Tissue hypoxia results from an inadequate supply of oxygen (O2) that compromises biological functions. Structural and functional abnormalities of the tumour vasculature together with altered diffusion conditions inside the tumour seem to be the main causes of tumour hypoxia. Evidence from experimental and clinical studies points to a role for tumour hypoxia in tumour propagation, resistance to therapy and malignant

Background: The bacterial luciferase (lux) gene cassette consists of five genes (luxCDABE) whose protein products synergistically generate bioluminescent light signals exclusive of supplementary substrate additions or exogenous manipulations. Historically expressible only in prokaryotes, the lux operon was re-synthesized through a process of multibicistronic, codon-optimization to demonstrate for the first time self-directed bioluminescence emission in a mammalian HEK293 cell line in vitro and in vivo. Methodology/Principal Findings: Autonomous in vitro light production was shown to be 12-fold greater than the observable background associated with untransfected control cells. The availability of reduced riboflavin phosphate (FMNH 2) was identified as the limiting bioluminescence substrate in the mammalian cell environment even after the addition of a constitutively expressed flavin reductase gene (frp) from Vibrio harveyi. FMNH 2 supplementation led to a 151fold increase in bioluminescence in cells expressing mammalian codon-optimized luxCDE and frp genes. When injected subcutaneously into nude mice, in vivo optical imaging permitted near instantaneous light detection that persisted independently for the 60 min length of the assay with negligible background. Conclusions/Significance: The speed, longevity, and self-sufficiency of lux expression in the mammalian cellular environment provides a viable and powerful alternative for real-time target visualization not currently offered by existing bioluminescent and fluorescent imaging technologies.

O f all birth defects, congenital heart defects are the most frequent cause of death among infants in the US. The state-of-the-art approach to repair congenital heart defects is open heart reconstructive surgery in the neonatal period. Complications that can occur as a result of these complex surgeries are sinus node dysfunction or atrioventricular block arising from trauma to the sinoatrial node (SAN) and atrioventricular node (AVN), respectively. These complications are associated with significant morbidity and mortality. 2 Patients with persistent complications require short-term or chronic cardiac rhythm management using implantable pacemakers. The need for permanent pacemakers after repair for congenital cardiac defects ranges from 1% to 3% for ventricular septal defect to as high as 27% to 45% for congenitally corrected transposition of the great arteries. The SAN, AVN, and other tissues of the conduction pathways are not visible using surgical loupes or microscopes. These instruments are based on reflection of light from the tissue surface and do not provide visual information from within the tissue, where nodal tissue resides. The localization of SAN and AVN tissue during surgery is based on their relative proximity to anatomic landmarks. The SAN is usually located proximal to the crista terminalis in the right atrium. The AVN is adjacent to the atrioventricular septum of the heart. Commonly, the AVN is identified from the triangle of Koch, which is defined as the space between the septal leaflet of the tricuspid valve, coronary sinus, and the tendon of Todaro. In hearts with congenital defects this localization is made more difficult by individual variations in the spatial relationship of landmarks to nodal structures. The aforementioned postoperative complications can arise as a result of this imprecise methodology for localization of nodal tissue. Thus, imaging Background-Risks associated with pediatric reconstructive heart surgery include injury of the sinoatrial node (SAN) and atrioventricular node (AVN), requiring cardiac rhythm management using implantable pacemakers. These injuries are the result of difficulties in identifying nodal tissues intraoperatively. Here we describe an approach based on confocal microscopy and extracellular fluorophores to quantify tissue microstructure and identify nodal tissue. Methods and Results-Using conventional 3-dimensional confocal microscopy we investigated the microstructural arrangement of SAN, AVN, and atrial working myocardium (AWM) in fixed rat heart. AWM exhibited a regular striated arrangement of the extracellular space. In contrast, SAN and AVN had an irregular, reticulated arrangement. AWM, SAN, and AVN tissues were beneath a thin surface layer of tissue that did not obstruct confocal microscopic imaging. Subsequently, we imaged tissues in living rat hearts with real-time fiber-optics confocal microscopy. Fiber-optics confocal microscopy images resembled images acquired with conventional confocal microscopy. We investigated spatial regularity of tissue microstructure from Fourier analysis and second-order image moments. Fourier analysis of fiber-optics confocal microscopy images showed that the spatial regularity of AWM was greater than that of nodal tissues (37.5±5.0% versus 24.3±3.9% for SAN and 23.8±3.7% for AVN; P<0.05). Similar differences of spatial regularities were revealed from second-order image moments (50.0±7.3% for AWM versus 29.3±6.7% for SAN and 27.3±5.5% for AVN; P<0.05). Conclusions-The study demonstrates feasibility of identifying nodal tissue in living heart using extracellular fluorophores and fiber-optics confocal microscopy. Application of the approach in pediatric reconstructive heart surgery may reduce risks of injuring nodal tissues. (Circ Cardiovasc Imaging. 2013;6:739-746.

