Connecticut Sea Grant

Mercury (Hg) is a global pollutant that affects human and ecosystem health. We synthesize understanding of sources, atmosphere-land-ocean Hg dynamics and health effects, and consider the implications of Hg-control policies. Primary anthropogenic Hg emissions greatly exceed natural geogenic sources, resulting in increases in Hg reservoirs and subsequent secondary Hg emissions that facilitate its global distribution. The ultimate fate of emitted Hg is primarily recalcitrant soil pools and deep ocean waters and sediments. Transfers of Hg emissions to largely unavailable reservoirs occur over the time scale of centuries, and are primarily mediated through atmospheric exchanges of wet/dry deposition and evasion from vegetation, soil organic matter and ocean surfaces. A key link between inorganic Hg inputs and exposure of humans and wildlife is the net production of methylmercury, which occurs mainly in reducing zones in freshwater, terrestrial, and coastal environments, and the subsurface ocean. Elevated human exposure to methylmercury primarily results from consumption of estuarine and marine fish. Developing fetuses are most at risk from this neurotoxin but health effects of highly exposed populations and wildlife are also a concern. Integration of Hg science with national and international policy efforts is needed to target efforts and evaluate efficacy.

SUMMARY The concept of ecosystem services (ES), the benefits humans derive from ecosystems, is increasingly applied to environmental conservation, human well-being and poverty alleviation, and to inform the development of interventions. Payments for ecosystem services (PES) implicitly recognize the unequal distribution of the costs and benefits of maintaining ES, through monetary compensation from ‘winners’ to ‘losers’. Some research into PES has examined how such schemes affect poverty, while other literature addresses trade-offs between different ES. However, much evolving ES literature adopts an aggregated perspective of humans and their well-being, which can disregard critical issues for poverty alleviation. This paper identifies four issues with examples from coastal ES in developing countries. First, different groups derive well-being benefits from different ES, creating winners and losers as ES, change. Second, dynamic mechanisms of access determine who can benefit. Third, individuals' contexts and needs determine how ES contribute to well-being. Fourth, aggregated analyses may neglect crucial poverty alleviation mechanisms such as cash-based livelihoods. To inform the development of ES interventions that contribute to poverty alleviation, disaggregated analysis is needed that focuses on who derives which benefits from ecosystems, and how such benefits contribute to the well-being of the poor. These issues present challenges in data availability and selection of how and at which scales to disaggregate. Disaggregation can be applied spatially, but should also include social groupings, such as gender, age and ethnicity, and is most important where inequality is greatest. Existing tools, such as stakeholder analysis and equity weights, can improve the relevance of ES research to poverty alleviation.

