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In JoVE (1)
Other Publications (13)
- Cancer Research
- Cancer Research
- Journal of Medical Entomology
- The Journal of Pathology
- Cancer Research
- Applied Physiology, Nutrition, and Metabolism = Physiologie Appliquée, Nutrition Et Métabolisme
- Clinical Cancer Research : an Official Journal of the American Association for Cancer Research
- The Journal of Pathology
- European Journal of Applied Physiology
- Advances in Physiology Education
- Clinics (São Paulo, Brazil)
- FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology
- The EMBO Journal
Articles by Scott Lyons in JoVE
JoVE Immunology and Infection
Finger-stick Blood Sampling Methodology for the Determination of Exercise-induced Lymphocyte Apoptosis
1Department of Kinesiology, Recreation, and Sport, Western Kentucky University, 2Department of Health and Human Performance, University of Houston
Exercise is capable of inducing apoptosis in immune cells. There are various measurement limitations, particularly relating to the amount of time required to isolate and treat a blood sample prior to the assessment. Demonstrated is a rapid and minimally invasive procedure for the analysis of exercise-induced lymphocyte apoptosis.
Other articles by Scott Lyons on PubMed
Noninvasive Imaging of Spontaneous Retinoblastoma Pathway-dependent Tumors in Mice
Identification of the critical pathways involved in tumorigenesis should ultimately lead to the design of better anticancer agents that target specific components of the disrupted pathways. Murine models of spontaneous cancer in which tumor formation is dependent on defined genetic alterations provide a powerful test system for evaluating the therapeutic efficacy of pathway-specific antineoplastics. We have generated a conditional mouse model for retinoblastoma-dependent sporadic cancer that permits noninvasive monitoring of pituitary tumor development in live animals via in vivo bioluminescence imaging of luciferase expression. We show that the high sensitivity of bioluminescence imaging can be used for noninvasive detection of luciferase expression in pituitary glands from tumor-free animals and for in vivo quantitation of tumor burden over a large dynamic range. This mouse model permits longitudinal monitoring of tumor onset, progression, and response to therapy and may be used effectively for testing cancer prevention and treatment strategies based on therapeutics that specifically target the retinoblastoma pathway.
The Generation of a Conditional Reporter That Enables Bioluminescence Imaging of Cre/loxP-dependent Tumorigenesis in Mice
The ability to noninvasively quantitate tumor burden from conditional (Cre/loxP-dependent) mouse cancer models would greatly increase their range of useful applications. We now report the generation of a reporter mouse that enables visualization of spontaneous tumor development from pre-existing conditional mouse tumor models via in vivo bioluminescence imaging. We demonstrate that bioluminescence can be "switched-on" in a Cre-dependent manner in every organ analyzed, and that this gives rise to between a 4 and 6-log increase in light emission per mg of wet tissue weight. Furthermore, we highlight the utility of this reporter by showing that it can be used as a sensitive means to measure spontaneous Kras2(v12)-induced lung tumorigenesis in a pre-existing mouse model of non-small cell lung cancer. Taken together, our results suggest that this reporter may be combined with a wide-range of other Cre/loxP tumor mouse models, irrespective of their tissue specificity and render them immediately amenable to longitudinal monitoring of tumor growth and therapeutic response with a noninvasive in vivo imaging approach.
Spatial Statistical Analysis of Adult Mosquito (Diptera: Culicidae) Counts: an Example Using Light Trap Data, in Redland Shire, Southeastern Queensland, Australia
Many mosquito control agencies use carbon dioxide-baited traps as surveillance tools for adult vector populations. However, decisions regarding the number and location of trap sites and the frequency of collections are often based on logistical issues, and not on the bionomics or spatial distribution of the target species. Therefore, with the aim of providing practical information for adult mosquito surveillance programs, we used an array of 81 carbon dioxide- and octenol-baited lights traps to obtain weekly samples of adult mosquitoes in Redland Shire in southeastern Queensland, Australia. The spatial patterns of four different mosquito species were examined, and positive spatial autocorrelation in trap counts was evident for Ochlerotatus vigilax (Skuse), Coquillettidia linealis (Skuse), and Culex annulirostris Skuse, but not for the container species Ochlerotatus notoscriptus (Skuse). Of the three species that exhibited spatially correlated trap counts, the autocorrelation was greatest in Oc. vigilax at a lag distance of 0-1.5 km, with Moran's I values of 0.30-0.64. Moran's I indices were also positive and statistically significant (P < 0.05) at lag distances of 1.5-3.0 and 3.0-4.5 km on each of the 15 sampling occasions. However, at 3.0-4.5 km the Moran's I values were low, which indicated only weak spatial autocorrelation in trap counts. Universal kriging was used to estimate the numbers of each species at unsampled locations throughout the study area, and leave-one-out cross validation analyses indicated that this was a robust method for Cq. linealis and Oc. vigilax. In contrast, trap counts for the container-breeding species Oc. notoscriptus were randomly distributed and the interpolated counts were not reliable. Comparisons of weekly contour maps of adult mosquito counts indicated a consistent spatial pattern for Oc. vigilax and Cq. linealis. Particular geographic areas had consistently high or low numbers of vectors, and these patterns were stable from year to year. Definition of geographic areas with consistently high or low numbers of vectors may allow control activities to be focused in areas with the greatest risk of arbovirus transmission.
