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Smokers develop metastatic prostate cancer more frequently than nonsmokers, suggesting that a tobacco-derived factor is driving metastatic progression. To identify smoking-induced alterations in human prostate cancer, we analyzed gene and protein expression patterns in tumors collected from current, past, and never smokers. By this route, we elucidated a distinct pattern of molecular alterations characterized by an immune and inflammation signature in tumors from current smokers that were either attenuated or absent in past and never smokers. Specifically, this signature included elevated immunoglobulin expression by tumor-infiltrating B cells, NF-κB activation, and increased chemokine expression. In an alternate approach to characterize smoking-induced oncogenic alterations, we also explored the effects of nicotine in human prostate cancer cells and prostate cancer–prone TRAMP mice. These investigations showed that nicotine increased glutamine consumption and invasiveness of cancer cells in vitro and accelerated metastatic progression in tumor-bearing TRAMP mice. Overall, our findings suggest that nicotine is sufficient to induce a phenotype resembling the epidemiology of smoking-associated prostate cancer progression, illuminating a novel candidate driver underlying metastatic prostate cancer in current smokers. Cancer Res; 76(5); 1055–65. ©2015 AACR.

Supplementary Table S1: Clinical characteristics of the study population by smoking status. Supplementary Table S2: Immunoglobulin expression in prostate tumors by smoking status of the patients. Supplementary Table S3: Genes differently expressed between current and never smokers in prostate tumors. Supplementary Table S4: Genes differently expressed between current and never/past smokers in 67 prostate tumors.

Supplementary Table S11: Differentially expressed genes comparing prostate tumors from nicotine-treated TRAMP mice with tumors from untreated mice (reference).

Supplementary Table 9: Genes differently expressed between nicotine-treated and untreated 22Rv1 cells.

Supplementary Table S6: Increased expression of genes regulating synaptic signal transduction in the cancerous prostate of nicotine�treated TRAMP mice.

Supplemetary Table 5: Genes differently expressed between current and never and between current and past/never smokers in the combined cohort of 67 prostate tumors

Supplementary Table 7: Genes differently expressed between current and never smokers in microdissected prostate tumors.

Supplementary Figure S1: Weight curves for untreated TRAMP mice and TRAMP mice treated with nicotine for 80 days. Supplementary Figure S2: Ig lambda� and Ig kappa�positive B lymphocytes in cancerous prostate tissue by in�situ hybridization. Supplementary Figure S3: qRT�PCR validation of six genes up�regulated among current smokers. Supplementary Figure S4: Analysis of prostate tumors using Bioconductor limma R. Supplementary Figure S5: Gene Set Enrichment Analysis (GSEA) highlighting common features between smoking�related gene signatures in prostate tumors, nicotine�induced gene signatures in human prostate cancer cells, and gene signatures archived in the GSEA database. Supplementary Figure S6: Expression of various nicotinic acetylcholine receptor (AChR) subunits in human prostate cancer cell lines, prostate tumors, and adjacent non�tumor prostate tissues. Supplementary Figure S7: Nicotine�mediated Akt activation in human immortalized prostate epithelial cells and prostate cancer cell lines. Supplementary Figure S8: Effect of nicotine on the mobility of human prostate cancer cell lines. Supplementary Figure S9: Nicotine enhances cell surface integrin expression and extracellular matrix binding (ECM) of 22Rv1 human prostate cancer cells. Supplementary Figure S10: Nicotine enhances proliferation of RWPE�1 in complete K�SFM medium but not in K�SFM medium without EGF and bovine pituitary extract. Supplementary Figure S11: Nicotine did not enhance proliferation of 22Rv1 and PC�3 human prostate cancer cells. Supplementary Figure S12: Lymphotoxin�β plasma levels in prostate cancer patients (Case) and population�based controls (Control) by smoking status.

Supplemetary Table 8: Genes differently expressed between current and never in microdissected tumors using the Bioconductor limma R package.

Carbon cycling in the coastal zone affects global carbon budgets and is critical for understanding the urgent issues of hypoxia, acidification, and tidal wetland loss. However, there are no regional carbon budgets spanning the three main ecosystems in coastal waters: tidal wetlands, estuaries, and shelf waters. Here, we construct such a budget for Eastern North America using historical data, empirical models, remote-sensing algorithms, and process-based models. Considering the net fluxes of total carbon at the domain boundaries, 59 ± 12% (± 2 standard errors) of the carbon entering is from rivers and 41 ± 12% is from the atmosphere, while 80 ± 9% of the carbon leaving is exported to the open ocean and 20 ± 9% is buried. Net lateral carbon transfers between the three main ecosystem types are comparable to fluxes at the domain boundaries. Each ecosystem type contributes substantially to exchange with the atmosphere, with CO2 uptake split evenly between tidal wetlands and shelf waters, and estuarine CO2 outgassing offsetting half of the uptake. Similarly, burial is about equal in tidal wetlands and shelf waters, while estuaries play a smaller but still substantial role. The importance of tidal wetlands and estuaries in the overall budget is remarkable given that they respectively make up only 2.4 and 8.9% of the study domain area. This study shows that coastal carbon budgets should explicitly include tidal wetlands, estuaries, shelf waters and the linkages between them; ignoring any of them may produce a biased picture of coastal carbon cycling.

Small ponds-farm ponds, detention ponds, or impoundments below 0.01 km(2)-serve important human needs throughout most large river basins. Yet the role of small ponds in regional nutrient and sediment budgets is essentially unknown, currently making it impossible to evaluate their management potential to achieve water quality objectives. Here we used new hydrography data sets and found that small ponds, depending on their spatial position within both their local catchments and the larger river network, can dominate the retention of nitrogen, phosphorus, and sediment compared to rivers, lakes, and reservoirs. Over 300,000 small ponds are collectively responsible for 34%, 69%, and 12% of the mean annual retention of nitrogen, phosphorus, and sediment in the Northeastern United States, respectively, with a dominant influence in headwater catchments (54%, 85%, and 50%, respectively). Small ponds play a critical role among the many aquatic features in long-term nutrient and sediment loading to downstream waters. Plain Language Summary Reservoirs created by river damming have extensive impacts on downstream water quality but are not necessarily the most important elements of a diverse aquatic landscape. Many more small ponds have been constructed to serve important human needs ranging from farm irrigation in agricultural areas to flood control and trapping of nutrients and fine sediment in urban areas. The number of human-influenced small ponds is projected to rise worldwide, yet their role in the delivery of nutrients and sediment from headwaters to oceans is currently unresolved. Here we used new data sets and found that small ponds are collectively responsible for trapping a substantial amount of the nutrients and sediment that are exported annually from headwaters. These findings support the need to jointly consider features such as urban detention ponds, farm ponds, and beaver ponds in managing headwaters to decrease long-term nutrient and sediment loading to downstream waters and sensitive coastal areas. U.S. Geological SurveyUnited States Geological Survey; National Science Foundation Hydrologic Sciences ProgramNational Science Foundation (NSF)NSF - Directorate for Geosciences (GEO); Water Mission Area The ideas for this work were formulated during meetings of the John Wesley Powell Center River Corridor Working Group, supported by U.S. Geological Survey and National Science Foundation Hydrologic Sciences Program. The work was carried out at the USGS where N. M. S. is a USGS Mendenhall postdoctoral fellow funded by the Water Mission Area. Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the U.S. Government. We used publicly available, nationally consistent data sets (see Table S2 for data sources). The SPARROW model source code is also publicly available (https://water.usgs.gov/nawqa/sparrow/). The authors declare no conflict of interest. Public domain – authored by a U.S. government employee

