Breaks in MODIS time series portend vegetation change: verification using long-term data in an arid grassland ecosystem.

Frequency and severity of extreme climatic events are forecast to increase in the 21st century. Predicting how managed ecosystems may respond to climatic extremes is intensified by uncertainty associated with knowing when, where, and how long effects of extreme events will be manifest in an ecosystem. In water-limited ecosystems with high inter-annual variability in rainfall, it is important to be able to distinguish responses that result from seasonal fluctuations in rainfall from long-term directional increases or decreases in precipitation. A tool that successfully distinguishes seasonal from directional biomass responses would allow land managers to make informed decisions about prioritizing mitigation strategies, allocating human resource monitoring efforts, and mobilizing resources to withstand extreme climatic events. We leveraged long-term observations (2000-2013) of quadrat-level plant biomass at multiple locations across a semiarid landscape in southern New Mexico to verify the use of Normalized Difference Vegetation Index (NDVI) time series derived from 250-m Moderate Resolution Imaging Spectroradiometer (MODIS) data as a proxy for changes in aboveground productivity. This period encompassed years of sustained drought (2000-2003) and record-breaking high rainfall (2006 and 2008) followed by subsequent drought years (2011 through 2013) that resulted in a restructuring of plant community composition in some locations. Our objective was to decompose vegetation patterns derived from MODIS NDVI over this period into contributions from (1) the long-term trend, (2) seasonal cycle, and (3) unexplained variance using the Breaks for Additive Season and Trend (BFAST) model. BFAST breakpoints in NDVI trend and seasonal components were verified with field-estimated biomass at 15 sites that differed in species richness, vegetation cover, and soil properties. We found that 34 of 45 breaks in NDVI trend reflected large changes in mean biomass and 16 of 19 seasonal breaks accompanied changes in the contribution to biomass by perennial and/or annual grasses. The BFAST method using satellite imagery proved useful for detecting previously reported ground-based changes in vegetation in this arid ecosystem. We demonstrate that time series analysis of NDVI data holds potential for monitoring landscape condition in arid ecosystems at the large spatial scales needed to differentiate responses to a changing climate from responses to seasonal variability in rainfall.

Errors in soil maps: The need for better on-site estimates and soil map predictions.

High-quality soil maps are urgently needed by diverse stakeholders, but errors in existing soil maps are often unknown, particularly in countries with limited soil surveys. To address this issue, we used field soil data to assess the accuracy of seven spatial soil databases (Digital Soil Map of the World, Namibian Soil and Terrain Digital Database, Soil and Terrain Database for Southern Africa, Harmonized World Soil Database, SoilGrids1km, SoilGrids250m, and World Inventory of Soil Property Estimates) using topsoil texture as an example soil property and Namibia as a case study area. In addition, we visually compared topsoil texture maps derived from these databases. We found that the maps showed the correct topsoil texture in only 13% to 42% of all test sites, with substantial confusion occurring among all texture categories, not just those in close proximity in the soil texture triangle. Visual comparisons of the maps moreover showed that the maps differ greatly with respect to the number, types, and spatial distribution of texture classes. The topsoil texture information provided by the maps is thus sufficiently inaccurate that it would result in significant errors in a number of applications, including irrigation system design and predictions of potential forage and crop productivity, water runoff, and soil erosion. Clearly, the use of these existing maps for policy- and decision-making is highly questionable and there is a critical need for better on-site estimates and soil map predictions. We propose that mobile apps, citizen science, and crowdsourcing can help meet this need.

Use of adenoviral vectors to target chemotherapy to tumor vascular endothelial cells suppresses growth of breast cancer and melanoma.

To target chemotherapy to tumor vascular endothelial cells (TVECs), we created the AdTie2RprCDFib(knob-RGD+) vector by inserting into an AdEasy adenoviral vector (Ad) backbone: (i) the cytosine deaminase (CD) gene driven by the Tie2 receptor promoter (Tie2Rpr) into the E1 region of Ad; (ii) mutations that reduce binding of the fiber knob to the Coxsackie adenovirus receptor (CAR); and (iii) the RGD peptide into the H1 loop of fiber for binding to the alpha(V)beta(3) integrin receptors on TVECs. To reduce uptake of the AdTie2RprCDFib(knob-RGD+) by reticuloendothelial (RE) and liver cells, we intravenously (i.v.) injected Hetastarch and low-dose Ad (one million vector particles (VPs)) prior to i.v. injection of a therapeutic dose (one billion VPs) of the AdTie2RprCDFib(knob-RGD+) vector. This treatment induced regressions of N202 breast cancer and B16 melanoma without toxicity to normal tissues. We showed that the tumor regression was induced by infection of the TVECs and not by the infection of tumor cells by the AdTie2RprCDFib(knob-RGD+) vector.