Advances in Imaging Mouse Tumour Models in Vivo
Significant progress has been made recently in the variety of ways that cancer can be non-invasively imaged in murine tumour models. The development and continued refinement of specialized hardware for an array of small animal imaging methodologies are only partly responsible. So too has been the development of new imaging techniques and materials that enable specific, highly sensitive and quantitative measurement of a wide range of tumour-related parameters. Included amongst these new materials are imaging probes that selectively accumulate in tumours, or that become activated by tumour-specific molecules in vivo. Other tumour imaging strategies have been developed that rely upon the detection of reporter transgene expression in vivo, and these too have made a significant impact on both the versatility and the specificity of tumour imaging in living mice. The biological implications resulting from these latest advances are presented here, with particular emphasis on those associated with MRI, PET, SPECT, BLI, and fluorescence-based imaging modalities. Taken together, these advances in tumour imaging are set to have a profound impact on our basic understanding of in vivo tumour biology and will radically alter the application of mouse tumour models in the laboratory.
Noninvasive Bioluminescence Imaging of Normal and Spontaneously Transformed Prostate Tissue in Mice
Several transgenic mouse models of prostate cancer have been developed recently that are able to recapitulate many key biological features of the human condition. It would, therefore, be desirable to employ these models to test the efficacy of new therapeutics before clinical trial; however, the variable onset and non-visible nature of prostate tumor development limit their use for such applications. We now report the generation of a transgenic reporter mouse that should obviate these limitations by enabling noninvasive in vivo bioluminescence imaging of normal and spontaneously transformed prostate tissue in the mouse. We used an 11-kb fragment of the human prostate-specific antigen (PSA) promoter to achieve specific and robust expression of firefly luciferase in the prostate glands of transgenic mice. Ex vivo bioluminescence imaging and in situ hybridization analysis confirmed that luciferase expression was restricted to the epithelium in all four lobes of the prostate. We also show that PSA-Luc mice exhibit decreased but readily detectable levels of in vivo bioluminescence over extended time periods following androgen ablation. These results suggest that this reporter should enable in vivo imaging of both androgen-dependent and androgen-independent prostate tumor models. As proof-of-principle, we show that we could noninvasively image SV40 T antigen-induced prostate tumorigenesis in mice with PSA-Luc. Furthermore, we show that our noninvasive imaging strategy can be successfully used to image tumor response to androgen ablation in transgenic mice and, as a result, that we can rapidly identify individual animals capable of sustaining tumor growth in the absence of androgen.
Excess Post-exercise Oxygen Consumption in Untrained Males: Effects of Intermittent Durations of Arm Ergometry
The purpose of this study was to investigate excess post-exercise oxygen consumption (EPOC) following a continuous 30 min bout of upper-body exercise (UBE) compared with 3 consecutive 10 min bouts of UBE. Ten male subjects (age (mean +/- standard deviation), 25.7 +/- 5.83 years; arm VO(2) (peak), 2.2 +/- 0.25 L x min(-1), on separate days (48 h between trials) and in counterbalanced order, performed a continuous 30 min bout of arm exercise at 60% of arm VO2 peak and 3 separate 10 min bouts of arm exercise at 60% of arm VO(2) (peak). Subjects reported to the laboratory rested and after a 12 h fast. Each test was preceded by a 30 min baseline test to determine resting metabolic rate. Post-exercise VO2 was continuously monitored until baseline was re-established. Results showed that the combined magnitude of the EPOCs from the intermittent exercise sessions was significantly (p > .05) greater (4.47 +/- 1.58 L O2) than that elicited from the continuous exercise session (1.54 +/- 1.25 L O2). These data indicate that separating a continuous 30 min arm exercise into 3 equal 10 min arm exercises will elicit a small but significantly higher EPOC, and thus result in greater post-exercise energy expenditure. This could be beneficial for those unable to perform lower-body exercise (LBE), or for those with limited exercise capacities.
A Spontaneous Acinar Cell Carcinoma Model for Monitoring Progression of Pancreatic Lesions and Response to Treatment Through Noninvasive Bioluminescence Imaging
We have generated an EL1-luc/TAg transgenic mouse model that develops spontaneous and bioluminescent acinar cell carcinomas. We applied this model to noninvasively monitor tumor development and drug response.
Bioluminescent Imaging: a Critical Tool in Pre-clinical Oncology Research
Bioluminescent imaging (BLI) is a non-invasive imaging modality widely used in the field of pre-clinical oncology research. Imaging of small animal tumour models using BLI involves the generation of light by luciferase-expressing cells in the animal following administration of substrate. This light may be imaged using an external detector. The technique allows a variety of tumour-associated properties to be visualized dynamically in living models. The increasing use of BLI as a small-animal imaging modality has led to advances in the development of xenogeneic, orthotopic, and genetically engineered animal models expressing luciferase genes. This review aims to provide insight into the principles of BLI and its applications in cancer research. Many studies to assess tumour growth and development, as well as efficacy of candidate therapeutics, have been performed using BLI. More recently, advances have also been made using bioluminescent imaging in studies of protein-protein interactions, genetic screening, cell-cycle regulators, and spontaneous cancer development. Such novel studies highlight the versatility and potential of bioluminescent imaging in future oncological research.