Downstream flow in rivers is repeatedly delayed by hydrologic exchange with off-channel storage zones where biogeochemical processing occurs. We present a dimensionless metric that quantifies river connectivity as the balance between downstream flow and the exchange of water with the bed, banks, and floodplains. The degree of connectivity directly influences downstream water quality - too little connectivity limits the amount of river water exchanged and leads to biogeochemically inactive water storage, while too much connectivity limits the contact time with sediments for reactions to proceed. Using a metric of reaction significance based on river connectivity, we provide evidence that intermediate levels of connectivity, rather than the highest or lowest levels, are the most efficient in removing nitrogen from Northeastern United States' rivers. Intermediate connectivity balances the frequency, residence time, and contact volume with reactive sediments, which can maximize the reactive processing of dissolved contaminants and the protection of downstream water quality. Our simulations suggest denitrification dominantly occurs in riverbed hyporheic zones of streams and small rivers, whereas vertical turbulent mixing in contact with sediments dominates in mid-size to large rivers. The metrics of connectivity and reaction significance presented here can facilitate scientifically based prioritizations of river management strategies to protect the values and functions of river corridors. U.S. Geological SurveyUnited States Geological Survey; National Science Foundation Hydrologic Sciences ProgramNational Science Foundation (NSF)NSF - Directorate for Geosciences (GEO); USGS National Water Quality Program; DOE Office of Biological and Environmental Research (BER) in the Subsurface Biogeochemistry Program (SBR) as part of SBR's Scientific Focus Area at the Pacific Northwest National Laboratory (PNNL) The work is a product of the John Wesley Powell Center River Corridor Synthesis Group, supported by U.S. Geological Survey and National Science Foundation Hydrologic Sciences Program. USGS authors received additional support from the USGS National Water Quality Program. Gomez-Velez received additional support from the DOE Office of Biological and Environmental Research (BER) in the Subsurface Biogeochemistry Program (SBR) as part of SBR's Scientific Focus Area at the Pacific Northwest National Laboratory (PNNL). The synthesis is based entirely on analysis of published information and publicly available data sources. Any use of trade, firm, or product is for descriptive purposes only and does not imply endorsement by the U.S. Government. Public domain – authored by a U.S. government employee

Despite progress in the implementation of conservation practices, related improvements in water quality have been challenging to measure in larger river systems. In this paper we quantify these downstream effects by applying the empirical U.S. Geological Survey water-quality model SPARROW to investigate whether spatial differences in conservation intensity were statistically correlated with variations in nutrient loads. In contrast to other forms of water quality data analysis, the application of SPARROW controls for confounding factors such as hydrologic variability, multiple sources and environmental processes. A measure of conservation intensity was derived from the USDA-CEAP regional assessment of the Upper Mississippi River and used as an explanatory variable in a model of the Upper Midwest. The spatial pattern of conservation intensity was negatively correlated (p = 0.003) with the total nitrogen loads in streams in the basin. Total phosphorus loads were weakly negatively correlated with conservation (p = 0.25). Regional nitrogen reductions were estimated to range from 5 to 34% and phosphorus reductions from 1 to 10% in major river basins of the Upper Mississippi region. The statistical associations between conservation and nutrient loads are consistent with hydrological and biogeochemical processes such as denitrification. The results provide empirical evidence at the regional scale that conservation practices have had a larger statistically detectable effect on nitrogen than on phosphorus loadings in streams and rivers of the Upper Mississippi Basin.

Large-domain hydrological models are increasingly needed to support water-resource assessment and management in large river basins. Here, we describe results for the first Brazilian application of the SPAtially Referenced Regression On Watershed attributes (SPARROW) model using a new open-source modeling and interactive decision support system tool (RSPARROW) to quantify the origin, flux, and fate of total nitrogen (TN) in two sub-basins of the Grande River Basin (GRB, 43,000 km2). Land under cultivation for sugar cane, urban land, and point source inputs from wastewater treatment plants was estimated to each contribute approximately 30% of the TN load at the outlet, with pasture land contributing about 10% of the load. Hypothetical assessments of wastewater treatment plant upgrades and the building of new facilities that could treat currently untreated urban runoff suggest that these management actions could potentially reduce loading at the outlet by as much as 20&ndash, 25%. This study highlights the ability of SPARROW and the RSPARROW mapping tool to assist with the development and evaluation of management actions aimed at reducing nutrient pollution and eutrophication. The freely available RSPARROW modeling tool provides new opportunities to improve understanding of the sources, delivery, and transport of water-quality contaminants in watersheds throughout the world.

River corridors store, convey, and process nutrients from terrestrial and upstream sources, regulating delivery from headwaters to estuaries. A consequence of chronic excess nitrogen loading, as supported by theory and field studies in specific watersheds, is saturation of the biogeochemically-mediated nitrogen removal processes that weakens the capacity of the river corridor to remove nitrogen. Regional nitrogen models typically assume that removal capacity exhibits first-order behavior, scaling positively and linearly with increasing concentration, which may bias the estimation of where and at what rate nitrogen is removed by river corridors. Here we estimate the nitrogen concentration saturation threshold and its effects on annual nitrogen export from the Northeastern United States, revealing an average 42% concentration-induced reduction in headwater removal capacity. The weakened capacity caused an average 10% increase in the predicted delivery of riverine nitrogen from urban and agricultural watersheds compared to estimates using first-order process assumptions. Our results suggest that nitrogen removal may fall below a first-order rate process as riverine concentration increases above a threshold of 0.5 mg N l(-1). Threshold behavior indicates that even modest mitigation of nitrogen concentration in river corridors above the threshold can cause a self-reinforcing boost to nitrogen removal. USGSUnited States Geological Survey; National Science Foundation Hydrologic Sciences ProgramNational Science Foundation (NSF)NSF - Directorate for Geosciences (GEO); USGS Water Resources Availability Program; USGS Mendenhall Postdoctoral FellowshipUnited States Geological Survey; US Department of Agriculture National Institute of Food and AgricultureUnited States Department of Agriculture (USDA); US Department of Energy Subsurface Biogeochemistry Research Program, Pacific Northwest National Laboratory's scientific focus area; Virginia Agricultural Experimental Station The US Geological Survey (USGS) River Corridor Working Group developed the project idea in their meetings at the JohnWesley Powell Center with support from the USGS and the National Science Foundation Hydrologic Sciences Program. NMS, JWH, RBA, GES, and CPK also received support from the USGS Water Resources Availability Program. NMS was additionally supported by a USGS Mendenhall Postdoctoral Fellowship. EWB and DS received support from the US Department of Agriculture National Institute of Food and Agriculture. JDGV received support from the US Department of Energy Subsurface Biogeochemistry Research Program as part of Pacific Northwest National Laboratory's scientific focus area. DS also received support from the Virginia Agricultural Experimental Station. The SPARROW source code is also publicly available (https://water.usgs.gov/nawqa/sparrow/). Expressions and equations in the Methods section can be used to reproduce the results. Any use of trade, firm, or product names is for descriptive purposes only and does not imply endorsement by the US Government. The authors declare no competing interests.

