Negative relationships between species richness and temporal variability are common but weak in natural systems.

Effects of species diversity on population and community stability (or more precisely, the effects of species richness on temporal variability) have been studied for several decades, but there have been no large-scale tests in natural communities of predictions from theory. We used 91 data sets including plants, fish, small mammals, zooplankton, birds, and insects, to examine the relationship between species richness and temporal variability in populations and communities. Seventy-eight of 91 data sets showed a negative relationship between species richness and population variability; 46 of these relationships were statistically significant. Only five of the 13 positive richness-population variability relationships were statistically significant. Similarly, 51 of 91 data sets showed a negative relationship between species richness and community variability; of these, 26 were statistically significant. Seven of the 40 positive richness-community-variability relationships were statistically significant. We were able to test transferability (i.e., the predictive ability of models for sites that are spatially distinct from sites that were used to build the models) for 69 of 91 data sets; 35 and 31 data sets were transferable at the population and community levels, respectively. Only four were positive at the population level, and two at the community level. We conclude that there is compelling evidence of a negative relationship between species richness and temporal variability for about one-half of the ecological communities we examined. However, species richness explained relatively little of the variability in population or community abundances and resulted in small improvements in predictive ability.

Carbon dating cancer: defining the chronology of metastatic progression in colorectal cancer.

Background: Patients often ask oncologists how long a cancer has been present before causing symptoms or spreading to other organs. The evolutionary trajectory of cancers can be defined using phylogenetic approaches but lack of chronological references makes dating the exact onset of tumours very challenging. Patients and methods: Here, we describe the case of a colorectal cancer (CRC) patient presenting with synchronous lung metastasis and metachronous thyroid, chest wall and urinary tract metastases over the course of 5 years. The chest wall metastasis was caused by needle tract seeding, implying a known time of onset. Using whole genome sequencing data from primary and metastatic sites we inferred the complete chronology of the cancer by exploiting the time of needle tract seeding as an in vivo 'stopwatch'. This approach allowed us to follow the progression of the disease back in time, dating each ancestral node of the phylogenetic tree in the past history of the tumour. We used a Bayesian phylogenomic approach, which accounts for possible dynamic changes in mutational rate, to reconstruct the phylogenetic tree and effectively 'carbon date' the malignant progression. Results: The primary colon cancer emerged between 5 and 8 years before the clinical diagnosis. The primary tumour metastasized to the lung and the thyroid within a year from its onset. The thyroid lesion presented as a tumour-to-tumour deposit within a benign Hurthle adenoma. Despite rapid metastatic progression from the primary tumour, the patient showed an indolent disease course. Primary cancer and metastases were microsatellite stable and displayed low chromosomal instability. Neo-antigen analysis suggested minimal immunogenicity. Conclusion: Our data provide the first in vivo experimental evidence documenting the timing of metastatic progression in CRC and suggest that genomic instability might be more important than the metastatic potential of the primary cancer in dictating CRC fate.

Complement inhibition prevents oncolytic vaccinia virus neutralization in immune humans and cynomolgus macaques.

Oncolytic viruses (OVs) have shown promising clinical activity when administered by direct intratumoral injection. However, natural barriers in the blood, including antibodies and complement, are likely to limit the ability to repeatedly administer OVs by the intravenous route. We demonstrate here that for a prototype of the clinical vaccinia virus based product Pexa-Vec, the neutralizing activity of antibodies elicited by smallpox vaccination, as well as the anamnestic response in hyperimmune virus treated cancer patients, is strictly dependent on the activation of complement. In immunized rats, complement depletion stabilized vaccinia virus in the blood and led to improved delivery to tumors. Complement depletion also enhanced tumor infection when virus was directly injected into tumors in immunized animals. The feasibility and safety of using a complement inhibitor, CP40, in combination with vaccinia virus was tested in cynomolgus macaques. CP40 pretreatment elicited an average 10-fold increase in infectious titer in the blood early after the infusion and prolonged the time during which infectious virus was detectable in the blood of animals with preexisting immunity. Capitalizing on the complement dependence of antivaccinia antibody with adjunct complement inhibitors may increase the infectious dose of oncolytic vaccinia virus delivered to tumors in virus in immune hosts.

Rapid biodiagnostic ex vivo imaging at 1 µm pixel resolution with thermal source FTIR FPA.