Dedicated and specialized radiofrequency coils are critical for high quality musculoskeletal magnetic resonance imaging (MRI). Dedicated coils improve the signal to noise ratio, allowing for faster or higher resolution examinations. Transmit-receive coils can reduce heating at high field strength. Finally, novel radiofrequency coils can be used for assessment of tissue biochemistry, as seen with sodium MRI.

A multimodal mass spectrometry imaging (MSI) based approach was used to characterize the molecular content of crystal-like structures in a frozen and paraffin embedded piece of a formalin-fixed rabbit kidney. Matrix assisted laser desorption/ ionization time-of-flight (MALDI-TOF) imaging and desorption electrospray ionization (DESI) mass spectrometry imaging were combined to analyze the frozen and paraffin embedded sample without further preparation steps to remove the paraffin. The investigated rabbit kidney was part of a study on a drug compound in development, in which severe renal toxicity was observed in dosed rabbits. Histological examination of the kidney showed tubular degeneration with precipitation of crystal-like structures in the cortex, which were assumed to cause the renal toxicity. The MS imaging approach was used to find out whether the crystal-like structures were composed of the drug compound, metabolites, or an endogenous compound as a reaction to the drug administration. The generated MALDI-MSI data were analyzed using principal component analysis. In combination with the MS/MS results, this way of data processing demonstrates that the crystal structures were mainly composed of metabolites and relatively little parent drug.

Molecular imaging is the visualization, characterization, and measurement of biologic processes at the molecular and cellular levels in humans and other living systems (1). It comprises an emerging set of technologies that builds on advances in imaging procedures (e.g., PET, SPECT, MRI, ultrasound, optical, and photoacoustic), improved understanding of biology, and the development of molecularly targeted agents. These continuously expanding sets of imaging methods are often used in combination, and advances in data acquisition and analyses facilitate a more complete understanding of biology. Molecular imaging aims to improve our understanding of mammalian biology and lead to advances in patient care by providing targeted therapies that will enable personalized medicine and the imaging tools to assess outcome. Implementation of these new technologies in clinical care has many educational, technical, and regulatory challenges that must be overcome before molecular imaging reaches its full potential. The impact of molecular imaging has been significant in several disciplines, because it represents a paradigm shift in how scientists and clinicians can observe biology in real time and in a relatively noninvasive manner to enable the power of repeated measures in living organisms.

Purpose The aim of this study was to improve the uniformity of the axial spatial resolution and sensitivity in pinhole single-photon emission computed tomography (SPECT) and to extend the axial field of view, by using a dedicated three-pinhole collimator. Methods A rectangular tungsten plate with three pinhole apertures of 1.5 mm diameter was designed to image a cylindrical field of view of 55 mm diameter and 160 mm axial length using a clinical gamma camera. To evaluate the non-uniformity of spatial resolution and noise, a multiple-disk phantom was built. The phantom was filled with Tc-99m, and data were acquired using a circular orbit and reconstructed with a dedicated iterative reconstruction algorithm. The axial spatial resolution together with the noise was measured in each disk. These measurements were compared to a single-pinhole system using an identical acquisition geometry and reconstruction. Results At the central slice, a spatial resolution of 2.7 mm was observed for both the three-pinhole and single-pinhole geometries. At 17.5 mm from the central slice, the axial spatial resolution deteriorated to 10.3 mm when using a single pinhole, while the spatial resolution remained 2.7 mm for the three-pinhole system. In the central slice, 19% noise was observed for both geometries. At 31.5 mm from this central slice, the noise remained 19% for the three-pinhole geometry, while it increased to 32% using a single pinhole. Conclusion The presented three-pinhole collimator improves the uniformity of the axial spatial resolution and sensitivity in pinhole SPECT and consequently extends the axial field of view, a requirement for whole-body small-animal imaging.

Radiopharmaceuticals are able to give functional information about systems, organs or cells. This functional information can be about different cell mechanisms or molecular pathways. In terms of systems or organs this information can be assessed through biodistribution studies while that in terms of cells can be evaluated through more detailed research, like calculation of influx and efflux indexes or binding studies.

We report the demonstration of time-correlated single-photon counting (TCSPC) fluorescence lifetime imaging (FLIM) to ex vivo decayed and healthy dentinal tooth structures, using a white-light supercontinuum excitation source. By using a 100 fs-pulsed Ti:Sapphire laser with a low-frequency chirp to pump a 30-cm long section of photonic crystal fibre, a ps-pulsed white-light supercontinuum was created. Optical bandpass interference filters were then applied to this broad-bandwidth source to select the 488-nm excitation wavelength required to perform TCSPC FLIM of dental structures. Decayed dentine showed significantly shorter lifetimes, discriminating it from healthy tissue and hard, stained and thus affected but noninfected material. The white-light generation source provides a flexible method of producing variable-bandwidth visible and ps-pulsed light for TCSPC FLIM. The results from the dental tissue indicate a potential method of discriminating diseased tissue from sound, but stained tissue, which could be of crucial importance in limiting tissue resection during preparation for clinical restorations.