Lakes, reservoirs, and other ponded waters are ubiquitous features of the aquatic landscape, yet their cumulative role in nitrogen removal in large river basins is often unclear. Here we use predictive modeling, together with comprehensive river water quality, land use, and hydrography datasets, to examine and explain the influences of more than 18,000 ponded waters on nitrogen removal through river networks of the Northeastern United States. Thresholds in pond density where ponded waters become important features to regional nitrogen removal are identified and shown to vary according to a ponded waters’ relative size, network position, and degree of connectivity to the river network, which suggests worldwide importance of these new metrics. Consideration of the interacting physical and biological factors, along with thresholds in connectivity, reveal where, why, and how much ponded waters function differently than streams in removing nitrogen, what regional water quality outcomes may result, and in what capacity management strategies could most effectively achieve desired nitrogen loading reduction., Lakes, reservoirs, and other ponded waters are common in large river basins yet their influence on nitrogen budgets is often indistinct. Here, the authors show how a ponded waters’ relative size, shape, and degree of connectivity to the river network control nitrogen removal.

The goal of this study was to determine if an alphavirus-based vaccine encoding human Prostate-Specific Antigen (PSA) could generate an effective anti-tumor immune response in a stringent mouse model of prostate cancer. DR2bxPSA F1 male mice expressing human PSA and HLA-DRB1(*)1501 transgenes were vaccinated with virus-like particle vector encoding PSA (VLPV-PSA) followed by the challenge with Transgenic Adenocarcinoma of Mouse Prostate cells engineered to express PSA (TRAMP-PSA). PSA-specific cellular and humoral immune responses were measured before and after tumor challenge. PSA and CD8 reactivity in the tumors was detected by immunohistochemistry. Tumor growth was compared in vaccinated and control groups. We found that VLPV-PSA could infect mouse dendritic cells in vitro and induce a robust PSA-specific immune response in vivo. A substantial proportion of splenic CD8 T cells (19.6 ± 7.4%) produced IFNγ in response to the immunodominant peptide PSA(65-73). In the blood of vaccinated mice, 18.4 ± 4.1% of CD8 T cells were PSA-specific as determined by the staining with H-2D(b)/PSA(65-73) dextramers. VLPV-PSA vaccination also strongly stimulated production of IgG2a/b anti-PSA antibodies. Tumors in vaccinated mice showed low levels of PSA expression and significant CD8+ T cell infiltration. Tumor growth in VLPV-PSA vaccinated mice was significantly delayed at early time points (p=0.002, Gehan-Breslow test). Our data suggest that TC-83-based VLPV-PSA vaccine can efficiently overcome immune tolerance to PSA, mediate rapid clearance of PSA-expressing tumor cells and delay tumor growth. The VLPV-PSA vaccine will undergo further testing for the immunotherapy of prostate cancer.

Tumor-associated macrophages (TAM) were shown to support the progression of many solid tumors. However, anti-tumor properties of TAM were also reported in several types of cancer. Here, we investigated the phenotype and functions of TAM in two transgenic mouse models of prostate cancer that display striking differences in tumor growth outcome. Mice expressing prostate-specific antigen (PSA) as a self-antigen specifically in prostate (PSAtg mice) rejected PSA-expressing transgenic adenocarcinoma of mouse prostate (TRAMP) tumors. However, the introduction of HLA-DRB1*1501 (DR2b) transgene presenting PSA-derived peptides in a MHC class II-restricted manner exacerbated the growth of TRAMP-PSA tumors in DR2bxPSA F 1 mice. Despite the difference in tumor growth outcome, tumors in both strains were equally and intensively infiltrated by macrophages on the first week after tumor challenge. TAM exhibited mixed M1/M2 polarization and simultaneously produced pro-inflammatory (TNFα, IL1β) and anti-inflammatory (IL10) cytokines. TAM from both mouse strains demonstrated antigen-presenting potential and pronounced immunostimulatory activity. Moreover, they equally induced apoptosis of tumor cells. In vivo depletion of macrophages in DR2bxPSA F 1 but not PSAtg mice aggravated tumor growth suggesting that macrophages more strongly contribute to anti-tumor immunity when specific presentation of PSA to CD4+ T cells is possible. In summary, we conclude that in the early stages of tumor progression, the phenotype and functional properties of TAM did not predict tumor growth outcome in two transgenic prostate cancer models. Furthermore, we demonstrated that during the initial stage of prostate cancer development, TAM have the potential to activate T cell immunity and mediate anti-tumor effects.

Net ecosystem production (NEP) and the overall organic carbon budget for the estuaries along the East Coast of the United States are estimated. We focus on the open estuarine waters, excluding the fringing wetlands. We developed empirical models relating NEP to loading ratios of dissolved inorganic nitrogen to total organic carbon, and carbon burial in the sediment to estuarine water residence time and total nitrogen input across the landward boundary. Output from a data-constrained water quality model was used to estimate inputs of total nitrogen and organic carbon to the estuaries across the landward boundary, including fluvial and tidal-wetland sources. Organic carbon export from the estuaries to the continental shelf was computed by difference, assuming steady state. Uncertainties in the budget were estimated by allowing uncertainties in the supporting model relations. Collectively, U.S. East Coast estuaries are net heterotrophic, with the area-integrated NEP of −1.5 (−2.8, −1.0) Tg C yr−1 (best estimate and 95% confidence interval) and area-normalized NEP of −3.2 (−6.1, −2.3) mol C m−2 yr−1. East Coast estuaries serve as a source of organic carbon to the shelf, exporting 3.4 (2.0, 4.3) Tg C yr−1 or 7.6 (4.4, 9.5) mol C m−2 yr−1. Organic carbon inputs from fluvial and tidal-wetland sources for the region are estimated at 5.4 (4.6, 6.5) Tg C yr−1 or 12 (10, 14) mol C m−2 yr−1 and carbon burial in the open estuarine waters at 0.50 (0.33, 0.78) Tg C yr−1 or 1.1 (0.73, 1.7) mol C m−2 yr−1. Our results highlight the importance of estuarine systems in the overall coastal budget of organic carbon, suggesting that in the aggregate, U.S. East Coast estuaries assimilate (via respiration and burial) ~40% of organic carbon inputs from fluvial and tidal-wetland sources and allow ~60% to be exported to the shelf.