A recent upgrade to the optics configuration of a thermal source FTIR microscope equipped with a focal plane array detector has enabled rapid acquisition of high magnification spectrochemical images, in transmission, with an effective geometric pixel size of ∼1 × 1 µm(2) at the sample plane. Examples, including standard imaging targets for scale and accuracy, as well as biomedical tissues and microorganisms, have been imaged with the new system and contrasted with data acquired at normal magnification and with a high magnification multi-beam synchrotron instrument. With this optics upgrade, one can now conduct rapid biodiagnostic ex vivo tissue imaging in-house, with images collected over larger areas, in less time (minutes) and with comparable quality and resolution to the best synchrotron source FTIR imaging capabilities.

Experimental study designs to improve the evaluation of road mitigation measures for wildlife.

An experimental approach to road mitigation that maximizes inferential power is essential to ensure that mitigation is both ecologically-effective and cost-effective. Here, we set out the need for and standards of using an experimental approach to road mitigation, in order to improve knowledge of the influence of mitigation measures on wildlife populations. We point out two key areas that need to be considered when conducting mitigation experiments. First, researchers need to get involved at the earliest stage of the road or mitigation project to ensure the necessary planning and funds are available for conducting a high quality experiment. Second, experimentation will generate new knowledge about the parameters that influence mitigation effectiveness, which ultimately allows better prediction for future road mitigation projects. We identify seven key questions about mitigation structures (i.e., wildlife crossing structures and fencing) that remain largely or entirely unanswered at the population-level: (1) Does a given crossing structure work? What type and size of crossing structures should we use? (2) How many crossing structures should we build? (3) Is it more effective to install a small number of large-sized crossing structures or a large number of small-sized crossing structures? (4) How much barrier fencing is needed for a given length of road? (5) Do we need funnel fencing to lead animals to crossing structures, and how long does such fencing have to be? (6) How should we manage/manipulate the environment in the area around the crossing structures and fencing? (7) Where should we place crossing structures and barrier fencing? We provide experimental approaches to answering each of them using example Before-After-Control-Impact (BACI) study designs for two stages in the road/mitigation project where researchers may become involved: (1) at the beginning of a road/mitigation project, and (2) after the mitigation has been constructed; highlighting real case studies when available.

Collaboration and engagement with decision-makers are needed to reduce evidence complacency in wildlife management.

There exists an extensive, diverse, and robust evidence base to support complex decisions that address the planetary biodiversity crisis. However, it is generally not sought or used by environmental decision-makers, who instead draw on intuition, experience, or opinion to inform important decisions. Thus, there is a need to examine evidence exchange processes in wildlife management to understand the multiple inputs to decisions. Here, we adopt a novel approach, fuzzy cognitive mapping (FCM), to examine perceptions of individuals from Indigenous and Western governments on the reliability of evidence which may influence freshwater fisheries management decisions in British Columbia, Canada. We facilitated four FCM workshops participants representing Indigenous or Western regulatory/governance groups of fisheries managers. Our results show that flows of evidence to decision-makers occur within a relatively closed governance network, constrained to the few well-connected decision-making organizations (i.e., wildlife management agencies) and their close partners. This implies that increased collaboration (i.e., knowledge co-production) and engagement (i.e., knowledge brokerage) with wildlife managers and decision-makers are needed to produce actionable evidence and increase evidence exchange.

Inhibiting ice recrystallization and optimization of cell viability after cryopreservation.

The ice recrystallization inhibition activity of various mono- and disaccharides has been correlated with their ability to cryopreserve human cell lines at various concentrations. Cell viabilities after cryopreservation were compared with control experiments where cells were cryopreserved with dimethylsulfoxide (DMSO). The most potent inhibitors of ice recrystallization were 220 mM solutions of disaccharides; however, the best cell viability was obtained when a 200 mM d-galactose solution was utilized. This solution was minimally cytotoxic at physiological temperature and effectively preserved cells during freeze-thaw. In fact, this carbohydrate was just as effective as a 5% DMSO solution. Further studies indicated that the cryoprotective benefit of d-galactose was a result of its internalization and its ability to mitigate osmotic stress, prevent intracellular ice formation and/or inhibit ice recrystallization. This study supports the hypothesis that the ability of a cryoprotectant to inhibit ice recrystallization is an important property to enhance cell viability post-freeze-thaw. This cryoprotective benefit is observed in three different human cell lines. Furthermore, we demonstrated that the ability of a potential cryoprotectant to inhibit ice recrystallation may be used as a predictor of its ability to preserve cells at subzero temperatures.

The longitudinal relationship between thigh muscle mass and the development of knee osteoarthritis.

