Sevuparin binds to multiple adhesive ligands and reduces sickle red blood cell-induced vaso-occlusion.

Sevuparin is a novel drug candidate in phase II development as a treatment for vaso-occlusive crises (VOC) in patients with sickle cell disease (SCD). As a heparin-derived polysaccharide, sevuparin has been designed to retain anti-adhesive properties, while the antithrombin-binding domains have been eliminated, substantially diminishing its anticoagulant activity. Here, we demonstrate that sevuparin inhibits the adhesion of human sickle red blood cells (SS-RBCs) to stimulated cultured endothelial cells in vitro. Importantly, sevuparin prevents vaso-occlusion and normalizes blood flow in an in vivo mouse model of SCD vaso-occlusion. Analyses by surface plasmon resonance (SPR) and fluorescence correlation spectroscopy (FCS) demonstrate that sevuparin binds to P- and L-selectins, thrombospondin, fibronectin and von Willebrand factor, all of which are thought to contribute to vaso-occlusion in SCD. Despite low anticoagulation activity, sevuparin has anti-adhesive efficacy similar to the low molecular weight heparin tinzaparin both in vitro and in vivo. These results suggest that the anti-adhesive properties rather than the anticoagulant effects of heparinoids are critical for the treatment of vaso-occlusion in SCD. Therefore, sevuparin is now being evaluated in SCD patients hospitalized for treatment of VOC.

Which Receives More Attention, Online Review Sentiment or Online Review Rating? Spillover Effect Analysis from JD.com.

Studies have found that competitive products' online review ratings (ORRs) have a spillover effect on the focal product's sales. However, the spillover effect of online review sentiment (ORS) as an essential component of online review analysis has yet to be studied. In this study, we analyze online review content from JD.com using the latent Dirichlet allocation to identify the product attribute topics that consumers are most concerned about. We then construct a baseline regression model of ORS and ORRs to explore the effects of online competitive product reviews on focal product sales. Moreover, we examine how the interaction between ORS and critical factors of online reviews affect sales. Our results indicate that the ORS of competitive products has a negative effect on focal product sales, and the effect is greater than the ORS and ORRs of focal products, respectively. In addition, the ORS of competitive products inhibits the sale of focal products as evaluations of product attributes become more positive or online review usefulness increases. We also find that the effect of ORRs of competitive products is not significant, which may be because clothing, as an experiential product, requires consumers to gain more information about specific usage scenarios before making a decision. This study provides a more accurate basis for consumer decision-making and offers retailers a novel approach to developing marketing strategies.

Automated measurement of blood flow velocity and direction and hemoglobin oxygen saturation in the rat lung using intravital microscopy.

Intravital microscopy of the pulmonary microcirculation in research animals is of great scientific interest for its utility in identifying regional changes in pulmonary microcirculatory blood flow. Although feasibility studies have been reported, the pulmonary window can be further refined into a practical tool for pharmaceutical research and drug development. We have established a method to visualize and quantify dynamic changes in three key features of lung function: microvascular red blood cell velocity, flow direction, and hemoglobin saturation. These physiological parameters were measured in an acute closed-chest pulmonary window, which allows real-time images to be captured by fluorescence and multispectral absorption microscopy; images were subsequently quantified using computerized analysis. We validated the model by quantifying changes in microcirculatory blood flow and hemoglobin saturation in two ways: 1) after changes in inspired oxygen content and 2) after pharmacological reduction of pulmonary blood flow via treatment with the ß1 adrenergic receptor blocker metoprolol. This robust and relatively simple system facilitates pulmonary intravital microscopy in laboratory rats for pharmacological and physiological research.

Social media-based urban disaster recovery and resilience analysis of the Henan deluge.

Measuring disaster resilience from the perspective of long-term recovery ability is important for the planning and construction of urban sustainability, whereas short-term resilient recovery can better reflect a city's ability to recover quickly after a disaster occurs. This study proposes an analytical framework for urban disaster recovery and resilience based on social media data that can analyze short-term disaster recovery and assess disaster resilience from the perspectives of infrastructure and people's psychological states. We consider the downpour in Henan, China, in July 2021. The results show that (1) social media data can effectively reflect short-term disaster recovery, (2) disaster resilience can be assessed using social media data combined with rainfall and damage data, and (3) the framework can quantitatively reflect the differences in disaster recovery and resilience across regions. The findings can facilitate better decision-making in disaster emergency management for precise and effective post-disaster reconstruction and psychological intervention, and provide references for cities to improve disaster resilience.