Chemotherapy targeted to cancer tissue potentiates antigen-specific immune response induced by vaccine for in vivo antigen loading and activation of dendritic cells.

Our laboratory has created an Ad-sig-TAA/ecdCD40L vaccine platform designed to activate dendritic cells (DCs). Two subcutaneous (s.c.) injections of the TAA/ecdCD40L protein following the s.c. injection of the Ad-sig-TAA/ecdCD40L vector (TAA/ecdCD40L VPP vaccine) further increases the levels of the tumor-associated antigen (TAA)-specific CD8 effector T cells induced by the vector. We tested the combined effect of chemotherapy-induced destruction of tumor cells and TAA/ecdCD40L VPP vaccine which further increases the levels of TAA available to the DCs at the time of vaccination. The chemotherapy was delivered selectively to the tumor cells using intratumoral (i.t.) injection of the AdCDIRESE1A vector followed by intraperitoneal (i.p.) 5-fluorocytosine (5FC). The 5-fluorouracil (5FU) produced in the vector infected the tumor cells, destroys them and releases the TAA for processing and presentation by the DCs. This mode of delivery spares the TAA CD8 effector T cells from the destructive effect of the 5FU when their proliferation is induced by the vaccine. Test mice treated with both the s.c. administered TAA/ecdCD40L VPP vaccine and the AdCDIRESE1A/5FC chemosensitization vector had the smallest tumor volumes and survived longer than mice treated with either of these agents alone (P < 0.001).

Bioavailability and fate of phosphorus in constructed wetlands receiving agricultural runoff in the San Joaquin Valley, California.

Elevated nutrient concentrations in agricultural runoff contribute to seasonal eutrophication and hypoxia in the lower portion of the San Joaquin River, California. Interception and filtration of agricultural runoff by constructed wetlands may improve water quality of return flows ultimately destined for major water bodies. This study evaluated the efficacy of two small flow-through wetlands (2.3 and 7.3 ha; hydraulic residence time = 11 and 31 h) for attenuating various forms of P from irrigation tailwaters during the 2005 irrigation season (May to September). Our goal was to examine transformations and removal efficiencies for bioavailable P in constructed wetlands. Inflow and outflow water volumes were monitored continuously and weekly water samples were collected to measure total P (TP), dissolved-reactive P (DRP), and bioavailable P (BAP). Suspended sediment was characterized and fractionated into five operationally-defined P fractions (i.e., NH4Cl, bicarbonate-dithionite, NaOH, HCl, residual) to evaluate particulate P (PP) transformations. DRP was the major source of BAP with the particulate fraction contributing from 11 to 26%. On a seasonal basis, wetlands removed 55 to 65% of PP, 61 to 63% of DRP, 57 to 62% of BAP, and 88 to 91% of TSS. Sequential fractionation indicated that the bioavailable fraction of PP was largely associated with clay-sized particles that remain in suspension, while less labile P forms preferentially settle with coarser sediment. Thus, removal of potentially bioavailable PP is dependent on factors that promote particle settling and allow for the removal of colloids. This study suggests that treatment of tailwaters in small, flow-through wetlands can effectively remove BAP. Wetland design and management strategies that enhance sedimentation of colloids can improve BAP retention efficiency.

Applying fingerprint FTIR spectroscopy and chemometrics to assess soil ecosystem disturbance and recovery.