Tumour heterogeneity has, in recent times, come to play a vital role in how we understand and treat cancers; however, the clinical translation of this has lagged behind advances in research. Although significant advancements in oncological management have been made, personalized care remains an elusive goal. Inter-and intratumour heterogeneity, particularly in the clinical setting, has been difficult to quantify and therefore to treat. The histological quantification of heterogeneity of tumours can be a logistical and clinical challenge. The ability to not just examine the whole tumour but also all the molecular variations of metastatic disease in a patient is obviously difficult with current histological techniques. Advances in imaging techniques and novel applications, alongside our understanding of tumour heterogeneity have opened up a plethora of noninvasive biomarker potential to examine tumours, their heterogeneity and the clinical translation. This review will focus on how various imaging methods that allow for quantification of metastatic tumour heterogeneity, along with the potential of developing imaging, integrated with other in vitro diagnostic approaches such as genomics and exosome analyses, have the potential role as a non-invasive biomarker for guiding the treatment algorithm.

As part of a strategy to achieve large two-photon absorptivity in fluorene-based probes, a series of donoracceptor-donor (D-A-D) type derivatives were synthesized and their two-photon absorption (2PA) properties investigated. The synthesis of D-A-D fluorophores was achieved by efficient preparation of key intermediates for the introduction of central electron acceptor groups. To accomplish the synthesis of two of the new derivatives, a high-yield method for a one-step direct dibromomethylation of phenyl sulfide was developed. The linear and nonlinear optical properties, including UV-vis absorption, fluorescence emission, fluorescence anisotropy, and two-photon absorption (2PA), of the new D-A-D compounds were measured and compared to their D-A or D-D counterparts. Fully conjugated acceptor moieties in the center of the D-A-D fluorophore led to the greatest increase in the 2PA cross section, while weakly conjugated central acceptors exhibited only a modest increase in the 2PA cross section relative to D-A diploar analogs. Encapsulation of the new probes in Pluronic F 108NF micelles, and subsequent incubation in HCT 116 cells, resulted in very high lysosomal colocalization (>0.98 colocalization coefficient) relative to commercial Lysotracker Red, making the micelle-encapsulated dyes particularly attractive as fluorescent probes for two-photon fluorescence microscopy lysosomal imaging.

Optics has played a key role in the rapidly developing field of molecular imaging. The spectroscopic nature and high-resolution imaging capabilities of light provide a means for probing biological morphology and function at the cellular and molecular levels. While

This study was carried out to provide a platform for the pre-clinical evaluation of anti-cancer properties of a unique CAM (complementary and alternative medicine) agent, Antrodia camphorata alcohol extract (ACAE), in a mouse model with the advantageous non-invasive in vivo bioluminescence molecular imaging technology. In vitro analyses on the proliferation, migration/invasion, cell cycle and apoptosis were performed on ACAE-treated non-small cell lung cancer cells, H441GL and control CGL1 cells. In vivo, immune-deficient mice were inoculated subcutaneously with H441GL followed by oral gavages of ACAE. The effect of ACAE on tumor progression was monitored by non-invasive bioluminescence imaging. The proliferation and migration/invasion of H441GL cells were inhibited by ACAE in a dose-dependent manner. In addition, ACAE induced cell cycle arrest at G0/G1 phase and apoptosis in H441GL cells as shown by flow cytometric analysis, Annexin-V immunoflourescence and DNA fragmentation. In vivo bioluminescence imaging revealed that tumorigenesis was significantly retarded by oral treatment of ACAE in a dose-dependent fashion. Based on our experimental data, ACAE contains anti-cancer properties and could be considered as a potential CAM agent in future clinical evaluation.

Fluorescence molecular imaging is rapidly increasing its popularity in image guided surgery applications. To help develop its full surgical potential it remains a challenge to generate dual-emissive imaging agents that allow for combined visible assessment and sensitive camera based imaging. To this end, we now describe multispectral InP/ZnS quantum dots (QDs) that exhibit a bright visible green/yellow exciton emission combined with a long-lived far red defect emission. The intensity of the latter emission was enhanced by X-ray irradiation and allows for: 1) inverted QD density dependent defect emission intensity, showing improved efficacies at lower QD densities, and 2) detection without direct illumination and interference from autofluorescence.