Preface Prologue Part I What Are You Getting Into? 1 Integrating Evaluation and Practice Introduction to Single-System Designs What Are Single-System Designs? Evidence-Based Practice Single-System Designs and Classical Research: The Knowledge-Building Context Single-System Evaluation, Qualitative Research, and Quantitative Research Advantages of Using Single-System Designs in Practice A Walk through the Evaluation Process Summary Part II Conceptualizing and Measuring Targets and Objectives/Goals 2 Basic Principles of Conceptualization and Measurement Introduction What Is Conceptualization? What Is Measurement? Definition as a First Step in Measurement Can Everything Be Measured? Key Characteristics of All Measures Summary 3 Specifying Problems and Goals Introduction: From General Problems to Specific Targets of Intervention Specifying Client Concerns: Identifying and Clarifying Problems and Potentials Specifying Goals and Objectives Using Goal Attainment Scaling (GAS) to Establish Goals Setting Goals in Groups Problems and Issues in Setting Goals Summary 4 Developing a Measurement and Recording Plan Introduction Steps in Developing a Recording Plan Charting: Putting Your Information on Graphs Problem-Oriented Records (POR) Use of Computers in Evaluation and Practice Summary Computer Assisted Social Services (CASS): A User's Guide Appendix: Installing CASS 5 Behavioral Observation Introduction General Guidelines for Behavioral Observation Sampling Behaviors Instruments for Recording Behaviors Ensuring Accurate Observations Methods of Recording Behavior Analog Situations Recording Behavior in Groups Summary 6 Individualized Rating Scales Introduction Uses of Individualized Rating Scales Constructing and Using Individualized Rating Scales Summary 7 Standardized Scales Introduction What Are Standardized Scales? Selecting a Standardized Scale Administering a Standardized Scale Some Available Standardized Self-Report Scales Some Available Standardized Scales for Practitioners Some Available Standardized Scales for Relevant Others Some Available Standardized Scales for Independent Observers Do-It-Yourself Scales Using Standardized Scales in Groups Computer Management of Standardized Scales Summary Appendix Computer Assisted Assessment Package (CAAP): A User's Guide 8 Logs Introduction Types of Client Logs Putting Qualitative and Quantitative Information Together Introducing Clients to Logs Practitioner Logs Maximizing and Verifying the Reliability and Validity of Logs Summary 9 Reactivity and Nonreactive Measures Introduction Reactivity of Measures Unobtrusive (Nonreactive) Measures Summary 10 Selecting a Measure Introduction Considerations in Deciding on a Measure Use of Multiple Measures Selecting a Measure Summary Part III Evaluation Designs 11 Basic Principles of Single-System Designs Introduction An Example Connecting Practice and Evaluation Designs Purposes of Single-System Designs Unique Characteristics of Single-System Designs Causality in Single-System Designs External Validity and Generalizability Overview of Single-System Designs Summary 12 Baselining Introduction Purposes of the Baseline Types of Baselines How Long Should Baselining Continue? When Are Baselines Not Necessary? Issues Regarding Baselining Summary 13 From the Case Study to the Basic Single-System Design: A-B Introduction Case Studies or Predesigns Design A-B: The Basic Single-System Design Summary 14 The Experimental Single-System Designs: A-B-A, A-B-A-B, B-A-B Introduction Basic Experimental Designs Summary 15 Multiple Designs for Single Systems Introduction Multiple-Baseline Designs: Across Problems, Clients, or Settings Multiple-Target Designs Variations on Multiple Designs Summary 16 Changing Intensity Designs and Successive Intervention Designs Introduction Changing Intensity Designs: A-B1-B2-B3 Successive Intervention Designs: A-B-C, A-B-A-C, A-B-A-C-A Summary 17 Designs for Comparing Interventions Introduction Alternating Intervention Design: A-B/C-(B orC) Interaction Design: A-B-A-B-BC-B-BC Summary 18 Selecting a Design Introduction Framework for Selecting a Design Needed: A Design for All Seasons Creativity in Single-System Designs: Making Your Own Designs Evaluation in Minimal-Contact Situations Single-System Designs in Managed Care: The Stretch Design Trouble-Shooting: Okay, I Understand Everything That You Said, but My Case Is Different. Summary Part IV Analyzing Your Results 19 Basic Principles of Analysis Introduction Distinguishing Effort, Effectiveness, and Efficiency Significance-Practical, Statistical, and Theoretical Evaluating Goal Achievement Issues in Analysis of Data Computer Analysis of Data for Single-System Designs The Issue of Autocorrelation Tools in Analysis of Data Summary 20 Visual Analysis of Single-System Design Data Introduction Definition of Terms Basic Patterns and Implications Visual Inspection of Raw Data Interpreting Ambiguous Patterns Problems of Visual Inspection Creating a Chart with SINGWIN Summary 21 Descriptive Statistics Introduction Measures of Central Tendency Measures of Variation Using Measures of Central Tendency and Variability to Graph Group Data Computing and Graphing Measures of Central Tendency and Variation with SINGWIN Measures of Trend Measures of Effect Size Optimal Uses and Cautions for Specific Descriptive Statistics Summary 22 Tests of Statistical Significance for Single-System Designs Introduction Proportion/Frequency Approach Three-Standard-Deviation-Band Approach (X-Moving-Range-Chart) Chi-Square Conservative Dual-Criteria (CDC) Approach t-Test General Considerations in Using Tests of Statistical Significance Optimal Uses and Cautions for Specific Analytic Procedures Summary 23 Computer Analysis of Single-System Design Data Chapter Overview Starting SINGWIN Exiting SINGWIN Getting the Big Picture Using Specific Procedures Appendix: Installing Singwin Summary 24 Selecting a Procedure for Analyzing Data Introduction Framework for Selecting a Procedure for Analyzing Data Other Statistical Considerations Nonstatistical Considerations Limitations Summary Part V The Challenge of Single-System Designs 25 Not for Practitioners Alone Introduction Special Applications of Single-System Designs Recent Criticisms of Single-System Evaluation For the Client For the Administrator For Educators and Students Summary References Name Index Subject Index

