

204

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

PubMed Central

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.

Leszczynski, Rafal; Stodolak, Ewa; Wieczorek, Jaroslaw; Orlowska-Heitzman, Jolanta; Gumula, Teresa

2010-01-01

205

Towards an inhalative in vivo application of immunomodulating gelatin nanoparticles in horse-related preformulation studies.  

PubMed

Delivering active ingredients using biocompatible and biodegradable carriers such as gelatin nanoparticles (GNPs) to the lung constitutes a promising non-invasive route of administration. However, the pulmonary delivery of nanoparticle-based immunotherapy is still a field that requires more clarification. In this study, GNPs loaded with cytosine-phosphate-guanine oligodeoxynucleotides (CpG-ODN)-loaded and plain GNPs were aerosolised either by a conventional pressured metered dose inhaler (pMDI) or by active or passive vibrating-mesh (VM) nebulisers. GNP sizes after nebulisation by active and passive VM nebulisers were 248.2?±?7.34 and 222.3?±?1.42?nm, respectively. GNP concentrations after aerosolisation were found consistent and second-stage particle deposition in an impinger was up to 65.68?±?11.2% of the nebulised dose. VM nebulisers produced high fine particle fractions, while pMDIs did not. Nebulised CpG-ODN-loaded GNPs remained capable to stimulate IL-10 release (225.2?±?56.3?pg/ml) in vitro from equine alveolar lymphocytes. Thus, a novel system for pulmonary GNP-mediated immunotherapy in vivo was established. PMID:22432849

Fuchs, Sebastian; Klier, John; May, Anna; Winter, Gerhard; Coester, Conrad; Gehlen, Heidrun

2012-03-20

206

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

PubMed Central

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.

Mudgil, Meetali; Pawar, Pravin K.

2013-01-01

207

Silver-nanolipid complex for application to atopic dermatitis skin: rheological characterization, in vivo efficiency and theory of action.  

PubMed

A skin care formulation was developed by incorporating microsilver, in combination with nanostructured lipid carriers (NLC) into an o/w cream and lotion. To increase skin adhesion of the NLC, and subsequent film formation and occlusion onto the skin, the NLC were produced with a size of about 200 nm. Production of NLC was performed by high-pressure homogenisation. Characterization was performed regarding size and charge (zeta potential), and for the cream and lotion also by rheology. Incorporation of NLC and/or microsilver into the cream or lotion led to pronounced changes in the thixotropic behaviour (shape of rheogram, yield point, viscosity). This was explained by specific interaction of the nanoparticles and/or the microsilver with the two formulations. In vivo studies revealed a high potential to remove not only symptoms of irritated sensitive skin, but also of light to medium atopic dermatitis. Based on zeta potential measurements, a silver ion-nanolipid complex seems to form which leads to a higher activity of the antimicrobial silver, e.g., increasing the silver ion concentration on skin and bacterial membranes. The antimicrobial effect in combination with restoration of normal skin condition (repair of stratum corneum lipid film by NLC) is obviously sufficient to replace in many cases medical therapy with glucocorticoids by a biological, natural skin care cosmetic nano formulation. PMID:20055090

Keck, Cornelia M; Schwabe, Kay

2009-08-01

208

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

PubMed Central

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.

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

209

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

PubMed

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

Mudgil, Meetali; Pawar, Pravin K

2013-02-04

210

Development and application of in vivo expression technology (IVET) for analysing microbial gene expression in complex environments.  

PubMed

Establishing the mechanisms by which microbes interact with their environment, including eukaryotic hosts, is a major challenge that is essential for the economic utilisation of microbes and their products. Techniques for determining global gene expression profiles of microbes, such as microarray analyses, are often hampered by methodological restraints, particularly the recovery of bacterial transcripts (RNA) from complex mixtures and rapid degradation of RNA. A pioneering technology that avoids this problem is In Vivo Expression Technology (IVET). IVET is a 'promoter-trapping' methodology that can be used to capture nearly all bacterial promoters (genes) upregulated during a microbe-environment interaction. IVET is especially useful because there is virtually no limit to the type of environment used (examples to date include soil, oomycete, a host plant or animal) to select for active microbial promoters. Furthermore, IVET provides a powerful method to identify genes that are often overlooked during genomic annotation, and has proven to be a flexible technology that can provide even more information than identification of gene expression profiles. A derivative of IVET, termed resolvase-IVET (RIVET), can be used to provide spatio-temporal information about environment-specific gene expression. More recently, niche-specific genes captured during an IVET screen have been exploited to identify the regulatory mechanisms controlling their expression. Overall, IVET and its various spin-offs have proven to be a valuable and robust set of tools for analysing microbial gene expression in complex environments and providing new targets for biotechnological development. PMID:16918484

Jackson, R W; Giddens, S R

2006-09-01

211

Choice of detectors for in vivo elemental analysis by counting natural and neutron-induced gamma rays for medical applications  

NASA Astrophysics Data System (ADS)

Body fat is measured by detecting C and O in vivo through fast neutron inelastic scattering. A sealed D-T neutron generator is used for the pulsed (4-10 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 12C and 16O. Large Bi4Ge3O12 (BGO) crystal detectors (127 × 76 mm) are used for the gamma ray detection during the 10 ?s neutron burst. BGO detectors improved the signal to background ratio for the carbon detection by a factor of six compared to 152 × 152 mm NaI (Tl) detectors. Exposure to scattered neutrons did not affect the gain stability of the BGOs. Thermal neutrons from a moderated 238Pu-Be source are used for the measurement of total body nitrogen (and thus protein). The resulting high energy prompt gamma rays from nitrogen (10.83 MeV) are detected simultaneously with the irradiation. BGO detectors have superior stability operating in an environment of variable neutron exposure and high counting rates. However, the presence of neutrons creates a 10.2 MeV gamma ray peak from 73Ge in the BGO detector which interferes with the nitrogen peak. Whole body gamma ray counters, consisting of NaI(Tl) crystal detectors in a shielded room, are used to measure the natural radioactivity of the body due to 40K. They are also used to measure body Ca, P, Na and Cl, following total body exposure to thermal neutrons.

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

1994-12-01

212

In vivo complementation of complex I by the yeast Ndi1 enzyme. Possible application for treatment of Parkinson disease.  

PubMed

Recent studies suggest that dysfunction of the NADH-quinone oxidoreductase (complex I) is associated with a number of human diseases, including neurodegenerative disorders such as Parkinson disease. We have shown previously that the single subunit rotenone-insensitive NADH-quinone oxidoreductase (Ndi1) of Saccharomyces cerevisiae mitochondria can restore NADH oxidation in complex I-deficient mammalian cells. The Ndi1 enzyme is insensitive to complex I inhibitors such as rotenone and 1-methyl-4-phenylpyridinium ion, known as a metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). To test the possible use of the NDI1 gene as a therapeutic agent in vivo, we chose a mouse model of Parkinson disease. The NDI1-recombinant adeno-associated virus particles (rAAV-NDI1) were injected unilaterally into the substantia nigra of mice. The animals were then subjected to treatment with MPTP. The degree of neurodegeneration in the nigrostriatal system was assessed immunohistochemically through the analysis of tyrosine hydroxylase and glial fibrillary acidic protein. It was evident that the substantia nigra neurons on the side used for injection of rAAV-NDI1 retained a high level of tyrosine hydroxylase-positive cells, and the ipsilateral striatum exhibited significantly less denervation than the contralateral striatum. Furthermore, striatal concentrations of dopamine and its metabolites in the hemisphere that received rAAV-NDI1 were substantially higher than those of the untreated hemisphere, reaching more than 50% of the normal levels. These results indicate that the expressed Ndi1 protein elicits resistance to MPTP-induced neuronal injury. The present study is the first successful demonstration of complementation of complex I by the Ndi1 enzyme in animals. PMID:16543240

Seo, Byoung Boo; Nakamaru-Ogiso, Eiko; Flotte, Terence R; Matsuno-Yagi, Akemi; Yagi, Takao

2006-03-16

213

Transcutaneous Application of Carbon Dioxide (CO2) Induces Mitochondrial Apoptosis in Human Malignant Fibrous Histiocytoma In Vivo  

PubMed Central

Mitochondria play an essential role in cellular energy metabolism and apoptosis. Previous studies have demonstrated that decreased mitochondrial biogenesis is associated with cancer progression. In mitochondrial biogenesis, peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1?) regulates the activities of multiple nuclear receptors and transcription factors involved in mitochondrial proliferation. Previously, we showed that overexpression of PGC-1? leads to mitochondrial proliferation and induces apoptosis in human malignant fibrous histiocytoma (MFH) cells in vitro. We also demonstrated that transcutaneous application of carbon dioxide (CO2) to rat skeletal muscle induces PGC-1? expression and causes an increase in mitochondrial proliferation. In this study, we utilized a murine model of human MFH to determine the effect of transcutaneous CO2 exposure on PGC-1? expression, mitochondrial proliferation and cellular apoptosis. PGC-1? expression was evaluated by quantitative real-time PCR, while mitochondrial proliferation was assessed by immunofluorescence staining and the relative copy number of mitochondrial DNA (mtDNA) was assessed by real-time PCR. Immunofluorescence staining and DNA fragmentation assays were used to examine mitochondrial apoptosis. We also evaluated the expression of mitochondrial apoptosis related proteins, such as caspases, cytochorome c and Bax, by immunoblot analysis. We show that transcutaneous application of CO2 induces PGC-1? expression, and increases mitochondrial proliferation and apoptosis of tumor cells, significantly reducing tumor volume. Proteins involved in the mitochondrial apoptotic cascade, including caspase 3 and caspase 9, were elevated in CO2 treated tumors compared to control. We also observed an enrichment of cytochrome c in the cytoplasmic fraction and Bax protein in the mitochondrial fraction of CO2 treated tumors, highlighting the involvement of mitochondria in apoptosis. These data indicate that transcutaneous application of CO2 may represent a novel therapeutic tool in the treatment of human MFH.

Onishi, Yasuo; Kawamoto, Teruya; Ueha, Takeshi; Kishimoto, Kenta; Hara, Hitomi; Fukase, Naomasa; Toda, Mitsunori; Harada, Risa; Minoda, Masaya; Sakai, Yoshitada; Miwa, Masahiko; Kurosaka, Masahiro; Akisue, Toshihiro

2012-01-01

214

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

PubMed Central

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.

Li, Tianshu; Takeoka, Shinji

2013-01-01

215

Intermolecular double-quantum coherence imaging without coherence selection gradients and its application in in vivo MRI.  

PubMed

In the COSY Revamped with Asymmetric Z-gradient Echo Detection (CRAZED) experiments, magnetization is modulated by the distant dipolar field (DDF) generated by coherence selection gradient (CSG) commonly in sinusoidal wave-form and results in detectable intermolecular multiple-quantum coherence (iMQC) signal. IMQCs have some attractive features, but their intrinsic weak signal intensity prevents their widespread applications. In this paper, a new phase cycling scheme was applied to obtain intermolecular double-quantum coherence (iDQC) signal. It is found that DDF can arise from nonspherical sample geometry or background inhomogeneous field in the absence of CSGs, which is more efficient than that created from CSGs. The experimental results show that the resulting DDF can refocus the ±iDQC signals simultaneously and thus enhance the signal intensity to about two folds of that from the conventional CRAZED sequence. Theoretical prediction and experiments give coincident results. PMID:23473838

Shen, Guiping; Cai, Congbo; Chen, Zhong; Cai, Shuhui

2013-03-06

216

[In vivo measurement of ocular circulation with the laser speckle method--development of apparatus and application in ophthalmological research].  

PubMed

We have developed an apparatus utilizing laser speckle phenomenon which can measure the peripheral circulation in the iris, choroid, retina and optic nerve head (ONH) and blood velocity through retinal vessels in the living eye non-invasively and quantitatively. A blue-component argon laser (wavelength 488 nm) was used for measurement of peripheral circulation in the retina and a diode laser (wavelength 808 nm) for measurements of peripheral circulation in the iris, posterior choroid and ONH, and measurement of centerline blood velocity through retinal vessels. A fundus camera (TRC-WT 3, Topcon) was equipped with a laser source and an image sensor where the speckle pattern from the fundus appears, and the data were analyzed with a personal computer to give a normalized blur (NB) value or a square blur rate (SBR) value, both quantitative indices of blood velocity. The NB value, whose computation requires much less time, was adopted to evaluate peripheral circulation because of non-linear correlation between the NB and actual blood velocity in the range above 20 mm/sec. The SBR value, whose computation requires a longer time, was adopted for measurement of blood velocity through retinal vessels. Measurement field in the living eye was 1.06 x 1.06 mm at its maximum and reproducibility index of the in vivo measurement in the rabbit iris, choroid, retina, and ONH was approximately 10%. When blood flow was changed by intraocular pressure (IOP) change in rabbit eyes, NB values obtained from the iris, choroid, and retina showed a significant correlation with the blood flow simultaneously determined with the colored microsphere technique in the same eye, and the NB obtained from the ONH also correlated with the blood flow determined with the H2 gas clearance method. Stepwise reduction in the ocular perfusion pressure (OPP) by stepwise increment of IOP resulted in proportional reduction in the iris- and choroid-NB. On the other hand, the retina- or ONH-NB remained almost unaltered at OPP levels above 50 mmHg, and decreased along with OPP at levels less than 50 mmHg. By monitoring NB values for 2 hours, presence or absence of autoregulatory mechanism against OPP change in the choroidal and ONH circulation was studied in rabbits. Throughout the experimental period of 2 hours, the choroidal NB was changed along with the OPP change, suggesting absence of blood flow autoregulation in this tissue. In the ONH, however, the NB returned to the baseline after its transient increase or decrease when the OPP was continuously increased or decreased, showing the presence of an autoregulatory mechanism in the ONH circulation. However, the time course of the NB resumption depended on the extent of OPP change. These results indicated that the laser speckle method can be useful in investigating the autoregulatory mechanism and processes of peripheral circulation in ocular tissues. Unilateral instillation of drugs with vasodilative activity (ifenprodil, betaxolol or nipradilol) in rabbit eyes significantly increased ONH and/or choroidal circulation. The extent in change in the ONH and/or choroidal circulation correlated with the number of doses, but not with the extent of IOP reduction, which suggested that the observed effects were attributable to the drug which penetrated locally. Intravenous administration of a Ca(2+)-antagonist (nicardipine, nilvadipine or pranidipine) significantly increased choroidal or retinal circulation in rabbits. The ONH circulation, however, was not affected by nicardipine, but affected by nilvadipine or pranidipine. Given the same effect on the ONH circulation, systemic hypotensive effect was stronger in pranidipine than in nilvadipine, which suggested that nilvadipine can be used in patients with ocular circulatory insufficiency. A modification of the laser speckle apparatus used for animal experiments was devised so that the NB or SBR values could be measured in human eyes every 0.12 sec on a real-time basis. (ABSTRACT TRUN PMID:10643292

Araie, M

1999-12-01

217

Multifocal animated imaging of changes in cellular oxygen and calcium concentrations and membrane potential within the intact adult mouse carotid body ex vivo.  

PubMed

Carotid body (CB) type I cell hypoxia-sensing function is assumed to be based on potassium channel inhibition. Subsequent membrane depolarization initiates an intracellular calcium increase followed by transmitter release for excitation of synapses with linked nerve endings. Several reports, however, contradict this generally accepted concept by showing that type I cell oxygen-sensing properties vary significantly depending on the method of their isolation. We report therefore for the first time noninvasive mapping of the oxygen-sensing properties of type I cells within the intact adult mouse CB ex vivo by using multifocal Nipkow disk-based imaging of oxygen-, calcium- and potential-sensitive cellular dyes. Characteristic type I cell clusters were identified in the compact tissue by immunohistochemistry because of their large cell nuclei combined with positive tyrosine hydroxylase staining. The cellular calcium concentrations in these cell clusters either increased or decreased in response to reduced tissue oxygen concentrations. Under control conditions, cellular potential oscillations were uniform at ?0.02 Hz. Under hypoxia-induced membrane depolarization, these oscillations ceased. Simultaneous increases and decreases in potential of these cell clusters resulted from spontaneous burstlike activities lasting ?1.5 s. type I cells, identified during the experiments by cluster formation in combination with large cell nuclei, seem to respond to hypoxia with heterogeneous kinetics. PMID:21525432

Wotzlaw, Christoph; Bernardini, André; Berchner-Pfannschmidt, Utta; Papkovsky, Dmitri; Acker, Helmut; Fandrey, Joachim

2011-04-27

218

Expanding the Versatility of Mesoporous Silica Nanoparticles towards Drug Delivery for In-vitro, In-vivo and Clinical Applications  

NASA Astrophysics Data System (ADS)

The work covered in this thesis focuses on research developments in the mesoporous silica nanoparticle platform as a drug delivery vehicle for containment and controlled release of therapeutic agents to inhibit disease. Mesoporous silica is a very versatile material with a very robust structure that is easily modified both internally and externally to change its physical properties. Once modified, the silica nanoparticies can be loaded with therapeutic agents that can be isolated from interacting with their surroundings until an on command delivery signal is received. In this dissertation, first, application of a noninvasive externally controlled means of activation such as light activation and magnetically based heating have been investigated and achieved. Next, by altering the structure of rotaxanes based on azobenzene, steps towards a self-sealing light activated full rotaxane system have been developed. Then, through the manipulation of the particle structure as well as the internal pore environment of silica particle, the interaction between guest drug molecules and the particles has been better understood towards optimizing drug loading and release efficiency. Finally, surface modification of silica nanoparticles with biomolcules has been achieved and observed to increase the efficacy of the silica nanoparticle system in the cellular environment. A combination of all these areas of research results in the advancement of the mesoporous silica nanoparticle drug delivery system towards utilization within living organisms.

Ferris, Daniel Patrick

219

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

NASA Astrophysics Data System (ADS)

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.

Cover, Keith S.

220

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

PubMed

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

Karvaly, G; Gachályi, A; Furész, J

2008-01-01

221

In vitro dentine permeability evaluation of HEMA-based (desensitizing) products using split-chamber model following in vivo application in the dog.  

PubMed

The aim of this study was to evaluate in vitro dentine permeability evaluation of 2-Hydroxyethyl methacrylate (HEMA)-based desensitizing products using split-chamber model following in vivo application in the dogs for three experimental time periods of 1 week, 1 month and 3 months. Buccal enamel of upper and lower canines of nine young dogs was removed and flat dentinal surface was obtained using a water-cooled diamond bur. The dentinal surface divided into four quarters, three experimental and a control. Health-Dent and Gluma Desensitizing agent and Single bond were applied to respective quarters of one tooth, according to manufacturers' instructions. The last quarters were left as control without any applications. For the each experimental period, 10 dentine samples from each group including control were used to measure the hydraulic conductance values. The dentine discs were placed pulp-side down in a split-chamber device in which the plastic spacers containing the rubber 0 rings have a surface area of 1 mm(2) and permeability was measured by fluid filtration. The data were expressed as hydraulic conductance (Lp). Differences in dentine permeability obtained for the desensitizing agents against the control were tested for statistical significance using Kruskal-Wallis one-way ANOVA and Bonferroni-adjusted Mann-Whitney U-tests. Differences in permeability in three time periods for each desensitizing agent were analysed using Friedman's anova and Wilcoxon signed rank tests. All desensitizing applications caused decreased Lp values compared with the control Lp value at the end of 1 week. In the 1-month samples, between the Lp values of Single bond and control, no statistical difference was found (P > 0.05). At the end of the 3-month period, the Gluma Desensitizing agent had the lowest the Lp value (P < 0.05). The result of this study is that the topical application of desensitizing agents led to decrease in dentine permeability in the dog model. The Gluma Desensitizing agent provide the more lasting tubule-occluding effect than the other material tested in this model. PMID:15634299

Duran, I; Sengun, A; Yildirim, T; Ozturk, B

2005-01-01

222

In vitro and in vivo application of active compounds with anti-yeast activity to improve the shelf life of ready-to-eat table grape.  

