A Beginner's Guide to Eddy Covariance: Methodology and Its Applications to Photosynthesis.

The eddy covariance technique, commonly applied using flux towers, enables the investigation of greenhouse gas (e.g., carbon dioxide, methane, nitrous oxide) and energy (latent and sensible heat) fluxes between the biosphere and the atmosphere. Through measuring carbon fluxes in particular, eddy covariance flux towers can give insight into how ecosystem scale photosynthesis (i.e., gross primary productivity) changes over time in response to climate and management. This chapter is designed to be a beginner's guide to understanding the eddy covariance method and how it can be applied in photosynthesis research. It introduces key concepts and assumptions that apply to the method, what materials are required to set up a flux tower, as well as practical advice for site installation, maintenance, data management, and postprocessing considerations. This chapter also includes examples of what can go wrong, with advice on how to correct these errors if they arise. This chapter has been crafted to help new users design, install, and manage the best towers to suit their research needs and includes additional resources throughout to further guide successful eddy covariance research activities.

Improved net carbon budgets in the US Midwest through direct measured impacts of enhanced weathering.

Terrestrial enhanced weathering (EW) through the application of Mg- or Ca-rich rock dust to soil is a negative emission technology with the potential to address impacts of climate change. The effectiveness of EW was tested over 4 years by spreading ground basalt (50 t ha-1 year-1 ) on maize/soybean and miscanthus cropping systems in the Midwest US. The major elements of the carbon budget were quantified through measurements of eddy covariance, soil carbon flux, and biomass. The movement of Mg and Ca to deep soil, released by weathering, balanced by a corresponding alkalinity flux, was used to measure the drawdown of CO2 , where the release of cations from basalt was measured as the ratio of rare earth elements to base cations in the applied rock dust and in the surface soil. Basalt application stimulated peak biomass and net primary production in both cropping systems and caused a small but significant stimulation of soil respiration. Net ecosystem carbon balance (NECB) was strongly negative for maize/soybean (-199 to -453 g C m-2 year-1 ) indicating this system was losing carbon to the atmosphere. Average EW (102 g C m-2 year-1 ) offset carbon loss in the maize/soybean by 23%-42%. NECB of miscanthus was positive (63-129 g C m-2 year-1 ), indicating carbon gain in the system, and EW greatly increased inorganic carbon storage by an additional 234 g C m-2 year-1 . Our analysis indicates a co-deployment of a perennial biofuel crop (miscanthus) with EW leads to major wins-increased harvested yields of 29%-42% with additional carbon dioxide removal (CDR) of 8.6 t CO2 ha-1 year-1 . EW applied to maize/soybean drives a CDR of 3.7 t CO2 ha-1 year-1 , which partially offsets well-established carbon losses from soil from this crop rotation. EW applied in the US Midwest creates measurable improvements to the carbon budgets perennial bioenergy crops and conventional row crops.

A 14-year-old Girl with Right-Sided Weakness and Speech Difficulty.

We herein report a case of Childhood Primary Central Nervous System (CNS) Angiitis. This case consisted of a 14-year-old girl who presented with right-sided weakness, aphasia, and a fever. A Magnetic Resonance Imaging of the brain showed multifocal areas of ischemia. Magnetic Resonance Angiography of the head and neck showed narrowing and irregularities of the left middle cerebral artery and right posterior cerebral artery. Cerebrospinal Fluid studies showed a lymphocytic pleocytosis and brain biopsy revealed leptomeningeal and perivascular inflammation. The epidemiology, presenting symptoms, work-up, pathophysiology, diagnostic criteria, and treatment of Childhood Primary CNS angiitis are discussed. This case serves as a reminder that when pediatric patients present with stroke-like symptoms inflammatory etiologies including Primary CNS Angiitis must be considered and treated appropriately in a time sensitive manner.

Bridge to the future: Important lessons from 20 years of ecosystem observations made by the OzFlux network.