OBJECTIVE: Greater quadriceps strength has been found to reduce risk for symptomatic knee osteoarthritis (SxKOA) and knee joint space narrowing (JSN). However, this finding could relate to muscle mass or activation pattern. The purpose of this study was to assess whether greater thigh muscle mass protects against (1) incident radiographic (RKOA), (2) incident SxKOA or (3) worsening of knee JSN by 30-month follow-up. DESIGN: Multicenter Osteoarthritis (MOST) study participants, who underwent dual-energy X-ray absorptiometry (DXA) at the Iowa site were included. Thigh muscle mass was calculated from DXA image sub-regions. Sex-stratified, knee-based analyses controlled for incomplete independence between limbs within subjects. The effect of thigh lean mass and specific strength as predictors of ipsilateral RKOA, SxKOA and worsening of JSN were assessed, while controlling for age, body mass index (BMI), and history of knee surgery. RESULTS: A total of 519 men (948 knees) and 784 women (1453 knees) were included. Mean age and BMI were 62 years and 30 kg/m(2). Thigh muscle mass was not associated with risk for RKOA, SxKOA or knee JSN. However, in comparison with the lowest tertile, those in the highest and middle tertiles of knee extensor specific strength had a lower risk for SxKOA and JSN [odds ratio (OR) 0.29-0.68]. CONCLUSIONS: Thigh muscle mass does not appear to confer protection against incident or worsening knee OA. These findings suggest that future studies of risk for knee OA should focus on the roles of knee extensor neuromuscular activation and muscle physiology, rather than the muscle mass.

Generation of priority research questions to inform conservation policy and management at a national level.

Integrating knowledge from across the natural and social sciences is necessary to effectively address societal tradeoffs between human use of biological diversity and its preservation. Collaborative processes can change the ways decision makers think about scientific evidence, enhance levels of mutual trust and credibility, and advance the conservation policy discourse. Canada has responsibility for a large fraction of some major ecosystems, such as boreal forests, Arctic tundra, wetlands, and temperate and Arctic oceans. Stressors to biological diversity within these ecosystems arise from activities of the country's resource-based economy, as well as external drivers of environmental change. Effective management is complicated by incongruence between ecological and political boundaries and conflicting perspectives on social and economic goals. Many knowledge gaps about stressors and their management might be reduced through targeted, timely research. We identify 40 questions that, if addressed or answered, would advance research that has a high probability of supporting development of effective policies and management strategies for species, ecosystems, and ecological processes in Canada. A total of 396 candidate questions drawn from natural and social science disciplines were contributed by individuals with diverse organizational affiliations. These were collaboratively winnowed to 40 by our team of collaborators. The questions emphasize understanding ecosystems, the effects and mitigation of climate change, coordinating governance and management efforts across multiple jurisdictions, and examining relations between conservation policy and the social and economic well-being of Aboriginal peoples. The questions we identified provide potential links between evidence from the conservation sciences and formulation of policies for conservation and resource management. Our collaborative process of communication and engagement between scientists and decision makers for generating and prioritizing research questions at a national level could be a model for similar efforts beyond Canada.

Research effort devoted to regulating and supporting ecosystem services by environmental scientists and economists.

The economic valuation of ecosystem services in part reflects the desire to use conventional economic tools (markets and economic instruments) to conserve ecosystem services. However, for regulating and supporting ecosystem services that depend on ecosystem structure and function, estimation of economic value requires estimates of the current level of underlying ecological functions first. This primary step is in principle, the job of environmental scientists, not economists. Here, we provide a coarse-level quantitative assessment of the relationship between the research effort expended by environmental scientists (on the biophysical values) and economists (on the monetary values) on 15 different regulating and supporting services in 32 ecosystem types using peer-reviewed article hits retrieved from bibliographic databases as a measure of research effort. We find a positive, moderately strong (r = 0.69) correlation between research efforts in the two domains, a result that, while encouraging, is likely to reflect serendipity rather than the deliberate design of integrated environmental science-economics research programs. Our results suggest that compared to environmental science research effort economic valuation is devoted to a smaller, less diverse set of ecosystem services but a broader, more diverse, set of ecosystem types. The two domains differed more with respect to the ecosystem services that are the major focus of research effort than they did with respect to the ecosystem types of principal research interest. For example, carbon sequestration, erosion regulation, and nutrient cycling receive more relative research effort in the environmental sciences; air quality regulation in economic valuations. For both domains, cultivated areas, wetlands, and urban/semi-urban ecosystem types received relatively large research effort, while arctic and mountain tundra, cave and subterranean, cryosphere, intertidal/littoral zone, and kelp forest ecosystem types received negligible research effort. We suggest ways and means by which the field of sustainability science may be improved by the design and implementation of a searchable database of environmental science and economic valuation literature as well as a global ecosystem service research network and repository that explicitly links research on the estimation and prediction of biophysical ecosystem functions with that of the social sciences and other knowledge systems. These suggestions would, at least in principle, facilitate a more efficient research agenda between economists and environmental scientists and aid management, regulatory and judicial decision-makers.