Monitoring the Relationship between Social Network Status and Influenza Based on Social Media Data.

BACKGROUND: This article aims to analyze the relationship between user characteristics on social networks and influenza. METHODS: Three specific research questions are investigated: (1) we classify Weibo updates to recognize influenza-related information based on machine learning algorithms and propose a quantitative model for influenza susceptibility in social networks; (2) we adopt in-degree indicator from complex networks theory as social media status to verify its coefficient correlation with influenza susceptibility; (3) we also apply the LDA topic model to explore users' physical condition from Weibo to further calculate its coefficient correlation with influenza susceptibility. From the perspective of social networking status, we analyze and extract influenza-related information from social media, with many advantages including efficiency, low cost, and real time. RESULTS: We find a moderate negative correlation between the susceptibility of users to influenza and social network status, while there is a significant positive correlation between physical condition and susceptibility to influenza. CONCLUSIONS: Our findings reveal the laws behind the phenomenon of online disease transmission, and providing important evidence for analyzing, predicting, and preventing disease transmission. Also, this study provides theoretical and methodological underpinnings for further exploration and measurement of more factors associated with infection control and public health from social networks.

Concerned or Apathetic? Using Social Media Platform (Twitter) to Gauge the Public Awareness about Wildlife Conservation: A Case Study of the Illegal Rhino Trade.

The illegal wildlife trade is resulting in worldwide biodiversity loss and species' extinction. It should be exposed so that the problems of conservation caused by it can be highlighted and resolutions can be found. Social media is an effective method of information dissemination, providing a real-time, low-cost, and convenient platform for the public to release opinions on wildlife protection. This paper aims to explore the usage of social media in understanding public opinions toward conservation events, and illegal rhino trade is an example. This paper provides a framework for analyzing rhino protection issues by using Twitter. A total of 83,479 useful tweets and 33,336 pieces of users' information were finally restored in our database after filtering out irrelevant tweets. With 2422 records of trade cases, this study builds up a rhino trade network based on social media data. The research shows important findings: (1) Tweeting behaviors are somewhat affected by the information of traditional mass media. (2) In general, countries and regions with strong negative sentiment tend to have high volume of rhino trade cases, but not all. (3) Social celebrities' participation in activities arouses wide public concern, but the influence does not last for more than a month. NGOs, GOs, media, and individual enterprises are dominant in the dissemination of information about rhino trade. This study contributes in the following ways: First, this paper conducts research on public opinions toward wildlife conservation using natural language processing technique. Second, this paper offers advice to governments and conservationist organizations, helping them utilize social media for protecting wildlife.

Suit the Remedy to the Case-The Effectiveness of COVID-19 Nonpharmaceutical Prevention and Control Policies Based on Individual Going-Out Behavior.

Nonpharmaceutical policies for epidemic prevention and control have been extensively used since the outbreak of COVID-19. Policies ultimately work by limiting individual behavior. The aim of this paper is to evaluate the effectiveness of policies by combining macro nonpharmaceutical policies with micro-individual going-out behavior. For different going out scenarios triggered by individual physiological safety needs, friendship needs, and family needs, this paper categorizes policies with significant differences in intensity, parameterizes the key contents of the policies, and simulates and analyzes the effectiveness of the policies in different going-out scenarios with simulation methods. The empirical results show that enhancing policy intensity can effectively improve policy effectiveness. Among different types of policies, restricting the times of going out is more effective. Further, the effect of controlling going out based on physiological safety needs is better than other needs. We also evaluate the policy effectiveness of 26 global countries or regions. The results show that the policy effectiveness varies among 26 countries or regions. The quantifiable reference provided by this study facilitates decision makers to establish policy and practices for epidemic prevention and control.

A Social Media Infodemic-Based Prediction Model for the Number of Severe and Critical COVID-19 Patients in the Lockdown Area.