PubMed

The anti-yeast effects of several compounds at different concentrations were screened in vitro against main table grape spoilage yeasts. The compounds showing the most significant anti-yeast activity were applied by dipping to table grape, to evaluate the sensory perception. In a subsequent final step, dipping treatments with potassium sorbate, eugenol, citrus extract and ethanol, were applied to ready-to-eat seedless table grape, packaged in air or under modified atmosphere packaging (MAP). The in vitro test highlights good effects of cinnamon bark oil and citrus extract, even at the lowest concentrations used in this work. From a sensory point of view, the preliminary panel test selected potassium sorbate, citrus extract, eugenol and ethanol as most suitable substances. The in vivo application of active compounds showed that dipping in eugenol solution and ethanol (20 and 50 %) in combination with MAP increased shelf life of fruit if compared to the control sample (24.08, 28.47, 35.79 and 14.26 days, respectively). PMID:23512208

Cristina, Costa; Annalisa, Lucera; Amalia, Conte; Francesco, Contò; Del Nobile, Matteo Alessandro

2013-03-20

223

Universally applicable methods for monitoring response regulator aspartate phosphorylation both in vitro and in vivo using Phos-tag-based reagents.  

PubMed

Recent development of the phosphate chelator, Phos-tag, together with Phos-tag pendant reagents, has provided new methods for detection of phosphorylated serine, threonine, tyrosine, and histidine residues in phosphoproteins. We have investigated the use of Phos-tag for detection and quantification of phospho-aspartate in response regulator proteins that function within two-component signaling systems. Alternative methods are especially important, because the labile nature of the acylphosphate bond in response regulator proteins has restricted the application of many traditional methods of phosphoprotein analysis. We demonstrate that Phos-tag gel stain can be used to detect phospho-Asp in response regulators and that Phos-tag acrylamide gel electrophoresis can be used to separate phosphorylated and unphosphorylated forms of response regulator proteins. The latter method, coupled to Western blot analysis, enables detection of specific phosphorylated proteins in complex mixtures such as cell lysates. Standards of phosphorylated proteins can be used to correct for hydrolysis of the labile phospho-Asp bond that invariably occurs during analysis. We have employed Phos-tag methods to characterize the phosphorylation state of the Escherichia coli response regulator PhoB both in vitro, using purified protein, and in vivo, by analyzing lysates of cells grown under different conditions of induction of the PhoR/PhoB phosphate assimilation pathway. PMID:18328252

Barbieri, Christopher M; Stock, Ann M

2008-02-13

224

Time-resolved singlet oxygen luminescence detection under photodynamic therapy relevant conditions: comparison of ex vivo application of two photosensitizer formulations  

NASA Astrophysics Data System (ADS)

Singlet oxygen plays a crucial role in photo-dermatology and photodynamic therapy (PDT) of cancer. Its direct observation by measuring the phosphorescence at 1270 nm, however, is still challenging due to the very low emission probability. It is especially challenging for the time-resolved detection of singlet oxygen kinetics in vivo which is of special interest for biomedical applications. Photosensitized generation of singlet oxygen, in pig ear skin as model for human skin, is investigated here. Two photosensitizers (PS) were topically applied to the pig ear skin and examined in a comparative study, which include the amphiphilic pheophorbide-a and the highly hydrophobic perfluoroalkylated zinc phthalocyanine (F64PcZn). Fluorescence microscopy indicates the exclusive accumulation of pheophorbide-a in the stratum corneum, while F64PcZn can also accumulate in deeper layers of the epidermis of the pig ear skin. The kinetics obtained with phosphorescence measurements show the singlet oxygen interaction with the PS microenvironment. Different generation sites of singlet oxygen correlate with the luminescence kinetics. The results show that singlet oxygen luminescence detection can be used as a diagnostic tool, not only for research, but also during treatment. The detection methodology is suitable for the monitoring of chemical quenchers' oxidation as well as O2 saturation at singlet oxygen concentration levels relevant to PDT treatment protocols.

Schlothauer, Jan C.; Hackbarth, Steffen; Jäger, Lutz; Drobniewski, Kai; Patel, Hemantbhai; Gorun, Sergiu M.; Röder, Beate

2012-11-01

225

In Vivo Imaging of Molecularly Targeted Phage  

Microsoft Academic Search

Rapid identification of in vivo affinity ligands would have far-reaching applications for imaging specific molecular targets, in vivo systems imaging, and medical use. We have developed a high-throughput method for identifying and optimizing ligands to map and image biologic targets of interest in vivo .W e directly labeled viable phage clones with far-red fluorochromes and comparatively imaged them in vivo

Kimberly A. Kelly; Peter Waterman; Ralph Weissleder

2006-01-01

226

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....

227

Alterations in a redox oxygen sensing mechanism in chronic hypoxia.  

PubMed

The mechanism of acute hypoxic pulmonary vasoconstriction (HPV) may involve the inhibition of several voltage-gated K+ channels in pulmonary artery smooth muscle cells. Changes in PO2 can either be sensed directly by the channel(s) or be transmitted to the channel via a redox-based effector mechanism. In control lungs, hypoxia and rotenone acutely decrease production of activated oxygen species, inhibit K+ channels, and cause constriction. Two-day and 3-wk chronic hypoxia (CH) resulted in a decrease in basal activated oxygen species levels, an increase in reduced glutathione, and loss of HPV and rotenone-induced constriction. In contrast, 4-aminopyridine- and KCl-mediated constrictions were preserved. After 3-wk CH, pulmonary arterial smooth muscle cell membrane potential was depolarized, K+ channel density was reduced, and acute hypoxic inhibition of whole cell K+ current was lost. In addition, Kv1.5 and Kv2.1 channel protein was decreased. These data suggest that chronic reduction of the cytosol occurs before changes in K+ channel expression. HPV may be attenuated in CH because of an impaired redox sensor. PMID:11356790

Reeve, H L; Michelakis, E; Nelson, D P; Weir, E K; Archer, S L

2001-06-01

228

HIF1 and oxygen sensing in the brain  

Microsoft Academic Search

Of all the chemical elements, oxygen is the most vital to the human body. The brain is the most sensitive organ to oxygen deprivation (hypoxia), which, over an extended period, can cause coma, seizures, cognitive impairment and other neurological disabilities, and even brain death. However, during mild hypoxia of short duration, the brain develops adaptative mechanisms that allow it to

Myriam Bernaudin; Frank R. Sharp

2004-01-01

229

FRET excited ratiometric oxygen sensing in living tissue.  

PubMed

Dynamic analysis of oxygen (O?) 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 [O?] with several hundred millisecond temporal resolution in varying biological microdomains of active brain tissue. PMID:23333398

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

2013-01-17

230

Application of the correlation of in vitro dissolution behavior and in vivo plasma concentration profile (IVIVC) for soft-gel capsules--a pointless pursuit?  

PubMed

Plasma concentration profiles of arundic acid ((R)-(-)-2-propyloctanoic acid), an oil-like medicine, administered as soft-gel capsules in human clinical tests were predicted from the dissolution test data of the soft-gel capsules with different storage terms (short- and long-term stored drugs) by applying the in vitro-in vivo correlation (IVIVC). We established two linear-regression IVIVCs, which were characterized by either the in vitro dissolution behaviors against the pH 8.0 dissolution medium or the pH 6.8 dissolution medium containing 2% sodium dodecyl sulfate (SDS), in this study. Also, the prediction accuracies for the in vivo plasma profiles in humans for these two IVIVCs were compared. Regarding dissolution from the long-term stored capsule in pH 8.0 dissolution medium without surfactant, the prediction accuracies of the in vivo plasma profiles in humans were not satisfactory for the obtained IVIVC. The use of pH 6.8 dissolution medium containing 2% SDS, according to the Japanese guideline, improved the dissolution of the long-term stored capsule. Furthermore, the prediction accuracies for the in vivo plasma profiles in humans for these two IVIVCs were compared. The IVIVC established by the in vitro dissolution data obtained with the dissolution medium containing surfactant more effectively predicted the plasma drug concentration profiles following oral administrations of the soft-gel capsules under both storage conditions. PMID:17978506

Nishimura, Hidekatsu; Hayashi, Chiaki; Aiba, Tetsuya; Okamoto, Ichiro; Miyamoto, Yuji; Nakade, Susumu; Takeda, Kazuhisa; Kurosaki, Yuji

2007-11-01

231

Diffuse optical tomography with an amplified ultrafast laser and a single-shot streak camera: application to real-time in vivo songbird neuro-imaging  

Microsoft Academic Search

A new ultrafast Diffuse Optical Tomography (DOT) has been developed for real time in vivo brain metabolism monitoring in songbird. The technique is based on space resolved time of flight measurements of the photons across the brain tissues. A three dimensional reconstruction of the brain activity is foreseeable by means of a double space and time sampling of the reflectance

Hugues Guillet de Chatellus; Clémentine Vignal; Stéphane Ramstein; Nicolas Verjat; Nicolas Mathevon; Stéphane Mottin

2005-01-01

232

EPR Spectroscopy of Function In Vivo  

Microsoft Academic Search

EPR can be used to study free radicals in vivo, environmental and biophysical parameters in cells and tissues, and to report metabolism, physiology, and biochemistry. The authors have attempted to judge which of these types of measurements will be productive for studies in animals and in humans. It is envisioned that a large number of in vivo applications of EPR

Harold M. Swartz; Nadeem Khan

233

Metal-Enhanced Fluorescence (MEF) Due to Silver Colloids on a Planar Surface:  Potential Applications of Indocyanine Green to in Vivo Imaging †  

Microsoft Academic Search

We examined the effects of metallic silver colloids on the fluorescence spectral properties of indocyanine green (ICG), which is a dye widely used for in vivo medical testing. Silver colloids from a suspension bind spontaneously to amine-coated surfaces. These colloid-coated surfaces were found to cause a 30-fold increase in the intensity of ICG, which was held close to the metal

Chris D. Geddes; Haishi Cao; Ignacy Gryczynski; Zygmunt Gryczynski; Jiyu Fang; Joseph R. Lakowicz

2003-01-01

234

Genetically engineered brain drug delivery vectors: cloning, expression and in vivo application of an anti-transferrin receptor single chain antibody-streptavidin fusion gene and protein.  

PubMed

A single chain Fv antibody-streptavidin fusion protein was expressed and purified from bacterial inclusion bodies following cloning of the genes encoding the variable region of the heavy chain and light chain of the murine OX26 monoclonal antibody to the rat transferrin receptor. The latter undergoes receptor mediated transcytosis through the brain capillary endothelial wall in vivo, which makes up the blood-brain barrier (BBB); therefore, the OX26 monoclonal antibody and its single chain Fv analog may act as brain drug delivery vectors in vivo. Attachment of biotinylated drugs to the antibody vector is facilitated by production of the streptavidin fusion protein. The bi-functionality of the OX26 single chain Fv antibody-streptavidin fusion protein was retained, as the product both bound biotin and the rat transferrin receptor in vitro and in vivo, based on pharmacokinetic and brain uptake analyses in anesthetized rats. The attachment of biotin-polyethyleneglycol-fluorescein to the OX26 single chain Fv antibody-streptavidin fusion protein resulted in illumination of isolated rat brain capillaries in confocal fluorescent microscopy. In conclusion, these studies demonstrate that genetically engineered single chain Fv antibody-streptavidin fusion proteins may be used for non-invasive neurotherapeutic delivery to the brain using endogenous BBB transport systems such as the transferrin receptor. PMID:10506289

Li, J Y; Sugimura, K; Boado, R J; Lee, H J; Zhang, C; Duebel, S; Pardridge, W M

1999-09-01

235

In vivo imaging flow cytometer  

PubMed Central

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.

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

2009-01-01

236

In vivo imaging flow cytometer.  

PubMed

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. PMID:19529148

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

2006-08-21

237

In vitro and in vivo analysis of endothelial progenitor cells from cryopreserved umbilical cord blood: are we ready for clinical application?  

PubMed

Umbilical cord blood (CB) represents a main source of circulating endothelial progenitor cells (cEPCs). In view of their clinical use, in either the autologous or allogeneic setting, cEPCs should likely be expanded from CB kept frozen in CB banks. In this study, we compared the expansion, functional features, senescence pattern over culture, and in vivo angiogenic potential of cEPCs isolated from fresh or cryopreserved CB (cryoCB). cEPCs could be isolated in only 59% of cryoCB compared to 94% for fresh CB, while CB units were matched in terms of initial volume, nucleated and CD34(+) cell number. Moreover, the number of endothelial colony-forming cells was significantly decreased when using cryoCB. Once cEPCs culture was established, the proliferation, migration, tube formation, and acetylated-LDL uptake potentials were similar in both groups. In addition, cEPCs derived from cryoCB displayed the same senescence status and telomeres length as that of cEPCs derived from fresh CB. Karyotypic aberrations were found in cells obtained from both fresh and cryoCB. In vivo, in a hind limb ischemia murine model, cEPCs from fresh and cryoCB were equally efficient to induce neovascularization. Thus, cEPCs isolated from cryoCB exhibited similar properties to those of fresh CB in vitro and in vivo. However, the low frequency of cEPCs colony formation after cryopreservation shed light on the need for specific freezing conditions adapted to cEPCs in view of their future clinical use. PMID:20447337

Vanneaux, Valérie; El-Ayoubi, Fida; Delmau, Catherine; Driancourt, Catherine; Lecourt, Séverine; Grelier, Aurore; Cras, Audrey; Cuccuini, Wendy; Soulier, Jean; Lataillade, Jean-Jacques; Lebousse-Kerdiles, Marie-Caroline; Oury, Jean François; Sibony, Olivier; Marolleau, Jean-Pierre; Benbunan, Marc; Uzan, Georges; Larghero, Jérôme

2010-05-04

238

[Frontiers in vitamin D; basic research and clinical application. Inhibitory effects of eldecalcitol, an active vitamin D derivative, on bone resorption in vivo].  

PubMed

Eldecalcitol is a new vitamin D(3) derivative recently approved for the treatment of osteoporosis in Japan. Previous studies showed that daily administration of eldecalcitol increased bone mineral density (BMD) by suppressing bone resorption in animals and in patients with osteoporosis. We examined how eldecalcitol suppresses bone resorption in vivo . Daily administration of eldecalcitol into mice did not affect properties of osteoclast precursors, but suppressed RANKL expression in bone. These results suggest that daily administration of eldecalcitol increase BMD by suppressing RANKL expression in bone. PMID:22040825

Harada, Suguru; Mizoguchi, Toshihide; Takahashi, Naoyuki

2011-11-01

239

Development of a disposable maglev centrifugal blood pump intended for one-month support in bridge-to-bridge applications: in vitro and initial in vivo evaluation.  

PubMed

MedTech Dispo, a disposable maglev centrifugal blood pump with two degrees of freedom magnetic suspension and radial magnetic coupling rotation, has been developed for 1-month extracorporeal circulatory support. As the first stage of a two-stage in vivo evaluation, 2-week evaluation of a prototype MedTech Dispo was conducted. In in vitro study, the pump could produce 5 L/min against 800 mm Hg and the normalized index of hemolysis was 0.0054 +/- 0.0008 g/100 L. In in vivo study, the pump, with its blood-contacting surface coated with biocompatible 2-methacryloyloxyethyl phosphorylcholine polymer, was implanted in seven calves in left heart bypass. Pump performance was stable with a mean flow of 4.49 +/- 0.38 L/min at a mean speed of 2072.1 +/- 64.5 rpm. The maglev control revealed its stability in rotor position during normal activity by the calves. During 2 weeks of operation in two calves which survived the intended study period, no thrombus formation was seen inside the pump and levels of plasma free hemoglobin were maintained below 4 mg/dL. Although further experiments are required, the pump demonstrated the potential for sufficient and reliable performance and biocompatibility in meeting the requirements for cardiopulmonary bypass and 1-week circulatory support. PMID:19775262

Someya, Takeshi; Kobayashi, Mariko; Waguri, Satoshi; Ushiyama, Tomohiro; Nagaoka, Eiki; Hijikata, Wataru; Shinshi, Tadahiko; Arai, Hirokuni; Takatani, Setsuo

2009-09-01

240

Simultaneous quantification of VX and its toxic metabolite in blood and plasma samples and its application for in vivo and in vitro toxicological studies.  

PubMed

The present study was initiated to develop a sensitive and highly selective method for the simultaneous quantification of the nerve agent VX (O-ethyl S-[2(diisopropylamino)ethyl] methylphosphonothioate) and its toxic metabolite (EA-2192) in blood and plasma samples in vivo and in vitro. For the quantitative detection of VX and EA-2192 the resolution was realized on a HYPERCARB HPLC phase. A specific procedure was developed to isolate both toxic analytes from blood and plasma samples. The limit of detection was 0.1 pg/ml and the absolute recovery of the overall sample preparation procedure was 74% for VX and 69% for EA-2192. After intravenous and percutaneous administration of a supralethal doses of VX in anaesthetised swine both VX and EA-2192 could be quantified over 540 min following exposure. This study is the first to verify the in vivo formation of the toxic metabolite EA-2192 after poisoning with the nerve agent VX. Further toxicokinetic and therapeutic studies are required in order to determine the impact of EA-2192 on the treatment of acute VX poisoning. PMID:21862421

Reiter, Georg; Mikler, John; Hill, Ira; Weatherby, Kendal; Thiermann, Horst; Worek, Franz

2011-07-30

241

3D Compressed Sensing for Highly Accelerated Hyperpolarized 13C MRSI With In Vivo Applications to Transgenic Mouse Models of Cancer  

PubMed Central

High polarization of nuclear spins in liquid state through hyperpolarized technology utilizing dynamic nuclear polarization has enabled the direct monitoring of 13C metabolites in vivo at a high signal-to-noise ratio. Acquisition time limitations due to T1 decay of the hyperpolarized signal require accelerated imaging methods, such as compressed sensing, for optimal speed and spatial coverage. In this paper, the design and testing of a new echo-planar 13C three-dimensional magnetic resonance spectroscopic imaging (MRSI) compressed sensing sequence is presented. The sequence provides up to a factor of 7.53 in acceleration with minimal reconstruction artifacts. The key to the design is employing x and y gradient blips during a fly-back readout to pseudorandomly undersample kf-kx-ky space. The design was validated in simulations and phantom experiments where the limits of undersampling and the effects of noise on the compressed sensing nonlinear reconstruction were tested. Finally, this new pulse sequence was applied in vivo in preclinical studies involving transgenic prostate cancer and transgenic liver cancer murine models to obtain much higher spatial and temporal resolution than possible with conventional echo-planar spectroscopic imaging methods.

Hu, Simon; Lustig, Michael; Balakrishnan, Asha; Larson, Peder E. Z.; Bok, Robert; Kurhanewicz, John; Nelson, Sarah J.; Goga, Andrei; Pauly, John M.; Vigneron, Daniel B.

2010-01-01

242

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)

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.

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

243

Targeted luminescent near-infrared polymer-nanoprobes for in vivo imaging of tumor hypoxia.  

PubMed

Polystyrene nanoparticles (PS-NPs) were doped with an oxygen-sensitive near-infrared (NIR)-emissive palladium meso-tetraphenylporphyrin and an inert reference dye which are both excitable at 635 nm. The nanosensors were characterized with special emphasis on fundamental parameters such as absolute photoluminescence quantum yield and fluorescence lifetime. The PS-NPs were employed for ratiometric dual-wavelength and lifetime-based photoluminescent oxygen sensing. They were efficiently taken up by cultured murine alveolar macrophages, yielding a characteristic and reversible change in ratiometric response with decreasing oxygen concentration. This correlated with the cellular hypoxic status verified by analysis of hypoxia inducible factor-1? (HIF-1?) accumulation. In addition, the surface of PS-NPs was functionalized with polyethylene glycol (PEG) and the monoclonal antibody herceptin, and their binding to HER2/neu-overexpressing tumor cells was confirmed in vitro. First experiments with tumor-bearing mouse revealed a distinctive ratiometric response within the tumor upon hypoxic condition induced by animal sacrifice. These results demonstrate the potential of these referenced NIR nanosensors for in vitro and in vivo imaging that present a new generation of optical probes for oncology. PMID:22007722

Napp, Joanna; Behnke, Thomas; Fischer, Lorenz; Würth, Christian; Wottawa, Marieke; Katschinski, Dörthe M; Alves, Frauke; Resch-Genger, Ute; Schäferling, Michael

2011-11-02

244

Application of a biphasic test for characterization of in vitro drug release of immediate release formulations of celecoxib and its relevance to in vivo absorption.  