Exercise-induced Immune Cell Apoptosis: Image-based Model for Morphological Assessment
Methods of assessing exercise-induced lymphocyte apoptosis have produced varying results. While morphological methods generally yield a significantly greater apoptotic index compared to those employing biochemical markers, benefits and limitations are associated with each methodology. Of interest in this report is the limitation of subjectivity associated with the morphological technique. To overcome the lack of objectivity associated with the morphological method, we describe an image-based approach by which computer software assesses the characteristics associated with lymphocyte apoptosis. A stochastic controlled deformable model can be employed to detect the classic morphological apoptotic changes induced by exercise, including membrane blebbing and the formation of apoptotic bodies. We propose that an objective evaluation of multiple points in the apoptotic process through the proposed model in conjunction with current biomarker methods has the potential to advance our understanding of the exercise-induced immune cell death response to a greater degree than current methods.
Student Peer Review Decisions on Submitted Manuscripts Are As Stringent As Faculty Peer Reviewers
The International Journal of Exercise Science is the only student-centered peer-reviewed journal in its field. Upon graduate student first author submissions, two student reviewers and one faculty reviewer are asked to review. On professionally submitted papers, two faculty peers are asked to assess the manuscript. The purpose of the present study was to determine whether graduate students returned similar decisions compared with faculty reviewers who evaluated the same manuscript. In addition, decisions of faculty peers reviewing graduate student- versus faculty-submitted manuscripts were compared. Mean comparisons between groups were evaluated using independent t-tests with significance at P ≤ 0.05. Graduate students (2.21 ± 0.69) and faculty peers (2.24 ± 0.66) returned similar decisions on student-submitted manuscripts (P = 0.84). Faculty decisions on manuscripts submitted by a professional primary author (1.86 ± 0.77) were not different compared with faculty peers reviewing student manuscripts (P = 0.06). Statistics revealed that graduate students are just as stringent in the peer review process as established reviewers. Additionally, faculty reviewers evaluated manuscripts equally regardless of submission type.
Cognitive Awareness of Carbohydrate Intake Does Not Alter Exercise-induced Lymphocyte Apoptosis
The purpose of this investigation was to determine whether cognitive awareness of carbohydrate beverage consumption affects exercise-induced lymphocyte apoptosis, independent of actual carbohydrate intake.
Imaging Sialylated Tumor Cell Glycans in Vivo
Cell surface glycans are involved in numerous physiological processes that involve cell-cell interactions and migration, including lymphocyte trafficking and cancer metastasis. We have used a bioorthogonal metabolic labeling strategy to detect cell surface glycans and demonstrate, for the first time, fluorescence and radionuclide imaging of sialylated glycans in a murine tumor model in vivo. Peracetylated azido-labeled N-acetyl-mannosamine, injected intraperitoneally, was used as the metabolic precursor for the biosynthesis of 5-azidoneuraminic, or azidosialic acid. Azidosialic acid-labeled cell surface glycans were then reacted, by Staudinger ligation, with a biotinylated phosphine injected intraperitoneally, and the biotin was detected by subsequent intravenous injection of a fluorescent or radiolabeled avidin derivative. At 24 h after administration of NeutrAvidin, labeled with either a far-red fluorophore or (111)In, there was a significant azido-labeled N-acetyl-mannosamine-dependent increase in tumor-to-tissue contrast, which was detected using optical imaging or single-photon-emission computed tomography (SPECT), respectively. The technique has the potential to translate to the clinic, where, given the prognostic relevance of altered sialic acid expression in cancer, it could be used to monitor disease progression.
The Androgen Receptor Fuels Prostate Cancer by Regulating Central Metabolism and Biosynthesis
The androgen receptor (AR) is a key regulator of prostate growth and the principal drug target for the treatment of prostate cancer. Previous studies have mapped AR targets and identified some candidates which may contribute to cancer progression, but did not characterize AR biology in an integrated manner. In this study, we took an interdisciplinary approach, integrating detailed genomic studies with metabolomic profiling and identify an anabolic transcriptional network involving AR as the core regulator. Restricting flux through anabolic pathways is an attractive approach to deprive tumours of the building blocks needed to sustain tumour growth. Therefore, we searched for targets of the AR that may contribute to these anabolic processes and could be amenable to therapeutic intervention by virtue of differential expression in prostate tumours. This highlighted calcium/calmodulin-dependent protein kinase kinase 2, which we show is overexpressed in prostate cancer and regulates cancer cell growth via its unexpected role as a hormone-dependent modulator of anabolic metabolism. In conclusion, it is possible to progress from transcriptional studies to a promising therapeutic target by taking an unbiased interdisciplinary approach.