Accurately predicting the number of severe and critical COVID-19 patients is critical for the treatment and control of the epidemic. Social media data have gained great popularity and widespread application in various research domains. The viral-related infodemic outbreaks have occurred alongside the COVID-19 outbreak. This paper aims to discover trustworthy sources of social media data to improve the prediction performance of severe and critical COVID-19 patients. The innovation of this paper lies in three aspects. First, it builds an improved prediction model based on machine learning. This model helps predict the number of severe and critical COVID-19 patients on a specific urban or regional scale. The effectiveness of the prediction model, shown as accuracy and satisfactory robustness, is verified by a case study of the lockdown in Hubei Province. Second, it finds the transition path of the impact of social media data for predicting the number of severe and critical COVID-19 patients. Third, this paper provides a promising and powerful model for COVID-19 prevention and control. The prediction model can help medical organizations to realize a prediction of COVID-19 severe and critical patients in multi-stage with lead time in specific areas. This model can guide the Centers for Disease Control and Prevention and other clinic institutions to expand the monitoring channels and research methods concerning COVID-19 by using web-based social media data. The model can also facilitate optimal scheduling of medical resources as well as prevention and control policy formulation.

Effects of PM2.5 on People's Emotion: A Case Study of Weibo (Chinese Twitter) in Beijing.

PM2.5 not only harms physical health but also has negative impacts on the public's wellbeing and cognitive and behavioral patterns. However, traditional air quality assessments may fail to provide comprehensive, real-time monitoring of air quality because of the sparse distribution of air quality monitoring stations. Overcoming some key limitations of traditional surface monitoring data, Web-based social media platforms, such as Twitter, Weibo, and Facebook, provide a promising tool and novel perspective for environmental monitoring, prediction, and evaluation. This study aims to investigate the relationship between PM2.5 levels and people's emotional intensity by observing social media postings. This study defines the "emotional intensity" indicator, which is measured by the number of negative posts on Weibo, based on Weibo data related to haze from 2016 and 2017. This study estimates sentiment polarity using a recurrent neural networks model based on LSTM (Long Short-Term Memory) and verifies the correlation between high PM2.5 levels and negative posts on Weibo using a Pearson correlation coefficient and multiple linear regression model. This study makes the following observations: (1) Taking the two-year data as an example, this study recorded the significant influence of PM2.5 levels on netizens' posting behavior. (2) Air quality, meteorological factors, the seasons, and other factors have a strong influence on netizens' emotional intensity. (3) From a quantitative viewpoint, the level of PM2.5 varies by 1 unit, and the number of negative Weibo posts fluctuates by 1.0168 units. Thus, it can be concluded that netizens' emotional intensity is significantly positively affected by levels of PM2.5. The high correlation between PM2.5 levels and emotional intensity and the sensitivity of social media data shows that social media data can be used to provide a new perspective on the assessment of air quality.

Infectious or Recovered? Optimizing the Infectious Disease Detection Process for Epidemic Control and Prevention Based on Social Media.

Detecting the period of a disease is of great importance to building information management capacity in disease control and prevention. This paper aims to optimize the disease surveillance process by further identifying the infectious or recovered period of flu cases through social media. Specifically, this paper explores the potential of using public sentiment to detect flu periods at word level. At text level, we constructed a deep learning method to classify the flu period and improve the classification result with sentiment polarity. Three important findings are revealed. Firstly, bloggers in different periods express significantly different sentiments. Blogger sentiments in the recovered period are more positive than in the infectious period when measured by the interclass distance. Secondly, the optimized disease detection process can substantially improve the classification accuracy of flu periods from 0.876 to 0.926. Thirdly, our experimental results confirm that sentiment classification plays a crucial role in accuracy improvement. Precise identification of disease periods enhances the channels for the disease surveillance processes. Therefore, a disease outbreak can be predicted credibly when a larger population is monitored. The research method proposed in our work also provides decision making reference for proactive and effective epidemic control and prevention in real time.

Angiostatin-like activity of a monoclonal antibody to the catalytic subunit of F1F0 ATP synthase.

The antiangiogenic protein angiostatin inhibits ATP synthase on the endothelial cell surface, blocking cellular proliferation. To examine the specificity of this interaction, we generated monoclonal antibodies (mAb) directed against ATP synthase. mAb directed against the beta-catalytic subunit of ATP synthase (MAb3D5AB1) inhibits the activity of the F(1) domain of ATP synthase and recognizes the catalytic beta-subunit of ATP synthase. We located the antibody recognition site of MAb3D5AB1 in domains containing the active site of the beta-subunit. MAb3D5AB1 also binds to purified Escherichia coli F(1) with an affinity 25-fold higher than the affinity of angiostatin for this protein. MAb3D5AB1 inhibits the hydrolytic activity of F(1) ATP synthase at lower concentrations than angiostatin. Like angiostatin, MAb3D5AB1 inhibits ATP generation by ATP synthase on the endothelial cell surface in acidic conditions, the typical tumor microenvironment where cell surface ATP synthase exhibits greater activity. MAb3D5AB1 disrupts tube formation and decreases intracellular pH in endothelial cells exposed to low extracellular pH. Neither angiostatin nor MAb3D5AB1 showed an antiangiogenic effect in the corneal neovascularization assay; however, both were effective in the low-pH environment of the chicken chorioallantoic membrane assay. Thus, MAb3D5AB1 shows angiostatin-like properties superior to angiostatin and may be exploited in cancer chemotherapy.