PubMed

A biphasic in vitro test method was used to examine release profiles of a poorly soluble model drug, celecoxib (CEB), from its immediate release formulations. Three formulations of CEB were investigated in this study, including a commercial Celebrex capsule, a solution formulation (containing cosolvent and surfactant) and a supersaturatable self-emulsifying drug delivery system (S-SEDDS). The biphasic test system consisted of an aqueous buffer and a water-immiscible organic solvent (e.g., octanol) with the use of both USP II and IV apparatuses. The aqueous phase provided a nonsink dissolution medium for CEB, while the octanol phase acted as a sink for CEB partitioning. For comparison, CEB concentration-time profiles of these formulations in the aqueous medium under either a sink condition or a nonsink condition were also explored. CEB release profiles of these formulations observed in the aqueous medium from either the sink condition test, the nonsink condition test, or the biphasic test have little relevance to the pharmacokinetic observations (e.g., AUC, C(max)) in human subjects. In contrast, a rank order correlation among the three CEB formulations is obtained between the in vitro AUC values of CEB from the octanol phase up to t = 2 h and the in vivo mean AUC (or C(max)) values. As the biphasic test permits a rapid removal of drug from the aqueous phase by partitioning into the organic phase, the amount of drug in the organic phase represents the amount of drug accumulated in systemic circulation in vivo. This hypothesis provides the scientific rationale for the rank order relationship among these CEB formulations between their CEB concentrations in the organic phase and the relative AUC or C(max). In addition, the biphasic test method permits differentiation and discrimination of key attributes among the three different CEB formulations. This work demonstrates that the biphasic in vitro test method appears to be useful as a tool in evaluating performance of formulations of poorly water-soluble drugs and to provide potential for establishing an in vitro-in vivo relationship. PMID:20704265

Shi, Yi; Gao, Ping; Gong, Yuchuan; Ping, Haili

2010-08-12

245

Patient-derived models of human breast cancer: protocols for in vitro and in vivo applications in tumor biology and translational medicine.  

PubMed

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 three-dimensional culturing of xenografted tissue, which enable use of bona fide uncultured human tissue in designing and validating cancer therapies. PMID:23456611

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

2013-03-01

246

Development of ultrafast laser-based x-ray in-vivo phase-contrast micro-CT beamline for biomedical applications at Advanced Laser Light Source (ALLS)  

NASA Astrophysics Data System (ADS)

We are developing and exploring the imaging performance of, an in vivo, in-line holography, x-ray phase-contrast, micro-CT system with an ultrafast laser-based x-ray (ULX) source. By testing and refining our system, and by performing computer simulations, we plan to improve system performance in terms of contrast resolution and multi-energy imaging to a level beyond what can be obtained using a conventional microfocal x-ray tube. Initial CT projection sets at single energy (Mo K? and K? lines) were acquired in the Fresnel regime and reconstructed for phantoms and a euthanized mouse. We also performed computer simulations of phase-contrast micro-CT scans for low-contrast, soft-tissue, tumor imaging. We determined that, in order to perform a phase-contrast, complete micro-CT scan using ULX, the following conditions must be met: (i) the x-ray source needs to be stable during the scan; (ii) the laser focal spot size needs to be less than 10 ?m for source-to-object distance greater than 30 cm; (iii) the laser light intensity on the target needs to be in the range of 5 × 1017 to 5 × 1019 W/cm2; (iv) the ablation protection system needs to allow uninterrupted scans; (v) the laser light focusing on the target needs to remain accurate during the entire scan; (vi) a fresh surface of the target must be exposed to consecutive laser shots during the entire scan; (vii) the effective detector element size must be less than 12 ?m. Based on the results obtained in this research project, we anticipate that the new 10 Hz, 200 TW laser with 50W average power that is being commissioned at ALLS will allow us practical implementation of in vivo x-ray phase-contrast micro-CT.

Kincaid, Russell; Krol, Andrzej; Fourmaux, Sylvain; Kieffer, Jean-Claude; Serbanescu, Cristina; Servol, Marina; Vogelsang, Levon; Wilkins, Steve; Stevenson, Andrew; Nesterets, Yakov; Lipson, Edward; Ye, Hongwei; Pogany, Andrew

2008-09-01

247

/sup 32/P-Postlabeling test for covalent DNA binding of chemicals in vivo: Application to a variety of aromatic carcinogens and methylating agents  

SciTech Connect

Carcinogen--DNA adducts were detected and determined by /sup 32/P-postlabeling assay after exposure of mouse or rat tissues in vivo to a total of 28 compounds comprising 7 arylamines and derivatives, 3 azo compounds, 2 nitroaromatics, 12 polycyclic aromatic hydrocarbons, and 4 methylating agents. DNA was isolated from mouse skin, mouse liver, and rat liver after treatment with the individual carcinogens, then digested enzymatically to deoxyribonucleoside 3'-monophosphates, which were converted to 5'-/sup 32/P-labeled deoxyribonucleoside 3',5'-bisphosphates by T4 polynucleotide kinase-catalyzed (/sup 32/P)phosphate transfer from (gamma-/sup 32/P)ATP. The nucleotides were resolved by anion-exchange t.l.c. on polyethyleneimine-cellulose and detected by autoradiography. The determination of low levels of DNA binding of the aromatic carcinogens entailed the removal of normal nucleotides prior to the resolution of adduct nucleotides. For this purpose, an alternative procedure employing reversed-phase t.l.c. was devised which offered advantages for the detection of quantitatively minor adducts. The procedures described enabled the detection of 1 aromatic DNA adduct in approximately 10(/sup 8/) normal nucleotides, while the limit of detection of methylated adducts was 1 adduct in approximately 6 X 10(/sup 5/) nucleotides. The results show that a great number of carcinogen-DNA adducts of diverse structure are substrates for /sup 32/P-labeling by polynucleotide kinase-catalyzed phosphorylation. Because covalent DNA adduct formation in vivo appears to be an essential property of the majority of chemical carcinogens, /sup 32/P-postlabeling analysis of carcinogen--DNA adducts in mammalian tissues may serve as a test for the screening of chemicals for potential carcinogenicity.

Reddy, M.V.; Gupta, R.C.; Randerath, E.; Randerath, K.

1984-02-01

248

Effects of In Vitro Low Oxygen Tension Preconditioning of Adipose Stromal Cells on Their In Vivo Chondrogenic Potential: Application in Cartilage Tissue Repair  

PubMed Central

Purpose Multipotent stromal cell (MSC)-based regenerative strategy has shown promise for the repair of cartilage, an avascular tissue in which cells experience hypoxia. Hypoxia is known to promote the early chondrogenic differentiation of MSC. The aim of our study was therefore to determine whether low oxygen tension could be used to enhance the regenerative potential of MSC for cartilage repair. Methods MSC from rabbit or human adipose stromal cells (ASC) were preconditioned in vitro in control or chondrogenic (ITS and TGF-?) medium and in 21 or 5% O2. Chondrogenic commitment was monitored by measuring COL2A1 and ACAN expression (real-time PCR). Preconditioned rabbit and human ASC were then incorporated into an Si-HPMC hydrogel and injected (i) into rabbit articular cartilage defects for 18 weeks or (ii) subcutaneously into nude mice for five weeks. The newly formed tissue was qualitatively and quantitatively evaluated by cartilage-specific immunohistological staining and scoring. The phenotype of ASC cultured in a monolayer or within Si-HPMC in control or chondrogenic medium and in 21 or 5% O2 was finally evaluated using real-time PCR. Results/Conclusions 5% O2 increased the in vitro expression of chondrogenic markers in ASC cultured in induction medium. Cells implanted within Si-HPMC hydrogel and preconditioned in chondrogenic medium formed a cartilaginous tissue, regardless of the level of oxygen. In addition, the 3D in vitro culture of ASC within Si-HPMC hydrogel was found to reinforce the pro-chondrogenic effects of the induction medium and 5% O2. These data together indicate that although 5% O2 enhances the in vitro chondrogenic differentiation of ASC, it does not enhance their in vivo chondrogenesis. These results also highlight the in vivo chondrogenic potential of ASC and their potential value in cartilage repair.

Gauthier, Olivier; Lesoeur, Julie; Sourice, Sophie; Masson, Martial; Fellah, Borhane Hakim; Geffroy, Olivier; Lallemand, Elodie; Weiss, Pierre

2013-01-01

249

The design of a double-tuned two-port surface resonator and its application to in vivo hydrogen- and sodium-MRI.  

PubMed

The design and construction of a two-port surface transceiver resonator for both (1)H-and (23)Na-MRI in the rodent brain at 7 T is described. Double-tuned resonators are required for accurately co-registering multi-nuclei data sets, especially when the time courses of (1)H and (23)Na signals are of interest as, for instance, when investigating the pathological progression of ischaemic stroke tissue in vivo. In the current study, a single-element two-port surface resonator was developed wherein both frequency components were measured with the same detector element but with each frequency signal routed along different output channels. This was achieved by using the null spot technique, allowing for optimal variable tuning and matching of each channel in situ within the MRI scanner. The (23)Na signal to noise ratio, measured in the ventricles of the rat brain, was increased by a factor of four compared to recent state-of-the-art rat brain studies reported in the literature. The resonator's performance was demonstrated in an in vivo rodent stroke model, where regional variations in (1)H apparent diffusion coefficient maps and the (23)Na signal were recorded in an interleaved fashion as a function of time in the acute phase of the stroke without having to exchange, re-adjust, or re-connect resonators between scans. Using the practical construction steps described in this paper, this coil design can be easily adapted for MRI of other X-nuclei, such as (17)O, (13)C, (39)K, and (43)Ca at various field strengths. PMID:22391488

Wetterling, Friedrich; Högler, Miroslav; Molkenthin, Ute; Junge, Sven; Gallagher, Lindsay; Mhairi Macrae, I; Fagan, Andrew J

2012-02-16

250

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)

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

Wollenberg, Lance A.

251

Tropisms of AAV for Subretinal Delivery to the Neonatal Mouse Retina and Its Application for In Vivo Rescue of Developmental Photoreceptor Disorders  

PubMed Central

Background Adeno-associated virus (AAV) is well established as a vehicle for in vivo gene transfer into the mammalian retina. This virus is promising not only for gene therapy of retinal diseases, but also for in vivo functional analysis of retinal genes. Previous reports have shown that AAV can infect various cell types in the developing mouse retina. However, AAV tropism in the developing retina has not yet been examined in detail. Methodology/Principal Findings We subretinally delivered seven AAV serotypes (AAV2/1, 2/2, 2/5, 2/8, 2/9, 2/10, and 2/11) of AAV-CAG-mCherry into P0 mouse retinas, and quantitatively evaluated the tropisms of each serotype by its infecting degree in retinal cells. After subretinal injection of AAV into postnatal day 0 (P0) mouse retinas, various retinal cell types were efficiently transduced with different AAVs. Photoreceptor cells were efficiently transduced with AAV2/5. Retinal cells, except for bipolar and Müller glial cells, were efficiently transduced with AAV2/9. Horizontal and/or ganglion cells were efficiently transduced with AAV2/1, AAV2/2, AAV2/8, AAV2/9 and AAV2/10. To confirm the usefulness of AAV-mediated gene transfer into the P0 mouse retina, we performed AAV-mediated rescue of the Cone-rod homeobox gene knockout (Crx KO) mouse, which exhibits an outer segment formation defect, flat electroretinogram (ERG) responses, and photoreceptor degeneration. We injected an AAV expressing Crx under the control of the Crx 2kb promoter into the neonatal Crx KO retina. We showed that AAV mediated-Crx expression significantly decreased the abnormalities of the Crx KO retina. Conclusion/Significance In the current study, we report suitable AAV tropisms for delivery into the developing mouse retina. Using AAV2/5 in photoreceptor cells, we demonstrated the possibility of gene replacement for the developmental disorder and subsequent degeneration of retinal photoreceptors caused by the absence of Crx.

Watanabe, Satoshi; Sanuki, Rikako; Ueno, Shinji; Koyasu, Toshiyuki; Hasegawa, Toshiaki; Furukawa, Takahisa

2013-01-01

252

Fibrin Gel-Immobilized Primary Osteoblasts in Calcium Phosphate Bone Cement: In vivo Evaluation with Regard to Application as Injectable Biological Bone Substitute  

Microsoft Academic Search

Osteogenic injectable bone substitutes may be useful for many applications. We developed a novel injectable bone substitute based on osteoblast-fibrin glue suspension and calcium phosphate bone cement (BC). Human osteoblasts were isolated from trabecular bone samples and cultured under standard conditions. Osteoblasts were suspended in fibrinogen solution (FS). BC was cured with thrombin solution. 8 × 4 mm injectable bone

U. Kneser; A. Voogd; J. Ohnolz; O. Buettner; L. Stangenberg; Y. H. Zhang; G. B. Stark; D. J. Schaefer

2005-01-01

253

Preliminary in vivo study on the potential application of a novel method of e-tracking to facilitate traceability in the poultry food chain  

Microsoft Academic Search

The feasibility of using GS1 DataMatrix (GS1 DM) barcodes laser printed onto the beaks of poultry as a possible method of identification and, therefore, traceability of the individual were examined in this study, including a preliminary live trial on layer hens. The optimal laser type and settings for this particular application had been selected during previous in vitro and in

Barry Mc Inerney; Gerard Corkery; Gashaw Ayalew; Shane Ward; Kevin Mc Donnell

2011-01-01

254

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

PubMed Central

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.

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

255

Isolation and ex vivo expansion of bone marrow-derived porcine mesenchymal stromal cells: potential for application in an experimental model of solid organ transplantation in large animals.  

PubMed

Pharmacological aspecific immunosuppression, despite being widely used in solid organ transplantation recipients, is unable to completely prevent allograft rejection. It promotes the occurrence of sometimes life-threatening infections. Due to their immunosuppressive and anti- inflammatory properties, there is great interest in the therapeutic use of bone marrow (BM)-derived mesenchymal stromal cells (MSC). Large animal models play a crucial role to investigate the biological and functional properties of MSCs as novel cellular therapy. In the current study we sought to isolate expand ex vivo, and phenotypically characterize MSC derived from BM of 4 Large White 6-month-old piglets. Porcine MSC (pMSC) were characterized for their in vitro differentiation capacity. pMSC were successfully isolated from all BM samples. They showed spindle-shaped morphology and a stable doubling time on culture. They were positive for CD90, CD29, CD105, and negative for CD45 and CD11b. Furthermore, they differentiated, upon specific in vitro conditions toward adipogenic and osteogenic lineages. The optimization of methods for the isolation and characterization of pMSC may be useful to elucidate their biological and functional properties. The anatomy and physiology of the pig, which is similar to humans, make this animal model more attractive than small animals to test the safety and efficacy of MSC in the context of solid organ transplantation. PMID:20534296

Comite, P; Cobianchi, L; Avanzini, M A; Zonta, S; Mantelli, M; Achille, V; De Martino, M; Cansolino, L; Ferrari, C; Alessiani, M; Maccario, R; Gandolfo, G M; Dionigi, P; Locatelli, F; Bernardo, M E

2010-05-01

256

In vivo application of sub-second spiral chemical shift imaging (CSI) to hyperpolarized 13C metabolic imaging: Comparison with phase-encoded CSI  

NASA Astrophysics Data System (ADS)

A fast spiral chemical shift imaging (CSI) has been developed to address the challenge of the limited acquisition window in hyperpolarized 13C metabolic imaging. The sequence exploits the sparsity of the spectra and prior knowledge of resonance frequencies to reduce the measurement time by undersampling the data in the spectral domain. As a consequence, multiple reconstructions are necessary for any given data set as only frequency components within a selected bandwidth are reconstructed “in-focus” while components outside that band are severely blurred (“spectral tomosynthesis”). A variable-flip-angle scheme was used for optimal use of the longitudinal magnetization. The sequence was applied to sub-second metabolic imaging of the rat in vivo after injection of hyperpolarized [1-13C]-pyruvate on a clinical 3T MR scanner. The comparison with conventional CSI based on phase encoding showed similar signal-to-noise ratio (SNR) and spatial resolution in metabolic maps for the substrate and its metabolic products lactate, alanine, and bicarbonate, despite a 50-fold reduction in scan time for the spiral CSI acquisition. The presented results demonstrate that dramatic reductions in scan time are feasible in hyperpolarized 13C metabolic imaging without a penalty in SNR or spatial resolution.

Mayer, Dirk; Yen, Yi-Fen; Levin, Yakir S.; Tropp, James; Pfefferbaum, Adolf; Hurd, Ralph E.; Spielman, Daniel M.

2010-06-01

257

Topical application of 17beta-estradiol increases extracellular matrix protein synthesis by stimulating tgf-Beta signaling in aged human skin in vivo.  

PubMed

To investigate the effects of topically applied 17beta-estradiol on the expression of extracellular matrix proteins in aged human skin, 17beta-estradiol (0.01%) and its vehicle (70% propylene glycol, 30% ethanol) were applied to aged (68-82 y, eight females and five males) human buttock skin under occlusion for 2 wk (three times per week). Topical 17beta-estradiol was found to increase the expression of type 1 procollagen mRNA and protein significantly in human aged skin in vivo. In addition, metalloproteinase (MMP-1 protein levels were reduced by topical 17beta-estradiol. The expressions of TGF-beta1, TGF-beta type II receptor, and Sma and Mad related (Smad)3 were increased by topical 17 beta-estradiol in aged human skin, and TGF-beta1 neutralizing antibody inhibited 17beta-estradiol-induced procollagen synthesis in cultured fibroblasts. We also found that the expressions of tropoelastin and fibrillin-1 mRNA and protein, and elastic fibers in aged skin were also increased by topical 17beta-estradiol. Topical 17beta-estradiol also increased keratinocyte proliferation and the epidermal thickness in aged human skin. We also observed the same effects of topical 17beta-estradiol in young skin. In conclusion, our results suggest that topical 17beta-estradiol treatment may improve the cutaneous function of aged human skin by improving the connective tissue and increasing epidermal thickness. PMID:15955089

Son, Eui Dong; Lee, Jin Young; Lee, Serah; Kim, Mi Sun; Lee, Byeong Gon; Chang, Ih Seoup; Chung, Jin Ho

2005-06-01

258

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

PubMed

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

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

2013-07-11

259

Cultivation of Keratinocytes and Fibroblasts in a Three-Dimensional Bovine Collagen-Elastin Matrix (Matriderm®) and Application for Full Thickness Wound Coverage in Vivo  

PubMed Central

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.

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

2013-01-01

260

Robust 3-D reconstruction of surfaces from image focus by local cross-sectional multivariate statistical analyses: application to human ex vivo corneal endotheliums.  

PubMed

The considered problem of 3-D reconstruction consists in computationally and passively recovering both topography and texture of a scene surface observed by optical sectioning with a limited depth-of-field imaging system (typically a conventional optical microscope). Throughout a sequence of registered 2-D images, the concepts of shape-from-focus and extended-depth-of-field involve recovering both topography (depth map) and texture image of the surface by researching in-focus information, respectively. Toward that aim, traditional approaches generally follow a 2-D sectional way and thereby fail to deal with noisy and disturbed acquisitions, quite frequent in transmitted light observations and of interest in this paper. Such examples are the acquisitions of human ex vivo corneal endotheliums from the medical issue addressed in this paper, which are mainly damaged by cellular fragments in the sample immersion medium and by emphasized contrast reversals. To achieve with such noisy and disturbed acquisitions, a new focus analysis is introduced that originally adopts a 3-D strategy throughout the image sequence. This method exploits simultaneously all available cross-sectional cues that effectively strengthens the robustness. More precisely, it locally performs multivariate statistical analyses over cross-sectional spatial windows so as to find sectional in-focus positions. Comparisons to state-of-the-art methods on both synthetic data and real acquisitions from the deal-with medical issue demonstrate the efficiency and the robustness of the proposed approach. PMID:22831775

Fernandes, Mathieu; Gavet, Yann; Pinoli, Jean-Charles

2012-06-01

261

Application of Retinol to Human Skin In Vivo Induces Epidermal Hyperplasia and Cellular Retinoid Binding Proteins Characteristic of Retinoic Acid but Without Measurable Retinoic Acid Levels or Irritation  

Microsoft Academic Search

We investigated the clinical, histologic, and molecular responses of normal human skin to all-trans-retinol (ROL) application, compared to those induced by topical all-trans-retinoic acid (RA), and measured ROL-derived metabolites. Up to 1.6% ROL, 0.025% RA in vehicle (70% ethanol\\/30% propylene glycol), or vehicle alone were applied in a double-blind fashion to normal buttock skin and occluded for 4 d. ROL

Sewon Kang; Elizabeth A Duell; Gary J Fisher; Subhash C Datta; Zeng-Quan Wang; Ambati P Reddy; Amir Tavakkol; Jong Y Yi; Christopher E M Griffiths; James T Elder; John J Voorhees

1995-01-01

262

Kinetic properties of the glucose-6-phosphate and 6-phosphogluconate dehydrogenases from Corynebacterium glutamicum and their application for predicting pentose phosphate pathway flux in vivo.  