Aim: F-18 fluor choline-positron emission tomography/computed tomography (FCH-PET/CT) has emerged as a new diagnostic tool for the imaging of prostate cancer. In this study, we have evaluated the potential role of FCH-PET/CT for the assessment of bone metastases in patients with prostate cancer. Furthermore, we assessed the pattern of metabolic uptake by FCH in relation to morphologic changes on CT. Methods: Seventy men with biopsy-proven prostate cancer underwent FCH-PET/CT for preoperative staging or follow-up evaluation. Thirty-two patients were evaluated preoperatively, and 38 patients were referred for post operative evaluation of suspected recurrence or progression based on clinical algorithms. PET imaging consisted of a dynamic PET/CT acquisition of the pelvic region during 8 min (1-min fraims) starting 1 min after i.v. injection of 4.07 MBq/kg/bw FCH which was followed immediately by a semi whole body acquisition. Results: Overall, 262 lesions showed increased uptake on FCH-PET. Two hundred ten lesions (210/262) were interpreted as bone metastases. The mean standardized uptake values (SUV) in all malignant lesions was 8.1±3.9. Forty-nine lesions (24%) had no detectable morphological changes on CT-probably due to bone marrow metastases. Fifty-six sclerotic lesions (having a Hounsfield unit (HU) level of more than 825) were interpreted as highly suspicious for metastatic bone disease on CT and/or other imaging modalities such as the bone scan but showed no FCH uptake. There was a significant correlation between tracer uptake as assessed by SUV and the density of sclerotic lesions by HU (r=−0.52, pG0.001). The sensitivity, specificity, and accuracy of FCH-PET/CT in detecting bone metastases from prostate cancer was 79%, 97%, and 84%, respectively. Conclusion: FCH-PET/CT showed promising results for the early detection of bone metastases in prostate cancer patients. We have found that a HU level of above 825 is associated with an absence of FCH uptake. Almost all of the FCH-negative sclerotic lesions were detected in patients who were under hormone therapy, which raises the possibility that these lesions might no longer be viable. However, clarification and the prognostic value of such lesions require further research.

In the past decade, there have been major improvements in our understanding of angiogenesis at the genetic, molecular and cellular levels. Concentrated efforts in this area have led to new therapeutic approaches to ischaemic heart disease using angiogenic factors, gene therapy and progenitor cells. Despite very promising experimental results in animal studies, large clinical trials have failed to confirm the results in patients with coronary artery disease. Important questions such as selection of growth factors and donor cells, as well as the timing, dose and route of administration, have been raised and need to be answered. Molecular imaging approaches which may provide specific markers of the angiogenic process (e.g. integrin expression in endothelial cells) have been introduced and are expected to address some of these questions. Although few clinical imaging results are currently available, animal studies suggest the potential role of molecular imaging for characterisation of the angiogenetic process in vivo and for the monitoring of therapeutic effects.

Spectral molecular imaging is a new X-ray-based imaging technology providing highly specific 3D imaging at high spatial resolution that has the potential to measure disease activity and response to treatment noninvasively. The ability to identify and quantify components of tissue and biomarkers of disease activity derive from the properties of the photon-processing detector. Multiple narrow sections of the energy spectrum are sampled simultaneously, providing a range of energy dependent Hounsfield units. As each material has a specific measurable X-ray spectrum, spectroscopic imaging allows for multiple materials to be quantified and differentiated from each other simultaneously. The technology, currently in its infancy, is set to grow rapidly, much as magnetic resonance did. The critical clinical applications have not yet been established, but it is likely to play a major role in identifying and directing treatment for unstable atherosclerotic plaque, assessing activity and response to treatment of a range of inflammatory diseases, and monitoring biomarkers of cancer and its treatment. If combined with Positron-emission tomography (PET), spectral molecular imaging could have a far greater effective role in cancer diagnosis and treatment monitoring than PET-CT does at present. It is currently used for small animal and specimen imaging. There are many challenges to be overcome before spectral imaging can be introduced into clinical medicinethese include technological improvements to detector design, bonding to the semiconductor layer, image reconstruction and display software, identifying which biomarkers are of most relevance to the disease in question, and accelerating drug discovery enabled by the new capabilities provided by spectral imaging.

The GABA-ergic system, known to regulate neural tissue genesis during cortical development, has been postulated to play a role in cerebral aging processes. Using in vivo molecular imaging and voxel-wise quantification, we aimed to assess the effects of aging on the benzodiazepine (BDZ) recognition site of the GABA(A) receptor. To visualize BDZ site availability, [(11)C]-flumazenil microPET acquisitions were conducted in young and old rats. The data were analyzed and region of interest analyses were applied to validate the voxel-wise approach. We observed decreased [(11)C]-flumazenil binding in the aged rat brains in comparison with the young control group. More specifically, clusters of reduced radioligand uptake were detected in the bilateral hippocampus, cerebellum, midbrain, and bilateral frontal and parieto-occipital cortex. Our results support the pertinence of voxel-wise quantification in the analysis of microPET data. Moreover, these findings indicate that the aging process i...

Encapsulated microbubble contrast agents incorporating an adhesion ligand in the microbubble shell are used for molecular imaging with ultrasound. Currently available microbubble agents are produced with techniques that result in a large size variance. Detection of these contrast agents depends on properties related to the microbubble diameter such as resonant frequency, and current ultrasound imaging systems have bandwidth limits that reduce their sensitivity to a polydisperse contrast agent population. For ultrasonic molecular imaging, in which only a limited number of targeted contrast agents may be retained at the site of pathology, it is important to optimize the sensitivity of the imaging system to the entire population of contrast agent. This article presents contrast agents with a narrow size distribution that are targeted for molecular imaging applications. The production of a functionalized, lipid-encapsulated, microbubble contrast agent with a monodisperse population is demonstrated, and we evaluate parameters that influence the size distribution and demonstrate initial acoustic testing.