Nitric oxide loading reduces sickle red cell adhesion and vaso-occlusion in vivo.

Sickle red blood cells (SSRBCs) are adherent to the endothelium, activate leukocyte adhesion, and are deficient in bioactive nitric oxide (NO) adducts such as S-nitrosothiols (SNOs), with reduced ability to induce vasodilation in response to hypoxia. All these pathophysiologic characteristics promote vascular occlusion, the hallmark of sickle cell disease (SCD). Loading hypoxic SSRBCs in vitro with NO followed by reoxygenation significantly decreased epinephrine-activated SSRBC adhesion to the endothelium, the ability of activated SSRBCs to mediate leukocyte adhesion in vitro, and vessel obstruction in vivo. Because transfusion is frequently used in SCD, we also determined the effects of banked (SNO-depleted) red blood cells (RBCs) on vaso-occlusion in vivo. Fresh or 14-day-old normal RBCs (AARBCs) reduced epinephrine-activated SSRBC adhesion to the vascular endothelium and prevented vaso-occlusion. In contrast, AARBCs stored for 30 days failed to decrease activated SSRBC adhesivity or vaso-occlusion, unless these RBCs were loaded with NO. Furthermore, NO loading of SSRBCs increased S-nitrosohemoglobin and modulated epinephrine's effect by upregulating phosphorylation of membrane proteins, including pyruvate kinase, E3 ubiquitin ligase, and the cytoskeletal protein 4.1. Thus, abnormal SSRBC NO/SNO content both contributes to the vaso-occlusive pathophysiology of SCD, potentially by affecting at least protein phosphorylation, and is potentially amenable to correction by (S)NO repletion or by RBC transfusion.

Bromelain treatment decreases neutrophil migration to sites of inflammation.

Bromelain, a mixture of proteases derived from pineapple stem, has been reported to have therapeutic benefits in a variety of inflammatory diseases, including murine inflammatory bowel disease. The purpose of this work was to understand potential mechanisms for this anti-inflammatory activity. Exposure to bromelain in vitro has been shown to remove a number of cell surface molecules that are vital to leukocyte trafficking, including CD128a/CXCR1 and CD128b/CXCR2 that serve as receptors for the neutrophil chemoattractant IL-8 and its murine homologues. We hypothesized that specific proteolytic removal of CD128 molecules by bromelain would inhibit neutrophil migration to IL-8 and thus decrease acute responses to inflammatory stimuli. Using an in vitro chemotaxis assay, we demonstrated a 40% reduction in migration of bromelain- vs. sham-treated human neutrophils in response to rhIL-8. Migration to the bacterial peptide analog fMLP was unaffected, indicating that bromelain does not induce a global defect in leukocyte migration. In vivo bromelain treatment generated a 50-85% reduction in neutrophil migration in 3 different murine models of leukocyte migration into the inflamed peritoneal cavity. Intravital microscopy demonstrated that although in vivo bromelain treatment transiently decreased leukocyte rolling, its primary long-term effect was abrogation of firm adhesion of leukocytes to blood vessels at the site of inflammation. These changes in adhesion were correlated with rapid re-expression of the bromelain-sensitive CD62L/L-selectin molecules that mediate rolling following in vivo bromelain treatment and minimal re-expression of CD128 over the time period studied. Taken together, these studies demonstrate that bromelain can effectively decrease neutrophil migration to sites of acute inflammation and support the specific removal of the CD128 chemokine receptor as a potential mechanism of action.

Observation of incipient tumor angiogenesis that is independent of hypoxia and hypoxia inducible factor-1 activation.