PubMed

The glucose-6-phosphate (Glc6P) and 6-phosphogluconate (6PG) dehydrogenases of the amino-acid-producing bacterium Corynebacterium glutamicum were purified to homogeneity and kinetically characterized. The Glc6P dehydrogenase was a heteromultimeric complex, which consists of Zwf and OpcA subunits. The product inhibition pattern of the Glc6P dehydrogenase was consistent with an ordered bi-bi mechanism. The 6PG dehydrogenase was found to operate according to a Theorell-Chance ordered bi-ter mechanism. Both enzymes were inhibited by NADPH and the 6PG dehydrogenase additionally by ATP, fructose 1,6-bisphosphate (Fru1,6P2), D-glyceraldehyde 3-phosphate (Gra3P), erythrose 4-phosphate and ribulose 5-phosphate (Rib5P). The inhibition by NADPH was considered to be most important, with inhibition constants of around 25 microM for both enzymes. Intracellular metabolite concentrations were determined in two isogenic strains of C. glutamicum with plasmid-encoded NAD- and NADP-dependent glutamate dehydrogenases. NADP+ and NADPH levels were between 130 microM and 290 microM, which is very much higher than the respective Km and Ki values. The Glc6P concentration was around 500 microM in both strains. The in vivo fluxes through the oxidative part of the pentose phosphate pathway calculated on the basis of intracellular metabolite concentrations and the kinetic constants of the purified enzymes determined in vitro were in agreement with the same fluxes determined by NMR after 13C-labelling. From the derived kinetic model thus validated, it is concluded that the oxidative pentose phosphate pathway in C. glutamicum is mainly regulated by the ratio of NADPH and NADP+ concentrations and the specific enzyme activities of both dehydrogenases. PMID:10848959

Moritz, B; Striegel, K; De Graaf, A A; Sahm, H

2000-06-01

263

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

PubMed Central

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.

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

2012-01-01

264

Silicate, borosilicate, and borate bioactive glass scaffolds with controllable degradation rate for bone tissue engineering applications. II. In vitro and in vivo biological evaluation.  

PubMed

In Part I, the in vitro degradation of bioactivAR52115e glass scaffolds with a microstructure similar to that of human trabecular bone, but with three different compositions, was investigated as a function of immersion time in a simulated body fluid. The glasses consisted of a silicate (13-93) composition, a borosilicate composition (designated 13-93B1), and a borate composition (13-93B3), in which one-third or all of the SiO2 content of 13-93 was replaced by B2O3, respectively. This work is an extension of Part I, to investigate the effect of the glass composition on the in vitro response of osteogenic MLO-A5 cells to these scaffolds, and on the ability of the scaffolds to support tissue infiltration in a rat subcutaneous implantation model. The results of assays for cell viability and alkaline phosphatase activity showed that the slower degrading silicate 13-93 and borosilicate 13-93B1 scaffolds were far better than the borate 13-93B3 scaffolds in supporting cell proliferation and function. However, all three groups of scaffolds showed the ability to support tissue infiltration in vivo after implantation for 6 weeks. The results indicate that the required bioactivity and degradation rate may be achieved by substituting an appropriate amount of SiO2 in 13-93 glass with B2O3, and that these trabecular glass scaffolds could serve as substrates for the repair and regeneration of contained bone defects. PMID:20540099

Fu, Qiang; Rahaman, Mohamed N; Bal, B Sonny; Bonewald, Lynda F; Kuroki, Keiichi; Brown, Roger F

2010-10-01

265

EDITORIAL: Nanotechnology in vivo Nanotechnology in vivo  

NASA Astrophysics Data System (ADS)

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

Demming, Anna

2010-04-01

266

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

PubMed

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

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

2012-11-20

267

Applications  

NASA Astrophysics Data System (ADS)

Economic incentives have spurred numerous applications of genetically engineered organisms in manufacture of pharmaceuticals and industrial chemicals. These successes, involving a variety of methods of genetic manipulation, have dispelled early fears that genetic engineering could not be handled safely, even in the laboratory. Consequently, the potential for applications in the wider environment without physical containment is being considered for agriculture, mining, pollution control, and pest control. These proposed applications range from modest extensions of current plant breeding techniques for new disease-resistant species to radical combinations of organisms (for example, nitrogen-fixing corn plants). These applications raise concerns about potential ecological impacts (see chapter 5), largely because of adverse experiences with both deliberate and inadvertent introductions of nonindigenous species.

Stern, Arthur M.

1986-07-01

268

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

NASA Astrophysics Data System (ADS)

Multi-functional rare-earth Yb3+ and Ln3+ (Ln = Er, Tm and Ho) ions doped one-dimensional (1-D) upconversion submicrocrystals (NaYF4 and NaGdF4) possessing upconversion luminescence, biocompatibility and magnetic properties have been synthesized by a one-pot hydrothermal method. Rare-earth Yb3+ and Ln3+ ions doped NaYF4 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 Yb3+ and Ln3+ ions doped NaGdF4 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 Yb3+ and Ln3+ ions doped 1-D upconversion submicrocrystals indicate they can be used for drug targeting under a magnetic field. Their unique upconversion emission (green for Yb3+/Er3+ and blue for Yb3+/Tm3+) 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.Multi-functional rare-earth Yb3+ and Ln3+ (Ln = Er, Tm and Ho) ions doped one-dimensional (1-D) upconversion submicrocrystals (NaYF4 and NaGdF4) possessing upconversion luminescence, biocompatibility and magnetic properties have been synthesized by a one-pot hydrothermal method. Rare-earth Yb3+ and Ln3+ ions doped NaYF4 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 Yb3+ and Ln3+ ions doped NaGdF4 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 Yb3+ and Ln3+ ions doped 1-D upconversion submicrocrystals indicate they can be used for drug targeting under a magnetic field. Their unique upconversion emission (green for Yb3+/Er3+ and blue for Yb3+/Tm3+) 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. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr32850j

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

2012-12-01

269

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

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.

Guzun, Rita; Saks, Valdur

2010-01-01

270

T*1e and T*2e maps derived in vivo from the rat using longitudinally detected electron spin resonance phase imaging: application to abdominal oxygen mapping.  

PubMed

A novel imaging modality is introduced which uses radiofrequency longitudinally detected electron spin resonance (RF-LODESR). It is capable of providing qualitative and semiquantitative information on a variety of parameters reflecting physiological function, the most significant being tissue oxygenation. Effective spin-lattice (T*1e) and spin-spin (T*2e) electronic relaxation time maps of the abdomen of living 200-g rats were generated after intravenous administration of a triarylmethyl free radical (TAM). These maps were used to evaluate oxygen distribution. Differences between the liver, kidneys, and bladder were noted. Conclusions were made regarding the distribution, perfusion, and excretion rate of the contrast medium. Ligature-induced anoxia in the kidney was also visualized. LODESR involves transverse ESR irradiation with a modulated excitation, and observing oscillations in the spin magnetization parallel to the main magnetic field. The T*1e and T*2e maps were calculated from a set of LODESR signal phase images collected at different detection frequencies. Each phase image also provides qualitative information on tissue oxygen levels without any further processing. This method presents an alternative to the conventional transverse ESR linewidth-based oximetry methods, particularly for animal whole-body imaging applications. PMID:11746590

Panagiotelis, I; Nicholson, I; Foster, M A; Hutchison, J M

2001-12-01

271

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

NASA Astrophysics Data System (ADS)

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.

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

2012-11-01

272

In vivo RNAi: Today and Tomorrow  

PubMed Central

SUMMARY RNA interference (RNAi) provides a powerful reverse genetics approach to analyze gene functions both in tissue culture and in vivo. Because of its widespread applicability and effectiveness it has become an essential part of the tool box kits of model organisms such as Caenorhabditis elegans, Drosophila, and the mouse. In addition, the use of RNAi in animals in which genetic tools are either poorly developed or nonexistent enables a myriad of fundamental questions to be asked. Here, we review the methods and applications of in vivo RNAi to characterize gene functions in model organisms and discuss their impact to the study of developmental as well as evolutionary questions. Further, we discuss the applications of RNAi technologies to crop improvement, pest control and RNAi therapeutics, thus providing an appreciation of the potential for phenomenal applications of RNAi to agriculture and medicine.

Perrimon, Norbert; Ni, Jian-Quan; Perkins, Lizabeth

2010-01-01

273

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

PubMed

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

Ahmed, Eilaf; Morton, Stephen W; Hammond, Paula T; Swager, Timothy M

2013-06-24

274

Radical protection in the visible and infrared by a hyperforin-rich cream--in vivo versus ex vivo methods.  

PubMed

The formation of radicals plays an important role in the development of atopic eczema or barrier-disrupted skin. We evaluated the radical scavenging effect of a cream containing a Hypericum perforatum extract rich in hyperforin in a double-blind placebo-controlled study on 11 healthy volunteers. Electron paramagnetic resonance spectroscopy was applied to determine radical formation during VIS/NIR irradiation of the inner forearm. The results were compared to ex vivo investigations on excised porcine ear skin after a single application of the creams. The non-treated skin was measured as control. The absolute values and the kinetics are not comparable for ex vivo and in vivo radical formation. Whereas in vivo, the radical production decreases with time, it remains stable ex vivo over the investigated timescale. Nevertheless, ex vivo methods could be developed to estimate the protection efficiency of creams. In vivo as well as ex vivo, the radical formation could be reduced by almost 80% when applying the hyperforin-rich cream onto the skin, whereas placebo resulted in about 60%. In vivo, a daylong protection effect could be validated after a 4-week application time of the cream indicating that a regular application is necessary to obtain the full effect. PMID:23614743

Arndt, Sophia; Haag, Stefan F; Kleemann, Anke; Lademann, Juergen; Meinke, Martina C

2013-05-01

275

In vivo fluorescence lifetime optical projection tomography  

PubMed Central

We demonstrate the application of fluorescence lifetime optical projection tomography (FLIM-OPT) to in vivo imaging of lysC:GFP transgenic zebrafish embryos (Danio rerio). This method has been applied to unambiguously distinguish between the fluorescent protein (GFP) signal in myeloid cells from background autofluorescence based on the fluorescence lifetime. The combination of FLIM, an inherently ratiometric method, in conjunction with OPT results in a quantitative 3-D tomographic technique that could be used as a robust method for in vivo biological and pharmaceutical research, for example as a readout of Förster resonance energy transfer based interactions.

McGinty, James; Taylor, Harriet B.; Chen, Lingling; Bugeon, Laurence; Lamb, Jonathan R.; Dallman, Margaret J.; French, Paul M. W.

2011-01-01

276

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

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

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

2013-09-01

277

Medial temporal cortices in ex vivo magnetic resonance imaging.  

PubMed

This review focuses on the ex vivo magnetic resonance imaging (MRI) modeling of medial temporal cortices and associated structures, the entorhinal verrucae and the perforant pathway. Typical in vivo MRI has limited resolution due to constraints on scan times and does not show laminae in the medial temporal lobe. Recent studies using ex vivo MRI have demonstrated lamina in the entorhinal, perirhinal, and hippocampal cortices. These studies have enabled probabilistic brain mapping that is based on the ex vivo MRI contrast, validated to histology, and subsequently mapped onto an in vivo spherically warped surface model. Probabilistic maps are applicable to other in vivo studies. J. Comp. Neurol. 521:4177-4188, 2013. © 2013 Wiley Periodicals, Inc. PMID:23881818

Augustinack, Jean C; van der Kouwe, André J W; Fischl, Bruce

2013-12-15

278

Bioluminescence imaging of myeloperoxidase activity in vivo  

PubMed Central

The myeloperoxidase (MPO) system of activated phagocytes is central to normal host defense mechanisms, and dysregulated MPO contributes to the pathogenesis of inflammatory disease states ranging from atherosclerosis to cancer. Here we show that upon systemic administration, the small molecule luminol enables noninvasive bioluminescence imaging (BLI) of MPO activity in vivo. Luminol-BLI allowed quantitative longitudinal monitoring of MPO activity in animal models of acute dermatitis, mixed allergic contact hypersensitivity, focal arthritis and spontaneous large granular lymphocytic tumors. Bioluminescence colocalized with histological sites of inflammation and was totally abolished in gene-deleted Mpo?/? mice, despite massive tissue infiltration of neutrophils and activated eosinophils, indicating that eosinophil peroxidase did not contribute to luminol-BLI in vivo. Thus, luminol-BLI provides a noninvasive, specific and highly sensitive optical readout of phagocyte-mediated MPO activity in vivo and may enable new diagnostic applications in a wide range of acute and chronic inflammatory conditions.

Gross, Shimon; Gammon, Seth T; Moss, Britney L; Rauch, Daniel; Harding, John; Heinecke, Jay W; Ratner, Lee; Piwnica-Worms, David

2010-01-01

279

Wireless implants for in vivo diagnosis and closed-loop treatment  

Microsoft Academic Search

In this presentation, we will review and discuss recent advances in the research of wireless telemetry for medical applications in our group at UT-Arlington, particularly those based on a similar platform for in vivo monitoring of physiological parameters. System for recording ECoG signals in brain, recording in vivo gastric myoelectric activities in stomach, sensing in vivo strain variations in bladder,

J.-C. Chiao; Aydin Farajidavar; Hung Cao; Philip McCorkle; Manthan Sheth; Young-sik Seo; Tim Wiggins; Shreyas Tharkar; Sanchali Deb; Smitha M. N. Rao

2011-01-01

280

In-vivo optical investigation of psoriasis  

NASA Astrophysics Data System (ADS)

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.

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

2011-02-01

281

In Vivo Bioluminescence Imaging  

Microsoft Academic Search

In vivo bioluminescent imaging (BLI) is a versatile and sensitive tool that is based on detection of light emission from cells or tissues. Bioluminescence, the biochemical generation of light by a living organism, is a naturally occur- ring phenomenon. Luciferase enzymes, such as that from the North American firefly (Photinus pyralis), catalyze the oxidation of a substrate (luciferin), and photons

Akiko Sato; Brenda Klaunberg; Ravi Tolwani

2004-01-01

282

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)

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.

Petrossians, Artin

283

Rubisco: Physiology in Vivo  

Microsoft Academic Search

A broad overview of the physiology and biochemistry of Rubisco is presented with a comparison of information obtained in vitro and in vivo. First a brief background to kinetic properties of Rubisco is given and Rubisco’s influence on photosynthetic metabolism is reviewed for C3, C4, CAM and C3-C4 species. The effect of environmental variables such as light and CO2 are

Susanne von Caemmerer; W. Paul Quick

284

In vivo effects of diadenosine polyphosphates on rat renal microcirculation  

Microsoft Academic Search

In vivo effects of diadenosine polyphosphates on rat renal microcirculation.BackgroundDiadenosine polyphosphates (APXA) are vasoactive nucleotides that elicit effects via purinoceptors. Recent data suggest differential effects of APXA on kidney vasculature.MethodsThe in vivo effects of AP3A, AP5A, and adenosine on renal microvessels and the role of purinoceptors were investigated by the application of agonists to the hydronephrotic rat kidney and preincubation

Gert Gabriëls; Karlhans Endlich; Karl Heinz Rahn; Eberhard Schlatter; Michael Steinhausen

2000-01-01

285

Protocol: ex vivo culture of mouse embryonic mammary buds.  

PubMed

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

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

2013-05-15

286

Communication Quantifying in vivo MR spectra with circles q  

Microsoft Academic Search

Accurate and robust quantification of in vivo magnetic resonance spectroscopy (MRS) data is essential to its application in research and medicine. The performance of existing analysis methods is problematic for in vivo studies where low signal-to-noise ratio, overlap- ping peaks and intense artefacts are endemic. Here, a new frequency-domain technique for MRS data analysis is introduced wherein the circular trajectories

Refaat E. Gabr; Ronald Ouwerkerk; Paul A. Bottomley

287

Quantifying in vivo MR spectra with circles ?  

PubMed Central

Accurate and robust quantification of in vivo magnetic resonance spectroscopy (MRS) data is essential to its application in research and medicine. The performance of existing analysis methods is problematic for in vivo studies where low signal-to-noise ratio, overlapping peaks and intense artefacts are endemic. Here, a new frequency-domain technique for MRS data analysis is introduced wherein the circular trajectories which result when spectral peaks are projected onto the complex plane, are fitted with active circle models. The use of active contour strategies naturally allows incorporation of prior knowledge as constraint energy terms. The problem of phasing spectra is eliminated, and baseline artefacts are dealt with using active contours-snakes. The stability and accuracy of the new technique, CFIT, is compared with a standard time-domain fitting tool, using simulated 31P data with varying amounts of noise and 98 real human chest and heart 31P MRS data sets. The real data were also analyzed by our standard frequency-domain absorption-mode technique. On the real data, CFIT demonstrated the least fitting failures of all methods and an accuracy similar to the latter method, with both these techniques outperforming the time-domain approach. Contrasting results from simulations argue that performance relative to Cramer-Rao Bounds may not be a suitable indicator of fitting performance with typical in vivo data such as these. We conclude that CFIT is a stable, accurate alternative to the best existing methods of fitting in vivo data.

Gabr, Refaat E.; Ouwerkerk, Ronald; Bottomley, Paul A.

2007-01-01

288

Acoustic radiation force impulse imaging of the mechanical properties of arteries: in vivo and ex vivo results.  

PubMed

We present results of a pilot study of ex vivo and in vivo acoustic radiation force impulse (ARFI) imaging demonstrating measurements of the mechanical properties of the carotid and popliteal arteries. The results were obtained on a modified commercial scanner, providing coregistered B-mode and color Doppler images. 2D and 1D through time images are formed from the measurements of tissues' response to very brief and localized applications of radiation force. The images show good correlation with B-mode and, in ex vivo studies, pathology-based characterizations of vessel geometry and plaque stiffness. In vivo measurements of arterial response during both systole and diastole are presented. We address implementation issues and discuss potential applications of this new vascular imaging method. PMID:15550320

Trahey, Gregg E; Palmeri, Mark L; Bentley, Rex C; Nightingale, Kathryn R

2004-09-01

289

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  

Microsoft Academic Search

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

Bich-Thuy Doan; Gwennhael Autret; Joël Mispelter; Philippe Méric; William Même; Céline Montécot-Dubourg; Jean-Loup Corrèze; Frédéric Szeremeta; Brigitte Gillet; Jean-Claude Beloeil

2009-01-01

290

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

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

D. B. Kintner; M. K. Anderson; J. H. Fitzpatrick; K. A. Sailor; D. D. Gilboe

2000-01-01

291

Multicolor core/shell silica nanoparticles for in vivo and ex vivo imaging  

NASA Astrophysics Data System (ADS)

Biocompatible highly bright silica nanoparticles were designed, prepared and tested in small living organisms for both in vivo and ex vivo imaging. The results that we report here demonstrate that they are suitable for optical imaging applications as a possible alternative to commercially available fluorescent materials including quantum dots. Moreover, the tunability of their photophysical properties, which was enhanced by the use of different dyes as doping agents, constitutes a very important added value in the field of medical diagnostics.Biocompatible highly bright silica nanoparticles were designed, prepared and tested in small living organisms for both in vivo and ex vivo imaging. The results that we report here demonstrate that they are suitable for optical imaging applications as a possible alternative to commercially available fluorescent materials including quantum dots. Moreover, the tunability of their photophysical properties, which was enhanced by the use of different dyes as doping agents, constitutes a very important added value in the field of medical diagnostics. Electronic supplementary information (ESI) available: Particle size distribution by DLS and TEM images. See DOI: 10.1039/c1nr11401h

Rampazzo, Enrico; Boschi, Federico; Bonacchi, Sara; Juris, Riccardo; Montalti, Marco; Zaccheroni, Nelsi; Prodi, Luca; Calderan, Laura; Rossi, Barbara; Becchi, Serena; Sbarbati, Andrea

2012-01-01

292

Hexokinase redistribution in vivo.  