In 2020, the Australian and New Zealand flux research and monitoring network, OzFlux, celebrated its 20th anniversary by reflecting on the lessons learned through two decades of ecosystem studies on global change biology. OzFlux is a network not only for ecosystem researchers, but also for those 'next users' of the knowledge, information and data that such networks provide. Here, we focus on eight lessons across topics of climate change and variability, disturbance and resilience, drought and heat stress and synergies with remote sensing and modelling. In distilling the key lessons learned, we also identify where further research is needed to fill knowledge gaps and improve the utility and relevance of the outputs from OzFlux. Extreme climate variability across Australia and New Zealand (droughts and flooding rains) provides a natural laboratory for a global understanding of ecosystems in this time of accelerating climate change. As evidence of worsening global fire risk emerges, the natural ability of these ecosystems to recover from disturbances, such as fire and cyclones, provides lessons on adaptation and resilience to disturbance. Drought and heatwaves are common occurrences across large parts of the region and can tip an ecosystem's carbon budget from a net CO2 sink to a net CO2 source. Despite such responses to stress, ecosystems at OzFlux sites show their resilience to climate variability by rapidly pivoting back to a strong carbon sink upon the return of favourable conditions. Located in under-represented areas, OzFlux data have the potential for reducing uncertainties in global remote sensing products, and these data provide several opportunities to develop new theories and improve our ecosystem models. The accumulated impacts of these lessons over the last 20 years highlights the value of long-term flux observations for natural and managed systems. A future vision for OzFlux includes ongoing and newly developed synergies with ecophysiologists, ecologists, geologists, remote sensors and modellers.

Clinical performance of a commercially available thymidine kinase 1 assay for diagnosis of lymphoma in 42 hospitalized horses (2017-2020).

BACKGROUND: Antemortem definitive diagnosis of lymphoma in horses is often difficult. Thymidine kinase 1 (TK1) assay is a potentially useful biomarker for lymphoma in horses. HYPOTHESIS/OBJECTIVES: To report the clinical performance of a commercially available TK1 assay for diagnosis of lymphoma in horses. We hypothesized that there would be no association between serum TK1 activity and a diagnosis of lymphoma in horses. ANIMALS: Forty-two hospitalized horses, 14 with a definitive diagnosis of lymphoma, 4 with other neoplasia, and 24 with inflammatory disease. METHODS: Retrospective medical record review, groups were compared via Kruskal-Wallis and Mann-Whitney tests, and logistic regression was performed. RESULTS: Median (range) TK1 was 3 U/L (0.4-17.7 U/L) in horses with lymphoma and 3.9 U/L (0.8-94 U/L) in horses without lymphoma (P = .59). There was no significant difference in total protein between horses with and without lymphoma (6.6 g/dL [5.5-8.3 g/dL] vs 6.6 g/dL [4.7-10.4 g/dL]; P = .83). There was no significant difference in fibrinogen between horses with and without lymphoma (447 [100-1364] mg/dL vs 433 [291-2004] mg/dL; P = .47). On logistic regression, serum TK1 activity was not associated with a diagnosis of lymphoma (odds ratio, 0.97; 95% confidence interval, 0.9-1.05, P = .48). CONCLUSION AND CLINICAL IMPORTANCE: Serum TK1 values were not predictive of lymphoma diagnosis in this cohort of horses.

A review of transformative strategies for climate mitigation by grasslands.

Grasslands can significantly contribute to climate mitigation. However, recent trends indicate that human activities have switched their net cooling effect to a warming effect due to management intensification and land conversion. This indicates an urgent need for strategies directed to mitigate climate warming while enhancing productivity and efficiency in the use of land and natural (nutrients, water) resources. Here, we examine the potential of four innovative strategies to slow climate change including: 1) Adaptive multi-paddock grazing that consists of mimicking how ancestral herds roamed the Earth; 2) Agrivoltaics that consists of simultaneously producing food and energy from solar panels on the same land area; 3) Agroforestry with a reverse phenology tree species, Faidherbia (Acacia) albida, that has the unique trait of being photosynthetically active when intercropped herbaceous plants are dormant; and, 4) Enhanced Weathering, a negative emission technology that removes atmospheric CO2 from the atmosphere. Further, we speculate about potential unknown consequences of these different management strategies and identify gaps in knowledge. We find that all these strategies could promote at least some of the following benefits of grasslands: CO2 sequestration, non-CO2 GHG mitigation, productivity, resilience to climate change, and an efficient use of natural resources. However, there are obstacles to be overcome. Mechanistic assessment of the ecological, environmental, and socio-economic consequences of adopting these strategies at large scale are urgently needed to fully assess the potential of grasslands to provide food, energy and environmental security.

Quantifying high-temperature stress on soybean canopy photosynthesis: The unique role of sun-induced chlorophyll fluorescence.