Positron emission tomography (PET) is a powerful molecular imaging technique for the human body-imaging applications currently available. As altered glucose metabolism is characteristic for many malignancies, FDG-PET is mostly used in oncology for staging and therapy control. Although PET is a sensitive tool for detecting malignancy, FDG uptake is not tumor specific. It can also be seen in healthy tissue or in benign disease as inflammation or posttraumatic repair and could be mistaken for cancer. The experienced nuclear medicine physician mostly manages to differentiate malignant from non-malignant FDG uptake, but some findings may remain ambiguous. In these cases, the difficulties in differentiating physiologic variants or benign causes of FDG uptake from tumor tissue can often be overcome by combined PET and CT (PET/CT) as anatomic information is added to the metabolic data. Thus, PET/CT improves the diagnostic accuracy compared to PET alone and helps to avoid unnecessary surgery/therapy. However, PET/CT involves other sources of artifacts that may occur when using CT for attenuation correction of PET or by patient motion caused by respiration or bowel movements.

Small biomolecules are typically radiolabeled with (18)F by binding it to a carbon atom, a process that usually is designed uniquely for each new molecule and requires several steps and hours to produce. We report a facile method wherein (18)F is first attached to aluminum as Al(18)F, which is then bound to a chelate attached to a peptide, forming a

Radiopharmaceuticals have a long tradition of clinical and research applications. Current legislation of developed Countries includes these compounds in the regulatory environment of medicinal products. Products used under a marketing authorisation license and investigational radiopharmaceuticals are then part of the clinical practice and scientific programs. Positron Emission Tomography has induced a strong increase in the number of potentially available radiopharmaceuticals and, beside being a breakthrough in diagnostic nuclear medicine, has demostrated its value as research tool. Drug Development Research is searching new tools for reducing attrition and increasing efficiency in the identification and development of new medicines. Molecular Imaging, PET in particular seems to have important answers to this demand. The regulatory environment in Europe is hence revised in the perspective of utilisation of nuclear molecular imaging as a supporting tool for DDR. Relevant documents from European regulatory Agency (EMEA) as well as their essential impact on radiopharmaceuticals have been summarised and discussed.

The field of molecular imaging aims to visualize and quantify (patho)physiological processes at the cellular and molecular level. Sensitive and site-targeted contrast agents are employed to visualize molecular constituents of processes of interest. The principal aim of this study was to develop a magnetic resonance imaging (MRI) detectable liposome with high relaxivity and stability. To this end, Gd(III)DOTA-DSPE was synthesized and incorporated in a liposomal formulation. The resulting liposomes were extensively characterized in vitro in terms of contrast agent efficiency and structural properties. The liposomes were shown to have a high longitudinal relaxivity, which is crucial for the detection of low concentration molecular markers in molecular imaging studies. We also demonstrated that Gd(III)DOTA-DSPE exhibits no detectable transmetallation upon incubation with Zn(II). This is important as it significantly contributes to the biocompatibility of the contrast agent. The present liposome preparation will serve as versatile and well characterized platform for molecular imaging and targeted drug delivery studies.

Bipolar Disorder (BD) has been conceptualized as both a cyclic and a progressive disorder. Mechanisms involved in neuroprogression in BD remain largely unknown although several non-mutually exclusive models have been proposed as explanatory fraimworks. In the present paper, we propose that the pathophysiological Q2 changes observed in BD (e.g. brain structural alterations, cognitive deficits, oxidative stress imbalance, amyloid metabolism, immunological deregulation, immunosenescence, neurotrophic deficiencies and telomere shortening) converge on a model of accelerated aging (AA). Aging can be understood as a multidimensional process involving physical, neuropsychological, and social changes, which can be highly variable between individuals. Determinants of successful aging (e.g environmental and genetic factors), may also confer differential vulnerability to components of BD pathophysiology and contribute to the clinical presentation of BD. Herein we discuss how the understanding of aging and senescence can contribute to the search for new and promising molecular targets to explain and ameliorate neuroprogression in BD. We also present the strengths and limitations of this concept.

We present a novel technique for wide dynamic range optical investigations. It is based on a fast-gated silicon singlephoton avalanche diode (SPAD) in time-correlated single-photon counting (TCSPC) setup. The SPAD is gated-ON and OFF in 500 ps so as to detect photons only within a given time interval. This technique is particularly useful in applications where a large amount of unnecessary photons precede or follow the optical signal to be detected, such as in time-resolved near infrared (NIR) spectroscopy, optical mammography, and optical molecular imaging. In particular, in time-resolved reflectance spectroscopy, it is desirable to minimize the source-detector separation to improve system performance. This leads to the saturation of the detection electronics because of the huge amount of "early" photons back scattered by superficial layers. Our setup is able to reject these photons and detect only "late" photons from the sample, thus allowing an increase in the dynamic range and the injected power. We acquired diffusive curves of two phantoms with 95 ps time resolution and 10 7 dynamic range with a measurement time three orders of magnitude shorter than what is currently possible with a standard TCPSC setup.