H; more recently, numerous protocols have achieved X-H insertion with novel non-ozone depleting difluorocarbene reagents. All together, these advances have streamlined access to molecules of pharmaceutical relevance, and generated interest for process chemistry.

) are among the most highly polychlorinated biphenyl (PCB)-contaminated mammals in the world, raising concern about the health consequences of current PCB exposures. Using an individual-based model framework and globally available data on PCB concentrations in killer whale tissues, we show that PCB-mediated effects on reproduction and immune function threaten the long-term viability of >50% of the world's killer whale populations. PCB-mediated effects over the coming 100 years predicted that killer whale populations near industrialized regions, and those feeding at high trophic levels regardless of location, are at high risk of population collapse. Despite a near-global ban of PCBs more than 30 years ago, the world's killer whales illustrate the troubling persistence of this chemical class.

Global atmospheric changes carry the potential to disrupt the normal cycling of mercury and its compounds. Acid rain may increase methylmercury levels in freshwater fish. Global warming and increased ultraviolet radiation may affect the global budget of methylmercury, including its formation and degradation in both biotic and abiotic environments. In this article we review current knowledge on mercury and monomethylmercury with regard to their environmental fate and the potential for human health effects. Recent findings indicate that atmospheric Hg deposition readily accounts for the total mass of Hg in fish, water, and sediment of Little Rock Lake, a representative temperate seepage lake in north-central Wisconsin. It is strikingly evident that modest increases in atmospheric Hg loading could lead directly to elevated levels in the fish stock. It is doubtful, given the experimental limitations in many recent studies, that the temporal pattern for Hg emissions, for background atmospheric Hg concentrations, and for changes in Hg depositional fluxes has been identified. Thus, the present and future questions of whether the environmental impact is of local, regional, or hemispheric significance remain. Contemporary investigations must address these important questions. Human exposure to methylmercury in the United States is probably increasing due to increased consumption of fish and fish products. A recent epidemiological investigation indicates high susceptibility to brain damage during prenatal exposures to Hg. An important objective for future investigation is to establish the lowest effect level for human exposure to methylmercury.

We develop an empirically constrained multicompartment box model for mercury cycling in open ocean regions to investigate changes in concentrations resulting from anthropogenic perturbations of the global mercury cycle. Using Monte Carlo simulations, we explicitly consider the effects of variability in measured parameters on modeled seawater concentrations. Our simulations show that anthropogenic enrichment in all surface (25%) and deep ocean waters (11%) is lower than global atmospheric enrichment (300–500%) and varies considerably among geographic regions, ranging from >60% in parts of the Atlantic and Mediterranean to <1% in the deep Pacific. Model results indicate that open ocean mercury concentrations do not rapidly equilibrate with atmospheric deposition and on average will increase if anthropogenic emissions remain at their present level. We estimate the temporal lag between changes in atmospheric deposition and ocean mercury concentrations will vary from decades in most of the Atlantic up to centuries in parts of the Pacific.