INTRODUCTION Recent studies suggest that the cancer immunotherapy based on the blockade of the CTLA-4-mediated inhibitory pathway is efficacious only in select populations, predominantly for immunogenic tumors or when delivered in combination with modalities that can break immunologic tolerance to tumor antigens. METHODS We studied the effect of CD25+ cell depletion and CTLA-4 blockade on the growth of Transgenic Mouse Adenocarcinoma of Prostate (TRAMP)-PSA tumor cells in DR2bxPSA F1 mice. In these mice, immunological tolerance to PSA was established in a context of the HLA-DRB1*1501(DR2b) allele. RESULTS In our model, single administration of anti-CD25 antibody prior to tumor inoculation significantly increased IFN-γ production in response to the CD8 T cell epitope PSA65–73, and delayed TRAMP-PSA tumor growth compared to mice treated with isotype control antibodies. In contrast, the anti-tumor effect of the anti-CTLA-4 antibody as a monotherapy was marginal. The combinatory treatment with anti-CD25/anti-CTLA-4 antibodies significantly enhanced anti-tumor immunity and caused more profound delay in tumor growth compared to each treatment alone. The proportion of tumor-free animals was higher in the group that received combination treatment (21%) compared to other groups (2–7%). The enhanced anti-tumor immunity in response to the CD25 depletion or CTLA-4 blockade was only seen in the immunogenic TRAMP-PSA tumor model, whereas the effect was completely absent in mice bearing poorly immunogenic TRAMP-C1 tumors. DISCUSSION Our data suggest that breaking immunological tolerance to “self” antigens is essential for the therapeutic effect of CTLA-4 blockade. Such combinatory treatment may be a promising approach for prostate cancer immunotherapy. Prostate 74:1423–1432, 2014. © 2014 Wiley Periodicals, Inc.

BK virus is a polyomavirus, which causes a ubiquitous infection with an asymptomatic course in immunocompetent individuals and is latent in the genitourinary tract. BK virus reactivation can occur with immunosuppression such as in patients after organ transplantation, which might lead to nephropathy, hemorrhagic cystitis, and has been associated with development of malignancy in the genitourinary tract and nervous system. We present a rare case of urothelial carcinoma with a prominent micropapillary component in a patient 8 years after a cardiac transplantation. Micropapillary urothelial carcinoma (MPUC) is a rare variant of bladder cancer with an aggressive course and poor prognosis. Because of its resistance to intravesical immunotherapy, radical cystectomy is the preferred treatment for nonmuscle-invasive MPUC. This case report adds to the growing body of evidence linking BK virus infection in immunosuppressed patients and the development of MPUC. This aggressive tumor should be suspected in this patient population who present with hematuria.

Cytomegalovirus (CMV) is a highly immunogenic virus that results in a persistent, life-long infection in the host typically with no ill effects. Certain unique features of CMV, including its capacity to actively replicate in the presence of strong host CMV-specific immunity, may give CMV an advantage compared with other virus-based vaccine delivery platforms. In the present study, we tested the utility of mouse CMV (mCMV)-based vaccines expressing human prostate-specific antigen (PSA) for prostate cancer immunotherapy in double-transgenic mice expressing PSA and HLA-DRB1*1501 (DR2bxPSA F1 mice). We assessed the capacity of 2 mCMV-based vectors to induce PSA-specific CD8 T-cell responses and affect the growth of PSA-expressing Transgenic Adenocarcinoma of the Mouse Prostate tumors (TRAMP-PSA). In the absence of tumor challenge, immunization with mCMV vectors expressing either a H2-D(b)-restricted epitope PSA(65-73) (mCMV/PSA(65-73)) or the full-length gene for PSA (mCMV/PSA(FL)) induced comparable levels of CD8 T-cell responses that increased (inflated) with time. Upon challenge with TRAMP-PSA tumor cells, animals immunized with mCMV/PSA(65-73) had delay of tumor growth and increased PSA-specific CD8 T-cell responses, whereas animals immunized with mCMV/PSA(FL) showed progressive tumor growth and no increase in number of splenic PSA(65-73)-specific T cells. The data show that a prototype CMV-based prostate cancer vaccine can induce an effective antitumor immune response in a "humanized" double-transgenic mouse model. The observation that mCMV/PSA(FL) is not effective against TRAMP-PSA is consistent with our previous findings that HLA-DRB1*1501-restricted immune responses to PSA are associated with suppression of effective CD8 T-cell responses to TRAMP-PSA tumors.

We previously reported that miR-1 is among the most consistently down-regulated miRs in primary human prostate tumors. In this follow-up study, we further corroborated this finding in an independent data set and made the novel observation that miR-1 expression is further reduced in distant metastasis and is a candidate predictor of disease recurrence. Moreover, we performed in vitro experiments to explore the tumor suppressor function of miR-1. Cell-based assays showed that miR-1 is epigenetically silenced in human prostate cancer. Overexpression of miR-1 in these cells led to growth inhibition and down-regulation of genes in pathways regulating cell cycle progression, mitosis, DNA replication/repair and actin dynamics. This observation was further corroborated with protein expression analysis and 3'-UTR-based reporter assays, indicating that genes in these pathways are either direct or indirect targets of miR-1. A gene set enrichment analysis revealed that the miR-1-mediated tumor suppressor effects are globally similar to those of histone deacetylase inhibitors. Lastly, we obtained preliminary evidence that miR-1 alters the cellular organization of F-actin and inhibits tumor cell invasion and filipodia formation. In conclusion, our findings indicate that miR-1 acts as a tumor suppressor in prostate cancer by influencing multiple cancer-related processes and by inhibiting cell proliferation and motility.

Preston, Stephen D., Richard B. Alexander, Gregory E. Schwarz, and Charles G. Crawford, 2011. Factors Affecting Stream Nutrient Loads: A Synthesis of Regional SPARROW Model Results for the Continental United States. Journal of the American Water Resources Association (JAWRA) 47(5):891-915. DOI: 10.1111/j.1752-1688.2011.00577.x Abstract: We compared the results of 12 recently calibrated regional SPARROW (SPAtially Referenced Regressions On Watershed attributes) models covering most of the continental United States to evaluate the consistency and regional differences in factors affecting stream nutrient loads. The models – 6 for total nitrogen and 6 for total phosphorus – all provide similar levels of prediction accuracy, but those for major river basins in the eastern half of the country were somewhat more accurate. The models simulate long-term mean annual stream nutrient loads as a function of a wide range of known sources and climatic (precipitation, temperature), landscape (e.g., soils, geology), and aquatic factors affecting nutrient fate and transport. The results confirm the dominant effects of urban and agricultural sources on stream nutrient loads nationally and regionally, but reveal considerable spatial variability in the specific types of sources that control water quality. These include regional differences in the relative importance of different types of urban (municipal and industrial point vs. diffuse urban runoff) and agriculture (crop cultivation vs. animal waste) sources, as well as the effects of atmospheric deposition, mining, and background (e.g., soil phosphorus) sources on stream nutrients. Overall, we found that the SPARROW model results provide a consistent set of information for identifying the major sources and environmental factors affecting nutrient fate and transport in United States watersheds at regional and subregional scales.