PubMed

Heterogenous stock mice in addition to mice selectively bred to maximally differ in their severity of alcohol withdrawal seizures (withdrawal seizure-resistant (WSR) and withdrawal seizure-prone (WSP] were used to provide evidence in favor of the importance of the rapidly changing distribution of brain hexokinase (ATP: D-hexose 6-phosphotransferase, EC 2.7.1.1) (HK). An ischemic response at 15, 30, 60 and 120 s after killing showed a decreasing cerebellar cytosolic HK concentration of 31%, 15%, 14% and 10% while the cerebral concentrations were 23%, 13%, 13% and 14%, respectively. WSR and WSP mice given an acute i.p. dose of 4 g/kg of alcohol showed opposite HK responses. Cytosolic HK in WSR mice decreased 18.5%, while WSP mice showed an increase of 20.3% over paired saline-injected controls. When ischemia was allowed to proceed in WSP mice following an in vivo alcohol treatment, cytosolic HK decreased in parallel to mice not given alcohol. These data suggest that alcohol can cause an HK redistribution in vivo which could play a role in the differing sensitivities of WSR and WSP mice to alcohol related seizures. PMID:2328257

Laursen, S E; Belknap, J K; Sampson, K E; Knull, H R

1990-04-23

293

In Vivo Imaging of Quantum Dots  

NASA Astrophysics Data System (ADS)

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.

Texier, Isabelle; Josser, Véronique

294

In vivo dosimetry for IMRT  

SciTech Connect

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.

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

295

In vivo optical imaging in arthritis--an enlightening future?  

PubMed

In vivo molecular optical imaging has significant potential to delineate and measure, at the macroscopic level, in vivo biological processes that are occurring at the cellular and molecular level. Optical imaging has already been developed for in vitro and ex vivo applications in molecular and cellular biology (e.g. fluorescence confocal microscopy), but is still at an early stage of development as a whole-animal in vivo imaging technique. Both sensitivity and spatial resolution remain incompletely defined. Rapid advances in hardware technology and highly innovative reporter probes and dyes will be expected to deliver significant insight into perturbations of molecular pathways that occur in disease, ultimately with the potential of translating into future molecular imaging techniques for patients with arthritis. This review will focus on currently available technologies for live in vivo animal optical imaging, including fluorescence reflectance imaging, potential novel tomographic techniques, bioluminescence reporter technology and potential novel labelling techniques, highlighting in particular the potential application of in vivo fluorescence imaging in arthritis. PMID:20338885

Gompels, Luke L; Lim, Ngee Han; Vincent, Tonia; Paleolog, Ewa M

2010-03-24

296

Usefulness of combined in vivo skin comet assay and in vivo skin micronucleus test.  

PubMed

We have already found that the in vivo skin comet assay is useful for the evaluation of primary DNA damage induced by genotoxic chemicals in epidermal skin cells. The aim of the present study was to evaluate the sensitivity and specificity of the combined in vivo skin comet assay and in vivo skin micronucleus (MN) test using the same animal to explore the usefulness of the new test method. The combined alkaline comet assay and MN test was carried out with three chemicals: 4-nitroquinoline-1-oxide (4NQO), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and benzo[a]pyrene (B[a]P). In the first experiment, we compared DNA- and chromosome-damaging effects of 3 [72, 24 and 3 hours (h) before sacrifice] and 4 applications (72, 48, 24 and 3h before sacrifice) of 4NQO, which induces dermal irritancy. The animals were euthanized and their skin was sampled for the combination test. As a result, the 4-application method was able to detect both DNA- and chromosome-damaging potential with a lower concentration; therefore, in the second experiment, MNNG and B[a]P were topically applied four times, respectively. The animals were euthanized, and then their skins were sampled for combination tests. In the alkaline comet assay, significant differences in the percent of DNA (%DNA) in the tail were observed in epidermal skin cells treated with MNNG and B[a]P. In the MN test, an increased frequency of MN cells (%MN) cells was observed by treatment with MNNG; however, there were no significant increases. In contrast, significant differences in %MN were observed by treatment with B[a]P. From these results, we conclude that the combined in vivo skin comet assay and in vivo MN test was useful because it can detect different genotoxicity with the same sampling time and reduce the number of animals used. PMID:22240394

Toyoizumi, Tomoyasu; Ohta, Ryo; Kawakami, Kumiko; Nakagawa, Yuzuki; Tazura, Yoshiyuki; Kuwagata, Makiko; Noguchi, Satoshi; Sui, Hajime; Yamakage, Kohji

2012-01-06

297

In vivo imaging of hydrogen peroxide with chemiluminescent nanoparticles.  

PubMed

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 (460-630 nm), 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. PMID:17704780

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

2007-08-19

298

Endocavitary in vivo Dosimetry for IMRT Treatments of Gynecologic Tumors  

SciTech Connect

The accuracy and reproducibility of endometrial carcinoma treatment with intensity-modulated radiotherapy (IMRT) was assessed by means of in vivo dosimetry. Six patients who had previously undergone radical hysterectomy for endometrial carcinoma were treated with IMRT using a vaginal applicator with radio-opaque fiducial markers. An ion-chamber inserted into the applicator supplied an endocavitary in vivo dosimetry for quality assurance purposes. The ratio R = D/D{sub TPS} between the in vivo measured dose D and the predicted dose by the treatment planning system D{sub TPS} was determined for every fraction of the treatment. Results showed that 90% and 100% of the ratios resulted equal to 1 within 5% and 10%, respectively. The mean value of the ratios distribution for the 6 patients was R = 0.995 and the SD = 0.034. The ratio R* between the measured and predicted total doses for each patient was near to 1, within 2%. The dosimetric results suggest that the use of a vaginal applicator in an image-guided approach could make the interfractions target position stable and reproducible, allowing a safe use of the IMRT technique in the treatment of postoperative vaginal vault. In vivo dosimetry may supply useful information about the discrimination of random vs. systematic errors. The workload is minimum and this in vivo dosimetry can be applied also in the clinical routine.

Cilla, Savino, E-mail: savinocilla@gmail.com [Medical Physics Unit, Radiotherapy Unit, John Paul II Center for High Technology Research and Education in Biomedical Sciences, Catholic University, Campobasso (Italy); Macchia, Gabriella; Digesu, Cinzia; Deodato, Francesco; Sabatino, Domenico; Morganti, Alessio G.; Piermattei, Angelo [Medical Physics Unit, Radiotherapy Unit, John Paul II Center for High Technology Research and Education in Biomedical Sciences, Catholic University, Campobasso (Italy)

2011-01-01

299

Optogenetic investigation of neural circuits in vivo  

PubMed Central

The recent development of light-activated optogenetic probes allows for the identification and manipulation of specific neural populations and their connections in awake animals with unprecedented spatial and temporal precision. This review describes the use of optogenetic tools to investigate neurons and neural circuits in vivo. We describe the current panel of optogenetic probes, methods of targeting these probes to specific cell types in the nervous system, and strategies of photostimulating cells in awake, behaving animals. Finally, we survey the application of optogenetic tools to studying functional neuroanatomy, behavior, and the etiology and treatment of various neurological disorders.

Carter, Matthew E.; de Lecea, Luis

2011-01-01

300

Quantification of carbon nanomaterials in vivo.  

PubMed

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

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

2012-10-04

301

Human skin levels of retinoic acid and cytochrome P-450-derived 4-hydroxyretinoic acid after topical application of retinoic acid in vivo compared to concentrations required to stimulate retinoic acid receptor-mediated transcription in vitro.  

PubMed Central

Metabolism of retinoic acid to a less active metabolite, 4-hydroxyretinoic acid, occurs via cytochrome P-450 isozyme(s). Effect of a pharmacological dose of retinoic acid on the level of retinoic acid in skin and on cytochrome P-450 activity was investigated. A cream containing 0.1% retinoic acid or cream alone was applied topically to adult human skin for four days under occlusion. Treated areas were removed by a keratome and a microsomal fraction was isolated from each biopsy. In vitro incubation of 3H-retinoic acid with microsomes from in vivo retinoic acid treated sites resulted in a 4.5-fold increase (P = 0.0001, n = 13) in its transformation to 4-hydroxyretinoic acid in comparison to in vitro incubations with microsomes from in vivo cream alone treated sites. This cytochrome P-450 mediated activity was oxygen- and NADPH-dependent and was inhibited 68% by 5 microM ketoconazole (P = 0.0035, n = 8) and 51% by carbon monoxide (P = 0.02, n = 6). Cotransfection of individual retinoic acid receptors (RARs) or retinoid X receptor-alpha (RXR-alpha) and a chloramphenicol acetyl transferase (CAT) reporter plasmid containing a retinoic acid responsive element into CV-1 cells was used to determine the ED50 values for stimulation of CAT activity by retinoic acid and its metabolites. Levels of all trans and 13-cis RA in RA-treated tissues were greater than the ED50 values determined for all three RARs with these compounds. Furthermore, the level of all trans RA was greater than the ED50 for RXR-alpha whereas the 4-OH RA level was greater than the ED50 for RAR-beta and RAR-gamma but less than for RAR-alpha and RXR-alpha. These data suggest that there are sufficient amounts of retinoic acid in treated skin to activate gene transcription over both RARs and RXR-alpha.

Duell, E A; Astrom, A; Griffiths, C E; Chambon, P; Voorhees, J J

1992-01-01

302

Simultaneous in vivo positron emission tomography and magnetic resonance imaging  

Microsoft Academic Search

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

Ciprian Catana; Daniel Procissi; Yibao Wu; Martin S. Judenhofer; Jinyi Qi; Bernd J. Pichler; Russell E. Jacobs; Simon R. Cherry

2008-01-01

303

Sparking New Frontiers: Using in Vivo Electroporation for Genetic Manipulations  

Microsoft Academic Search

In vivo electroporation is a fascinating new approach by which gene expression, regulation, and function can be studied in developmental systems. This technique offers new opportunities for manipulations in animal models that lack genetic approaches, including avians. Furthermore, this approach is applicable to other embryo populations including mice, ascidians, zebrafish, Xenopus, and Drosophila. In this review, we discuss technical aspects

M. Swartz; J. Eberhart; G. S. Mastick; C. E. Krull

2001-01-01

304

A new method to measure oxygen solubility in organic solvents through optical oxygen sensing.  

PubMed

A new method to measure oxygen concentration in air-saturated organic solvents and binary mixtures has been developed. The methodology relies on the ability of HPLC columns to retain the molecular oxygen contained in different types of solvents which are injected into the system at 298.15 K. The outlet of the HPLC is coupled with an optical oxygen sensor which continuously measures changes in oxygen partial pressure. PMID:23803965

Quaranta, Michela; Murkovic, Michael; Klimant, Ingo

2013-09-30

305

Oxidant and Redox Signaling in Vascular Oxygen Sensing: Implications for Systemic and Pulmonary Hypertension  

PubMed Central

Abstract It has been well known for >100 years that systemic blood vessels dilate in response to decreases in oxygen tension (hypoxia; low Po2), and this response appears to be critical to supply blood to the stressed organ. Conversely, pulmonary vessels constrict to a decrease in alveolar Po2 to maintain a balance in the ventilation-to-perfusion ratio. Currently, although little question exists that the Po2 affects vascular reactivity and vascular smooth muscle cells (VSMCs) act as oxygen sensors, the molecular mechanisms involved in modulating the vascular reactivity are still not clearly understood. Many laboratories, including ours, have suggested that the intracellular calcium concentration ([Ca2+ ]i), which regulates vasomotor function, is controlled by free radicals and redox signaling, including NAD(P)H and glutathione (GSH) redox. In this review article, therefore, we discuss the implications of redox and oxidant alterations seen in pulmonary and systemic hypertension, and how key targets that control [Ca2+ ]i, such as ion channels, Ca2+ release from internal stores and uptake by the sarcoplasmic reticulum, and the Ca2+ sensitivity to the myofilaments, are regulated by changes in intracellular redox and oxidants associated with vascular Po2 sensing in physiologic or pathophysiologic conditions. Antioxid. Redox Signal. 10, 1137–1152.

Wolin, Michael S.

2008-01-01

306

Luminescence and oxygen sensing properties of ORMOSILs covalently grafted with a novel ruthenium(II) complex  

Microsoft Academic Search

A series of VTES\\/TEOS composite xerogels covalently grafted with a novel complex Ru(phen)2(Dppz-Si)Cl2 were prepared, using the alkoxysilane-modified dipyrido[3,2-a:2?,3?-c]phenazine compound (denoted as Dppz-Si) as the second\\u000a ligand of the Ru(phen)2Cl2 (phen = 1,10-phenanthroline) complex and a precursor of the sol–gel process. Bulk xerogels were obtained by co-hydrolyzing\\u000a and co-condensation from a mixture of triethoxysilane (TEOS), Ru(phen)2(Dppz-Si)Cl2 and Vinyltriethoxysilane (VTES). The luminescence intensity

Xiudong Wu; Yan Cong; Yanhong Liu; Jun Ying; Bin Li

2009-01-01

307

Fiber Optical Micro-detectors for Oxygen Sensing in Power Plants  

SciTech Connect

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.

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

2003-09-30

308

Subfunctionalization of cyprinid hypoxia-inducible factors for roles in development and oxygen sensing.  

PubMed

Among vertebrates, teleost fishes have evolved the most impressive adaptations to variable oxygen tensions in water (Shoubridge and Hochachka 1980; Nilsson and Randall 2010). Under conditions of oxygen deprivation (hypoxia), major changes in gene expression are mediated by hypoxia-inducible factors (HIF alpha). Here we show that hif alpha genes were duplicated in the teleost specific whole-genome duplication. Although one of each paralogous gene pair was lost in most teleosts, both copies were retained in cyprinids. Computational analyses suggest that these duplicates have become subfunctionalized with complementary changes in coding and regulatory sequences within each paralogous gene pair. We tested our predictions with comparisons of hif alpha transcription in zebrafish, a cyprinid, and sturgeon, an outgroup that diverged from teleosts before the duplication event. Our experiments revealed distinct transcriptional profiles in the cyprinid duplicates: while one of each paralogous pair maintained the ancestral developmental response, the other was more sensitive to changes in oxygen tension. These results demonstrate the subfunctionalization of cyprinid hif alpha paralogs for specialized roles in development and the hypoxic stress response. PMID:23461336

Rytkönen, Kalle T; Akbarzadeh, Arash; Miandare, Hamed K; Kamei, Hiroyasu; Duan, Cunming; Leder, Erica H; Williams, Tom A; Nikinmaa, Mikko

2012-10-29

309

ZnO nanowire field-effect transistor and oxygen sensing property  

Microsoft Academic Search

Single-crystal ZnO nanowires are synthesized using a vapor trapping chemical vapor deposition method and configured as field-effect transistors. Electrical transport studies show n-type semiconducting behavior with a carrier concentration of ~107 cm-1 and an electron mobility of ~17 cm2\\/V s. The contact Schottky barrier between the Au\\/Ni electrode and nanowire is determined from the temperature dependence of the conductance. Thermionic

Zhiyong Fan; Dawei Wang; Pai-Chun Chang; Wei-Yu Tseng; Jia G. Lu

2004-01-01

310

The regulation, localization, and functions of oxygen-sensing prolyl hydroxylase PHD3.  

PubMed

The prolyl 4-hydroxylase domain protein 3 (PHD3) belongs to 2-oxoglutarate and iron-dependent dioxygenases. Together with the two closest paralogues, PHD1 and PHD2, these enzymes have been identified as cellular oxygen sensors that can mark the hypoxia-inducible factor ? (HIF-?) for von Hippel-Lindau protein-mediated proteasomal destruction. Although having overlapping functions with PHD1 and PHD2, PHD3 markedly differs from the two isoforms. PHD3 shows a different expression pattern and subcellular localization as well as activity under low oxygen tension. Moreover, it has the widest range of non-HIF targets underlying its diverse functions. The functions of PHD3 differ depending on the cell type and also partially on the microenvironmental conditions it is expressed at. Under normoxia, PHD3 has been shown to be proapoptotic, but under hypoxia, it can have cell survival or proliferation-supporting functions. Here we discuss the regulation, targets, and functions of PHD3. PMID:23380539

Jaakkola, Panu M; Rantanen, Krista

2013-04-01

311

Imaging of the hypoxia-inducible factor pathway: insights into oxygen sensing  

Microsoft Academic Search

The transcription factor complex hypoxia-inducible factor (HIF)-1 controls the expression of most genes involved in adaptation to hypoxic conditions. HIF-1 is a heterodimer composed of oxygen-labile HIF-a and constitutively expressed HIF-b subunits. The oxygen- dependent regulation of HIF-a is a multistep process that includes degradation under normoxia but stabilisation, translocation into the nucleus and activation under hypoxic conditions. The present

U. Berchner-Pfannschmidt; S. Frede; C. Wotzlaw; J. Fandrey

2008-01-01

312

HIF-1? in Epidermis: Oxygen Sensing, Cutaneous Angiogenesis, Cancer, and Non-Cancer Disorders  

Microsoft Academic Search

Besides lung, postnatal human epidermis is the only epithelium in direct contact with atmospheric oxygen. Skin epidermal oxygenation occurs mostly through atmospheric oxygen rather than tissue vasculature, resulting in a mildly hypoxic microenvironment that favors increased expression of hypoxia-inducible factor-1? (HIF-1?). Considering the wide spectrum of biological processes, such as angiogenesis, inflammation, bioenergetics, proliferation, motility, and apoptosis, that are regulated

Hamid R Rezvani; Nsrein Ali; Lars J Nissen; Ghida Harfouche; Hubert de Verneuil; Alain Taïeb; Frédéric Mazurier

2011-01-01

313

In Vivo Study of Intradermal Focusing for Tattoo Removal  

Microsoft Academic Search

.\\u000a \\u000a Delivery of intradermally focused nanosecond laser pulses with small energy as an alternate technique applicable to clinical\\u000a procedures in dermatological and plastic surgery is an area of relatively new interest with multiple potential applications.\\u000a We assessed this approach on common tattoo pigments in dermis in an in vivo study using a wavelength of 1064?nm. Paired micropigs\\u000a were tattooed with

X. H. Hu; W. A. Wooden; S. J. Vore; M. J. Cariveau; Q. Fang; G. W. Kalmus

2002-01-01

314

Controlled cobalt doping of magnetosomes in vivo  

NASA Astrophysics Data System (ADS)

Magnetotactic bacteria biomineralize iron into magnetite (Fe3O4) nanoparticles that are surrounded by lipid vesicles. These `magnetosomes' have considerable potential for use in bio- and nanotechnological applications because of their narrow size and shape distribution and inherent biocompatibility. The ability to tailor the magnetic properties of magnetosomes by chemical doping would greatly expand these applications; however, the controlled doping of magnetosomes has so far not been achieved. Here, we report controlled in vivo cobalt doping of magnetosomes in three strains of the bacterium Magnetospirillum. The presence of cobalt increases the coercive field of the magnetosomes-that is, the field necessary to reverse their magnetization-by 36-45%, depending on the strain and the cobalt content. With elemental analysis, X-ray absorption and magnetic circular dichroism, we estimate the cobalt content to be between 0.2 and 1.4%. These findings provide an important advance in designing biologically synthesized nanoparticles with useful highly tuned magnetic properties.