Species listing under Canada's Species at Risk Act.

In a preliminary analysis of listing decisions under Canada's Species at Risk Act (SARA), Mooers et al. (2007)demonstrated an apparent bias against marine and northern species. As a follow-up, we expanded the set of potential explanatory variables, including information on jurisdictional and administrative elements of the listing process, and considered an additional 16 species recommended for listing by SARA's scientific advisory committee as of 15 August 2006. Logistic model selection based on Akaike differences suggested that species were less likely to be listed if they were harvested or had commercial or subsistence harvesting as an explicitly identified threat; had Department of Fisheries and Oceans (DFO) as a responsible authority (RA); were located in Canada's north generally, and especially in Nunavut; or were found mostly or entirely within Canada. Subsequent model validation with an independent set of 50 species for which a listing decision was handed down in December 2007 showed an overall misclassification rate of <0.10, indicating reasonable predictive power. In light of these results, we recommend that RAs under SARA adopt a two-track listing approach to address problems of delays arising from extended consultations and the inconsistent use by the RAs of socioeconomic analysis; consider revising SARA so that socioeconomic analysis occurs during decisions about protecting species and their habitats rather than at the listing stage; and maintain an integrated database with information on species' biology, threats, and agency actions to enable future evaluation of SARA's impact.

Variation in large-bodied fish-community structure and abundance in relation to water-management regime in a large regulated river.

Variation in life-history traits (growth, condition, mortality and recruitment) and relative abundance of 11 large-bodied fish species was investigated among three water-management regimes (unimpounded, run-of-the-river and winter reservoirs) in the large regulated Ottawa River, Canada. If waterpower management had an effect on fishes, then (1) would be expected community structuring among water-management regimes and (2) species with similar life-history traits should be affected in a similar manner. Large-bodied fish communities were assessed using two different standard index-netting techniques, one using trap nets and the other gillnets. Community structure could be discriminated based on species caught in nets using holographic neural networks (78.8% correct overall classification rate using trap nets and 76.0% using gillnets); therefore, water-management regimes affected community structure in the Ottawa River. Littoral zone benthivores were significantly lower in abundance (P < 0.001) or absent in winter reservoirs, whereas the abundance of planktivores or species that were planktivorous at young ages were significantly greater than in unimpounded river reaches. Growth, condition and mortality did not vary among reach types except smallmouth bass Micropterus dolomieu were in better condition in winter reservoirs than unimpounded reaches. Lake sturgeon Acipenser fulvescens recruitment was impaired in run-of-the-river reaches, whereas recruitment for other species that spawn in fast water was not affected.

Exploring aboriginal views of health using fuzzy cognitive maps and transitive closure. A case study of the determinants of diabetes.

OBJECTIVES: To demonstrate how fuzzy cognitive maps may be used to extract, present and compare Aboriginal perspectives, using the determinants of diabetes as a case study. METHODS: Participants from the Mohawk Community of Akwesasne and the Miawpukek First Nation in Conne River created fuzzy cognitive maps (FCMs) (N=3) detailing their views of "the causes of diabetes in their community", through a facilitated, group mapping session. For each FCM, the net causal effect of every determinant (direct or indirect) on diabetes was calculated from its transitive closure. The net causal effects were then compared across the set of FCMs to identify strong, weak and controversial determinants. RESULTS: Comparison of FCMs revealed significant heterogeneity in the perspectives of diabetes. The Akwesasne participants focused heavily on social, traditional and spiritual factors, while Conne River participants placed more importance on direct personal and lifestyle factors. There was, however, a core of strong, validated determinants related primarily to healthy diet and physical activity. CONCLUSIONS: This work demonstrates how FCM may be used to extract and represent different perspectives of complex issues allowing for comparisons among stakeholders or knowledge groups. Comparison of multiple FCM employing the transitive closure may then be used to identify areas of agreement and controversy.