Abstract Purpose: Utomilumab (PF-05082566) is an agonistic mAb that engages the immune costimulatory molecule 4-1BB/CD137. In this first-in-human, phase I, open-label, multicenter, multiple-dose study (NCT01307267) we evaluated safety, tolerability, pharmacokinetics, preliminary clinical activity, and pharmacodynamics of single-agent utomilumab in patients with advanced malignancies. Experimental Design: Dose escalation was based on a standard 3+3 design for doses of utomilumab from 0.006 to 0.3 mg/kg every 4 weeks and a time-to-event continual reassessment method for utomilumab 0.6 to 10 mg/kg every 4 weeks. The primary study endpoint was dose-limiting toxicity (DLT) in the first two cycles. Results: Utomilumab demonstrated a well-tolerated safety profile (N = 55). None of the patients experienced a DLT at the dose levels evaluated. The most common treatment-related adverse events were fatigue, pyrexia, decreased appetite, dizziness, and rash (<10% of patients). Only one (1.8%) patient experienced a grade 3–4 treatment-related adverse event (fatigue), and no clinically relevant elevations in transaminases were noted. Utomilumab demonstrated linear pharmacokinetics at doses ranging from 0.006 to 10 mg/kg, with similar safety and pharmacokinetics in anti-drug antibody (ADA)-negative and ADA-positive patients. The overall objective response rate was 3.8% (95% CI, 0.5%–13.0%) in patients with solid tumors and 13.3% in patients with Merkel cell carcinoma, including a complete response and a partial response. Circulating biomarkers support 4-1BB/CD137 engagement by utomilumab and suggest that circulating lymphocyte levels may influence probability of clinical benefit. Conclusions: The favorable safety profile and preliminary antitumor activity demonstrated by utomilumab warrant further evaluation in patients with advanced malignancies. Clin Cancer Res; 24(8); 1816–23. ©2018 AACR.

We review the molecular and epidemiological characteristics of cetacean morbillivirus (CeMV) and the diagnosis and pathogenesis of associated disease, with six different strains detected in cetaceans worldwide. CeMV has caused epidemics with high mortality in odontocetes in Europe, the USA and Australia. It represents a distinct species within the Morbillivirus genus. Although most CeMV strains are phylogenetically closely related, recent data indicate that morbilliviruses recovered from Indo-Pacific bottlenose dolphins (Tursiops aduncus), from Western Australia, and a Guiana dolphin (Sotalia guianensis), from Brazil, are divergent. The signaling lymphocyte activation molecule (SLAM) cell receptor for CeMV has been characterized in cetaceans. It shares higher amino acid identity with the ruminant SLAM than with the receptors of carnivores or humans, reflecting the evolutionary history of these mammalian taxa. In Delphinidae, three amino acid substitutions may result in a higher affinity for the virus. Infection is diagnosed by histology, immunohistochemistry, virus isolation, RT-PCR, and serology. Classical CeMV-associated lesions include bronchointerstitial pneumonia, encephalitis, syncytia, and lymphoid depletion associated with immunosuppression. Cetaceans that survive the acute disease may develop fatal secondary infections and chronic encephalitis. Endemically infected, gregarious odontocetes probably serve as reservoirs and vectors. Transmission likely occurs through the inhalation of aerosolized virus but mother to fetus transmission was also reported.

Abstract Purpose: This phase Ib study (NCT02179918) evaluated the safety, antitumor activity, pharmacokinetics, and pharmacodynamics of utomilumab, a fully human IgG2 mAb agonist of the T-cell costimulatory receptor 4-1BB/CD137 in combination with the humanized, PD-1–blocking IgG4 mAb pembrolizumab in patients with advanced solid tumors. Experimental Design: Utomilumab (0.45–5.0 mg/kg) and pembrolizumab (2 mg/kg) were administered intravenously every 3 weeks. Utomilumab dose escalation was conducted using the time-to-event continual reassessment method. Results: Twenty-three patients received combination treatment with no dose-limiting toxicities. Treatment-emergent adverse events were mostly grades 1 to 2, without any treatment-related discontinuations. Six patients (26.1%) had confirmed complete or partial responses. Pharmacokinetics and immunogenicity of utomilumab and pembrolizumab were similar when administered alone or in combination. A trend toward higher levels of activated memory/effector peripheral blood CD8+ T cells was observed in responders versus nonresponders. Conclusions: The safety, tolerability, and clinical activity demonstrated by utomilumab in combination with pembrolizumab support further investigation in patients with advanced solid tumors. Clin Cancer Res; 23(18); 5349–57. ©2017 AACR. See related commentary by Pérez-Ruiz et al., p. 5326