The assessment and monitoring of soil disturbance and its effect on soil quality (i.e., ability to support a range of ecosystem services) has been hindered due to the shortcomings of many traditional analytical techniques (e.g., soil enzyme activities, microbial incubations), including: high cost, long-term time investment and difficulties with data interpretation. Consequently, there is a critical need to develop a rapid and repeatable approach for quantifying changes in soil quality that will provide an assessment of the current status, condition and trend of natural and managed ecosystems. Here we report on a rapid, high-throughput approach to develop an ecological 'fingerprint' of a soil using Fourier transformed infrared (FTIR) spectroscopy and chemometric modeling, and its application to assess soil ecosystem status and trend. This methodology was applied in a highly disturbed forest ecosystem over a 19-year sampling period to detect changes in soil quality (detected via changes in spectral properties), resulting from changes in dynamic soil properties (e.g., soil organic matter, reactive mineralogy). Two chemometric statistical techniques (i.e., hierarchical clustering analysis and discriminate analysis of principal components) were evaluated for interpreting and quantifying similarities/dissimilarities between samples utilizing the entire FTIR spectra (i.e., fingerprint) from each sample. We found that this approach provided a means for clearly discriminating between degraded soils, soils in recovery and reference soils. Results from fingerprint FTIR analysis illustrate its power and potential for the monitoring and assessment of soil quality and soil landscape change.

Endothelin-3 growth factor levels decreased in cervical cancer compared with normal cervical epithelial cells.

We used cDNA microarray analysis of RNA extracted from normal, dysplastic, and cancerous cervical tissues to identify the changes in gene expression during the procession from normal to cancerous cervical epithelial cells. We found the expression of 5 genes in cancerous cervical epithelial cells that were not found in normal cervical epithelial cells, among which were lymphoid-restricted membrane protein, protease serine 2, WD repeat domain 59, thyrotropin-releasing hormone degrading enzyme, and the endothelin-3 growth factor. We then analyzed the expression levels of endothelin growth factors 1, 2, and 3 (ET-1, ET-2, and ET-3) and their receptors A and B (ETR-A and ETR-B) by reverse transcriptase-polymerase chain reaction in 3 cervical cancer cell lines and by immunohistochemical staining in cervical normal, dysplastic, and cancer tissues. ET-1, ET-2, and ET-3 growth factor levels were detectable in the maturing layer of cervical epithelium but not in the germinal layer. All 3 growth factors (ET-1, ET-2, and ET-3) were detected in the cytoplasm of the maturing normal cervical epithelial cells. In addition, there were decreased levels of ET-3 and increased levels of ET-1, ET-2, ETR-A, and ETR-B in cancerous cervical epithelial cells compared with normal cervical epithelial cells. These results suggest that the reduction of ET-3 growth factor levels may be important in the transition from normal to cancerous cervical epithelium.

Antitumor immune response induced by i.t. injection of vector-activated dendritic cells and chemotherapy suppresses metastatic breast cancer.

S.c. injection of the Ad-sig-tumor-associated antigen (TAA)/ecdCD40L vector vaccine has been shown to induce a CD8 immune response against TAA for up to 1 year. The first goal of this article is to test if the injection of autologous dendritic cells infected ex vivo with the Ad-sig-TAA/ecdCD40L can increase the immune response induced against TAA. The second goal is to test the effect of adding local chemotherapy in the form of i.t. injection of the AdCDIRESE1A vector-directed chemotherapy on the immune response induced by i.t. injection of adenoviral vector-activated dendritic cells. The results show that the i.t. injection of the AdCDIRESE1A chemotherapy sensitization vector, which encodes the cytosine deaminase chemotherapy sensitization transcription unit, to the i.t. injection of Ad-sig-ecdCD40L vector-infected dendritic cells increased the level of suppression of the growth of the CCL-51 breast cancer cells. The combination of i.t. injection of the AdCDIRESE1A chemotherapy sensitization vector and Ad-sig-ecdCD40L vector-infected dendritic cells into s.c. CCL-51 breast cancer nodules suppressed the growth of uninjected metastatic tumor nodules in the lung. Finally, adding the i.t. injection of the AdCDIRESE1A chemotherapy sensitization vector to the i.t. administration of dendritic cells infected with a rat HER-2/neu (rH2N)-expressing vector (Ad-sig-rH2N/ecdCD40L) led to the induction of rH2N-specific antitumoral immunity in rH2N transgenic mice (which are anergic to the rH2N antigen). This anti-rH2N immune response suppressed the growth of established H2N-positive NT2 breast cancer more efficiently than did the vector-targeted chemotherapy or Ad-sig-rH2N/ecdCD40L-infected dendritic cell vaccine alone.