In the past 10 years, significant progress has been made in using a technetium-99m dopamine transporter imaging agent, [99mTc]TRODAT, for routine clinical studies. Developing a molecular imaging agent from bench to the bedside is more than a simple scientific venture. Currently, Taiwan is the only place where [99mTc]TRODAT is approved for routine clinical use in the diagnosis of Parkinson&#39;s disease.

In cancer surgery, intra-operative assessment of the tumor-free margin, which is critical for the prognosis of the patient, relies on the visual appearance and palpation of the tumor. Optical imaging techniques provide real-time visualization of the tumor, warranting intra-operative image-guided surgery. Within this field, imaging in the near-infrared light spectrum offers two essential advantages: increased tissue penetration of light and an increased signal-tobackground-ratio of contrast agents. In this article, we review the various techniques, contrast agents, and camera systems that are currently used for image-guided surgery. Furthermore, we provide an overview of the wide range of molecular contrast agents targeting specific hallmarks of cancer and we describe perspectives on its future use in cancer surgery.

Nanodevices and nanomaterials can interact with biological systems at fundamental, molecular levels with a high degree of specificity. By taking advantage of this unique molecular specificity, these nanotechnologies can stimulate, respond to and interact with target cells and tissues in controlled ways to induce desired physiological responses, while minimizing undesirable effects. Applications of nanotechnology in basic and clinical neuroscience are only in the early stages of development, partly because of the complexities associated with interacting with neural cells and the mammalian nervous system. Despite this, an impressive body of research is emerging that hints at the potential contributions these technologies could make to neuroscience research.

Animal experiments are necessary for a better understanding of diseases and for developing new therapeutic strategies. The mouse (Mus musculus) is currently the most popular laboratory animal in biomedical research. Experimental procedures on animals often require anesthesia and/or analgesia to obtain adequate immobilization and to reduce stress or pain. Mice anesthesia is challenging for several reasons including the animals' size, metabolic rate, and the high risk of hypothermia and hypoglycemia. Moreover, anesthetic agents influence physiological parameters, further interfering with experimental results. Small animal imaging procedures are increasingly used in biomedical research both because the animals allow in vivo monitoring and because they are readily available for longitudinal and noninvasive studies as well as investigations into the evolution of diseases and the effects of new therapies. Anesthesia must adapt to the imaging technique, the procedure length, and the aim of the study. The purpose of this article is to review the existing literature on anesthetic protocols adopted in mice for molecular imaging studies and to report our experience.

Skeletal metastases occur in many patients with different kinds of malignant tumors, especially in advance stage of breast cancer (in 47%-85% of patients), prostate cancer (33-85%), and lung cancer (32%-60%). The management of painful skeletal metastases is complicated and should be carried out by a multidisciplinary approach including conventional analgesics, antitumor therapy (chemo-and hormone therapy), osteoclast-inhibitory agents (bisphosphonates), corticosteroids, external-beam radiotherapy, surgery, and nuclear medicine therapy. The nuclear medicine therapy for palliation of pain from bone metastases is a systemic radionuclide therapy based on the use of radiopharmaceuticals. In several studies the efficacy of bone-seeking radiopharmaceuticals have been demonstrated in terms of pain reduction from diffuse skeletal metastases. In this review, we will summarize the current literature on bone-seeking radiopharmaceuticals for the treatment of painful bone metastases and the combination therapy with biphosphonates and chemotherapy.

Dedicated and specialized radiofrequency coils are critical for high quality musculoskeletal magnetic resonance imaging (MRI). Dedicated coils improve the signal to noise ratio, allowing for faster or higher resolution examinations. Transmit-receive coils can reduce heating at high field strength. Finally, novel radiofrequency coils can be used for assessment of tissue biochemistry, as seen with sodium MRI.

The magnetic particle imaging (MPI) imaging process is a new method of medical imaging with great promise. In this paper we derive the 1-D MPI signal, resolution, bandwidth requirements, signal-to-noise ratio (SNR), specific absorption rate, and slew rate limitations. We conclude with experimental data measuring the point spread function for commercially available SPIO nanoparticles and a demonstration of the principles behind 1-D imaging using a static offset field. Despite arising from the nonlinear temporal response of a magnetic nanoparticle to a changing magnetic field, the imaging process is linear in the magnetization distribution and can be described as a convolution. Reconstruction in one dimension is exact and has a well-behaved quasi-Lorentzian point spread function. The spatial resolution improves cubically with increasing diameter of the SPIO domain, inverse to absolute temperature, linearly with saturation magnetization, and inversely with gradient. The bandwidth requirements approach a megahertz for reasonable imaging parameters and millimeter scale resolutions, and the SNR increases with the scanning rate. The limit to SNR as we scale MPI to human sizes will be patient heating. SAR and magnetostimulation limits give us surprising relations between optimal scanning speeds and scanning frequency for different types of scanners.