Total disc replacement versus anterior cervical discectomy and fusion: a systematic review with meta-analysis of data from a total of 3160 patients across 14 randomized controlled trials with both short- and medium- to long-term outcomes.

Aims: The aim of this study was to determine how the short- and medium- to long-term outcome measures after total disc replacement (TDR) compare with those of anterior cervical discectomy and fusion (ACDF), using a systematic review and meta-analysis. Patients and Methods: Databases including Medline, Embase, and Scopus were searched. Inclusion criteria involved prospective randomized control trials (RCTs) reporting the surgical treatment of patients with symptomatic degenerative cervical disc disease. Two independent investigators extracted the data. The strength of evidence was assessed using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) criteria. The primary outcome measures were overall and neurological success, and these were included in the meta-analysis. Standardized patient-reported outcomes, including the incidence of further surgery and adjacent segment disease, were summarized and discussed. Results: A total of 22 papers published from 14 RCTs were included, representing 3160 patients with follow-up of up to ten years. Meta-analysis indicated that TDR is superior to ACDF at two years and between four and seven years. In the short-term, patients who underwent TDR had better patient-reported outcomes than those who underwent ACDF, but at two years this was typically not significant. Results between four and seven years showed significant differences in Neck Disability Index (NDI), 36-Item Short-Form Health Survey (SF-36) physical component scores, dysphagia, and satisfaction, all favouring TDR. Most trials found significantly less adjacent segment disease after TDR at both two years (short-term) and between four and seven years (medium- to long-term). Conclusion: TDR is as effective as ACDF and superior for some outcomes. Disc replacement reduces the risk of adjacent segment disease. Continued uncertainty remains about degeneration of the prosthesis. Long-term surveillance of patients who undergo TDR may allow its routine use. Cite this article: Bone Joint J 2018;100-B:991-1001.

Mapping widespread and increasing underwater noise pollution from acoustic deterrent devices.

Acoustic deterrent devices (ADDs) are used in attempts to mitigate pinniped depredation on aquaculture sites through the emission of loud and pervasive noise. This study quantified spatio-temporal changes in underwater ADD noise detections along western Scotland over 11 years. Acoustic point data ('listening events') collected during cetacean line-transect surveys were used to map ADD presence between 2006 and 2016. A total of 19,601 listening events occurred along the Scottish west coast, and ADD presence was recorded during 1371 listening events. Results indicated a steady increase in ADD detections from 2006 (0.05%) to 2016 (6.8%), with the highest number of detections in 2013 (12.6%), as well as substantial geographic expansion. This study demonstrates that ADDs are a significant and chronic source of underwater noise on the Scottish west coast with potential adverse impacts on target (pinniped) and non-target (e.g. cetaceans) species, which requires further study and improved monitoring and regulatory strategies.

Integrating conventional science and aboriginal perspectives on diabetes using fuzzy cognitive maps.

There is concern among Aboriginal communities in Canada that conventional approaches to the treatment of diabetes are ineffective in part because they fail to recognize the local Aboriginal perspective on the causal determinants of diabetes. While this shortcoming has been recognized, there have been no explicit attempts to practically define these perspectives and prescribe how conventional health management might be altered to incorporate them. In part, this may be due to difficulties in communicating Aboriginal perspectives in a manner which permits incorporation into conventional science-based frameworks. Here we use fuzzy cognitive mapping (FCM) as a technique to represent and compare Canadian Aboriginal and conventional science perspectives on the determinants of diabetes as contained in published articles drawn from a search of Medline and PubMed (1966-2005). The FCM allows for a detailed description of the complex system of culture, spirituality and balance at the root of the Aboriginal view. It also highlights how, for these less tangible factors, it is possible to identify more concrete stressors and outcomes which are amenable to management and monitoring. This preliminary comparison of conventional and Aboriginal views also demonstrates the potential for FCM as a technique to extract, compare and integrate perspectives of different knowledge systems into health management and policy development.

How Effective Is Road Mitigation at Reducing Road-Kill? A Meta-Analysis.