A hyper-temporal remote sensing protocol for high-resolution mapping of ecological sites.

Ecological site classification has emerged as a highly effective land management framework, but its utility at a regional scale has been limited due to the spatial ambiguity of ecological site locations in the U.S. or the absence of ecological site maps in other regions of the world. In response to these shortcomings, this study evaluated the use of hyper-temporal remote sensing (i.e., hundreds of images) for high spatial resolution mapping of ecological sites. We posit that hyper-temporal remote sensing can provide novel insights into the spatial variability of ecological sites by quantifying the temporal response of land surface spectral properties. This temporal response provides a spectral 'fingerprint' of the soil-vegetation-climate relationship which is central to the concept of ecological sites. Consequently, the main objective of this study was to predict the spatial distribution of ecological sites in a semi-arid rangeland using a 28-year time series of normalized difference vegetation index from Landsat TM 5 data and modeled using support vector machine classification. Results from this study show that support vector machine classification using hyper-temporal remote sensing imagery was effective in modeling ecological site classes, with a 62% correct classification. These results were compared to Gridded Soil Survey Geographic database and expert delineated maps of ecological sites which had a 51 and 89% correct classification, respectively. An analysis of the effects of ecological state on ecological site misclassifications revealed that sites in degraded states (e.g., shrub-dominated/shrubland and bare/annuals) had a higher rate of misclassification due to their close spectral similarity with other ecological sites. This study identified three important factors that need to be addressed to improve future model predictions: 1) sampling designs need to fully represent the range of both within class (i.e., states) and between class (i.e., ecological sites) spectral variability through time, 2) field sampling protocols that accurately characterize key soil properties (e.g., texture, depth) need to be adopted, and 3) additional environmental covariates (e.g. terrain attributes) need to be evaluated that may help further differentiate sites with similar spectral signals. Finally, the proposed hyper-temporal remote sensing framework may provide a standardized approach to evaluate and test our ecological site concepts through examining differences in vegetation dynamics in response to climatic variability and other drivers of land-use change. Results from this study demonstrate the efficacy of the hyper-temporal remote sensing approach for high resolution mapping of ecological sites, and highlights its utility in terms of reduced cost and time investment relative to traditional manual mapping approaches.

Vector targeting makes 5-fluorouracil chemotherapy less toxic and more effective in animal models of epithelial neoplasms.

PURPOSE: 5-fluorouracil (5-FU) has been combined in the past with other drugs for the combination chemotherapy for cancers of the breast, ovary, and colon. These drug regimens were limited by the fact that 5-FU fails to kill nondividing cancer cells at the doses that are safe to deliver. The goal of the present study is to test the feasibility of replacing 5-FU in established 5-FU combination chemotherapy with the Ad-LpCDIRESE1A/5-fluorocytosine (5-FC) system for the purpose of reducing toxicity and increasing efficacy. EXPERIMENTAL DESIGN: We have replaced 5-FU in the weekly combination of CPT-11, folinic acid (FA) and 5-FU chemotherapy by 5-FC and an adenoviral vector that carries the L-plastin (Lp) tumor-specific promoter-driven transcription unit encoding the cytosine deaminase gene linked to the E1A gene by an internal ribosomal entry site element. This combination is called "genetic combination therapy." The goal of using the vector was to decrease the toxicity to normal tissue and to increase the efficacy of therapy in the cancer cells by increasing the concentration of 5-FU sufficiently high that even nondividing cancer cells would be killed by 5-FU through its incorporation into mRNA and consequent inhibition of synthesis of functional proteins. We compared the in vivo efficacy of the genetic combination therapy with the conventional combination chemotherapy in a mouse colon cancer model. RESULTS: Both replication-competent and -noncompetent adenoviral vectors carrying an L-plastin-driven cytosine deaminase transcription unit when combined with 5-FC, CPT-11, and FA showed increased in vitro therapeutic activity that was significantly higher than that of the conventional chemotherapy combination. Tumor-bearing mice treated with the genetic combination therapy showed a statistically significant advantage in terms of increased response rate, response duration, survival, and reduced toxicity when compared with tumor-bearing mice treated with the conventional combination chemotherapy. CONCLUSIONS: Replacement of 5-FU in 5-FU-based combination chemotherapy with the Ad-LpCDIRESE1A vector and 5-FU reduces toxicity and increases efficacy. This is a concept that could be potentially applied widely for many forms of cancer treatment.

