Tear Proteomics in Infants at Risk of Retinopathy of Prematurity: A Feasibility Study.

Purpose: This feasibility study investigated the practicability of collecting and analyzing tear proteins from preterm infants at risk of retinopathy of prematurity (ROP). We sought to identify any tear proteins which might be implicated in the pathophysiology of ROP as well as prognostic markers. Methods: Schirmer's test was used to obtain tear samples from premature babies, scheduled for ROP screening, after parental informed consent. Mass spectrometry was used for proteomic analysis. Results: Samples were collected from 12 infants, which were all adequate for protein analysis. Gestational age ranged from 25 + 6 to 31 + 1 weeks. Postnatal age at sampling ranged from 19 to 66 days. One infant developed self-limiting ROP. Seven hundred one proteins were identified; 261 proteins identified in the majority of tear samples, including several common tear proteins, were used for analyses. Increased risk of ROP as determined by the postnatal growth ROP (G-ROP) criteria was associated with an increase in lactate dehydrogenase B chain in tears. Older infants demonstrated increased concentration of immunoglobulin complexes within their tear samples and two sets of twins in the cohort showed exceptionally similar proteomes, supporting validity of the analysis. Conclusions: Tear sampling by Schirmer test strips and subsequent proteomic analysis by mass spectrometry in preterm infants is feasible. A larger study is required to investigate the potential use of tear proteomics in identification of ROP. Translational Relevance: Tear sampling and subsequent mass spectrometry in preterm infants is feasible. Investigation of the premature tear proteome may increase our understanding of retinal development and provide noninvasive biomarkers for identification of treatment-warranted ROP.

Quantifying the land and population risk of sewage spills overland using a fine-scale, DEM-based GIS model.

Accidental releases of untreated sewage into the environment, known as sewage spills, may cause adverse gastrointestinal stress to exposed populations, especially in young, elderly, or immune-compromised individuals. In addition to human pathogens, untreated sewage contains high levels of micropollutants, organic matter, nitrogen, and phosphorus, potentially resulting in aquatic ecosystem impacts such as algal blooms, depleted oxygen, and fish kills in spill-impacted waterways. Our Geographic Information System (GIS) model, Spill Footprint Exposure Risk (SFER) integrates fine-scale elevation data (1/3 arc-second) with flowpath tracing methods to estimate the expected overland pathways of sewage spills and the locations where they are likely to pool. The SFER model can be integrated with secondary measures tailored to the unique needs of decision-makers so they can assess spatially potential exposure risk. To illustrate avenues to assess risk, we developed risk measures for land and population health. The land risk of sewage spills is calculated for subwatershed regions by computing the proportion of the subwatershed's area that is affected by one modeled footprint. The population health risk is assessed by computing the estimated number of individuals who are within the modeled footprint using fine-scale (90 square meters) population estimates data from LandScan USA. In the results, with a focus on the Atlanta metropolitan region, potential strategies to combine these risk measures with the SFER model are outlined to identify specific areas for intervention.

Omentum-derived matrix enables the study of metastatic ovarian cancer and stromal cell functions in a physiologically relevant environment.

High-grade serous (HGS) ovarian cancer is the most lethal gynaecological disease in the world and metastases is a major cause. The omentum is the preferential metastatic site in HGS ovarian cancer patients and in vitro models that recapitulate the original environment of this organ at cellular and molecular level are being developed to study basic mechanisms that underpin this disease. The tumour extracellular matrix (ECM) plays active roles in HGS ovarian cancer pathology and response to therapy. However, most of the current in vitro models use matrices of animal origin and that do not recapitulate the complexity of the tumour ECM in patients. Here, we have developed omentum gel (OmGel), a matrix made from tumour-associated omental tissue of HGS ovarian cancer patients that has unprecedented similarity to the ECM of HGS omental tumours and is simple to prepare. When used in 2D and 3D in vitro assays to assess cancer cell functions relevant to metastatic ovarian cancer, OmGel performs as well as or better than the widely use Matrigel and does not induce additional phenotypic changes to ovarian cancer cells. Surprisingly, OmGel promotes pronounced morphological changes in cancer associated fibroblasts (CAFs). These changes were associated with the upregulation of proteins that define subsets of CAFs in tumour patient samples, highlighting the importance of using clinically and physiologically relevant matrices for in vitro studies. Hence, OmGel provides a step forward to study the biology of HGS omental metastasis. Metastasis in the omentum are also typical of other cancer types, particularly gastric cancer, implying the relevance of OmGel to study the biology of other highly lethal cancers.