High temperature and accompanying high vapor pressure deficit often stress plants without causing distinctive changes in plant canopy structure and consequential spectral signatures. Sun-induced chlorophyll fluorescence (SIF), because of its mechanistic link with photosynthesis, may better detect such stress than remote sensing techniques relying on spectral reflectance signatures of canopy structural changes. However, our understanding about physiological mechanisms of SIF and its unique potential for physiological stress detection remains less clear. In this study, we measured SIF at a high-temperature experiment, Temperature Free-Air Controlled Enhancement, to explore the potential of SIF for physiological investigations. The experiment provided a gradient of soybean canopy temperature with 1.5, 3.0, 4.5, and 6.0°C above the ambient canopy temperature in the open field environments. SIF yield, which is normalized by incident radiation and the fraction of absorbed photosynthetically active radiation, showed a high correlation with photosynthetic light use efficiency (r = 0.89) and captured dynamic plant responses to high-temperature conditions. SIF yield was affected by canopy structural and plant physiological changes associated with high-temperature stress (partial correlation r = 0.60 and -0.23). Near-infrared reflectance of vegetation, only affected by canopy structural changes, was used to minimize the canopy structural impact on SIF yield and to retrieve physiological SIF yield (ΦF ) signals. ΦF further excludes the canopy structural impact than SIF yield and indicates plant physiological variability, and we found that ΦF outperformed SIF yield in responding to physiological stress (r = -0.37). Our findings highlight that ΦF sensitively responded to the physiological downregulation of soybean gross primary productivity under high temperature. ΦF , if reliably derived from satellite SIF, can support monitoring regional crop growth and different ecosystems' vegetation productivity under environmental stress and climate change.

The effect of increasing temperature on crop photosynthesis: from enzymes to ecosystems.

As global land surface temperature continues to rise and heatwave events increase in frequency, duration, and/or intensity, our key food and fuel cropping systems will likely face increased heat-related stress. A large volume of literature exists on exploring measured and modelled impacts of rising temperature on crop photosynthesis, from enzymatic responses within the leaf up to larger ecosystem-scale responses that reflect seasonal and interannual crop responses to heat. This review discusses (i) how crop photosynthesis changes with temperature at the enzymatic scale within the leaf; (ii) how stomata and plant transport systems are affected by temperature; (iii) what features make a plant susceptible or tolerant to elevated temperature and heat stress; and (iv) how these temperature and heat effects compound at the ecosystem scale to affect crop yields. Throughout the review, we identify current advancements and future research trajectories that are needed to make our cropping systems more resilient to rising temperature and heat stress, which are both projected to occur due to current global fossil fuel emissions.

Emerging approaches to measure photosynthesis from the leaf to the ecosystem.

Measuring photosynthesis is critical for quantifying and modeling leaf to regional scale productivity of managed and natural ecosystems. This review explores existing and novel advances in photosynthesis measurements that are certain to provide innovative directions in plant science research. First, we address gas exchange approaches from leaf to ecosystem scales. Leaf level gas exchange is a mature method but recent improvements to the user interface and environmental controls of commercial systems have resulted in faster and higher quality data collection. Canopy chamber and micrometeorological methods have also become more standardized tools and have an advanced understanding of ecosystem functioning under a changing environment and through long time series data coupled with community data sharing. Second, we review proximal and remote sensing approaches to measure photosynthesis, including hyperspectral reflectance- and fluorescence-based techniques. These techniques have long been used with aircraft and orbiting satellites, but lower-cost sensors and improved statistical analyses are allowing these techniques to become applicable at smaller scales to quantify changes in the underlying biochemistry of photosynthesis. Within the past decade measurements of chlorophyll fluorescence from earth-orbiting satellites have measured Solar Induced Fluorescence (SIF) enabling estimates of global ecosystem productivity. Finally, we highlight that stronger interactions of scientists across disciplines will benefit our capacity to accurately estimate productivity at regional and global scales. Applying the multiple techniques outlined in this review at scales from the leaf to the globe are likely to advance understanding of plant functioning from the organelle to the ecosystem.

High-throughput field phenotyping using hyperspectral reflectance and partial least squares regression (PLSR) reveals genetic modifications to photosynthetic capacity.