Staniland, Sarah; Williams, Wyn; Telling, Neil; van der Laan, Gerrit; Harrison, Andrew; Ward, Bruce

2008-03-01

315

Survival of thefittest: in vivo selection and stem cell gene therapy  

Microsoft Academic Search

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,

Tobias Neff; Brian C. Beard; Hans-Peter Kiem

2006-01-01

316

In Vivo Proton Relaxation Times Analysis of the Skin Layers by Magnetic Resonance Imaging  

Microsoft Academic Search

If in vivo magnetic resonance imaging is nowadays a powerful non-invasive method in medical diagnosis, its application in order to study the skin in vivo is not yet in common use because skin imaging requires a high resolution, at least in the direction perpendicular to the skin surface. We have therefore designed a specific imaging module, which, connected to a

Stéphanie Richard; Bernard Querleux; Jacques Bittoun; Ilana Idy-Peretti; Odile Jolivet; Eva Cermakova; Jean-Luc Lévêque

1991-01-01

317

In Vivo Microdialysis in an Animal Model of Neurological Disease: Thiamine Deficiency (Wernicke) Encephalopathy  

Microsoft Academic Search

In vivo microdialysis allows for the constant monitoring of brain neurotransmitters in the extracellular fluid of awake and freely moving animals. Considerations including factors affecting probe recoveries, the blood-brain barrier, and tissue reactions to probe implantation are discussed in this paper. Details of the application of in vivo microdialysis to an animal model of encephalopathy are then presented. Thiamine deficiency

Kathryn G. Todd; Roger F. Butterworth

2001-01-01

318

In Vivo Treg Suppression Assays  

PubMed Central

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.

Workman, Creg J.; Collison, Lauren W.; Bettini, Maria; Pillai, Meenu R.; Rehg, Jerold E.; Vignali, Dario A.A.

2011-01-01

319

In vivo photoacoustic mapping of lymphatic systems with plasmon-resonant nanostars  

PubMed Central

Plasmon-resonant nanostars (NSTs) provide excellent contrast enhancement for photoacoustic tomography. The high photoacoustic sensitivity of NSTs at near-infrared wavelengths enable their in vivo detection in rat sentinel lymph nodes and vessels, with direct application toward lymphangiography.

Kim, Chulhong; Song, Hyon-Min; Cai, Xin; Yao, Junjie; Wei, Alexander; Wang, Lihong V.

2011-01-01

320

Defining human mesenchymal stem cell efficacy in vivo  

PubMed Central

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.

2010-01-01

321

Metal Nanoshells for Biosensing Applications.  

National Technical Information Service (NTIS)

The present invention provides nanoshell particles ('nanoshells') for use in biosensing applications, along with their manner of making and methods of using the nanoshells for in vitro and in vivo detection of chemical and biological analytes, preferably ...

J. L. West N. J. Halas R. D. Averitt S. J. Oldenburg

2004-01-01

322

Investigation of ex vivo stability of fesoterodine in human plasma and its simultaneous determination together with its active metabolite 5-HMT by LC-ESI-MS/MS: Application to a bioequivalence study.  

PubMed

Fesoterodine is a non-selective muscarinic-receptor antagonist, used in the treatment of overactive bladder syndrome. A highly sensitive, selective and rapid method has been developed for the simultaneous determination of fesoterodine and its active metabolite, 5-hydroxymethyl tolterodine (5-HMT) in human plasma by liquid chromatography-tandem mass spectrometry (LC-ESI-MS/MS). Due to rapid conversion of parent drug to 5-HMT, ex vivo stability of fesoterodine in human plasma was extensively studied to optimize the extraction protocol. The analytes and their deuterated analogs were quantitatively extracted from 100?L human plasma by liquid-liquid extraction in methyl tert-butyl ether: n-hexane. The chromatographic separation of analytes was achieved on a Kromasil C18 (100mm×4.6mm, 5?m) column under isocratic conditions. The method was validated over a dynamic concentration range of 0.01-10ng/mL for both the analytes. Ion-suppression effects were investigated by post-column infusion of analytes. The precision (% CV) values for the calculated slopes of calibration curves, which would reflect the relative matrix effect, were less than 1.5% for both the analytes. The intra-batch and inter-batch precision (% CV) across quality control levels varied from 1.82 to 3.73% and the mean extraction recovery was >96% for both the analytes. The method was successfully applied to a bioequivalence study of 8mg fesoterodine tablet formulation (test and reference) in 12 healthy Indian subjects under fasted and fed condition. The assay reproducibility estimated by reanalysis of incurred samples showed a change of ±12.0%. PMID:23266359

Parekh, Jignesh M; Sanyal, Mallika; Yadav, Manish; Shrivastav, Pranav S

2012-11-23

323

Tailoring vessel morphology in vivo  

NASA Astrophysics Data System (ADS)

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.

Gould, Daniel Joseph

324

Progress Toward In Vivo Use of siRNAs-II  

PubMed Central

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.

Rettig, Garrett R; Behlke, Mark A

2012-01-01

325

Quantum dot conjugated hydroxylapatite nanoparticles for in vivo imaging  

NASA Astrophysics Data System (ADS)

Hydroxylapatite (HA) nanoparticles were conjugated with quantum dots (QDs) for in vivo imaging. The surface structures of HA nanoparticles with conjugated quantum dots (HA-QD) were studied by transmission electron microscopy (TEM) and laser fluorescent spectroscopy. The TEM data showed that the quantum dots were well conjugated on the HA nanoparticle surfaces. The laser fluorescent spectroscopy results indicated that the HA-QD exhibited promising luminescent emission in vitro. The initial in vivo experiments revealed clear images of HA-QD from the hypodermic injected area at the emission of 600 nm. Furthermore, the optimized in vivo images of HA-QD with near-infrared emission at 800 nm were visualized after intravenous injection. These luminescent HA-QD nanoparticles may find important applications as biodegradable substrates for biomarkers and in drug delivery.

Guo, Yan; Shi, Donglu; Lian, Jie; Dong, Zhongyun; Wang, Wei; Cho, Hoonsung; Liu, Guokui; Wang, Lumin; Ewing, Rodney C.

2008-04-01

326

Tracking immune cells in vivo using magnetic resonance imaging.  

PubMed

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 (19)F-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

Ahrens, Eric T; Bulte, Jeff W M

2013-09-10

327

Progress toward in vivo use of siRNAs-II.  

PubMed

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

Rettig, Garrett R; Behlke, Mark A

2011-12-20

328

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)

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 mm3) 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.

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

329

Experimental Cryosurgery Investigations In Vivo  

PubMed Central

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.

Gage, A.A.; Baust, J.M.; Baust, J.G.

2009-01-01

330

In-vivo morphologic and spectroscopic investigation of Psoriasis  

NASA Astrophysics Data System (ADS)

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.

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

2011-06-01

331

Quantifying drug-protein binding in vivo.  

SciTech Connect

Accelerator mass spectrometry (AMS) provides precise quantitation of isotope labeled compounds that are bound to biological macromolecules such as DNA or proteins. The sensitivity is high enough to allow for sub-pharmacological (''micro-'') dosing to determine macromolecular targets without inducing toxicities or altering the system under study, whether it is healthy or diseased. We demonstrated an application of AMS in quantifying the physiologic effects of one dosed chemical compound upon the binding level of another compound in vivo at sub-toxic doses [4].We are using tissues left from this study to develop protocols for quantifying specific binding to isolated and identified proteins. We also developed a new technique to quantify nanogram to milligram amounts of isolated protein at precisions that are comparable to those for quantifying the bound compound by AMS.

Buchholz, B; Bench, G; Keating III, G; Palmblad, M; Vogel, J; Grant, P G; Hillegonds, D

2004-02-17

332

Transepidermal Water Loss in vivo  

Microsoft Academic Search

In vivo, noninvasive, transepidermal water loss measurements were taken from 26- to 41-week gestational age infants with a Meeco® electrolytic water analyzer to yield information on the development of the function of the skin barrier. Infants 28 weeks or less have significantly greater transepidermal water loss than older gestational aged infants, thus indicating a less efficient barrier to water. Until

Donald R. Wilson; Howard I. Maibach

1980-01-01

333

Imaging cell death in vivo  

Microsoft Academic Search

A technique to image programmed cell death would be useful both in clinical care and in drug development. The most widely studied agent for the in vivo study of apop- tosis is radiolabeled annexin V, an endogenous protein labeled with technectium-99m, now undergoing clinical trials in both Europe and the United States. While annex- in V has been studied extensively

F. BLANKENBERG; C. MARI; H. W. STRAUSS

2003-01-01

334

Characterization and application of temperature micro-optodes for use in aquatic biology  

NASA Astrophysics Data System (ADS)

Benthic aquatic environments like biofilms or sediments are often investigation by measuring profiles of chemical or physical parameters at a high spatial resolution (< 50 micrometers ). This is necessary to understand e.g. transport processes and the biogeochemistry of the sediment water interface. A variety of electrochemical and optical microsensors has been developed and used for this purpose. In most of these applications the temperature of the investigated biofilms or sediments is assumed to be constant. However measurements with thermocouples of an appr. diameter of 300 micrometers have shown that this is not always the case for illuminated shallow water sediments and biofilms. We developed new microoptodes for measuring temperature distributions at a high spatial (< 50 micrometers ) and thermal (< 0.2 degree(s)C) resolution in aquatic systems. The new sensors are based on a fluorophore that is well known for its application in oxygen sensing-Ruthenium(II)- tris-1,10-phenantroline. Demas et al. (1992) discussed the possible use of highly luminescent transition metal complexes as temperature indicators. We have approached this idea from our experiences with ruthenium complexes as oxygen indicators. The first realized sensor consists of a closed microcapillary filled with an indicator solution and in inserted tapered optical fiber. The principle uses the temperature dependence of the fluorescence lifetime in the solution. To keep the solution oxygen free an oxygen scavenger is added to it. The change of the lifetime is detected by a special measuring device that uses a phase modulation technique.

Holst, Gerhard A.; Kuehl, Michael; Klimant, Ingo; Liebsch, Gregor; Kohls, Oliver

1997-05-01

335

Extradiscal ultrasound thermal therapy (ExDUSTT): evaluation in ex vivo and in vivo spine models  

NASA Astrophysics Data System (ADS)

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.

Diederich, Chris J.; Kinsey, Adam; Nau, William H.; Shu, Richard; Lotz, Jeffrey C.

2005-04-01

336

Silver Nanoplate Contrast Agents for In Vivo Molecular Photoacoustic Imaging  

PubMed Central

Silver nanoplates are introduced as a new photoacoustic contrast agent that can be easily functionalized for molecular photoacoustic imaging in vivo. Methods are described for synthesis, functionalization, and stabilization of silver nanoplates using biocompatible (“green”) reagents. Directional antibody conjugation to the nanoplate surface is presented along with proof of molecular sensitivity in vitro with pancreatic cancer cells. Cell viability tests show the antibody-conjugated silver nanoplates to be nontoxic at concentrations up to 1 mg/ml. Furthermore, the silver nanoplates' potential for in vivo application as a molecularly sensitive photoacoustic contrast agent is demonstrated using an orthotopic mouse model of pancreatic cancer. Results of these studies suggest that the synthesized silver nanoplates are well suited for a host of biomedical imaging and sensing applications.

Homan, Kimberly A.; Souza, Michael; Truby, Ryan; Luke, Geoffrey P.; Green, Christopher; Vreeland, Erika; Emelianov, Stanislav

2012-01-01

337

Near-infrared Molecular Probes for In Vivo Imaging  

PubMed Central

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.

Zhang, Xuan; Bloch, Sharon; Akers, Walter; Achilefu, Samuel

2012-01-01

338

In vivo photoacoustic imaging of osteosarcoma in a rat model  

NASA Astrophysics Data System (ADS)

Osteosarcoma is one of the most common primary malignant tumors of the bone and the second leading cause of cancer-related deaths in the pediatric age group. Confirmed diagnosis and prompt treatment of osteosarcoma are critical for effective prognosis. In this study, we investigate the application of photoacoustic imaging (PAI) for the detection of osteosarcoma in an animal model. Cross-section images of a normal rat leg and a tumorous rat leg were successfully reconstructed in vivo. Morphological changes and the development of the implanted osteosarcoma were accurately mapped with time-dependent photoacoustic images. Furthermore, we evaluate the use of gold nanorods as contrast agents for imaging osteosarcoma with PAI. This is the first study that uses PAI to detect osteosarcoma in vivo, and the results suggest that PAI has the potential clinical application for detecting osteosarcoma in the early stage.

Hu, Jun; Yu, Menglei; Ye, Fei; Xing, Da

2011-02-01

339

Silver nanoplate contrast agents for in vivo molecular photoacoustic imaging.  

PubMed

Silver nanoplates are introduced as a new photoacoustic contrast agent that can be easily functionalized for molecular photoacoustic imaging in vivo. Methods are described for synthesis, functionalization, and stabilization of silver nanoplates using biocompatible ("green") reagents. Directional antibody conjugation to the nanoplate surface is presented along with proof of molecular sensitivity in vitro with pancreatic cancer cells. Cell viability tests show the antibody-conjugated silver nanoplates to be nontoxic at concentrations up to 1 mg/mL. Furthermore, the silver nanoplates' potential for in vivo application as a molecularly sensitive photoacoustic contrast agent is demonstrated using an orthotopic mouse model of pancreatic cancer. Results of these studies suggest that the synthesized silver nanoplates are well suited for a host of biomedical imaging and sensing applications. PMID:22188516

Homan, Kimberly A; Souza, Michael; Truby, Ryan; Luke, Geoffrey P; Green, Christopher; Vreeland, Erika; Emelianov, Stanislav

2012-01-06

340

19F MRI for quantitative in vivo cell tracking  

PubMed Central

Cellular therapy, including stem cell transplants and dendritic cell vaccines, is typically monitored for dosage optimization, accurate delivery and localization using non-invasive imaging, of which magnetic resonance imaging (MRI) is a key modality. 19F MRI retains the advantages of MRI as an imaging modality, while allowing direct detection of labelled cells for unambiguous identification and quantification, unlike typical metal-based contrast agents. Recent developments in 19F MRI-based in vivo cell quantification, the existing clinical use of 19F compounds and current explosive interest in cellular therapeutics have brought 19F imaging technology closer to clinical application. We review the application of 19F MRI to cell tracking, discussing intracellular 19F labels, cell labelling and in vivo quantification, as well as the potential clinical use of 19F MRI.

Srinivas, Mangala; Heerschap, Arend; Ahrens, Eric T.; Figdor, Carl G.; de Vries, I. Jolanda M.

2010-01-01

341

Development and evaluation of colloidal modified nanolipid carrier: application to topical delivery of tacrolimus, Part II - in vivo assessment, drug targeting, efficacy, and safety in treatment for atopic dermatitis.  

PubMed

In atopic dermatitis (AD), topical anti-inflammatory therapy with skin barrier restoration to prevent repeated inflammatory episodes leads to long-term therapeutic success. Tacrolimus, although effective against AD, is a challenging molecule due to low solubility, low-penetration, poor-bioavailability, and toxicity. Part I of this paper, reported novel modified nanolipid carrier system for topical delivery of tacrolimus (T-MNLC), offering great opportunity to load low-solubility drug with improved entrapment efficiency, enhanced stability and improved skin deposition. Present investigation focused on restoration of skin barrier, site-specific delivery, therapeutic effectiveness, and safety of novel T-MNLC. T-MNLC greatly enhanced occlusive properties, skin hydration potential and reduced transepidermal water loss. This might help to reduce the number of flares and better control the disease. Cutaneous uptake and drug deposition in albino rats by HPLC and confocal laser scanning microscopy revealed prominently elevated drug levels in all skin strata with T-MNLC as compared to reference. T-MNLC demonstrated efficient suppression of inflammatory responses in BALB/c mice model of AD. Safety assessment by acute and repeated-dose dermal toxicity demonstrated mild keratosis and collagenous mass infiltration at the treatment area with repeated application of reference. Interestingly, T-MNLC showed no evident toxicity exhibiting safe drug delivery. Thus, novel T-MNLC would be a safe, effective, and esthetically appealing alternative to conventional vehicles for treatment for AD. PMID:23246619

Pople, Pallavi V; Singh, Kamalinder K

2012-12-12

342

In vivo high-resolution structural imaging of large arteries in small rodents using two-photon laser scanning microscopy  

Microsoft Academic Search

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,

Remco T. A. Megens; Sietze Reitsma; Lenneke Prinzen; Mirjam G. A. Oude Egbrink; Wim Engels; Peter J. A. Leenders; Ellen J. L. Brunenberg; Koen D. Reesink; Ben J. A. Janssen; Bart M. Ter Haar Romeny; Dick W. Slaaf; Marc A. M. J. van Zandvoort

2010-01-01

343

Positron emitters for {ital in vivo} plant studies  

SciTech Connect

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.}

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

344

In vivo imaging with oligonucleotides for diagnosis and drug development  

PubMed Central

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.

Tavitian, B

2003-01-01

345

Keratinocyte Growth Factor Accelerates Corneal Epithelial Wound Healing In Vivo  

Microsoft Academic Search

Purpose. To examine whether the topical application of keratinocyte growth factor (KGF) can enhance corneal epithelial healing in vivo. In addition, the distribution of S-phase cells in KGF-treated and control corneas was investigated during regeneration and under normal conditions. Methods. A 10-mm diameter epithelial defect was made in the center of rabbit corneas. A 50- fA aliquot of 10 \\/\\/g\\/ml

Chie Sotozono; Tsutomu Inatomi; Masatsugu Nakamura; Shigeru Kinoshita

346

In vivo whole-field blood velocity measurement techniques  

Microsoft Academic Search

In this article a number of whole-field blood velocity measurement techniques are concisely reviewed. We primarily focus on\\u000a optical measurement techniques for in vivo applications, such as laser Doppler velocimetry (including time varying speckle),\\u000a laser speckle contrast imaging and particle image velocimetry (including particle tracking). We also briefly describe nuclear\\u000a magnetic resonance and ultrasound particle image velocimetry, two techniques that

Peter Vennemann; Ralph Lindken; Jerry Westerweel

2007-01-01

347

Systems for in vivo site-directed mutagenesis using oligonucleotides  

US Patent & Trademark Office Database

This disclosure provides several methods to generate nucleic acid mutations in vivo, for instance in such a way that no heterologous sequence is retained after the mutagenesis is complete. The methods employ integrative recombinant oligonucleotides (IROs). Specific examples of the described mutagenesis methods enable site-specific point mutations, deletions, and insertions. Also provided are methods that enable multiple rounds of mutation and random mutagenesis in a localized region. The described methods are applicable to any organism that has a homologous recombination system.

2008-01-01

348

Pharmaceutical applications of chitosan  

Microsoft Academic Search

Considerable research efforts have been directed towards the development of safe and efficient chitosan-based drug delivery systems. In this review, the authors outline the major new approaches to the pharmaceutical applications of chitosan and discuss its mechanisms of action in various in vitro and in vivo models.

Valérie Dodane; Vinod D Vilivalam

1998-01-01

349

Quantification of siRNAs in vitro and in vivo.  

PubMed

RNA interference (RNAi) is a regulatory mechanism of eukaryotic cells that uses small interfering RNAs (siRNA) to direct homology-dependent control of gene activity. Applications of RNAi include functional genomics, in vivo target validation, and gene-specific medicines. A key to in vivo application of siRNA is the advancement of efficient delivery to organs, tissues, or cell types of interest. There is a need to develop reliable and easy-to-use assays to evaluate siRNA delivery efficiency and distribution, study pathways, and stability of siRNAs in cells (post-transfection) and in animals (post- injection). We have adopted the Applied Biosystems TaqMan(®) based stem-loop RT-PCR technology, originally developed for quantification of endogenous microRNAs in cells, to fulfill these needs. In this chapter, application protocols are described, which enable robust quantification of siRNA, including chemically modified molecules, in vitro and in vivo. PMID:21748641

Cheng, Angie; Vlassov, Alexander V; Magdaleno, Susan

2011-01-01

350

In vivo biofilm composition of Aspergillus fumigatus.  

PubMed

The in vivo composition of the mycelial extracellular matrix (ECM) of Aspergillus fumigatus during host invasion is reported here for the first time. A new galactosaminogalactan and the galactomannan were the major polysaccharides of the in vivo ECM. The composition of the ECM in vivo varied with the aspergillosis pathologies. PMID:19889082

Loussert, Céline; Schmitt, Christine; Prevost, Marie-Christine; Balloy, Viviane; Fadel, Elie; Philippe, Bruno; Kauffmann-Lacroix, Catherine; Latgé, Jean Paul; Beauvais, Anne

2009-11-04

351

In vivo biofilm composition of Aspergillus fumigatus  

Microsoft Academic Search

Summary The in vivo composition of the mycelial extracellu- lar matrix (ECM) of Aspergillus fumigatus during host invasion is reported here for the first time. A new galactosaminogalactan and the galactoman- nan were the major polysaccharides of the in vivo ECM. The composition of the ECM in vivo varied with the aspergillosis pathologies.