Cascading effects of COVID-19 on population mobility and air quality: An exploration including place characteristics using geovisualization.

This study hypothesizes that public health responses to coronavirus disease 2019 (COVID-19), including a mandated restriction of activity (commonly called a 'lockdown') resulted in reduced transportation activities and changes in air quality in Texas, USA. This presented a natural experiment where population mobility and air quality before and after the lockdown could be compared. Changes in mobility were measured by SafeGraph mobility data (from opt-in smart phone applications that transmit location data) and air quality changes were based on NO2 concentrations measured by the European Space Agency's Sentinel-5 Precursor satellite (from the TROPOspheric Monitoring Instrument). The changes in population mobility and NO2 concentration between mid-March 2020 (lockdown initiated) and the end of 2020, as compared to the same time window in 2019, were the basis of exploring the lockdown hypothesis. Additionally, numerous socio-economic (place based) indicators were hypothesized to follow public health vulnerability assumptions based on COVID- 19 incidence patterns. This hypothesis was subjected to geovisualization techniques in order to find potential patterns and insights into the complex combinations of these place-based data. Our results suggest that simultaneously visualizing COVID-19, mobility, air quality and socio-economic data yields insights in underlying spatial processes related to public health policy decisions. The hypothesis that the lockdown resulted in reduced mobility and NO2 concentrations was found partially correct - this trend was observed in highly urbanized areas, but not in less populated areas. Data related public health vulnerability assumptions (e.g. a region's age, poverty, education, etc.) were agreed with in part, but disagreed with in part.

Exploring the socio-economic and environmental components of infectious diseases using multivariate geovisualization: West Nile Virus.

BACKGROUND: This study postulates that underlying environmental conditions and a susceptible population's socio-economic status should be explored simultaneously to adequately understand a vector borne disease infection risk. Here we focus on West Nile Virus (WNV), a mosquito borne pathogen, as a case study for spatial data visualization of environmental characteristics of a vector's habitat alongside human demographic composition for understanding potential public health risks of infectious disease. Multiple efforts have attempted to predict WNV environmental risk, while others have documented factors related to human vulnerability to the disease. However, analytical modeling that combines the two is difficult due to the number of potential explanatory variables, varying spatial resolutions of available data, and differing research questions that drove the initial data collection. We propose that the use of geovisualization may provide a glimpse into the large number of potential variables influencing the disease and help distill them into a smaller number that might reveal hidden and unknown patterns. This geovisual look at the data might then guide development of analytical models that can combine environmental and socio-economic data. METHODS: Geovisualization was used to integrate an environmental model of the disease vector's habitat alongside human risk factors derived from socio-economic variables. County level WNV incidence rates from California, USA, were used to define a geographically constrained study area where environmental and socio-economic data were extracted from 1,133 census tracts. A previously developed mosquito habitat model that was significantly related to WNV infected dead birds was used to describe the environmental components of the study area. Self-organizing maps found 49 clusters, each of which contained census tracts that were more similar to each other in terms of WNV environmental and socio-economic data. Parallel coordinate plots permitted visualization of each cluster's data, uncovering patterns that allowed final census tract mapping exposing complex spatial patterns contained within the clusters. RESULTS: Our results suggest that simultaneously visualizing environmental and socio-economic data supports a fuller understanding of the underlying spatial processes for risks to vector-borne disease. Unexpected patterns were revealed in our study that would be useful for developing future multilevel analytical models. For example, when the cluster that contained census tracts with the highest median age was examined, it was determined that those census tracts only contained moderate mosquito habitat risk. Likewise, the cluster that contained census tracts with the highest mosquito habitat risk had populations with moderate median age. Finally, the cluster that contained census tracts with the highest WNV human incidence rates had unexpectedly low mosquito habitat risk.

NRP2 as an Emerging Angiogenic Player; Promoting Endothelial Cell Adhesion and Migration by Regulating Recycling of α5 Integrin.