Quantitative real-time PCR (qPCR) has become a gold standard for the quantification of nucleic acids and microorganism abundances, in which plasmid DNA carrying the target genes are most commonly used as the standard. A recent study showed that supercoiled circular confirmation of DNA appeared to suppress PCR amplification. However, to what extent to which different structural types of DNA (circular versus linear) used as the standard may affect the quantification accuracy has not been evaluated. In this study, we quantitatively compared qPCR accuracies based on circular plasmid (mostly in supercoiled form) and linear DNA standards (linearized plasmid DNA or PCR amplicons), using proliferating cell nuclear gene (pcna), the ubiquitous eukaryotic gene, in five marine microalgae as a model gene. We observed that PCR using circular plasmids as template gave 2.65-4.38 more of the threshold cycle number than did equimolar linear standards. While the documented genome sequence of the diatom Thalassiosira pseudonana shows a single copy of pcna, qPCR using the circular plasmid as standard yielded an estimate of 7.77 copies of pcna per genome whereas that using the linear standard gave 1.02 copies per genome. We conclude that circular plasmid DNA is unsuitable as a standard, and linear DNA should be used instead, in absolute qPCR. The serious overestimation by the circular plasmid standard is likely due to the undetected lower efficiency of its amplification in the early stage of PCR when the supercoiled plasmid is the dominant template.

PURPOSE: To estimate the maximum tolerated dose (MTD) for continuous oral administration of the γ-secretase inhibitor PF-03084014, determine the recommended phase II dose (RP2D), and evaluate safety and preliminary activity in patients with advanced solid tumors. EXPERIMENTAL DESIGN: This open-label, phase I study consisted of a dose-finding portion based on a 3+3 design, followed by an expansion cohort. PF-03084014 was administered orally, twice daily (BID) for 21 continuous days. Tested doses ranged from 20 to 330 mg BID. In the expansion cohort, patients were to receive the estimated MTD or a lower dose of PF-03084014. RESULTS: A total of 64 patients received treatment. The MTD was estimated to be 220 mg BID. The RP2D was determined to be 150 mg BID, based on the better safety profile versus the 220-mg BID dose, given comparable NOTCH-related target inhibition. The most common treatment-related adverse events were diarrhea, nausea, fatigue, hypophosphatemia, vomiting, rash, and decreased appetite, which were generally mild to moderate in severity. One patient with advanced thyroid cancer had a complete response, and five of seven response-evaluable patients with desmoid tumor achieved a partial response (71.4% objective response rate). Tumor responses were mostly durable, ranging from 1.74+ to 24+ months. PF-03084014 demonstrated a generally dose-dependent pharmacokinetic profile at doses ranging from 20 to 330 mg BID. Consistent downmodulation of NOTCH-related HES4 gene expression was observed in peripheral blood from all evaluable patients. CONCLUSION: Further development of PF-03084014 for the treatment of patients with advanced solid tumors is warranted and currently under evaluation.

Abstract Trastuzumab emtansine (T-DM1) is an antibody–drug conjugate (ADC) that has demonstrated clinical benefit for patients with HER2+ metastatic breast cancer; however, its clinical activity is limited by inherent or acquired drug resistance. The molecular mechanisms that drive clinical resistance to T-DM1, especially in HER2+ tumors, are not well understood. We used HER2+ cell lines to develop models of T-DM1 resistance using a cyclical dosing schema in which cells received T-DM1 in an “on-off” routine until a T-DM1–resistant population was generated. T-DM1–resistant N87 cells (N87-TM) were cross-resistant to a panel of trastuzumab-ADCs (T-ADCs) with non–cleavable-linked auristatins. N87-TM cells do not have a decrease in HER2 protein levels or an increase in drug transporter protein (e.g., MDR1) expression compared with parental N87 cells. Intriguingly, T-ADCs using auristatin payloads attached via an enzymatically cleavable linker overcome T-DM1 resistance in N87-TM cells. Importantly, N87-TM cells implanted into athymic mice formed T-DM1 refractory tumors that remain sensitive to T-ADCs with cleavable-linked auristatin payloads. Comparative proteomic profiling suggested enrichment in proteins that mediate caveolae formation and endocytosis in the N87-TM cells. Indeed, N87-TM cells internalize T-ADCs into intracellular caveolin-1 (CAV1)–positive puncta and alter their trafficking to the lysosome compared with N87 cells. T-DM1 colocalization into intracellular CAV1-positive puncta correlated with reduced response to T-DM1 in a panel of HER2+ cell lines. Together, these data suggest that caveolae-mediated endocytosis of T-DM1 may serve as a novel predictive biomarker for patient response to T-DM1. Mol Cancer Ther; 17(1); 243–53. ©2017 AACR.