Schwarz, Gregory E., Richard B. Alexander, Richard A. Smith, and Stephen D. Preston, 2011. The Regionalization of National-Scale SPARROW Models for Stream Nutrients. Journal of the American Water Resources Association (JAWRA) 47(5):1151-1172. DOI: 10.1111/j.1752-1688.2011.00581.x Abstract: This analysis modifies the parsimonious specification of recently published total nitrogen (TN) and total phosphorus (TP) national-scale SPAtially Referenced Regressions On Watershed attributes models to allow each model coefficient to vary geographically among three major river basins of the conterminous United States. Regionalization of the national models reduces the standard errors in the prediction of TN and TP loads, expressed as a percentage of the predicted load, by about 6 and 7%. We develop and apply a method for combining national-scale and regional-scale information to estimate a hybrid model that imposes cross-region constraints that limit regional variation in model coefficients, effectively reducing the number of free model parameters as compared to a collection of independent regional models. The hybrid TN and TP regional models have improved model fit relative to the respective national models, reducing the standard error in the prediction of loads, expressed as a percentage of load, by about 5 and 4%. Only 19% of the TN hybrid model coefficients and just 2% of the TP hybrid model coefficients show evidence of substantial regional specificity (more than ±100% deviation from the national model estimate). The hybrid models have much greater precision in the estimated coefficients than do the unconstrained regional models, demonstrating the efficacy of pooling information across regions to improve regional models.

Preston, Stephen D., Richard B. Alexander, and David M. Wolock, 2011. SPARROW Modeling to Understand Water-Quality Conditions in Major Regions of the United States: A Featured Collection Introduction. Journal of the American Water Resources Association (JAWRA) 47(5):887-890. DOI: 10.1111/j.1752-1688.2011.00585.x

BACKGROUND Transgenic mice engineered to express human leukocyte antigen (HLA) alleles are widely used for identification of immunogenic and naturally processed epitopes. Using HLA-DRB1*1501 (DR2b) transgenic mice, we have previously identified epitopes from two prostatic antigens, prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP). These antigens are implicated in the development of autoimmunity in the prostate and also are considered promising targets for prostate cancer immunotherapy. HLA-DRB1*1501 is the most common DR15 allele in Caucasians, while HLA-DRB1*1503 is the most common in African Americans. Hence characterization of peptide immunogenicity for these alleles is important for the development of prostate cancer immunotherapy in white and black patients. METHODS HLA-DRB1*1501 or HLA-DRB1*1503 transgenic mice were immunized with human PSA or PAP. Libraries of overlapping 20-mer peptides spanning the entire sequences of these proteins were screened by IFN-γ ELISPOT assay. RESULTS PSA and PAP peptides that were previously identified in HLA-DRB1*1501 tg mice were immunogenic in HLA-DR1503 tg mice and induced CD4 T-cell response against whole processed PSA or PAP respectively. However, the hierarchy of the immunodominance among the peptides differed significantly between strains. Using HLA-DRB1*1503 tg mice, a novel immunogenic and naturally processed 20-mer peptide, PAP (233-252) has been identified that showed no reactivity in HLA-DRB1*1501 tg mice. CONCLUSIONS Our data demonstrate a disparity in CD4 T-cell immune reactivity to PSA and PAP between HLA-DRB1*1501 and -DRB1*1503 alleles in HLA transgenic mouse models. It is possible that such immunological differences could contribute to racial disparity in prostate cancer outcome. Prostate 71:561–566, 2011. © 2010 Wiley-Liss, Inc.

Excessive loads of nutrients transported by tributary rivers have been linked to hypoxia in the Gulf of Mexico. Management efforts to reduce the hypoxic zone in the Gulf of Mexico and improve the water quality of rivers and streams could benefit from targeting nutrient reductions toward watersheds with the highest nutrient yields delivered to sensitive downstream waters. One challenge is that most conventional watershed modeling approaches (e.g., mechanistic models) used in these management decisions do not consider uncertainties in the predictions of nutrient yields and their downstream delivery. The increasing use of parameter estimation procedures to statistically estimate model coefficients, however, allows uncertainties in these predictions to be reliably estimated. Here, we use a robust bootstrapping procedure applied to the results of a previous application of the hybrid statistical/mechanistic watershed model SPARROW (Spatially Referenced Regression On Watershed attributes) to develop a statistically reliable method for identifying “high priority” areas for management, based on a probabilistic ranking of delivered nutrient yields from watersheds throughout a basin. The method is designed to be used by managers to prioritize watersheds where additional stream monitoring and evaluations of nutrient-reduction strategies could be undertaken. Our ranking procedure incorporates information on the confidence intervals of model predictions and the corresponding watershed rankings of the delivered nutrient yields. From this quantified uncertainty, we estimate the probability that individual watersheds are among a collection of watersheds that have the highest delivered nutrient yields. We illustrate the application of the procedure to 818 eight-digit Hydrologic Unit Code watersheds in the Mississippi/Atchafalaya River basin by identifying 150 watersheds having the highest delivered nutrient yields to the Gulf of Mexico. Highest delivered yields were from watersheds in the Central Mississippi, Ohio, and Lower Mississippi River basins. With 90% confidence, only a few watersheds can be reliably placed into the highest 150 category; however, many more watersheds can be removed from consideration as not belonging to the highest 150 category. Results from this ranking procedure provide robust information on watershed nutrient yields that can benefit management efforts to reduce nutrient loadings to downstream coastal waters, such as the Gulf of Mexico, or to local receiving streams and reservoirs.

We studied the growth of transgenic adenocarcinoma of mouse prostate (TRAMP)-C1 tumor cells expressing human prostate-specific Ag (PSA) in HLA-DRB1*1501 (DR2b) transgenic mice. TRAMP-PSA tumors were frequently rejected by HLA-DR2b− mice but had increased incidence in HLA-DR2b+ littermates. The levels of PSA-specific CD8 T cell responses were significantly higher in the HLA-DR2b− mice that rejected TRAMP-PSA tumors compared with HLA-DR2b+ tumor-bearing littermates. In contrast, Ab responses to PSA were strong in HLA-DR2b+ mice bearing TRAMP-PSA tumors and were virtually undetectable in HLA-DR2b− littermates. The analysis of CD4 T cell responses to PSA revealed the presence of several CD4 T cell epitopes in HLA-DR2b+ mice but failed to identify strong I-Ab-restricted epitopes in HLA-DR2b− mice. Our data demonstrate that the expression of a permissive HLA class II allele can change the pattern of the immune response to a tumor Ag, resulting in the failure of tumor rejection.