Angiogenesis relies on the ability of endothelial cells (ECs) to migrate over the extracellular matrix via integrin receptors to respond to an angiogenic stimulus. Of the two neuropilin (NRP) orthologs to be identified, both have been reported to be expressed on normal blood and lymphatic ECs, and to play roles in the formation of blood and lymphatic vascular networks during angiogenesis. Whilst the role of NRP1 and its interactions with integrins during angiogenesis has been widely studied, the role of NRP2 in ECs is poorly understood. Here we demonstrate that NRP2 promotes Rac-1 mediated EC adhesion and migration over fibronectin (FN) matrices in a mechanistically distinct fashion to NRP1, showing no dependence on ß3 integrin (ITGB3) expression, or VEGF stimulation. Furthermore, we highlight evidence of a regulatory crosstalk between NRP2 and α5 integrin (ITGA5) in ECs, with NRP2 depletion eliciting an upregulation of ITGA5 expression and disruptions in ITGA5 cellular organization. Finally, we propose a mechanism whereby NRP2 promotes ITGA5 recycling in ECs; NRP2 depleted ECs were found to exhibit reduced levels of total ITGA5 subunit recycling compared to wild-type (WT) ECs. Our findings expose NRP2 as a novel angiogenic player by promoting ITGA5-mediated EC adhesion and migration on FN.

Prioritizing riparian corridors for ecosystem restoration in urbanizing watersheds.

BACKGROUND: Riparian corridors can affect nutrient, organic matter, and sediment transport, all of which shape water quality in streams and connected downstream waters. When functioning riparian corridors remain intact, they provide highly valued water quality ecosystem services. However, in rapidly urbanizing watersheds, riparian corridors are susceptible to development modifications that adversely affect those ecosystem services. Protecting high quality riparian corridors or restoring low quality corridors are widely advocated as watershed level water quality management options for protecting those ecosystem services. The two approaches, protection or restoration, should be viewed as complementary by watershed managers and provide a foundation for targeting highly functioning riparian corridors for protection or for identifying poorly functioning corridors for restoration. Ascertaining which strategy to use is often motivated by a specific ecosystem service, for example water quality, upon which watershed management is focused. We have previously reported on a spatially explicit model that focused on identifying riparian corridors that have specific characteristics that make them well suited for purposes of preservation and protection focused on water quality. Here we hypothesize that focusing on restoration, rather than protection, can be the basis for developing a watershed level strategy for improving water quality in urbanizing watersheds. METHODS: The model described here represents a geographic information system (GIS) based approach that utilizes riparian characteristics extracted from 40-meter wide corridors centered on streams and rivers. The model focuses on drinking water reservoir watersheds that can be analyzed at the sub-watershed level. Sub-watershed riparian data (vegetation, soil erodibility and surface slope) are scaled and weighted based on watershed management theories for water quality, and riparian restoration scores are assigned. Those scores are used to rank order riparian zones -the lower the score the higher the priority for riparian restoration. RESULTS: The model was applied to 90 sub-watersheds in the watershed of an important drinking water reservoir in north central Texas, USA. Results from this study area suggest that corridor scores were found to be most correlated to the amount of: forested vegetation, residential land use, soils in the highest erodibility class, and highest surface slope (r 2 = 0.92, p < 0.0001). Scores allow watershed managers to rapidly focus on riparian corridors most in need of restoration. A beneficial feature of the model is that it also allows investigation of multiple scenarios of restoration strategies (e.g.,  revegetation, soil stabilization, flood plain leveling), giving watershed managers a tool to compare and contrast watershed level management plans.

Hypoxic cancer-associated fibroblasts increase NCBP2-AS2/HIAR to promote endothelial sprouting through enhanced VEGF signaling.

Intratumoral hypoxia causes the formation of dysfunctional blood vessels, which contribute to tumor metastasis and reduce the efficacy of therapeutic treatments. Blood vessels are embedded in the tumor stroma of which cancer-associated fibroblasts (CAFs) constitute a prominent cellular component. We found that hypoxic human mammary CAFs promoted angiogenesis in CAF-endothelial cell cocultures in vitro. Mass spectrometry-based proteomic analysis of the CAF secretome unraveled that hypoxic CAFs contributed to blood vessel abnormalities by altering their secretion of various pro- and anti-angiogenic factors. Hypoxia induced pronounced remodeling of the CAF proteome, including proteins that have not been previously related to this process. Among those, the uncharacterized protein NCBP2-AS2 that we renamed HIAR (hypoxia-induced angiogenesis regulator) was the protein most increased in abundance in hypoxic CAFs. Silencing of HIAR abrogated the pro-angiogenic and pro-migratory function of hypoxic CAFs by decreasing secretion of the pro-angiogenic factor VEGFA and consequently reducing VEGF/VEGFR downstream signaling in the endothelial cells. Our study has identified a regulator of angiogenesis and provides a map of hypoxia-induced molecular alterations in mammary CAFs.