It is well established that hypoxia potently stimulates tumor angiogenesis by activating hypoxia inducible factor-1 (HIF-1)-induced proangiogenic factors, such as vascular endothelial growth factor. However, very little is known about the role of hypoxia in incipient angiogenesis in avascular tumors during their early stages of growth. To noninvasively investigate the functional significance of hypoxia and HIF-1 activation in incipient tumor angiogenesis, we genetically engineered HCT116 human colon carcinoma cells and 4T1 mouse mammary carcinoma cells with constitutively expressed red fluorescence protein as a tumor marker and green fluorescence protein (GFP) as a reporter for hypoxia and HIF-1 activation. The accuracy of GFP fluorescence in reporting hypoxia was confirmed by flow cytometry analysis and by immunohistochemical comparison with pimonidazole, a well-established hypoxia marker drug. Mouse dorsal skin-fold window chambers showed that incipient angiogenesis preceded a detectable level of hypoxia. The detectable levels of hypoxia were spatially and temporally related with more intensive secondary angiogenesis following the initial onset of new vessel formation. Selective killing of hypoxic cells by tirapazamine efficiently eliminated or delayed the detection of hypoxic cells, but it did not significantly delay the onset of incipient angiogenesis. These findings provide the first in vivo evidence that incipient tumor angiogenesis may not depend on hypoxia or HIF-1 activation. This is in contrast to the clear role of hypoxia in driving angiogenesis once initial tumor microvessel formation has occurred.

Melanoma, a tumor based on a mutant stem cell?

Stem cells play a critical role in normal tissue maintenance, and mutations in these stem cells may give rise to cancer. We hypothesize that melanoma develops from a mutated stem cell and therefore residual stem cell characteristics should be able to be identified in melanoma cell lines. We studied three metastatic melanoma cell lines that exhibited multiple morphologic forms in culture and demonstrated the capacity to pigment. We used the ability to efflux Hoechst 33342 dye, a technique known to enrich for stem cells in many tissues, to segregate cell populations. The cells with the greatest ability to efflux the dye were (1) small in size, (2) had the capacity to give rise to larger cell forms, and (3) had the greatest ability to expand in culture. The small cells were found to have a decreased proliferative rate and were less melanized. Large dendritic cells that appeared to be nonproliferative were identified in cultures. Treatment with cytosine beta-D-arabinofuranoside hydrochloride (Ara-C) expanded the large cell population but the residual proliferative capacity, both in vitro and in vivo, remained concentrated in the smaller cell fraction. Antigenic staining patterns were variable and heterogeneous. Nestin (a neural stem cell marker) and gp100 (premelanosomal marker) favored the smaller cell population, while nerve growth factor receptor often labeled larger cells. Morphologic and antigenic heterogeneity remained intact after clonal purification. These findings are consistent with the behavior expected for a tumor based on stem cell biology; this finding has diagnostic and therapeutic implications for melanocytic neoplasias.

Preferential extravasation and accumulation of liposomal vincristine in tumor comparing to normal tissue enhances antitumor activity.

To quantitatively evaluate the extravasation, accumulation and selectivity to tumor tissues of liposomal vincristine (LV), dorsal skin-fold window chambers on athymic mice with or without LX-1, a human small cell lung cancer, xenograft implants and fluorescent intravital microscopy imaging were used. In vitro studies show that minimal loss of fluorescence marker DiI from liposomes occurs after 4 days of inoculation in murine plasma, and the release profiles of DiI-LV and LV were essentially the same with approximately 40% of the encapsulated vincristine sulfate (VCR) released after 26 h. Significantly faster extravasation of DiI-LV from tumor vessels was shown compared to non-tumor tissue after single dose i.v. administration. The relative interstitial amounts at 60 min (RIA(60)) for tumor and non-tumor tissues were 0.837+/-0.314 and 0.012+/-0.091, respectively (P=0.01). DiI-LV accumulation was significantly higher in tumor than in normal tissue, which continued beyond 48 h. Both DiI-LV and LV showed significant antitumor effects in window chambers and in flank tumors, compared with controls and VLS alone. The preferential extravasation of DiI-LV from tumor vasculature as well as its differential retention in tumor tissue provides the basis for the enhancement in antitumor activity of LV over VCR.

Responses of vascular endothelial cells to angiogenic signaling are important for tumor cell survival.