Spectroscopy is becoming an increasingly powerful tool to alleviate the challenges of traditional measurements of key plant traits at the leaf, canopy, and ecosystem scales. Spectroscopic methods often rely on statistical approaches to reduce data redundancy and enhance useful prediction of physiological traits. Given the mechanistic uncertainty of spectroscopic techniques, genetic modification of plant biochemical pathways may affect reflectance spectra causing predictive models to lose power. The objectives of this research were to assess over two separate years, whether a predictive model can represent natural and imposed variation in leaf photosynthetic potential for different crop cultivars and genetically modified plants, to assess the interannual capabilities of a partial least square regression (PLSR) model, and to determine whether leaf N is a dominant driver of photosynthesis in PLSR models. In 2016, a PLSR analysis of reflectance spectra coupled with gas exchange data was used to build predictive models for photosynthetic parameters including maximum carboxylation rate of Rubisco (V c,max ), maximum electron transport rate (J max ) and percentage leaf nitrogen ([N]). The model was developed for wild type and genetically modified plants that represent a wide range of photosynthetic capacities. Results show that hyperspectral reflectance accurately predicted V c,max, J max and [N] for all plants measured in 2016. Applying these PLSR models to plants grown in 2017 resulted in a strong predictive ability relative to gas exchange measurements for V c,max, but not for J max, and not for genotypes unique to 2017. Building a new model including data collected in 2017 resulted in more robust predictions, with R2 increases of 17% for V c,max . and 13% J max . Plants generally have a positive correlation between leaf nitrogen and photosynthesis, however, tobacco with reduced Rubisco (SSuD) had significantly higher [N] despite much lower V c,max. The PLSR model was able to accurately predict both lower V c,max and higher leaf [N] for this genotype suggesting that the spectral based estimates of V c,max and leaf nitrogen [N] are independent. These results suggest that the PLSR model can be applied across years, but only to genotypes used to build the model and that the actual mechanism measured with the PLSR technique is not directly related to leaf [N]. The success of the leaf-scale analysis suggests that similar approaches may be successful at the canopy and ecosystem scales but to use these methods across years and between genotypes at any scale, application of accurately populated physical based models based on radiative transfer principles may be required.

Small-molecule allosteric activators of PDE4 long form cyclic AMP phosphodiesterases.

Cyclic AMP (cAMP) phosphodiesterase-4 (PDE4) enzymes degrade cAMP and underpin the compartmentalization of cAMP signaling through their targeting to particular protein complexes and intracellular locales. We describe the discovery and characterization of a small-molecule compound that allosterically activates PDE4 long isoforms. This PDE4-specific activator displays reversible, noncompetitive kinetics of activation (increased Vmax with unchanged Km), phenocopies the ability of protein kinase A (PKA) to activate PDE4 long isoforms endogenously, and requires a dimeric enzyme assembly, as adopted by long, but not by short (monomeric), PDE4 isoforms. Abnormally elevated levels of cAMP provide a critical driver of the underpinning molecular pathology of autosomal dominant polycystic kidney disease (ADPKD) by promoting cyst formation that, ultimately, culminates in renal failure. Using both animal and human cell models of ADPKD, including ADPKD patient-derived primary cell cultures, we demonstrate that treatment with the prototypical PDE4 activator compound lowers intracellular cAMP levels, restrains cAMP-mediated signaling events, and profoundly inhibits cyst formation. PDE4 activator compounds thus have potential as therapeutics for treating disease driven by elevated cAMP signaling as well as providing a tool for evaluating the action of long PDE4 isoforms in regulating cAMP-mediated cellular processes.

Seasonal, interannual and decadal drivers of tree and grass productivity in an Australian tropical savanna.

Tree-grass savannas are a widespread biome and are highly valued for their ecosystem services. There is a need to understand the long-term dynamics and meteorological drivers of both tree and grass productivity separately in order to successfully manage savannas in the future. This study investigated the interannual variability (IAV) of tree and grass gross primary productivity (GPP) by combining a long-term (15 year) eddy covariance flux record and model estimates of tree and grass GPP inferred from satellite remote sensing. On a seasonal basis, the primary drivers of tree and grass GPP were solar radiation in the wet season and soil moisture in the dry season. On an interannual basis, soil water availability had a positive effect on tree GPP and a negative effect on grass GPP. No linear trend in the tree-grass GPP ratio was observed over the 15-year study period. However, the tree-grass GPP ratio was correlated with the modes of climate variability, namely the Southern Oscillation Index. This study has provided insight into the long-term contributions of trees and grasses to savanna productivity, along with their respective meteorological determinants of IAV.

Cholinesterase inhibitory activity of chlorophenoxy derivatives-Histamine H3 receptor ligands.