Road traffic kills hundreds of millions of animals every year, posing a critical threat to the populations of many species. To address this problem there are more than forty types of road mitigation measures available that aim to reduce wildlife mortality on roads (road-kill). For road planners, deciding on what mitigation method to use has been problematic because there is little good information about the relative effectiveness of these measures in reducing road-kill, and the costs of these measures vary greatly. We conducted a meta-analysis using data from 50 studies that quantified the relationship between road-kill and a mitigation measure designed to reduce road-kill. Overall, mitigation measures reduce road-kill by 40% compared to controls. Fences, with or without crossing structures, reduce road-kill by 54%. We found no detectable effect on road-kill of crossing structures without fencing. We found that comparatively expensive mitigation measures reduce large mammal road-kill much more than inexpensive measures. For example, the combination of fencing and crossing structures led to an 83% reduction in road-kill of large mammals, compared to a 57% reduction for animal detection systems, and only a 1% for wildlife reflectors. We suggest that inexpensive measures such as reflectors should not be used until and unless their effectiveness is tested using a high-quality experimental approach. Our meta-analysis also highlights the fact that there are insufficient data to answer many of the most pressing questions that road planners ask about the effectiveness of road mitigation measures, such as whether other less common mitigation measures (e.g., measures to reduce traffic volume and/or speed) reduce road mortality, or to what extent the attributes of crossing structures and fences influence their effectiveness. To improve evaluations of mitigation effectiveness, studies should incorporate data collection before the mitigation is applied, and we recommend a minimum study duration of four years for Before-After, and a minimum of either four years or four sites for Before-After-Control-Impact designs.

Directed mutation of the basic domain of v-Jun alters DNA binding specificity and abolishes its oncogenic activity in chicken embryo fibroblasts.

Overexpression of v-Jun in chicken embryo fibroblasts (CEF) leads to oncogenic transformation phenotypically characterized by anchorage independent growth and release from contact inhibition (focus formation). The mechanisms involved in this oncogenic conversion however, are not yet clear. Because Jun is a transcription factor, it has been assumed that oncogenic transformation results directly from deregulated AP-1 target gene expression. However, a number of experimental observations in avian cell culture models fail to correlate oncogenesis with AP-1 activity suggesting that transformation induced by v-Jun may occur through an indirect mechanism. To test this possibility, we introduced point mutations into the basic DNA binding domain of v-Jun and created mutants that exhibit altered binding specificity. When expressed in CEF, these mutants fail to deregulate three known v-Jun target genes (JTAP-1, apolipoprotein A1, c-Jun) thus demonstrating in vivo specificity changes. Each of the binding specificity mutants was also tested for its ability to induce oncogenic transformation. Interestingly, expression of these mutants in CEF results in a phenotype indistinguishable from the vector control with respect to growth rate, focus formation and the ability to form colonies in soft agar. These results are consistent with a model requiring direct AP-1 target deregulation as a prerequisite of v-Jun induced cell transformation. With this in mind, we generated a series of additional mutants that retain the ability to bind AP-1 sequence elements, but vary in their oncogenic potential. We demonstrate the use of these mutants to screen v-Jun induced gene targets for a functional role in cell transformation.

The modulation of the human sodium iodide symporter activity by Graves' disease sera.

The transport of iodide into the thyroid, catalyzed by the Na+/I- symporter (NIS), is the initial and rate-limiting step in the formation of thyroid hormones. To study the basic characteristics of the human (h) NIS, we have established a Chinese hamster ovary cell line stably expressing the hNIS (CHO-NIS9). In agreement with previous work on the rat NIS, iodide uptake in these cells was initiated within 2 min of the addition of 131I, reaching a plateau after 30 min. Both perchlorate and thiocyanate inhibited iodide uptake in a dose-dependent manner, with inhibition evident at concentrations of 0.01 and 0.1 micromol/L, respectively, and reaching complete inhibition at 20 micromol/L and 500 micromol/L, respectively. Ouabain, which blocks the activity of the Na+/K+ adenosine triphosphatase, also inhibited iodide uptake in a dose-dependent manner, starting at concentrations of 100 micromol/L and reaching maximum inhibition at 1600 micromol/L, indicating that iodide uptake in these cells is sodium dependent. CHO-NIS9 cells were further used to study 88 sera from patients with Graves' disease, for iodide uptake inhibitory activity, which were compared with sera from 31 controls. Significant iodide uptake inhibition was taken as any inhibition in excess of the mean + 3 SD of the results with the control sera. On this basis, 27 (30.7%) of the Graves' sera, but none of the controls, inhibited iodide uptake in CHO-NIS9. IgGs from these patients also inhibited iodide uptake, indicating that this inhibitory activity was antibody mediated. In summary, we have established a CHO cell line stably expressing the hNIS and shown that antibodies in GD sera can inhibit iodide uptake in these cells. This further emphasizes the role of NIS as a novel autoantigen in thyroid immunity.