The partitioning of gaseous mercury between the atmosphere and surface waters was determined in the equatorial Pacific Ocean. The highest concentrations of dissolved gaseous mercury occurred in cooler, nutrient-rich waters that characterize equatorial upwelling and increased biological productivity at the sea surface. The surface waters were supersaturated with respect to elemental mercury; a significant flux of elemental mercury to the atmosphere is predicted for the equatorial Pacific. When normalized to primary production on a global basis, the ocean effluxes of mercury may rival anthropogenic emissions of mercury to the atmosphere.

Short-term effects of bottom trawling on a 'hard-bottom' (pebble, cobble, and boulder) seafloor were studied on the outer continental shelf in the eastern Gulf of Alaska. Eight sites were trawled in August 1996; then, from a research submersible we videotaped each trawl path and a nearby reference transect to obtain quantitative data. Boulders were displaced, and large epifaunal invertebrates were removed or damaged by a single trawl pass. These structural components of habitat were the dominant features on the seafloor. There was a significant decrease in density, and an increase in damage, to sponges and anthozoans in trawled versus reference transects. Changes in density, or damage to most motile invertebrates were not detected. Delayed mortality, of apparently undamaged invertebrates, may have resulted in greater impact than we detected. Alternatively, over time, some invertebrates may have recovered from any damage previously suffered. A subsequent survey at these sites will address these questions.

Phytoplankton biomass and productivity have been continuously monitored from ocean color satellites for over a decade. Yet, the most widely used empirical approach for estimating chlorophyll a (Chl) from satellites can be in error by a factor of 5 or more. Such variability is due to differences in absorption and backscattering properties of phytoplankton and related concentrations of colored-dissolved organic matter (CDOM) and minerals. The empirical algorithms have built-in assumptions that follow the basic precept of biological oceanography—namely, oligotrophic regions with low phytoplankton biomass are populated with small phytoplankton, whereas more productive regions contain larger bloom-forming phytoplankton. With a changing world ocean, phytoplankton composition may shift in response to altered environmental forcing, and CDOM and mineral concentrations may become uncoupled from phytoplankton stocks, creating further uncertainty and error in the empirical approaches. Hence, caution is warranted when using empirically derived Chl to infer climate-related changes in ocean biology. The Southern Ocean is already experiencing climatic shifts and shows substantial errors in satellite-derived Chl for different phytoplankton assemblages. Accurate global assessments of phytoplankton will require improved technology and modeling, enhanced field observations, and ongoing validation of our “eyes in space.”

Suspension-feeding bivalve molluscs are foundation species in coastal intertidal systems. The selective feeding capabilities of these animals can have a large influence on phytoplankton communities and nutrient flow to the benthos. Particle selection, including the types of particles chosen for ingestion and the possible mechanisms mediating selection, has been studied extensively and reported in the literature. To date, however, the possible mechanisms mediating these selective processes have remained elusive. Generally, the focus on a few key commercial species, and their demonstrated range of selective capabilities, has made it difficult to design studies that elucidate the mechanisms behind particle selection. This review focuses on key research that has been carried out in the last 20 y toward better understanding the mechanism that underlays selective capture and ingestion of particles in this important group of animals. Recently, work has been completed which has advanced the field in pointing to a passive mechanism as a mediator of selection, with the interactions between the physicochemical properties of particles and the mucus covering the pallial organs most likely mediating food choice. Although no strong evidence for an immediate, active mechanism which underlies particle selection was found, avenues for future research are suggested in this review. The possible mechanisms that control capture, including qualitative precapture selection, are also summarized and discussed in depth. Methodological considerations for rigorous experiments to advance the field are also discussed, including suggestions of general guidelines for experimental designs, which will allow better comparison of findings across studies.