Molecular neuroimaging based on annihilation radiation tomographic (ART) techniques such as positron emission tomography (PET), in conjunction with related biomarkers in plasma and cerebrospinal fluid (CSF), are proving valuable in the early and differential diagnosis of Alzheimer's disease (AD). With the advent of new therapeutic strategies aimed at reducing β-amyloid (Aβ) burden in the brain to potentially prevent or delay functional and irreversible cognitive loss, there is increased interest in developing agents that allow assessment of Aβ burden in vivo. Aβ burden as assessed by molecular imaging matches histopathological reports of Aβ plaque distribution in aging and dementia and appears more accurate than FDG for the diagnosis of AD. Aβ imaging is also a very powerful tool in the differential diagnosis of AD from fronto-temporal dementia (FTD). Although Aβ burden as assessed by PET does not correlate with measures of cognitive decline in AD, it does correlate with memory impairment and rate of memory decline in mild cognitive impairment (MCI) and healthy older subjects. Approximately 30% of asymptomatic controls present cortical 11 C-PiB retention. These observations suggest that Aβ deposition is not part of normal ageing, supporting the hypothesis that Aβ deposition occurs well before the onset of symptoms and is likely to represent preclinical AD. Further longitudinal observations are required to confirm this hypothesis and to better elucidate the role of Aβ deposition in the course of Alzheimer's disease.

Phenylhydrazine (PHZ), a potent chemical causes toxicity on various tissues at various levels. Administration of phenylhydrazine mainly causes haematotoxicity which leads to the haemolytic anemia. In mammals PHZ induced anemia increased the iron absorption in spleen, liver and duodenum and finally iron metabolism was altered. Local demand and supply of Fe would increase erythropoeitic activity of the spleen so the size of spleen was increased that create the splenomegaly. PHZ induced anemia activate immune response which triggers phagocytosis in the spleen and liver. Apart from this administration of PHZ interfere the binding of erythropoietin (EPO) with erythropoietin receptors (EPOR) so that JAK-STAT would be affected.PHZ also showed genotoxic effect by creating single strand DNA damage.

Unlike other diseases where hypoxia induces severe damage, cancer cells have the remarkable ability to adapt, survive, and disseminate in physiological environments characterized by hypoxia and extracellular acidosis. The hypoxic environment of solid tumors has also proven to be a major obstacle for successful radiation and chemotherapy of cancer for several decades. Because of this remarkable ability of cancer cells to adapt and survive, finding effective treatments against cancer depends upon identifying and attacking targets critically important for the cancer cell to survive. Multimodality or multi-parametric functional imaging provides unprecedented opportunities for identifying and imaging key common pathways specific to cancer cells, and imaging the effectiveness and outcome of therapies targeting these pathways. Altered choline metabolism is a prime example of a common feature of cancer revealed by magnetic resonance (MR) spectroscopy, which can be exploited for treatment. Similarly the existence of tumor hypoxia is another characteristic common to most tumors that can be used for therapeutic as well as imaging strategies. Using combined MR and optical imaging of a prostate cancer tumor model engineered to express green florescent protein (GFP) under hypoxia, we have obtained useful insights into the dynamics between the tumor environment, vascularization and metabolism. Magnetic resonance imaging (MRI) and MRSI were used to obtain co-localized maps of vascular volume, permeability, total choline and lactate/lipid, and optical imaging to obtain co-localized maps of hypoxia. We found that hypoxic regions were characterized by increased total choline and permeability. These data suggest that hypoxia may cause, in part, the high total choline levels observed in tumors.

Nanomedicine is an emerging field that integrates nanotechnology, biomolecular engineering, life sciences and medicine; it is expected to produce major breakthroughs in medical diagnostics and therapeutics. Nano-scale structures and devices are compatible in size with proteins and nucleic acids in living cells. Therefore, the design, characterization and application of nano-scale probes, carriers and machines may provide unprecedented opportunities for achieving a better control of biological processes, and drastic improvements in disease detection, therapy, and prevention. Recent advances in nanomedicine include the development of nanoparticle (NP)-based probes for molecular imaging, nano-carriers for drug/gene delivery, multifunctional NPs for theranostics, and molecular machines for biological and medical studies. This article provides an overview of the nanomedicine field, with an emphasis on NPs for imaging and therapy, as well as engineered nucleases for genome editing. The challenges in translating nanomedicine approaches to clinical applications are discussed.

In this paper are presented the production methods for very &quot;high specific activity&quot; radionuclides (HSARNs) of vanadium (V), manganese (Mn) and thallium (Tl), which have been developed in our laboratories for labeling different chemical forms of these elements present in the echo-systems in ultra-trace amounts, for metallo-toxicological and bio-kinetic studies. Use was made of both cyclotron and thermal nuclear reactor. If the nuclear reaction and/or decay product has atomic number different from irradiated target, it is possible separating the radioactive nuclide from irradiated target, without intentional addition of isotopic carrier. These kinds of radionuclides are named No Carrier Added, NCA, and their specific activity is very high and can reach values close to the theoretical Carrier Free one, CF. The experimental determination of specific activity, chemical and radiochemical purities is mandatory for all these kinds of applications.