Human activities have greatly increased the transport of biologically available nitrogen (N) through watersheds to potentially sensitive coastal ecosystems. Lentic water bodies (lakes and reservoirs) have the potential to act as important sinks for this reactive N as it is transported across the landscape because they offer ideal conditions for N burial in sediments or permanent loss via denitrification. However, the patterns and controls on lentic N removal have not been explored in great detail at large regional to global scales. In this paper we describe, evaluate, and apply a new, spatially explicit, annual-scale, global model of lentic N removal called NiRReLa (Nitrogen Retention in Reservoirs and Lakes). The NiRReLa model incorporates small lakes and reservoirs than have been included in previous global analyses, and also allows for separate treatment and analysis of reservoirs and natural lakes. Model runs for the mid-1990s indicate that lentic systems are indeed important sinks for N and are conservatively estimated to remove 19.7 Tg N year−1 from watersheds globally. Small lakes (

Chronic prostatitis/chronic pelvic pain syndrome remains an enigmatic medical condition. Creation of the National Institutes of Health-funded Chronic Prostatitis Collaborative Research Network (CPCRN) has stimulated a renewed interest in research on and clinical aspects of chronic prostatitis/chronic pelvic pain syndrome. Landmark publications of the CPCRN document a decade of progress. Insights from these CPCRN studies have improved our management of chronic prostatitis/chronic pelvic pain syndrome and offer hope for continued progress.

We synthesize and update the science supporting the Action Plan for Reducing, Mitigating, and Controlling Hypoxia in the Northern Gulf of Mexico (Mississippi River/Gulf of Mexico Watershed Nutrient Task Force 2001) with a focus on the spatial and temporal discharge and patterns of nutrient and organic carbon delivery to the northern Gulf of Mexico, including data through 2006. The discharge of the Mississippi River watershed over 200 years varies but is not demonstrably increasing or decreasing. About 30% of the Mississippi River was shunted westward to form the Atchafalaya River, which redistributed water and nutrient loads on the shelf. Data on nitrogen concentrations from the early 1900s demonstrate that the seasonal and annual concentrations in the lower river have increased considerably since then, including a higher spring loading, following the increase in fertilizer applications after World WarII. The loading of total nitrogen (TN) fell from 1990 to 2006, but the loading of total phosphorus (TP) has risen slightly, resulting in a decline in the TN:TP ratios. The present TN:TP ratios hover around an average indicative of potential nitrogen limitation on phytoplankton growth, or balanced growth limitation, but not phosphorus limitation. The dissolved nitrogen:dissolved silicate ratios are near the Redfield ratio indicative of growth limitations on diatoms. Although nutrient concentrations are relatively high compared to those in many other large rivers, the water quality in the Mississippi River is not unique in that nutrient loads can be described by a variety of land-use models. There is no net removal of nitrogen from water flowing through the Atchafalaya basin, but the concentrations of TP and suspended sediments are lower at the exit point (Morgan City, Louisiana) than in the water entering the Atchafalaya basin. The removal of nutrients entering offshore waters through diversion of river water into wetlands is presently less than 1% of the total loadings going directly offshore, and would be less than 8% if the 10,093 km2 of coastal wetlands were successfully engineered for that purpose. Wetland loss is an insignificant contribution to the carbon loading offshore, compared to in situ marine production. The science-based conclusions in the Action Plan about nutrient loads and sources to the hypoxic zone off Louisiana are sustained by research and monitoring occurring in the subsequent 10 years.

Knowledge of headwater influences on the water-quality and flow conditions of downstream waters is essential to water-resource management at all governmental levels; this includes recent court decisions on the jurisdiction of the Federal Clean Water Act (CWA) over upland areas that contribute to larger downstream water bodies. We review current watershed research and use a water-quality model to investigate headwater influences on downstream receiving waters. Our evaluations demonstrate the intrinsic connections of headwaters to landscape processes and downstream waters through their influence on the supply, transport, and fate of water and solutes in watersheds. Hydrological processes in headwater catchments control the recharge of subsurface water stores, flow paths, and residence times of water throughout landscapes. The dynamic coupling of hydrological and biogeochemical processes in upland streams further controls the chemical form, timing, and longitudinal distances of solute transport to downstream waters. We apply the spatially explicit, mass-balance watershed model SPARROW to consider transport and transformations of water and nutrients throughout stream networks in the northeastern United States. We simulate fluxes of nitrogen, a primary nutrient that is a water-quality concern for acidification of streams and lakes and eutrophication of coastal waters, and refine the model structure to include literature observations of nitrogen removal in streams and lakes. We quantify nitrogen transport from headwaters to downstream navigable waters, where headwaters are defined within the model as first-order, perennial streams that include flow and nitrogen contributions from smaller, intermittent and ephemeral streams. We find that first-order headwaters contribute approximately 70% of the mean-annual water volume and 65% of the nitrogen flux in second-order streams. Their contributions to mean water volume and nitrogen flux decline only marginally to about 55% and 40% in fourth- and higher-order rivers that include navigable waters and their tributaries. These results underscore the profound influence that headwater areas have on shaping downstream water quantity and water quality. The results have relevance to water-resource management and regulatory decisions and potentially broaden understanding of the spatial extent of Federal CWA jurisdiction in U.S. waters.

Quantifying where, when, and how much denitrification occurs on the basis of measurements alone remains particularly vexing at virtually all spatial scales. As a result, models have become essential tools for integrating current understanding of the processes that control denitrification with measurements of rate-controlling properties so that the permanent losses of N within landscapes can be quantified at watershed and regional scales. In this paper, we describe commonly used approaches for modeling denitrification and N cycling processes in terrestrial and aquatic ecosystems based on selected examples from the literature. We highlight future needs for developing complementary measurements and models of denitrification. Most of the approaches described here do not explicitly simulate microbial dynamics, but make predictions by representing the environmental conditions where denitrification is expected to occur, based on conceptualizations of the N cycle and empirical data from field and laboratory investigations of the dominant process controls. Models of denitrification in terrestrial ecosystems include generally similar rate-controlling variables, but vary in their complexity of the descriptions of natural and human-related properties of the landscape, reflecting a range of scientific and management perspectives. Models of denitrification in aquatic ecosystems range in complexity from highly detailed mechanistic simulations of the N cycle to simpler source-transport models of aggregate N removal processes estimated with empirical functions, though all estimate aquatic N removal using first-order reaction rate or mass-transfer rate expressions. Both the terrestrial and aquatic modeling approaches considered here generally indicate that denitrification is an important and highly substantial component of the N cycle over large spatial scales. However, the uncertainties of model predictions are large. Future progress will be linked to advances in field measurements, spatial databases, and model structures.