Neoplastic cells overexpress several angiogenic cytokines, which stimulate neovascularization. Whether the responses of the host endothelial cells to these signaling molecules affect tumor cells during early tumorigenesis has not been investigated. We investigated pre-angiogenic tumor cell survival and angiogenesis initiation by two murine tumor lines (4T1 mammary carcinoma and B16 melanoma), which constitutively expressed GFP, in dorsal skin-fold window chambers of mice treated with extracellular domain of Tie-2 (ExTek) or bFGF. ExTek reduced tumor cell survival, retarded tumor growth, and inhibited angiogenesis onset compared with controls. bFGF increased tumor cell survival and promoted earlier angiogenesis and tumor growth. Neither bFGF nor ExTek affected cell proliferation in vitro. RT-PCR showed mRNA expression of bFGF receptor 2 (FGFR2) IIIb, which does not bind bFGF efficiently, by 4T1 cells and B16 cells express FGFR1 but not FGFR2. B16 cells expressed angiopoietin (Ang) 2, but neither cell line expresses Ang1. Both tumor lines express VEGF. These findings suggested that effects of bFGF and ExTek on tumor cell survival and angiogenesis were not due to direct action but were instead a result of paracrine factors secreted by endothelial cells. These subsequent signals from endothelial cells promote early survival and proliferation of disseminated tumor cells before onset of angiogenesis.

High-resolution in vivo imaging of fluorescent proteins using window chamber models.

Fluorescent proteins enable in vivo characterization of a wide and growing array of morphological and functional biomarkers. To fully capitalize on the spatial and temporal information afforded by these reporter proteins, a method for imaging these proteins at high resolution longitudinally is required. This chapter describes the use of window chamber models as a means of imaging fluorescent proteins and other optical parameters. Such models essentially involve surgically implanting a window through which tumor or normal tissue can be imaged using existing microscopy techniques. This enables acquisition of high-quality images down to the cellular or subcellular scale, exploiting the diverse array of optical contrast mechanisms, while also maintaining the native microenvironment of the tissue of interest. This makes these techniques applicable to a wide array of problems in the biomedical sciences.

The combination of theophylline and endothelin receptor antagonism improves exercise performance of rats under simulated high altitude.

Decreased physical performance is a well-known consequence of rapid ascent to high altitude. Hypoxic pulmonary vasoconstriction (HPV) potentially limits cardiac output and systemic blood flow, thus preventing successful adaptation to rapid ascent. We hypothesized that pharmacological enhancement of the heart rate with theophylline, combined with reversal of HPV via endothelin blockade, could increase exercise performance at high altitude. Female Sprague-Dawley rats were treated with combinations of 1) theophylline, 2) the endothelin receptor antagonists sitaxsentan/ambrisentan, and/or 3) phosphodiesterase-5 inhibitor sildenafil and exposed to either a simulated high altitude (4,267 m) or 12% oxygen. Exercise capacity, peripheral blood flow, hemodynamics, and pulmonary leak were examined. Combination treatment with theophylline and endothelin blockade, but not with the respective single compounds, significantly prolonged run-to-fatigue time under simulated high altitude. No such efficacy was found when theophylline was combined with sildenafil. Neither theophylline nor sitaxsentan or their combination influenced breathing rates and hemoglobin oxygen saturation. Whereas under hypoxia, theophylline significantly increased muscular blood flow, and sitaxsentan increased tissue oxygenation, the combination improved both parameters but in a reduced manner. Under hypoxia, the combination treatment but not the single compounds significantly enhanced pulmonary arterial pressure compared with controls (13.1 ± 6.3 vs. 11.9 ± 5.2 mmHg), whereas mean arterial pressure remained unaffected. Pulmonary wet-to-dry weight ratios were unaffected by combination treatment. We conclude that concomitant dosing with a cardiac stimulant and endothelin antagonist can partially reverse loss of physical performance capacity under hypobaric hypoxia, independent from improving blood oxygen saturation.

In vivo optical molecular imaging and analysis in mice using dorsal window chamber models applied to hypoxia, vasculature and fluorescent reporters.

Optical techniques for functional imaging in mice have a number of key advantages over other common imaging modalities such as magnetic resonance imaging, positron emission tomography or computed tomography, including high resolution, low cost and an extensive library of available contrast agents and reporter genes. A major challenge to such work is the limited penetration depth imposed by tissue turbidity. We describe a window chamber technique by which these limitations can be avoided. This facilitates the study of a wide range of processes, with potential endpoints including longitudinal gene expression, vascular remodeling and angiogenesis, and tumor growth and invasion. We further describe several quantitative imaging and analysis techniques for characterizing in vivo fluorescence properties and functional endpoints, including vascular morphology and oxygenation. The procedure takes ∼2 h to complete, plus up to several weeks for tumor growth and treatment procedures.