OBJECTIVE: To determine if n-3 polyunsaturated fatty acid (PUFA) supplementation (versus treatment with n-6 polyunsaturated or other fatty acid supplements) affects the metabolism of osteoarthritic (OA) cartilage. METHODS: The metabolic profile of human OA cartilage was determined at the time of harvest and after 24-hour exposure to n-3 PUFAs or other classes of fatty acids, followed by explant culture for 4 days in the presence or absence of interleukin-1 (IL-1). Parameters measured were glycosaminoglycan release, aggrecanase and matrix metalloproteinase (MMP) activity, and the levels of expression of messenger RNA (mRNA) for mediators of inflammation, aggrecanases, MMPs, and their natural tissue inhibitors (tissue inhibitors of metalloproteinases [TIMPs]). RESULTS: Supplementation with n-3 PUFA (but not other fatty acids) reduced, in a dose-dependent manner, the endogenous and IL-1-induced release of proteoglycan metabolites from articular cartilage explants and specifically abolished endogenous aggrecanase and collagenase proteolytic activity. Similarly, expression of mRNA for ADAMTS-4, MMP-13, and MMP-3 (but not TIMP-1, -2, or -3) was also specifically abolished with n-3 PUFA supplementation. In addition, n-3 PUFA supplementation abolished the expression of mRNA for mediators of inflammation (cyclooxygenase 2, 5-lipoxygenase, 5-lipoxygenase-activating protein, tumor necrosis factor alpha, IL-1alpha, and IL-1beta) without affecting the expression of message for several other proteins involved in normal tissue homeostasis. CONCLUSION: These studies show that the pathologic indicators manifested in human OA cartilage can be significantly altered by exposure of the cartilage to n-3 PUFA, but not to other classes of fatty acids.

Mechanisms of dimethyl sulphide (DMS) and methanethiol (MT) production and consumption were determined in moderately hypersaline mats, Guerrero Negro, Mexico. Biological pathways regulated the net flux of DMS and MT as revealed by increases in flux resulting from decreased salinity, increased temperature and the removal of oxygen. Dimethylsulphoniopropionate (DMSP) was not present in these microbial mats and DMS and MT are probably formed by the reaction of photosynthetically produced low-molecular weight organic carbon and biogenic hydrogen sulphide derived from sulphate reduction. These observations provide an alternative to the notion that DMSP or S-containing amino acids are the dominant precursors of DMS in intertidal sediment systems. The major sink for DMS in the microbial mats was biological consumption, whereas photochemical oxidation to dimethylsulphoxide was the major sink for DMS in the overlying water column. Diel flux measurements demonstrated that significantly more DMS is released from the system during the night than during the day. The major consumers of DMS in the presence of oxygen were monooxygenase-utilizing bacteria, whereas under anoxic conditions, DMS was predominantly consumed by sulphate-reducing bacteria and methanethiol was consumed by methanogenic bacteria. Aerobic and anaerobic consumption rates of DMS were nearly identical. Mass balance estimates suggest that the consumption in the water column is likely to be smaller than net the flux from the mats. Volatile organic sulphur compounds are thus indicators of high rates of carbon fixation and sulphate reduction in these laminated sediment ecosystems, and atmospheric sulphur can be generated as a biogenic signature of the microbial mat community.

Tremelimumab (CP-675,206) is a fully human monoclonal antibody specific for human cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, CD152) in clinical development for patients with cancer. Blocking the CTLA-4 negative costimulatory receptor with the antagonistic antibody tremelimumab results in immune activation. Administration of tremelimumab to patients with locally advanced and metastatic melanoma has resulted in a subset of patients with durable objective tumor regressions. Its IgG(2) isotype minimizes the possibility of cytotoxic effects on activated T lymphocytes and cytokine release syndrome. Preclinical testing in vitro and in large animal models predicted the target concentrations of circulating antibody in humans necessary for a pharmacodynamic effect. Phase I clinical trials provided evidence of dose- or exposure-related effects consistent with the anticipated mechanism of action. Further clinical development has led to two ongoing registration trials in patients with metastatic melanoma: a phase III randomized trial of tremelimumab versus dacarbazine or temozolomide in previously untreated patients with advanced melanoma and a phase II trial of tremelimumab in previously treated patients with advanced melanoma.