Cognitive/behavioral and environmental variables are significant predictors of patient adjustment in chronic pain. Using a biopsychosocial template and selecting several pain-relevant constructs from physical, cognitive/behavioral, and environmental predictors, outcomes of pain and disability in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) were explored. Men (n = 253) from a North American multi-institutional NIH-funded Chronic Prostatitis Cohort Study in 6 US and 1 Canadian centers participated in a survey examining pain and disability. Measures included demographics, urinary symptoms, depression, pain, disability, catastrophizing, control over pain, pain-contingent rest, social support, and solicitous responses from a significant other. Regressions showed that urinary symptoms (beta = .20), depression (beta = .24), and helplessness catastrophizing (beta = .29) predicted overall pain. Further, affective pain was predicted by depression (beta = .39) and helplessness catastrophizing (beta = .44), whereas sensory pain was predicted by urinary symptoms (beta = .25) and helplessness catastrophizing (beta = .37). With regard to disability, urinary symptoms (beta = .17), pain (beta = .21), and pain-contingent rest (beta = .33) were the predictors. These results suggest cognitive/behavioral variables (ie, catastrophizing, pain-contingent rest) may have significant impact on patient adjustment in CP/CPPS. Findings support the need for greater research of such pain-related variables in CP/CPPS.This article explores predictors of patient adjustment in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). Cognitive/behavioral variables of catastrophizing and pain-contingent rest respectively predicted greater pain and disability. Catastrophic helplessness was a prominent pain predictor. These findings inform clinicians and researchers on several new variables in CP/CPPS outcomes and suggest future research.

Objectives: The NIH-Chronic Prostatitis Symptom Index (NIH-CPSI) was developed to assess symptoms and quality of life in men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). We assessed the responsiveness of the NIH-CPSI to change over time and defined thresholds for changes perceptible to patients. Methods: We studied 174 men with CP/CPPS who participated in a placebo-controlled randomized clinical trial. Changes from baseline to six weeks in the NIH-CPSI total score and pain, urinary, and quality of life subscores were compared to a global response assessment (GRA). Effect sizes and Guyatt statistics were calculated to evaluate responsiveness; 95% confidence intervals were produced using bootstrapping. Results: All scores decreased over time with the largest decrease in subjects who reported on the GRA that they were markedly improved. The NIH-CPSI total, pain, and quality of life scores were highly responsive in the improved groups; the urinary score showed minimal responsiveness. There was no evidence of responsiveness among those subjects who worsened on the trial. ROC curves identified a 6-point decline in the NIH-CPSI total score as the optimal threshold to predict treatment response. Conclusions: The NIH-CPSI total score and pain and quality of life subscores are responsive to change over␣time.

Objectives To determine whether prostate-specific antigen (PSA), the percent free PSA, or free PSA isoforms may be used as diagnostic markers for chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS; National Institutes of Health category IIIa and IIIb). Methods We evaluated 421 patients enrolled in the Chronic Prostatitis Cohort Study and 112 age-matched controls. Subjects were stratified by the number of white blood cells (WBCs) in their expressed prostatic secretions and pain as determined by the National Institutes of Health Chronic Prostatitis Symptom Index. Results Total PSA, free PSA, and [−2]proPSA ([−2]pPSA) were significantly elevated in those with CP/CPPS compared with controls (mean PSA 1.97 ng/mL versus 1.72 ng/mL, P = 0.03; mean free PSA 0.76 ng/mL versus 0.70 ng/mL, P = 0.01; and [−2]pPSA 2.38 ng/mL versus 1.80 ng/mL, P = 0.04). The percent free PSA was not significantly different between the patients and controls. For those with CP/CPPS, the percent free PSA was significantly lower as the WBC count rose in the expressed prostatic secretions (0 WBCs = 43.29 versus more than 25 WBCs = 26.52; P Conclusions Men with elevated PSA values and CP/CPPS should be treated as one would any other patient screened for prostate cancer with an elevated PSA level. Although PSA, free PSA, and [−2]pPSA were slightly elevated in men with CP/CPPS, the low sensitivity and specificity do not warrant using them as biomarkers for CP/CPPS.

Purpose: To determine whether mesothelin, a cell surface protein highly expressed in mesothelioma and ovarian cancer, is shed into serum and if so to accurately measure it. Experimental Design: We developed a sandwich ELISA using antibodies reacting with two different epitopes on human mesothelin. To quantitate serum mesothelin levels, a standard curve was generated using a mesothelin-Fc fusion protein. Sera from 24 healthy volunteers, 95 random hospital patients, 56 patients with mesothelioma, and 21 patients with ovarian cancer were analyzed. Serum mesothelin levels were also measured before and after surgical cytoreduction in six patients with peritoneal mesothelioma. Results: Elevated serum mesothelin levels were noted in 40 of 56 (71%) patients with mesothelioma and in 14 of 21 (67%) patients with ovarian cancer. Serum mesothelin levels were increased in 80% and 75% of the cases of mesothelioma and ovarian cancer, respectively, in which the tumors expressed mesothelin by immunohistochemistry. Out of the six patients with peritoneal mesothelioma who underwent surgery, four had elevated serum mesothelin levels before surgery. Out of these four patients, three had cytoreductive surgery and the serum mesothelin level decreased by 71% on postoperative day 1 and was undetectable by postoperative day 7. Conclusions: We developed a serum mesothelin assay that shows that mesothelin is elevated in patients with mesothelioma and ovarian cancer. The rapid decrease in mesothelin levels after surgery in patients with peritoneal mesothelioma suggests that serum mesothelin may be a useful test to monitor treatment response in mesothelin-expressing cancers.

We estimated trends in concentrations of total phosphorus (TP) and total nitrogen (TN) and the related change in the probabilities of trophic conditions from 1975 to 1994 at 250 nationally representative riverine monitoring locations in the U.S. with drainage areas larger than about 1,000 km 2 . Statistically significant (p , 0.05) declines were detected in TP and TN concentrations at 44% and 37% of the monitoring sites, and significant increases were detected at 3% and 9% of the sites, respectively. We used a statistical model to assess changes in the probable trophic-state classification of the sites after adjusting for climate-related variability in nutrient concentrations. The probabilistic assessment accounts for current knowledge of the trophic response of streams to nutrient enrichment, based on a recently proposed definition of ‘‘eutrophic,’’ ‘‘mesotrophic,’’ and ‘‘oligotrophic’’ conditions in relation to total nutrient concentrations. Based on these trophic definitions, we found that the trophic state improved at 25% of the monitoring sites and worsened at fewer than 5% of the sites; about 70% of the sites were unchanged. Improvements in trophic-state related to declines in TP were more common in predominantly forested and shrub‐ grassland watersheds, whereas the trophic state of predominantly agricultural sites was unchanged. Despite the declines in TP concentrations at many sites, about 50% of all monitoring sites, and more than 60% of the sites in predominantly agricultural and urban watersheds, were classified as eutrophic in 1994 based on TP concentrations. Contemporaneous reductions in major nutrient sources to streams, related to wastewater treatment upgrades, phosphate detergent bans, and declines in some agricultural sources, may have contributed to the declines in riverine nutrient concentrations and associated improvements in trophic conditions. Cultural sources of nutrients to streams changed appreciably during the last three decades of the 20th century in the U.S. Advances in wastewater treatment, enabled by the 1972 Clean Water Act, and state-level phosphate detergent bans greatly reduced municipal point sources of nutrients to streams (Litke 1999). Diffuse nutrient sources, including fertilizers and fossil-fuel emissions, rose to peak levels in the late 1970s and early 1980s and have varied considerably since that time (Howarth et al. 2002). Soil conservation and government-sponsored retirements of farmlands have received increasing attention since the 1980s (USDA 1997), although knowledge of their effects on stream nutrients are limited.