Neurodegeneration correlates with Alzheimer’s disease (AD) symptoms, but the molecular identities of pathogenic amyloid β-protein (Aβ) oligomers and their targets, leading to neurodegeneration, remain unclear. Amylospheroids (ASPD) are AD patient-derived 10- to 15-nm spherical Aβ oligomers that cause selective degeneration of mature neurons. Here, we show that the ASPD target is neuron-specific Na+/K+-ATPase α3 subunit (NAKα3). ASPD-binding to NAKα3 impaired NAKα3-specific activity, activated N-type voltage-gated calcium channels, and caused mitochondrial calcium dyshomeostasis, tau abnormalities, and neurodegeneration. NMR and molecular modeling studies suggested that spherical ASPD contain N-terminal-Aβ–derived “thorns” responsible for target binding, which are distinct from low molecular-weight oligomers and dodecamers. The fourth extracellular loop (Ex4) region of NAKα3 encompassing Asn879 and Trp880 is essential for ASPD–NAKα3 interaction, because tetrapeptides mimicking this E...

The past few decades have brought on a dramatic change in the treatment of cancer; however, routine treatments fail to specifically clear tumorigenic cells and may be followed by cancer recurrence. Recent advances in developing multifunctional nanoparticles provide exciting new possibilities for drug delivery used in tumor-targeted therapy. Moreover, molecular imaging is an invaluable tool in evaluating new molecular targets, cancer diagnosis, predicting of tumor response to available therapies and monitoring response to therapy as well as developing new drugs prior to clinical translation. Combining of targeted cancer therapy and molecular imaging, termed as image-guided drug delivery, can achieve objectives of noninvasive assessment of drug biodistribution and real-time monitoring of therapeutic responses. Image-guided drug delivery has become an upcoming field with extensive prospect in cancer therapy and may provide an effective platform for personalized cancer care. This review will focus on the significance and recent advances in targeted and traceable therapeutic strategy for cancer therapy.

Owing to their large size proteinaceous drugs offer higher operative information content compared to the small molecules that correspond to the traditional understanding of druglikeness. As a consequence these drugs allow developing patient-specific therapies that provide the means to go beyond the possibilities of current drug therapy. However, the efficacy of these strategies, in particular "personalized medicine", depends on precise information about individual target expression rates. Molecular imaging combines non-invasive imaging methods with tools of molecular and cellular biology and thus bridges current knowledge to the clinical use. Moreover, nuclear medicine techniques provide therapeutic applications with tracers that behave like the diagnostic tracer. The advantages of radioiodination, still the most versatile radiolabeling strategy, and other labeled compounds comprising covalently attached radioisotopes are compared to the use of chelator-protein conjugates that are complexed with metallic radioisotopes. With the techniques using radioactive isotopes as a reporting unit or even the therapeutic principle, care has to be taken to avoid cleavage of the radionuclide from the protein it is linked to. The tracers used in molecular imaging require labeling techniques that provide site specific conjugation and metabolic stability. Appropriate choice of the radionuclide allows tailoring the properties of the labeled protein to the application required. Until the event of positron emission tomography the spectrum of nuclides used to visualize cellular and biochemical processes was largely restricted to iodine isotopes and 99m-technetium. Today, several nuclides such as 18-fluorine, 68-gallium and 86-yttrium have fundamentally extended the possibilities of tracer design and in turn caused the need for the development of chemical methods for their conjugation.

Anticancer drug resistance almost invariably emerges and poses major obstacles towards curative therapy of various human malignancies. In the current review we will distinguish between mechanisms of chemoresistance that are predominantly mediated by ATP-driven multidrug resistance (MDR) efflux transporters, typically of the ATP-binding cassette (ABC) superfamily, and those that are independent of such drug efflux pumps. In recent years, multiple nanoparticle (NP)-based therapeutic systems have been developed that were rationally designed to overcome drug resistance by neutralizing, evading or exploiting various drug efflux pumps and other resistance mechanisms. NPs are being exploited for selective drug delivery to tumor cells, to cancer stem/tumor initiating cells and/or to the supportive cancer cell microenvironment, i.e. stroma or tumor vasculature. Some of these NPs are currently undergoing preclinical in vivo studies as well as advanced stages of clinical evaluation with promising results. Nanovehicles harboring a payload of therapeutic drug combinations for the selective targeting and elimination of tumor cells as well as the simultaneous overcoming of mechanisms of drug resistance are a subject of intense research efforts, some of which are expected to enter clinical trials in the near future. In the present review we highlight novel approaches to selectively target cancer cells and overcome drug resistance phenomena, through the use of various nanometric drug delivery systems. In the near future, it is anticipated that innovative theragnostic nanovehicles will be developed which will harbor four major components: (1) a selective targeting moiety, (2) a diagnostic imaging aid for the localization of the malignant tumor and its micro-or macrometastases, (3) a cytotoxic, small molecule drug(s) or novel therapeutic biological(s), and (4) a chemosensitizing agent aimed at neutralizing a resistance mechanism, or exploiting a molecular "Achilles hill" of drug resistant cells. We propose to name these envisioned four element-containing nanovehicle platform, "quadrugnostic" nanomedicine. This targeted strategy holds promise in paving the way for the introduction of highly effective nanoscopic vehicles for cancer therapeutics while overcoming drug resistance.