Céline Loussert; Christine Schmitt; Marie-Christine Prevost; Viviane Balloy; Elie Fadel; Bruno Philippe; Catherine Kauffmann-Lacroix; Jean Paul Latgé; Anne Beauvais

2009-01-01

352

In vivo imaging of sulfotransferases  

DOEpatents

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.

Barrio, Jorge R; Kepe, Vladimir; Small, Gary W; Satyamurthy, Nagichettiar

2013-02-12

353

Receptor microscopic autoradiography for the study of percutaneous absorption, in vivo skin penetration, and cellular–intercellular deposition  

Microsoft Academic Search

Introduction: Microscopic autoradiography with cellular resolution and preservation of in vivo conditions is potentially the method of choice to gain detailed information about sites of deposition and retention in the epidermis and of penetration to the dermis after topical application of drugs. We tested this using 3H-Maxacalcitol. Methods: Dorsal skin of adult rats was treated in vivo with ointment containing

Naohiko Hayakawa; Naoki Kubota; Nobuo Imai; Walter E. Stumpf

2004-01-01

354

Intervertebral disc regeneration in an ex vivo culture system using mesenchymal stem cells and platelet-rich plasma  

Microsoft Academic Search

An ex vivo degenerative intervertebral disc (IVD) organ culture system was established for the screening of disc regeneration agents. Its application was demonstrated by a stem cell and growth factor-based therapeutic approach for the amelioration of IVD. An ex vivo culture system using chymopapain to partially digest nucleus proposus tissue was established to mimic human IVD degeneration. This system was

Wei-Hong Chen; Hen-Yu Liu; Wen-Cheng Lo; Shinn-Chih Wu; Chau-Hwa Chi; Hsueh-Yuan Chang; Shih-Hsiang Hsiao; Chih-Hsiung Wu; Wen-Ta Chiu; Bao-Ji Chen; Win-Ping Deng

2009-01-01

355

Protection against TNF?-Dependent Liver Toxicity by Intraperitoneal Liposome Delivered DsiRNA Targeting TNF? In Vivo  

Microsoft Academic Search

Tumor necrosis factor alpha (TNF?) is a classic proinflammatory cytokine implicated in the pathogenesis of several autoimmune and inflammatory diseases including viral encephalitis. Macrophages being major producers of TNF? are thus attractive targets for in vivo RNA interference (RNAi) mediated down regulation of TNF?. The application of RNAi technology to in vivo models however presents obstacles, including rapid degradation of

Patric Lundberg; Hui-Jung Yang; Seung-Jae Jung; Mark A. Behlke; Scott D. Rose; Edouard M. Cantin

356

In vivo fluorescence lifetime tomography  

NASA Astrophysics Data System (ADS)

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.

Nothdurft, Ralph E.; Patwardhan, Sachin V.; Akers, Walter; Ye, Yunpeng; Achilefu, Samuel; Culver, Joseph P.

2009-03-01

357

An Antibody Delivery System for Regulated Expression of Therapeutic Levels of Monoclonal Antibodies In Vivo  

Microsoft Academic Search

Monoclonal antibody (mAb) delivery by gene transfer in vivo may be an attractive alternative to current mAb therapies for applications that require long-term therapy. This article describes a transfer system that allows inducible high-level expression of unmodified mAbs in vivo. A recombinant adeno-associated viral (rAAV) vector is used that comprises an expression cassette consisting of a dimerizer-regulated promoter that drives

Jianmin Fang; Saili Yi; Andrew Simmons; Guang Huan Tu; Minh Nguyen; Thomas C Harding; Melinda VanRoey; Karin Jooss

2007-01-01

358

Following protein association in vivo with fluorescence fluctuation spectroscopy  

NASA Astrophysics Data System (ADS)

The combination of fluorescence correlation spectroscopy and two-photon excitation provides us with a powerful spectroscopic technique. Its submicron resolution and single molecule sensitivity make it an attractive technique for in vivo applications. Experiments have demonstrated that quantitative in vivo fluorescence fluctuation measurements are feasible, despite the presence of autofluorescence and the heterogeneity of the cellular environment. I will demonstrate that molecular brightness of proteins tagged with green fluorescent protein (GFP) is a useful and robust parameter for in vivo studies. Knowledge of photon statistics is crucial for the interpretation of fluorescence fluctuation experiments. I will describe photon counting histogram (PCH) analysis, which determines the molecular brightness and complements autocorrelation analysis. Non-ideal detector effects and their influence on the photon statistics will be discussed. The goal of in vivo fluorescence fluctuation experiments is to address functional properties of biomolecules. We will focus on retinoid X receptor (RXR), a nuclear receptor, which is crucial for the regulation of gene expression. The fluorescence brightness of RXR tagged with EGFP will be used to probe the oligomerization state of RXR.

Muller, Joachim D.

2003-07-01

359

Widefield quantitative multiplex surface enhanced Raman scattering imaging in vivo  

NASA Astrophysics Data System (ADS)

In recent years numerous studies have shown the potential advantages of molecular imaging in vitro and in vivo using contrast agents based on surface enhanced Raman scattering (SERS), however the low throughput of traditional point-scanned imaging methodologies have limited their use in biological imaging. In this work we demonstrate that direct widefield Raman imaging based on a tunable filter is capable of quantitative multiplex SERS imaging in vivo, and that this imaging is possible with acquisition times which are orders of magnitude lower than achievable with comparable point-scanned methodologies. The system, designed for small animal imaging, has a linear response from (0.01 to 100 pM), acquires typical in vivo images in <10 s, and with suitable SERS reporter molecules is capable of multiplex imaging without compensation for spectral overlap. To demonstrate the utility of widefield Raman imaging in biological applications, we show quantitative imaging of four simultaneous SERS reporter molecules in vivo with resulting probe quantification that is in excellent agreement with known quantities (R2>0.98).

McVeigh, Patrick Z.; Mallia, Rupananda J.; Veilleux, Israel; Wilson, Brian C.

2013-04-01

360

Optical and imaging techniques for in-vivo sunscreens investigation  

NASA Astrophysics Data System (ADS)

The methods available for testing the efficacy of topical sunscreens have improved considerably in recent years. Nevertheless, so far no simple and rapid test has been proposed to measure in vivo transmission spectra of sunscreens in the UVA region. Spectral changes that occur after sunscreen application were measured with a fluorescence spectrometer (LS 50B, Perkin Elmer, UK) equipped with a Y-shape quartz guide for in vivo measurements. Three sunscreens with different protection factors in the UVA range were tested. The excitation-emission maps of human collagen, skin, and sunscreens were analyzed. Visual demonstrations of the protective effects of sunscreens were also performed with photo- and video imaging techniques. As a consequence of the human skin and sunscreen's fluorescence map analysis, the optimal spectral regions (both for direct and indirect fluorescence measurements) were detected. In vivo fluorescence and remittance spectroscopy were used to investigate the time dependence in transmission spectra of epidermis with applied sunscreens. We also evaluate the feasibility of in vivo fluorescence measurements for the investigation of the sunscreen's water-resistance. The procedure is simple, and values obtained can be used to predict UVA protection on the basis of the mathematical algorithms.

Utz, Sergei R.; Knuschke, Peter; Sinichkin, Yuri P.

1996-01-01

361

Cavitation enhances treatment depth when combined with thermal effect using a plane ultrasound transducer: an in vivo study  

Microsoft Academic Search

Minimally invasive techniques using miniaturised ultrasound applicators have been developed for surgical and medical applications. However, the volume of treatment remains limited. Therefore, we propose to combine cavitation with a thermal effect to enhance the treatment depth. This study was conducted in vivo on pig liver, 51 exposures were performed on the surface of the liver, with a miniaturised applicator

C. Goldenstedt; D. Melodelima; F. Mithieux; S. Chesnais; Y. Theillere; D. Cathignol

2004-01-01

362

Design of meloxicam and lornoxicam transdermal patches: Preparation, physical characterization, ex vivo and in vivo studies.  

PubMed

Transdermal patches of meloxicam (MX) and lornoxicam (LX) were aimed to be prepared in order to overcome their side effects by oral application. The strategy was formulation of optimized films to prepare transdermal patches by determination of physical properties and investigation of drug-excipient compatibility. As the next step, in vitro drug release, assesment of anti-inflammatory effect on Wistar Albino rats, ex vivo skin penetration and investigation of factors on drug release from transdermal patches were studied. Hydroxypropyl methylcellulose (HPMC) was concluded to be suitable polymer for formulation of MX and LX transdermal films indicating pharmaceutical quality required. MX and LX transdermal patches gave satisfactory results regarding to the edema inhibition in the assessment of anti-inflammatory effect. MX was found out to be more effective compared to LX on relieving of edema and swelling. These results were supported by data obtained from ex vivo penetration experiments of drug through rat skin. Indicative parameters like log P, molecular weight and solubility constraint on penetration rate of drugs also indicated good skin penetration. Transdermal patches of MX and LX can be suggested to be used especially for the immediate treatment of inflammated area since it displays anti-inflammatory effect, soon. PMID:21048338

Yener, Gülgün; Üner, Melike; Gönüllü, Ümit; Yildirim, Sinem; Kiliç, P?nar; Aslan, Serap Sa?lik; Barla, Asl?

2010-11-01

363

In-vivo biodegradation of extruded lipid implants in rabbits.  

PubMed

In-vitro studies with twin-screw extruded triglyceride based implants showed promising erosion behavior. However, little is known about the behavior of such systems in-vivo and therefore a degradability study was performed in a rabbit model. Four formulations were used to prepare tsc-extrudates and implanted in rabbits and mass loss of the implants was measured after one, three and six months. Samples were additionally assessed by differential scanning calorimetry (DSC) and scanning electron microscopy (SEM). A time dependent in-vivo erosion of the triglyceride matrices was detected and only 24% (± 17%) of the matrix material was recovered after 6 months of incubation (n=9). The application of a pore forming agent resulted in higher mass loss and an accelerated degradation rate. Exchange of the type of low melting triglyceride during preparation resulted in a better preservation of the porous implant structure, explained by a higher melting point of the alternate low-melting lipid. The good in-vivo degradability of tsc-extrudates was explained by the composition of the implants and the application of a low melting lipid which allowed partial melting of the implant at body temperature and rendered the implants more susceptible to degradation processes (e.g. lipolyses). PMID:22964393

Sax, Gerhard; Kessler, Barbara; Wolf, Eckhard; Winter, Gerhard

2012-09-01

364

In Vivo Delivery of Nucleic Acids via Glycopolymer Vehicles Affords Therapeutic Infarct Size Reduction In Vivo  

PubMed Central

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.

Tranter, Michael; Liu, Yemin; He, Suiwen; Gulick, James; Ren, Xiaoping; Robbins, Jeffrey; Jones, W. Keith; Reineke, Theresa M.

2012-01-01

365

Evolution of BRET Biosensors from Live Cell to Tissue-Scale In vivo Imaging  

PubMed Central

Development of bioluminescence resonance energy transfer (BRET) based genetic sensors for sensing biological functions such as protein–protein interactions (PPIs) in vivo has a special value in measuring such dynamic events at their native environment. Since its inception in the late nineties, BRET related research has gained significant momentum in terms of adding versatility to the assay format and wider applicability where it has been suitably used. Beyond the scope of quantitative measurement of PPIs and protein dimerization, molecular imaging applications based on BRET assays have broadened its scope for screening pharmacologically important compounds by in vivo imaging as well. In this mini-review we focus on an in-depth analysis of engineered BRET systems developed and their successful application to cell-based assays as well as in vivo non-invasive imaging in live subjects.

De, Abhijit; Jasani, Akshi; Arora, Rohit; Gambhir, Sanjiv S.

2013-01-01

366

Application of nuclear resonance scattering for in vivo measurements  

SciTech Connect

Nuclear resonance scattering is applied in our laboratory to measure hepatic and cardiac iron overload. For iron analysis, a gaseous source of 4 mg MnCl/sub 2/ is introduced into an evacuated quartz vial. Following irradiation in a nuclear reactor, /sup 56/Mn decays by beta emission to the 847-keV level of /sup 56/Fe, which subsequently decays to the ground state of /sup 56/Fe with a 7 ps half-life. The principal aim of this work is to evaluate the efficacy of the iron chelation therapy. Serial measurements over a time period of 6 to 12 months of a given patient will enable us to see how the iron is removed from the critical organs.

Wielopolski, L.; Vartsky, D.; Cohn, S.H.

1983-01-01

367

Fishing Pluripotency Mechanisms In Vivo  

PubMed Central

To understand the molecular mechanisms that regulate the biology of embryonic stem cells (ESCs) it is necessary to study how they behave in vivo in their natural environment. It is particularly important to study the roles and interactions of the different proteins involved in pluripotency and to use this knowledge for therapeutic purposes. The recent description of key pluripotency factors like Oct4 and Nanog in non-mammalian species has introduced other animal models, such as chicken, Xenopus, zebrafish and medaka, to the study of pluripotency in vivo. These animal models complement the mouse model and have provided new insights into the evolution of Oct4 and Nanog and their different functions during embryonic development. Furthermore, other pluripotency factors previously identified in teleost fish such as Klf4, STAT3, Sox2, telomerase and Tcf3 can now be studied in the context of a functional pluripotency network. The many experimental advantages of fish will fuel rapid analysis of the roles of pluripotency factors in fish embryonic development and the identification of new molecules and mechanisms governing pluripotency.

Sanchez-Sanchez, Ana V.; Camp, Esther; Mullor, Jose L.

2011-01-01

368

In vivo recordings of brain activity using organic transistors.  

PubMed

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

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

369

Nanoparticle PEBBLE sensors in live cells and in vivo  

PubMed Central

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.

Smith, Ron

2009-01-01

370

The zebrafish: scalable in vivo modeling for systems biology  

PubMed Central

The zebrafish offers a scalable vertebrate model for many areas of biologic investigation. There is substantial conservation of genetic and genomic features and, at a higher order, conservation of intermolecular networks, as well as physiologic systems and phenotypes. We highlight recent work demonstrating the extent of this homology, and efforts to develop high-throughput phenotyping strategies suited to genetic or chemical screening on a scale compatible with in vivo validation for systems biology. We discuss the implications of these approaches for functional annotation of the genome, elucidation of multicellular processes in vivo, and mechanistic exploration of hypotheses generated by a broad range of ‘unbiased’ ‘omic technologies such as expression profiling and genome-wide association. Finally, we outline potential strategies for the application of the zebrafish to the systematic study of phenotypic architecture, disease heterogeneity and drug responses.

Deo, Rahul C.; MacRae, Calum A.

2011-01-01

371

In vivo imaging of light-emitting probes  

NASA Astrophysics Data System (ADS)

In vivo imaging of cells tagged with light-emitting probes, such as firefly luciferase or fluorescent proteins, is a powerful technology that enables a wide range of biological studies in small research animals. Reporters with emission in the red to infrared (> 600 nm) are preferred due to the low absorption in tissue at these wavelengths. Modeling of photon diffusion through tissue indicates that bioluminescent cell counts as low as a few hundred can be detected subcutaneously, while approximately106 cells are required to detect signals at approximately 2 cm depth in tissue. Signal-to- noise estimates show that cooled back-thinned integrating charge coupled devices (CCDs) are preferred to image-intensified CCDs for this application, mainly due to their high quantum efficiency (approximately 85%) at wavelengths > 600 nm where tissue absorption is low. Instrumentation for in vivo imaging developed at Xenogen is described and several examples of images of mice with bioluminescent cells are presented.

Rice, Bradley W.; Cable, Michael D.; Nelson, Michael B.

2001-10-01

372

Chemiluminescence detection from sonodynamic action in vitro and in vivo  

NASA Astrophysics Data System (ADS)

In this work, chemiluminescence method was engaged for the first time to detect the active oxygen spices during sonocynamic action both in vitro and in vivo. We used CLA derivatives, which can efficiently react with singlet oxygen (1O2) or superoxide anion (O2-) to emit light, and luminol, which can be oxidized by a variety of free radicals to emit photons, to real-timely detect oxygen free radical formation in the sonosensitization of two sonosensitizer ATX-70 and HpD. The results show that 1O2 is involved in the sonosensitization. The mechanism of sonosensitizing was discussed. In vivo experiments, tumor imaging by sonodynamic chemiluminescence detection methods was established. This method could have potential applications in clinics for early-stage tumor diagnosis.

He, Yonghong; Xing, Da; Yan, Guihong; Ueda, Kenichi

2001-10-01

373

Chemotherapy with hybrid liposomes for lymphoma without drugs in vivo.  

PubMed

Hybrid liposomes composed of dimyristoylphosphatidylcholine (DMPC) and polyoxyethylene (n) dodecyl ether (C(12)(EO)(n), n=21 and 25) were prepared with the method of sonication. Clear solution of hybrid liposomes having hydrodynamic diameter of 80-100 nm could be maintained over 3 weeks. Hybrid liposomes induced apoptosis for human lymphoma (MOLT-4 and RAJI) cells in vitro. No toxicity was observed in the rats after intravenously injecting hybrid liposomes in vivo. We clearly demonstrated that a mouse model of lymphoma was established and prolonged survival was obtained in mice models of lymphoma after the treatment with hybrid liposomes without drugs in vivo. The results in this study should support the prolonged survival for patients with lymphoma in clinical applications. PMID:16574356

Nagami, Hideaki; Matsumoto, Yoko; Ueoka, Ryuichi

2006-03-06

374

In Vivo Enzyme Immobilization by Inclusion Body Display? †  

PubMed Central

A novel strategy for in vivo immobilization of enzymes on the surfaces of inclusion bodies has been established. It relies on expression in Escherichia coli of the polyhydroxybutyrate synthase PhaC from Cupriavidus necator, which carries at its amino terminus an engineered negatively charged ?-helical coil (Ecoil) and forms inclusion bodies upon high-level expression. Coexpression in the same cell of galactose oxidase (GOase) from Fusarium spp. carrying a carboxy-terminal positively charged coil (lysine-rich coil [Kcoil]) sequence results in heterodimeric coiled-coil formation in vivo and in the capture of the enzyme in active form on the surface of the inclusion body particle. These round-shaped enzyme-decorated microparticles, with sizes of approximately 0.7 ?m, can be isolated from lysed cells simply by centrifugation. The cost-effective one-step generation and isolation of enzymes immobilized on inclusion body particles may become useful for various applications in bioprocessing and biotransformation.

Steinmann, Bjorn; Christmann, Andreas; Heiseler, Tim; Fritz, Janine; Kolmar, Harald

2010-01-01

375

Assessing ex vivo dental biofilms and in vivo composite restorations using cross-polarization optical coherence tomography  

NASA Astrophysics Data System (ADS)

A cross-polarization 1310-nm optical coherence tomography system (CP-OCT), using a beam splitter based design, was used to assess ex vivo growth of complex multi-species dental biofilms. These biofilm microcosms were derived from plaque samples along the interface of composite or amalgam restoration in children with a history of early childhood caries. This paper presents a method of measuring the mean biofilm height of mature biofilms using CP-OCT. For our in vivo application, the novel swept source based CP-OCT intraoral probe (Santec Co. Komaki, Japan) dimensions and system image acquisition speed (20 image frames/second) allowed imaging pediatric subjects as young as 4 years old. The subsurface enamel under the interface of composite resin restorations of pediatric subjects were imaged using CP-OCT. Cavitated secondary caries is clearly evident from sound resin composite restorations.

Jones, R.; Aparicio, C.; Chityala, R.; Chen, R.; Fok, A.; Rudney, J.

2012-02-01

376

Optical stimulation of peripheral nerves in vivo  

NASA Astrophysics Data System (ADS)

This dissertation documents the emergence and validation of a new clinical tool that bridges the fields of biomedical optics and neuroscience. The research herein describes an innovative method for direct neurostimulation with pulsed infrared laser light. Safety and effectiveness of this technique are first demonstrated through functional stimulation of the rat sciatic nerve in vivo. The Holmium:YAG laser (lambda = 2.12 mum) is shown to operate at an optimal wavelength for peripheral nerve stimulation with advantages over standard electrical neural stimulation; including contact-free stimulation, high spatial selectivity, and lack of a stimulation artifact. The underlying biophysical mechanism responsible for transient optical nerve stimulation appears to be a small, absorption driven thermal gradient sustained at the axonal layer of nerve. Results explicitly prove that low frequency optical stimulation can reliably stimulate without resulting in tissue thermal damage. Based on the positive results from animal studies, these optimal laser parameters were utilized to move this research into the clinic with a combined safety and efficacy study in human subjects undergoing selective dorsal rhizotomy. The clinical Holmium:YAG laser was used to effectively stimulate human dorsal spinal roots and elicit functional muscle responses recorded during surgery without evidence of nerve damage. Overall these results predict that this technology can be a valuable clinical tool in various neurosurgical applications.