The ß3-integrin endothelial adhesome regulates microtubule-dependent cell migration.

Integrin ß3 is seen as a key anti-angiogenic target for cancer treatment due to its expression on neovasculature, but the role it plays in the process is complex; whether it is pro- or anti-angiogenic depends on the context in which it is expressed. To understand precisely ß3's role in regulating integrin adhesion complexes in endothelial cells, we characterised, by mass spectrometry, the ß3-dependent adhesome. We show that depletion of ß3-integrin in this cell type leads to changes in microtubule behaviour that control cell migration. ß3-integrin regulates microtubule stability in endothelial cells through Rcc2/Anxa2-driven control of active Rac1 localisation. Our findings reveal that angiogenic processes, both in vitro and in vivo, are more sensitive to microtubule targeting agents when ß3-integrin levels are reduced.

Sulforaphane exerts anti-angiogenesis effects against hepatocellular carcinoma through inhibition of STAT3/HIF-1α/VEGF signalling.

Angiogenesis plays an important role in hepatocellular carcinoma (HCC), the inhibition of which is explored for cancer prevention and treatment. The dietary phytochemical sulforaphane (SFN) is known for its anti-cancer properties in vitro and in vivo; but until now, no study has focused on the role of SFN in HCC tumor angiogenesis. In the present study, in vitro cell models using a HCC cell line, HepG2, and human endothelial cells, HUVECs, as well as ex vivo and in vivo models have been used to investigate the anti-tumor and anti-angiogenic effect of SFN. The results showed that SFN decreased HUVEC cell viability, migration and tube formation, all of which are important steps in angiogenesis. More importantly, SFN markedly supressed HepG2-stimulated HUVEC migration, adhesion and tube formation; which may be due to its inhibition on STAT3/HIF-1α/VEGF signalling in HepG2 cells. In addition, SFN significantly reduced HepG2 tumor growth in a modified chick embryo chorioallantoic membrane (CAM) assay, associated with a decrease of HIF-1α and VEGF expression within tumors. Collectively, these findings provide new insights into the inhibitory effect of SFN on HCC tumor angiogenesis as well as tumor growth, and indicate that SFN has potential for the prevention and treatment of HCC.

Rapid, Secure Drug Testing Using Fingerprint Development and Paper Spray Mass Spectrometry.

BACKGROUND: Paper spray mass spectrometry (PS-MS) is a technique that has recently emerged and has shown excellent analytical sensitivity to a number of drugs in blood. As an alternative to blood, fingerprints have been shown to provide a noninvasive and traceable sampling matrix. Our goal was to validate the use of fingerprint samples to detect cocaine use. METHODS: Samples were collected on triangular pieces (168 mm2) of washed Whatman Grade I chromatography paper. Following application of internal standard, spray solvent and a voltage were applied to the paper before mass spectrometry detection. A fingerprint visualization step was incorporated into the analysis procedure by addition of silver nitrate solution and exposing the sample to ultraviolet light. RESULTS: Limits of detection for cocaine, benzoylecgonine, and methylecgonine were 1, 2, and 31 ng/mL respectively, with relative standard deviations < 33%. No matrix effects were observed. Analysis of 239 fingerprint samples yielded a 99% true-positive rate and a 2.5% false-positive rate, based on the detection of cocaine, benzoylecgonine, or methylecgonine with use of a single fingerprint. CONCLUSIONS: The method offers a qualitative and noninvasive screening test for cocaine use. The analysis method developed is rapid (4 min/sample) and requires no sample preparation.

An updated understanding of Texas bumble bee (Hymenoptera: Apidae) species presence and potential distributions in Texas, USA.