Conditionally replication-competent adenoviral vectors with enhanced infectivity for use in gene therapy of melanoma.

To generate vector Ad.Tyr-E1A, which is cytolytic for tyrosinase-positive melanoma cells, we replaced the adenoviral E1A promoter with a human tyrosinase enhancer/promoter. To overcome the low transduction efficiency in populations of melanoma cells that exhibit a low level of the coxsackievirus-adenovirus receptor (CAR), we inserted an RGD-4C peptide into the HI loop of the fiber knob domain of the Ad.Tyr-E1A vector. The resulting vector was named Ad.Tyr-E1A(RGD). As a result of these changes, the transduction efficiency of the RGD-modified vector was increased both in vitro and in vivo. Western blot analysis proved that infection of cells with the Ad.Tyr-E1A(RGD) vector led to expression of the E1A gene selectively in tyrosinase-positive melanoma cell lines, but not in tyrosinase-negative cell lines. The Ad.Tyr-E1A(RGD) vector was as potent in its cytotoxic effect as a tumor nonselective vector (Ad.CMV-E1A) in tyrosinase-positive melanoma cell lines. The Ad.Tyr-E1A(RGD) vector produced a higher vector particle yield in tumor cells than did the Ad.Tyr-E1A vector. Intratumoral injection of the Ad.Tyr-E1A(RGD) vector into xenotransplanted human melanoma tumors led to tumor regression in vivo. The combination of tumor-specific replication and enhanced infectivity generates a more potent CRAD vector for gene therapy of melanoma.

Vector prime/protein boost vaccine that overcomes defects acquired during aging and cancer.

We showed that the Ad-sig-TAA/ecdCD40L vaccine induces a tumor suppressive immune response to the hMUC-1 and rH2N tumor-associated self Ags (TAA) and to the Annexin A1 tumor vascular Ag, even in mice in which anergy exists to these Ags. When the TAA/ecdCD40L protein is given s.c. as a boost following the Ad-sig-TAA/ecdCD40L vector, the levels of the TAA-specific CD8 T cells and Abs increase dramatically over that seen with vector alone, in young (2-mo-old) as well as old (18-mo-old) mice. The Abs induced against hMUC-1 react with human breast cancer. This vaccine also induces a 4-fold decrement of negative regulatory CD4CD25FOXP3-T cells in the tumor tissue of 18-mo-old mice. These results suggest that the Ad-sig-TAA/ecdCD40L vector prime-TAA/ecdCD40L protein boost vaccine platform may be valuable in reducing postsurgery recurrence in a variety of epithelial neoplasms.

Multistep process through which adenoviral vector vaccine overcomes anergy to tumor-associated antigens.

Our goal in the present work was to characterize the multiple steps involved in overcoming the anergy that exists in tumor hosts to tumor-associated antigen (TAA). Our studies showed that the subcutaneous injection of the Ad-sig-TAA/ecdCD40L vector resulted in secretion of the TAA/ecdCD40L protein for at least 10 days from infected cells. Binding of the TAA/ecdCD40L protein to dendritic cells (DCs) resulted in the induction of CCR-7 chemokine receptor expression and cytokine release. This was followed by migration of the DCs to regional lymph nodes. Tetramer staining, enzyme-linked immunospot (ELISPOT) assay, and cytotoxicity assay all showed that the Ad-sig-TAA/ecdCD40L vector increased the levels of splenic CD8(+) T cells specific for the 2 TAAs (human MUC1 [hMUC1] and HPV E7) tested. Vaccination with the Ad-sighMUC1/ecdCD40L vector suppressed the growth of hMUC1 antigen-positive tumor cells in 100% of the test mice that were previously anergic to the hMUC1 antigen. These data suggest that Ad-sig-TAA-ecd/ecdCD40L vector injections may be of value in treating the many epithelial malignancies in which TAA-like hMUC1 is overexpressed.