In recent years, multitarget-directed ligands have become an interesting strategy in a search for a new treatment of Alzheimer's disease. Combination of both: a histamine H3 receptor antagonist/inverse agonist and a cholinesterases inhibitor in one molecule could provide a new therapeutic opportunity. Here, we present biological evaluation of histamine H3 receptor ligands-chlorophenoxyalkylamine derivatives against cholinesterases: acetyl- and butyrylcholinesterase. The target compounds showed cholinesterase inhibitory activity in a low micromolar range. The most potent in this group was 1-(7-(4-chlorophenoxy)heptyl)homopiperidine (18) inhibiting the both enzymes (EeAChE IC50=1.93µM and EqBuChE IC50=1.64µM). Molecular modeling studies were performed to explain the binding mode of 18 with histamine H3 receptor as well as with cholinesterases.

IL-2Rα mediates temporal regulation of IL-2 signaling and enhances immunotherapy.

Interleukin-2 (IL-2) is a lymphocyte growth factor that is an important component of many immune-based cancer therapies. The efficacy of IL-2 is thought to be limited by the expansion of T regulatory cells, which express the high-affinity IL-2 receptor subunit IL-2Rα. IL-15 is under investigation as an alternative to IL-2. Although both cytokines signal through IL-2Rßγ, IL-15 does not bind IL-2Rα and therefore induces less T regulatory cell expansion. However, we found that transferred effector CD8(+) T cells induced curative responses in lymphoreplete mice only with IL-2-based therapy. Although conventional in vitro assays showed similar effector T cell responsiveness to IL-2 and IL-15, upon removal of free cytokine, IL-2 mediated sustained signaling dependent on IL-2Rα. Mechanistically, IL-2Rα sustained signaling by promoting a cell surface IL-2 reservoir and recycling of IL-2 back to the cell surface. Our results demonstrate that IL-2Rα endows T cells with the ability to compete temporally for limited IL-2 via mechanisms beyond ligand affinity. These results suggest that strategies to enhance IL-2Rα expression on tumor-reactive lymphocytes may facilitate the development of more effective IL-2-based therapies.

Effect of long-term oral administration of a low dosage of clenbuterol on body fat percentage in working and nonworking adult horses.

OBJECTIVE: To determine the anabolic and lipolytic effects of a low dosage of clenbuterol administered orally in working and nonworking equids. ANIMALS: 8 nonworking horses and 47 polo ponies in active training. PROCEDURES: Each polo pony continued training and received either clenbuterol (0.8 µg/kg) or an equal volume of corn syrup (placebo) orally twice daily for 21 days, and then was evaluated for another 21-day period. Nonworking horses received clenbuterol or placebo at the same dosage for 21 days in a crossover trial (2 treatments/horse). For working and nonworking horses, percentage body fat (PBF) was estimated before treatment and then 2 and 3 times/wk, respectively. Body weight was measured at intervals. RESULTS: Full data sets were not available for 8 working horses. For working horses, a significant treatment effect of clenbuterol was detected by day 3 and continued through the last day of treatment; at day 21, the mean change in PBF from baseline following clenbuterol or placebo treatment was -0.80% (representing a 12% decrease in PBF) and -0.32%, respectively. By day 32 through 42 (without treatment), PBF change did not differ between groups. When treated with clenbuterol, the nonworking horses had a similar mean change in PBF from baseline from day 6 onward, which peaked at -0.75% on day 18 (an 8% decrease in PBF). Time and treatment had no significant effect on body weight in either experiment. CONCLUSIONS AND CLINICAL RELEVANCE: Among the study equids, long-term low-dose clenbuterol administration resulted in significant decreases in body fat with no loss in body weight.

Fire in Australian savannas: from leaf to landscape.

Savanna ecosystems comprise 22% of the global terrestrial surface and 25% of Australia (almost 1.9 million km2) and provide significant ecosystem services through carbon and water cycles and the maintenance of biodiversity. The current structure, composition and distribution of Australian savannas have coevolved with fire, yet remain driven by the dynamic constraints of their bioclimatic niche. Fire in Australian savannas influences both the biophysical and biogeochemical processes at multiple scales from leaf to landscape. Here, we present the latest emission estimates from Australian savanna biomass burning and their contribution to global greenhouse gas budgets. We then review our understanding of the impacts of fire on ecosystem function and local surface water and heat balances, which in turn influence regional climate. We show how savanna fires are coupled to the global climate through the carbon cycle and fire regimes. We present new research that climate change is likely to alter the structure and function of savannas through shifts in moisture availability and increases in atmospheric carbon dioxide, in turn altering fire regimes with further feedbacks to climate. We explore opportunities to reduce net greenhouse gas emissions from savanna ecosystems through changes in savanna fire management.