Texas is the second largest state in the United States of America, and the largest state in the contiguous USA at nearly 700,000 sq. km. Several Texas bumble bee species have shown evidence of declines in portions of their continental ranges, and conservation initiatives targeting these species will be most effective if species distributions are well established. To date, statewide bumble bee distributions for Texas have been inferred primarily from specimen records housed in natural history collections. To improve upon these maps, and help inform conservation decisions, this research aimed to (1) update existing Texas bumble bee presence databases to include recent (2007-2016) data from citizen science repositories and targeted field studies, (2) model statewide species distributions of the most common bumble bee species in Texas using MaxEnt, and (3) identify conservation target areas for the state that are most likely to contain habitat suitable for multiple declining species. The resulting Texas bumble bee database is comprised of 3,580 records, to include previously compiled museum records dating from 1897, recent field survey data, and vetted records from citizen science repositories. These data yielded an updated state species list that includes 11 species, as well as species distribution models (SDMs) for the most common Texas bumble bee species, including two that have shown evidence of range-wide declines: B. fraternus (Smith, 1854) and B. pensylvanicus (DeGeer, 1773). Based on analyses of these models, we have identified conservation priority areas within the Texas Cross Timbers, Texas Blackland Prairies, and East Central Texas Plains ecoregions where suitable habitat for both B. fraternus and B. pensylvanicus are highly likely to co-occur.

A comparison of least squares regression and geographically weighted regression modeling of West Nile virus risk based on environmental parameters.

BACKGROUND: The primary aim of the study reported here was to determine the effectiveness of utilizing local spatial variations in environmental data to uncover the statistical relationships between West Nile Virus (WNV) risk and environmental factors. Because least squares regression methods do not account for spatial autocorrelation and non-stationarity of the type of spatial data analyzed for studies that explore the relationship between WNV and environmental determinants, we hypothesized that a geographically weighted regression model would help us better understand how environmental factors are related to WNV risk patterns without the confounding effects of spatial non-stationarity. METHODS: We examined commonly mapped environmental factors using both ordinary least squares regression (LSR) and geographically weighted regression (GWR). Both types of models were applied to examine the relationship between WNV-infected dead bird counts and various environmental factors for those locations. The goal was to determine which approach yielded a better predictive model. RESULTS: LSR efforts lead to identifying three environmental variables that were statistically significantly related to WNV infected dead birds (adjusted R2 = 0.61): stream density, road density, and land surface temperature. GWR efforts increased the explanatory value of these three environmental variables with better spatial precision (adjusted R2 = 0.71). CONCLUSIONS: The spatial granularity resulting from the geographically weighted approach provides a better understanding of how environmental spatial heterogeneity is related to WNV risk as implied by WNV infected dead birds, which should allow improved planning of public health management strategies.

Suppression of ß3-integrin in mice triggers a neuropilin-1-dependent change in focal adhesion remodelling that can be targeted to block pathological angiogenesis.

Anti-angiogenic treatments against αvß3-integrin fail to block tumour growth in the long term, which suggests that the tumour vasculature escapes from angiogenesis inhibition through αvß3-integrin-independent mechanisms. Here, we show that suppression of ß3-integrin in mice leads to the activation of a neuropilin-1 (NRP1)-dependent cell migration pathway in endothelial cells via a mechanism that depends on NRP1's mobilisation away from mature focal adhesions following VEGF-stimulation. The simultaneous genetic targeting of both molecules significantly impairs paxillin-1 activation and focal adhesion remodelling in endothelial cells, and therefore inhibits tumour angiogenesis and the growth of already established tumours. These findings provide a firm foundation for testing drugs against these molecules in combination to treat patients with advanced cancers.

A determination of the spatial concordance between Lyme disease incidence and habitat probability of its primary vector Ixodes scapularis (black-legged tick).

The spatial distribution of Ixodes scapularis, the most common tick vector of the bacterium Borrelia burgdorferi, the cause of Lyme disease in humans, has not been studied previously in Texas, United States of America. It has only rarely been reported in this state, so its local, spatial relationship to the distribution of this disease is unknown. From an epidemiological perspective, one would tend to hypothesise that there should be a high degree of spatial concordance between habitat suitability for the tick and incidence of the disease. Both maximum-entropy modelling of the tick's habitat probability and modelling of human incidence of Lyme disease using spatially adaptive filters provide reliable portrayals of the spatial distributions of these phenomena. Even though rates of human cases of Lyme disease as well as rates of Ixodes ticks infected with Borrelia bacteria are both relatively low in Texas, the best data currently available indicate that the assumption of high levels of spatial concordance would not be correct in Texas (Kappa coefficient of agreement = 0.039). It will take substantially more data to provide conclusive findings and to understand the results reported here, but this study provides an approach to begin understanding the discrepancy.