Wells, Jonathon D.

377

Feasibility of in vivo cardiac HIFU ablation  

NASA Astrophysics Data System (ADS)

The potential for cardiac applications of HIFU remains unexamined. In order to create reproducible lesions in a beating heart, it is necessary to maintain focusing at a certain position within moving myocardial tissue. One technique is to use multiple short HIFU exposures (0.2 s) and synchronize them with an EKG signal and respiration. The left ventricular free wall (LVFW) of calf hearts were cut into 4-cm cubes, degassed in phosphate buffer saline (PBS), and heated to 37°C. An 80-mm-diam spherical-cap transducer with a focus of 90 mm was operated at a frequency of 4.67 MHz and a nominal focal point intensity of 26.9 kW/cm2. The transducer was coupled to the LVFW using degassed PBS. First, the effect of pericardial fat, focal depth, and temperature on lesion size was individually evaluated. Then the 0.2-s HIFU exposure was applied 10 to 30 times at 4-s intervals. The same HIFU transducer was applied to an open-chest canine LVFW with the same triggering protocol. Dimensions of all lesions were measured by visual examination of the fresh, unstained tissue. A histopathological examination of the lesion was also performed. The in vivo lesions were created in similar size to those in vitro.

Fujikura, Kana; Otsuka, Ryo; Homma, Shunichi; Muratore, Robert; Ketterling, Jeffrey A.; Lizzi, Frederic L.

2001-05-01

378

In Vivo EPR For Dosimetry  

PubMed Central

As a result of terrorism, accident, or war, populations potentially can be exposed to doses of ionizing radiation that could cause direct clinical effects within days or weeks. There is a critical need to determine the magnitude of the exposure to individuals so that those with significant risk have appropriate procedures initiated immediately, while those without a significant probability of acute effects can be reassured and removed from the need for further consideration in the medical/emergency system. In many of the plausible scenarios there is an urgent need to make the determination very soon after the event and while the subject is still present. In vivo EPR measurements of radiation-induced changes in the enamel of teeth is a method, perhaps the only such method, which can differentiate among doses sufficiently for classifying individuals into categories for treatment with sufficient accuracy to facilitate decisions on medical treatment. In its current state, the in vivo EPR dosimeter can provide estimates of absorbed dose with an error approximately ± 50 cGy over the range of interest for acute biological effects of radiation, assuming repeated measurements of the tooth in the mouth of the subject. The time required for acquisition, the lower limit, and the precision are expected to improve, with improvements in the resonator and the algorithm for acquiring and calculating the dose. The magnet system that is currently used, while potentially deployable, is somewhat large and heavy, requiring that it be mounted on a small truck or trailer. Several smaller magnets, including an intraoral magnet are under development, which would extend the ease of use of this technique.

Swartz, Harold M.; Burke, Greg; Coey, M.; Demidenko, Eugene; Dong, Ruhong; Grinberg, Oleg; Hilton, James; Iwasaki, Akinori; Lesniewski, Piotr; Kmiec, Maciej; Lo, Kai-Ming; Nicolalde, R. Javier; Ruuge, Andres; Sakata, Yasuko; Sucheta, Artur; Walczak, Tadeusz; Williams, Benjamin B.; Mitchell, Chad; Romanyukha, Alex; Schauer, David A.

2007-01-01

379

Reprogramming in vivo produces teratomas and iPS cells with totipotency features.  

PubMed

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

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

2013-09-11

380

In vivo and ex vivo Diffusion Tensor Imaging of Cuprizone Induced Demyelination in the Mouse Corpus Callosum  

PubMed Central

Diffusion tensor imaging (DTI) has been widely used in studying rodent models of white matter diseases. In this study, we examined the differences between in vivo and ex vivo fractional anisotropy (FA) and diffusivity measurements in the mouse cuprizone model. In the control mouse corpus callosum (CC), ex vivo diffusivities were significantly lower than in vivo measurements, but ex vivo FA values were not significantly different from in vivo FA values. With cuprizone induced demyelination and accompanying pathology in the CC, changes in in vivo and ex vivo FA and diffusivity measurements were not always in agreement. Our results suggest that ex vivo ?? was a more reliable indicator of white matter demyelination than in vivo ?? and in vivo ?? was a more reliable indicator of axonal injury than ex vivo ?? in this model. When comparing in vivo and ex vivo DTI results of axon and myelin pathology in the rodent models, potential changes in tissue microstructures associated with perfusion fixation should be considered.

Zhang, Jiangyang; Jones, Melina V.; McMahon, Michael T.; Mori, Susumu; Calabresi, Peter A.

2011-01-01

381

Rotator cuff repair with bioabsorbable screws: An in vivo and ex vivo investigation  

Microsoft Academic Search

Purpose: The purpose of this study was to evaluate in vivo the clinical outcomes of rotator cuff repairs with bioabsorbable screws compared with metal suture anchors, and to compare the ex vivo initial load to failure of rotator cuff repairs using 3 different bioabsorbable screws, suture anchors, and transosseous sutures. Type of Study: In vivo clinical outcomes investigation, and ex

Craig A. Cummins; Sabrina Strickland; Richard C. Appleyard; Zoltan L. Szomor; Jeanette Marshall; George A. C. Murrell

2003-01-01

382

A Comparison of General Case In Vivo and General Case Simulation Plus In Vivo Training.  

ERIC Educational Resources Information Center

The study examined the relative effectiveness and efficiency of general case in vivo and general case simulation plus in vivo training in teaching six students with moderate and severe disabilities to purchase items in fast-food restaurants. Although both strategies led to reliable performance in nontrained settings, the in vivo instruction…

McDonnell, John J.; Ferguson, Brad

1988-01-01

383

A Comparison of General Case In Vivo and General Case Simulation Plus In Vivo Training.  

ERIC Educational Resources Information Center

|The study examined the relative effectiveness and efficiency of general case in vivo and general case simulation plus in vivo training in teaching six students with moderate and severe disabilities to purchase items in fast-food restaurants. Although both strategies led to reliable performance in nontrained settings, the in vivo instruction…

McDonnell, John J.; Ferguson, Brad

1988-01-01

384

Optimal ultraviolet wavelength for in vivo photoacoustic imaging of cell nuclei  

NASA Astrophysics Data System (ADS)

In order to image noninvasively cell nuclei in vivo without staining, we have developed ultraviolet photoacoustic microscopy (UV-PAM), in which ultraviolet light excites nucleic acids in cell nuclei to produce photoacoustic waves. Equipped with a tunable laser system, the UV-PAM was applied to in vivo imaging of cell nuclei in small animals. We found that 250 nm was the optimal wavelength for in vivo photoacoustic imaging of cell nuclei. The optimal wavelength enables UV-PAM to image cell nuclei using as little as 2 nJ laser pulse energy. Besides the optimal wavelength, application of a wavelength between 245 and 275 nm can produce in vivo images of cell nuclei with specific, positive, and high optical contrast.

Yao, Da-Kang; Chen, Ruimin; Maslov, Konstantin; Zhou, Qifa; Wang, Lihong V.

2012-05-01

385

Functionalized Magnetic Nanoparticles as an In Vivo Delivery System  

NASA Astrophysics Data System (ADS)

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.

Taira, Shu; Moritake, Shinji; Hatanaka, Takahiro; Ichiyanagi, Yuko; Setou, Mitsutoshi

386

Fructation in vivo: detrimental and protective effects of fructose.  

PubMed

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

Semchyshyn, H M

2013-07-24

387

Fructation In Vivo: Detrimental and Protective Effects of Fructose  

PubMed Central

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.

Semchyshyn, H. M.

2013-01-01

388

The measurement of oxygen in vivo using EPR techniques  

NASA Astrophysics Data System (ADS)

The measurement of in vivo using EPR has some features which have already led to very useful applications and this approach is likely to have increasingly wide and effective use. It is based on the effect of oxygen on EPR spectra which provides a sensitive and accurate means to measure quantitatively. The development of oxygen-sensitive paramagnetic materials which are very stable, combined with instrumental developments, has been crucial to the in vivo applications of this technique. The physical basis and biological applications of in vivo EPR oximetry are reviewed, with particular emphasis on the use of EPR spectroscopy at 1 GHz using particulate paramagnetic materials for the repetitive and non-invasive measurement of in tissues. In vivo EPR has already produced some very useful results which have contributed significantly to solving important biological problems. The characteristics of EPR oximetry which appear to be especially useful are often complementary to existing techniques for measuring oxygen in tissues. These characteristics include the capability of making repeated measurements from the same site, high sensitivity to low levels of oxygen, and non-invasive options. The existing techniques are especially useful for studies in small animals, where the depth of measurements is not an overriding issue. In larger animals and potentially in human subjects, non-invasive techniques seem to be immediately applicable to study phenomena very near the surface (within 10 mm) while invasive techniques have some very promising uses. The clinical uses of EPR oximetry which seem especially promising and likely to be undertaken in the near future are long-term monitoring of the status and response to treatment of peripheral vascular disease and optimizing cancer therapy by enabling it to be modified on the basis of the measured in the tumour.

Swartz, Harold M.; Clarkson, Robert B.

1998-07-01

389

Intervention against the Maillard reaction in vivo  

Microsoft Academic Search

The field of Maillard\\/glycation reactions in vivo has grown enormously during the past 20 years, going from 25 to 500 publications per year. It is now well recognized that many of the “advanced” products form oxidatively or anaerobically and can have deleterious effects on macromolecular and biological function. The feasibility of developing pharmacological agents with beneficial in vivo properties, based

Vincent M. Monnier

2003-01-01

390

In Vitro\\/In Vivo Correlations  

Microsoft Academic Search

In vitro\\/in vivo correlations can and should be developed in many cases. This paper provides one approach to developing in vitro\\/in vivo correlations, along with examples, based upon the position of the United States Pharmacopeia (USP) Subcommittee on Bioavailability, Bioequivalence, and Dissolution (DBA). Although correlations should not be required, the Food and Drug Administration (FDA) must take a definitive position

Lewis J. Leeson

1995-01-01

391

In vivo correlates of altered blood rheology  

Microsoft Academic Search

It is has been known for more than 80 years that compared to in vitro determinations, blood behaves as a less viscous fluid under in vivo flow conditions. The experiments of Whittaker and Winton were among the first dealing with the in vivo effects of altered blood rheology, and experimental studies during the second half of 20th century have provided

Oguz K. Baskurt

2008-01-01

392

In vivo evaluations of retinal prostheses  

Microsoft Academic Search

In this article, the in vivo evaluation of retinal prostheses were achieved either in animals with normal vision or in blind persons. To assess the stimulating electrodes in vivo on an animal with photoreceptor degeneration, we have implanted a P23H rhodopsin rats. The present study describes the implant design and our surgical strategy. The experiments indicate clearly that the implant

J. Salzmann; P. Linderholm; M. Paques; M. Simonutti; J. A. Chiappore; A. B. Safran; P. Renaud; J. Sahel; S. Picaud

2004-01-01

393

In Vivo Models of Biofilm Infection  

Microsoft Academic Search

\\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.

Kendra P. Rumbaugh; Nancy L. Carty

394

Elastographic image quality vs. tissue motion in vivo.  

PubMed

Elastography is a noninvasive method of imaging tissue elasticity using standard ultrasound equipment. In conventional elastography, axial strain elastograms are generated by cross-correlating pre- and postcompression digitized radio frequency (RF) echo frames acquired from the tissue before and after a small uniaxial compression, respectively. The time elapsed between the pre- and the postcompression frames is referred to as the interframe interval. For in vivo elastography, the interframe interval is critical because uncontrolled physiologic motion such as heartbeat, muscle motion, respiration and blood flow introduce interframe decorrelation that reduces the quality of elastograms. To obtain a measure of this decorrelation, in vivo experimental data (from human livers and thyroids) at various interframe intervals were obtained from 20 healthy subjects. To further examine the effect of the different interframe intervals on the elastographic image quality, the experimental data were also used in combination with elastographic simulation data. The deterioration of elastographic image quality was objectively evaluated by computing the area under the strain filter (SF) at a given resolution. The experimental results of this study demonstrate a statistical exponential behavior of the temporal decay of the echo signal cross-correlation amplitudes from the in vivo tissues due to uncontrollable motion. The results also indicate that the dynamic range and height of the SF are reduced at increased interframe intervals, suggesting that good objective image quality may be achieved provided only that a high frame rate is maintained in elastographic applications. PMID:16785007

Chandrasekhar, R; Ophir, J; Krouskop, T; Ophir, K

2006-06-01

395

2D luminescence imaging of pH in vivo.  

PubMed

Luminescence imaging of biological parameters is an emerging field in biomedical sciences. Tools to study 2D pH distribution are needed to gain new insights into complex disease processes, such as wound healing and tumor metabolism. In recent years, luminescence-based methods for pH measurement have been developed. However, for in vivo applications, especially for studies on humans, biocompatibility and reliability under varying conditions have to be ensured. Here, we present a referenced luminescent sensor for 2D high-resolution imaging of pH in vivo. The ratiometric sensing scheme is based on time-domain luminescence imaging of FITC and ruthenium(II)tris-(4,7-diphenyl-1,10-phenanthroline). To create a biocompatible 2D sensor, these dyes were bound to or incorporated into microparticles (aminocellulose and polyacrylonitrile), and particles were immobilized in polyurethane hydrogel on transparent foils. We show sensor precision and validity by conducting in vitro and in vivo experiments, and we show the versatility in imaging pH during physiological and chronic cutaneous wound healing in humans. Implementation of this technique may open vistas in wound healing, tumor biology, and other biomedical fields. PMID:21262842

Schreml, Stephan; Meier, Robert J; Wolfbeis, Otto S; Landthaler, Michael; Szeimies, Rolf-Markus; Babilas, Philipp

2011-01-24

396

[In vivo single molecular fluorescence imaging for analysis of pharmacokinetics].  

PubMed

Nano-materials are expected for research on molecular imaging of pharmacokinetics. We measured in vivo migration of CdSe nano-particles(Quantum Dots(QDs))conjugated with monoclonal anti-HER2 antibody(trastuzumab)in tumor vessel to breast cancer cells. We established a high resolution in vivo 3D microscopic system for a novel imaging method at single molecular level. The HER2 protein expressed in cancer cells and its dynamics were visualized by QDs in vivo at the spatial resolution of 30 nm. It suggests future utilization of the system in medical applications to improve the drug delivery system to target primary and metastatic tumors for made-to-order treatment. Future innovation in cancer imaging by nano-technology and novel measurement technology will provide great improvement, not only in the clinical field, but also in basic medical science. Advances in nano-biotechnology have great potential to improve prevention, diagnosis and treatment of human disease. PMID:18701837

Takeda, Motohiro; Gonda, Kohsuke; Higuchi, Hideo; Ohuchi, Noriaki

2008-08-01

397

In vivo gene transfer into muscle via electro-sonoporation.  

PubMed

Among the nonviral techniques for gene transfer in vivo, electroporation is simple, potent, inexpensive, and safe. To upregulate the expression levels of the transferred gene, we investigated the applicability of in vivo electro-sonoporation, which consists of a combination of electric pulse and ultrasound, for gene transfer using plasmid DNA encoding luciferase and mouse interleukin-12 (mIL-12). The quadriceps muscles of mice were injected with plasmid DNA, then sonoporated for 5 min, and electroporated by a pair of electrode plates at the middle of the duration of sonoporation. Three days later, mice that had undergone electro-sonoporation demonstrated twofold higher luciferase activity and low tissue damage in quadriceps muscle compared to mice having undergone electroporation alone. Serum mIL-12 levels in mice that had undergone electro-sonoporation (peaking at 25.5 ng/ml) were twofold higher after gene transfer than were those in mice having undergone electroporation alone (peaking at 14.3 ng/ml), and maintained high serum level of 13.9 ng/ml at 28 days after gene transfer. The efficacy of gene transfer via electro-sonoporation was superior when the plasmid DNA solution was 0.85% NaCl compared to albumin microbubble echo-contrast material. These results demonstrated that gene transfer into muscle via electro-sonoporation could provide a new potent nonviral technique for gene transfer in vivo. PMID:12490002

Yamashita, Yo-ichi; Shimada, Mitsuo; Tachibana, Katsuro; Harimoto, Norifumi; Tsujita, Eiji; Shirabe, Ken; Miyazaki, Jun-ichi; Sugimachi, Keizo

2002-11-20

398

In vivo infrared and Raman spectroscopy of human stratum corneum  

NASA Astrophysics Data System (ADS)

ATR-FTIR spectroscopy and Raman spectroscopy were employed to obtain information about the molecular composition and hydration of skin in vivo. Both techniques enable the in vivo acquisition of high quality spectra within 10-30s at a spectral resolution of 8cm-1. The penetration depth of ATR-FTIR is about 1.5 (Mu) m. Raman spectra could be obtained with a resolution of about 5 micrometers . ATR-FTIR spectra of hydrated stratum corneum were analyzed using a band fitting algorithm. By means of this algorithm the signal contributions of water relative to protein signal contributions could be determined. The results of Raman microspectroscopic experiments on frozen sections and isolated skin components were used for the interpretation of Raman spectra obtained in vivo. Information was obtained about lipid components present in the stratum corneum. These were shown to vary widely between individuals and between different locations on the body. The combination of these spectroscopic techniques may prove to be valuable for applications in dermatology and skin care.

Lucassen, Gerald W.; Caspers, Peter J.; Puppels, Gerwin J.

1998-04-01

399

In vivo brain microdialysis: advances in neuropsychopharmacology and drug discovery  

PubMed Central

Introduction Microdialysis is an important in vivo sampling technique, useful in the assay of extracellular tissue fluid. The technique has both pre-clinical and clinical applications but is most widely used in neuroscience. The in vivo microdialysis technique allows measurement of neurotransmitters such as acetycholine (ACh), the biogenic amines including dopamine (DA), norepinephrine (NE) and serotonin (5-HT), amino acids such as glutamate (Glu) and gamma aminobutyric acid (GABA), as well as the metabolites of the aforementioned neurotransmitters, and neuropeptides in neuronal extracellular fluid in discrete brain regions of laboratory animals such as rodents and non-human primates. Areas covered In this review we present a brief overview of the principles and procedures related to in vivo microdialysis and detail the use of this technique in the pre-clinical measurement of drugs designed to be used in the treatment of chemical addiction, neurodegenerative diseases such as Alzheimer’s disease (AD), Parkinson’s disease (PD) and as well as psychiatric disorders such as attention-deficit/hyperactivity disorder (ADHD) and schizophrenia. This review offers insight into the tremendous utility and versatility of this technique in pursuing neuropharmacological investigations as well its significant potential in rational drug discovery. Expert opinion In vivo microdialysis is an extremely versatile technique, routinely used in the neuropharmacological investigation of drugs used for the treatment of neurological disorders. This technique has been a boon in the elucidation of the neurochemical profile and mechanism of action of several classes of drugs especially their effects on neurotransmitter systems. The exploitation and development of this technique for drug discovery in the near future will enable investigational new drug candidates to be rapidly moved into the clinical trial stages and to market thus providing new successful therapies for neurological diseases that are currently in demand.

Darvesh, Altaf S.; Carroll, Richard T.; Geldenhuys, Werner J.; Gudelsky, Gary A.; Klein, Jochen; Meshul, Charles K.; Van der Schyf, Cornelis J.

2010-01-01

400

Comparative in vivo biocompatibility study of single- and multi-wall carbon nanotubes  

Microsoft Academic Search

Carbon nanotubes are expected to be of use in both genetic engineering and biomaterials engineering. In each of these potential areas of application, nanoparticles are introduced into a living organism either in the form of active biomolecule carriers or as a result of the degradation process of an implant. In the present study we focus on the in vivo behavior

Aneta Fraczek; Elzbieta Menaszek; Czeslawa Paluszkiewicz; Marta Blazewicz

2008-01-01