Granulomatous prostatitis is characterized by a pattern of granulomatous inflammation in the prostate. In most cases the etiology is unknown. Based on the hypothesis that granulomatous prostatitis may be an autoimmune disease we performed intermediate and selective high resolution typing of HLA-DR in a group of patients with the disease and compared the frequency of class II HLA phenotypes to that in a control group of volunteer marrow donors in the military.Histological records from 1 institution from 1990 to 2000 revealed 12 patients with diffuse granulomatous prostatitis. Three patients were dead and 1 refused blood drawing. Peripheral blood from the remaining 8 patients was typed along with blood from an additional 3 identified at the practice of one of us from 1999 through 2002. All slides were reviewed by 1 pathologist. Intermediate resolution typing of HLA-A, B and DR was performed by polymerase chain reaction-sequence specific oligonucleotide probe. High resolution, allele specific identification of HLA DR15 was performed if patients were DR15 positive by intermediate resolution typing.There were 3 black and 8 white individuals identified with diffuse nonspecific granulomatous prostatitis. Six of 8 white patients (75%) were HLA-DR15 by intermediate resolution typing. One of the 3 black American patients (33%) was HLA-DR15. In the control group 127 of 451 white (28.2%) and 23 of 89 black (25.8%) volunteer marrow donors were HLA-DR15. The case-control comparison of white patients was significantly different (Fisher's exact test p = 0.0086). There were no statistically significant differences between case-control comparisons for any other HLA-DR phenotype. High resolution DR15 typing showed that the white patients were HLA-DRB1*1501 and the black patient was HLA-DRB1*1503.The data suggest an association between HLA-DRB1*1501 and granulomatous prostatitis. HLA-DR15 is strongly associated with other autoimmune diseases, notably multiple sclerosis. The data are consistent with an autoimmune etiology for nonspecific granulomatous prostatitis.

In order to develop immunotherapies for prostate cancer, many groups are exploring vaccination strategies to induce an immune response against prostate specific antigen (PSA). To determine if T-cell recognition of PSA might be a feature of a naturally occurring human disease, we have studied patients with prostatitis, a poorly understood clinical syndrome of men in which there is evidence that an immune response directed against the prostate may be occurring. We wished to determine if a T-cell response to PSA might be occurring in these patients. We generated long-term T-cell lines from peripheral blood mononuclear cells (PBMC) of one patient with granulomatous prostatitis using purified PSA as an antigen. Several CD4+ and CD8+ TcR alpha/beta+ T-cell lines were selected for PSA reactivity as measured by at least a threefold increase in IFN-gamma secretion in response to PSA presented by irradiated autologous PBMC. CD4 and CD8 T-cell lines recognized PSA in the context of HLA-DRbeta1*1501 and HLA-B*0702, respectively. The specificity and HLA restriction of the lines was confirmed using EBV-B cell lines infected with a recombinant PSA-expressing vaccinia virus and also engineered to express PSA by retroviral transfection. HLA-matched targets infected by control vector as well as HLA-mismatched PSA-expressing targets did not induce the response. The data demonstrate that PSA-specific T cells are present in the PBMC of this patient with granulomatous prostatitis, who may be manifesting naturally the type of immune response directed at the prostate that is the goal of prostate cancer immunotherapy. However, the Class I-restricted epitope has not yet been demonstrated to be expressed on the surface of prostate cancer cells. To our knowledge, this is the first demonstration of HLA-DRB1*1501- or HLA-B*0702-restricted responses to PSA and extends the number of HLA molecules accommodating the use of PSA antigen as a candidate vaccine for prostate cancer immunotherapy.

Chronic prostatitis has been traditionally characterized by inflammation and/or infection of the prostate gland, objectively categorized by white blood cells and cultured bacteria in prostate specific specimens. We compared leukocyte counts and localization rates for bacterial cultures of segmented urine samples (VB1, VB2, VB3), expressed prostatic secretion (EPS) and semen in men diagnosed with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) to men without pelvic pain (controls).A total of 463 men enrolled in the National Institutes of Health Chronic Prostatitis Cohort study and 121 age matched men without urinary symptoms had leukocyte counts performed and 5-day bacterial cultures on specimens obtained from a standard 4-glass test (VB1, VB2, EPS, VB3) and semen. All risk factor comparisons between case and control analyses were tested using generalized Mantel-Haenszel methods, and multivariable models were developed using logistic regression methods, adjusting for clustering by clinical center within both methods.Men with CP/CPPS had statistically higher leukocyte counts in all segmented urine samples and EPS, but not in semen compared to asymptomatic control men. However, the control population also had a high prevalence of leukocytes. Of the men with CP/CPPS 50% and 32% had 5 or more, or 10 or more white blood cells (WBCs) per high power field, respectively, in EPS compared to 40% and 20% of the control population. Similarly, 32% and 14% of the patients with CP/CPPS had 5 or more, or 10 or more WBCs per high power field in VB3 compared to 19% and 11% in the control population. Localization of uropathogenic bacteria in EPS, VB3 and/or semen was similar in men with CP/CPPS (8.0%) and asymptomatic men (8.3%).Men with CP/CPPS have significantly higher leukocyte counts in all segmented urine samples and EPS but not in semen as compared to controls. There is no difference in rates of localization of bacterial cultures for men with CP/CPPS compared to control men. The high prevalence of WBCs and positive bacterial cultures in the asymptomatic control population raises questions about the clinical usefulness of the standard 4-glass test as a diagnostic tool in men with CP/CPPS.

The spatially referenced regressions on watershed attributes modeling approach, as applied to predictions of total nitrogen flux in three North Carolina river basins, addresses several information needs identified by a National Research Council evaluation of the total maximum daily load program. The model provides reach-level predictions of the probability of exceeding water-quality criteria, and estimates of total nitrogen budgets. Model estimates of point- and diffuse-source contributions and nitrogen loss rates in streams and reservoirs compared moderately well with literature estimates. Maps of reach-level predictions of nutrient inputs and delivery provide an intuitive and spatially detailed summary of the origins and fate of nutrients within a basin.