Redefining the role(s) of endothelial αvß3-integrin in angiogenesis.

For nearly two decades now, the RGD (Arg-Gly-Asp)-binding αvß3-integrin has been a focus of anti-angiogenic drug design. These inhibitors are well-tolerated, but have shown only limited success in patients. Over the years, studies in ß3-integrin-knockout mice have shed some light on possible explanations for disappointing clinical outcomes. However, studying angiogenesis in ß3-integrin-knockout mice is a blunt tool to investigate ß3-integrin's role in pathological angiogenesis. Since establishing our laboratory at University of East Anglia (UEA), we have adopted more refined models of genetically manipulating the expression of the ß3-integrin subunit. The present review will highlight some of our findings from these models and describe how data from them have forced us to rethink how targeting αvß3-integrin expression affects tumour angiogenesis and cancer progression. Revisiting the fundamental biology behind how this integrin regulates tumour growth and angiogenesis, we believe, is the key not only to understanding how angiogenesis is normally co-ordinated, but also in success with drugs directed against it.

Modelling spatial concordance between Rocky Mountain spotted fever disease incidence and habitat probability of its vector Dermacentor variabilis (American dog tick).

The spatial distribution of Dermacentor variabilis, the most commonly identified vector of the bacterium Rickettsia rickettsii which causes Rocky Mountain spotted fever (RMSF) in humans, and the spatial distribution of RMSF, have not been previously studied in the south central United States of America, particularly in Texas. From an epidemiological perspective, one would tend to hypothesise that there would be a high degree of spatial concordance between the habitat suitability for the tick and the incidence of the disease. Both maximum-entropy modelling of the tick's habitat suitability and spatially adaptive filters modelling of the human incidence of RMSF disease provide reliable portrayals of the spatial distributions of these phenomenons. Even though rates of human cases of RMSF in Texas and rates of Dermacentor ticks infected with Rickettsia bacteria are both relatively low in Texas, the best data currently available allows a preliminary indication that the assumption of high levels of spatial concordance would not be correct in Texas (Kappa coefficient of agreement = 0.17). It will take substantially more data to provide conclusive findings, and to understand the results reported here, but this study provides an approach to begin understanding the discrepancy.

Borrelia, Ehrlichia, and Rickettsia spp. in ticks removed from persons, Texas, USA.

Data regarding the type, frequency, and distribution of tick-borne pathogens and bacterial agents are not widely available for many tick species that parasitize persons in the southern United States. We therefore analyzed the frequency and identity of pathogens and bacterial agents in ticks removed from humans and subsequently submitted to the Texas Department of State Health Services, Zoonosis Control Program, from October 1, 2004, through September 30, 2008. The data showed associations of bacterial agents and potential vectors. Tick-related illnesses may pose unidentified health risks in areas such as Texas, where incidence of human disease related to tick bites is low but well above zero and where ticks are not routinely suspected as the cause of disease. Cause, treatment, and prevention strategies can be better addressed through collecting sufficient data to establish baseline assessments of risk.

Near real-time monitoring and mapping of specific conductivity levels across Lake Texoma, USA.

A submersible sonde equipped with a specific conductivity probe, linked with a global positioning satellite receiver was developed, deployed on a small boat, and used to map spatial and temporal variations in specific conductivity in a large reservoir. 7,695 sample points were recorded during 8 sampling trips. Specific conductivity ranged from 442,uS/cm to 3,378,uS/cm over the nine-month study. The data showed five statistically different zones in the reservoir: 2 different riverine zones, 2 different riverine transition zones, and a lacustrine zone (the main lake zone). These data were imported to a geographic information system where they were spatially interpolated to generate 8 maps showing specific conductivity levels across the entire surface of the lake. The highly dynamic nature of water quality, due to the widely differing nature of the rivers that flow into the reservoir and the effect of large inflows of fresh water during winter storms is easily captured and visualized using this approach.