Erectile dysfunction, vascular risk, and cognitive performance in late middle age.

Vascular disease is the most common etiology of erectile dysfunction (ED). Men with ED are at a 65% increased relative risk of developing coronary heart disease and a 43% increased risk of stroke within 10 years. Vascular disease is associated with cognitive impairment; ED-an overt manifestation of vascular dysfunction-could also signal early compromised cognition. We sought to determine whether cognitive differences existed between men with ED and healthy peers. Our sample consisted of 651 men (ages 51-60 years) from the Vietnam Era Twin Study of Aging. ED was associated with poorer cognitive performance, particularly on attention-executive-psychomotor speed tasks. ED remained significantly associated with cognition after inclusion of other cardiovascular risk factors (including hypertension, high cholesterol, body mass index, and smoking). These findings underscore the importance of further study of ED as a predictor of cognitive and cardiovascular health. (PsycINFO Database Record (c) 2014 APA, all rights reserved).

G-CSF/anti-G-CSF antibody complexes drive the potent recovery and expansion of CD11b+Gr-1+ myeloid cells without compromising CD8+ T cell immune responses.

BACKGROUND: Administration of recombinant G-CSF following cytoreductive therapy enhances the recovery of myeloid cells, minimizing the risk of opportunistic infection. Free G-CSF, however, is expensive, exhibits a short half-life, and has poor biological activity in vivo. METHODS: We evaluated whether the biological activity of G-CSF could be improved by pre-association with anti-G-CSF mAb prior to injection into mice. RESULTS: We find that the efficacy of G-CSF therapy can be enhanced more than 100-fold by pre-association of G-CSF with an anti-G-CSF monoclonal antibody (mAb). Compared with G-CSF alone, administration of G-CSF/anti-G-CSF mAb complexes induced the potent expansion of CD11b+Gr-1+ myeloid cells in mice with or without concomitant cytoreductive treatment including radiation or chemotherapy. Despite driving the dramatic expansion of myeloid cells, in vivo antigen-specific CD8+ T cell immune responses were not compromised. Furthermore, injection of G-CSF/anti-G-CSF mAb complexes heightened protective immunity to bacterial infection. As a measure of clinical value, we also found that antibody complexes improved G-CSF biological activity much more significantly than pegylation. CONCLUSIONS: Our findings provide the first evidence that antibody cytokine complexes can effectively expand myeloid cells, and furthermore, that G-CSF/anti-G-CSF mAb complexes may provide an improved method for the administration of recombinant G-CSF.

Ex vivo interleukin-12-priming during CD8(+) T cell activation dramatically improves adoptive T cell transfer antitumor efficacy in a lymphodepleted host.

BACKGROUND: Clinical application of adoptive T cell therapy has been hindered by an inability to generate adequate numbers of nontolerized, functionally active, tumor-specific T cells, which can persist in vivo. In order to address this, we evaluated the impact of interleukin (IL)-12 signaling during tumor-specific CD8(+) T cell priming in terms of persistence and antitumor efficacy using an established B16 melanoma tumor adoptive therapy model. STUDY DESIGN: B6 mice were injected subcutaneously with B16 melanoma tumor cells. On day 12 of tumor growth, mice were preconditioned with cyclophosphamide (4mg dose, intraperitoneally), and 1 day later were treated by adoptive transfer of tumor-specific pmel-1 CD8(+) T cells primed ex vivo 3 days earlier with both IL-12 and antigen (hGP100(25-33) peptide) or antigen only. Tumors were measured biweekly, and infused donor T cells were analyzed for persistence, localization to the tumor, phenotype, and effector function. RESULTS: Adoptive transfer of tumor-specific CD8(+) T cells primed with IL-12 was significantly more effective in reducing tumor burden in mice preconditioned with cyclophosphamide compared with transfer of T cells primed without IL-12. This enhanced antitumor response was associated with increased frequencies of infused T cells in the periphery and tumor as well as elevated expression of effector molecules including granzyme B and interferon-γ (IFNγ). CONCLUSIONS: Our findings demonstrate that ex vivo priming of tumor-specific CD8(+) T cells with IL-12 dramatically improves their in vivo persistence and therapeutic ability on transfer to tumor-bearing mice. These findings can be directly applied as novel clinical trial strategies.