Predictors of fuel poverty and the equity of local fuel poverty support: secondary analysis of data from Bradford, England.

AIMS: Addressing fuel poverty is a critical public health issue given its recent rise in prevalence across Europe. Although previous research identifies national risk markers of fuel poverty, evidence is lacking on whether these are consistent across local geographies, and the equity of local interventions. In the UK's current economic climate, it is more crucial than ever that services benefit households in greatest need. This study aimed to determine significant predictors of fuel poverty among households in Bradford, England, comparing them to national-level predictors, and evaluate if households possessing significant fuel poverty predictors were equitably referred to a local fuel poverty service (Warm Homes Healthy People, WHHP). METHODS: A multivariate logistic regression model determined significant fuel poverty predictors in Bradford using household-level data from the Energy Saving Trust and the Low Income High Costs fuel poverty definition. Statistical testing highlighted significant differences in predictors of fuel poverty between households referred to WHHP and all Bradford households. RESULTS: Significant (p < .05) predictors of fuel poverty included: living in an area with lower average household incomes and higher proportion of ethnic minority individuals, and living in a property with a lower energy efficiency rating. Households living in a detached or older property, and homeowners were more likely to be fuel poor. Differences in the direction of the relationship with fuel poverty were identified between some national and local predictors. Most predictors were significantly (p < .05) overrepresented among WHHP households, suggesting equitable service reach. Ethnic minorities, younger people, and multiperson households were underrepresented. CONCLUSIONS: Local fuel poverty predictors were similar to many national-level predictors, but identified differences in the direction of the relationship between some national and local predictors reaffirm the value of locally focused research. WHHP successfully targeted households possessing key predictors, but should ensure that ethnic minorities, younger people, and multiperson households are equitably referred.

Socio-Ecological Mechanisms Supporting High Densities of Aedes albopictus (Diptera: Culicidae) in Baltimore, MD.

Social, ecological, and climatic factors interact creating a heterogeneous matrix that determines the spatiotemporal distribution of mosquitoes and human risks of exposure to the diseases they transmit. We explore linkages between the social and institutional processes behind residential abandonment, urban ecology, and the interactions of socio-ecological processes with abiotic drivers of mosquito production. Specifically, we test the relative roles of infrastructure degradation and vegetation for explaining the presence of Aedes albopictus Skuse 1894 to better predict spatial heterogeneity in mosquito exposure risk within urban environments. We further examine how precipitation interacts with these socially underpinned biophysical variables. We use a hierarchical statistical modeling approach to assess how environmental and climatic conditions over 3 years influence mosquito ecology across a socioeconomic gradient in Baltimore, MD. We show that decaying infrastructure and vegetation are important determinants of Ae. albopictus infestation. We demonstrate that both precipitation and vegetation influence mosquito production in ways that are mediated by the level of infrastructural decay on a given block. Mosquitoes were more common on blocks with greater abandonment, but when precipitation was low, mosquitoes were more likely to be found in higher-income neighborhoods with managed container habitat. Likewise, although increased vegetation was a negative predictor of mosquito infestation, more vegetation on blocks with high abandonment was associated with the largest mosquito populations. These findings indicate that fine spatial scale modeling of mosquito habitat within urban areas is needed to more accurately target vector control.

Diversity of Serratia marcescens Strains Associated with Cucurbit Yellow Vine Disease in Georgia.

Cucurbit yellow vine disease (CYVD), caused by the squash bug (Anasa tristis)-transmitted bacterium Serratia marcescens, was first identified in Oklahoma and Texas in 1988 and in Georgia in 2012. S. marcescens is a highly diverse species found in many ecological niches. In previous studies, CYVD strains of S. marcescens formed a closely related group separate from non-CYVD strains based on biological and molecular characterization techniques. Multilocus sequence analysis (MLSA) of six housekeeping genes and repetitive elements-based polymerase chain reaction (rep-PCR) using the BOX and ERIC primers were used to assess the genetic diversity of CYVD strains of S. marcescens collected in Georgia together with a strain from Texas and seven non-CYVD strains of S. marcescens. rep-PCR results revealed genetic diversity among CYVD strains while MLSA results showed a 100% similarity across the six loci for all but one of the CYVD strains, which differed at the icd locus by five polymorphisms. For both methods, CYVD strains clustered separately from nonplant-pathogenic S. marcescens strains and were most similar to a rice endophyte strain. One CYVD strain isolated from a squash bug shared genetic similarities with non-CYVD strains, and may be the result of a recombination event between CYVD and non-CYVD strains.

Degree of corneal anaesthesia after topical application of 0.4% oxybuprocaine hydrochloride and 0.5% proparacaine hydrochloride ophthalmic solution in clinically normal cattle.

OBJECTIVES: The use of corneal anaesthesia is necessary for a range of clinical purposes. Therefore, we assessed and compared the efficacy of corneal anaesthesia after application of 0.4% oxybuprocaine hydrochloride and 0.5% proparacaine hydrochloride ophthalmic solution in clinically normal cattle. METHODS: The 24 clinically normal cows were allocated into two groups. Cows in group 1 (n = 12) received 0.2 mL of 0.4% oxybuprocaine hydrochloride with fluorescein ophthalmic solution in one eye and 0.2 mL of sterile saline (0.9% NaCl) with fluorescein in the contralateral eye (control). Group 2 (n = 12) received 0.2 mL of 0.4% oxybuprocaine hydrochloride with fluorescein ophthalmic solution in one eye and 0.2 mL of 0.5% proparacaine hydrochloride with fluorescein in the contralateral eye (control). In each group, corneal touch threshold was determined by Cochet-Bonnet aesthesiometer for both eyes immediately prior to topical administration of solutions, at 1 min and 5 min after administration of topical solutions and every 5 min thereafter for a total of 75 min. RESULTS: Significant corneal anaesthesia was noted immediately following topical application of both oxybuprocaine and proparacaine as compared with controls, with maximal corneal anaesthesia noted 1 min after administration. Both oxybuprocaine and proparacaine produced significant corneal anaesthesia for the duration of the 75-min study. Neither oxybuprocaine hydrochloride nor proparacaine hydrochloride treatment resulted in visible adverse effects. CONCLUSION: There are limited data available demonstrating the efficacy and duration of corneal anaesthetic agents in cattle. Both oxybuprocaine hydrochloride and proparacaine hydrochloride should be considered practical options for providing corneal anaesthesia in cattle in a clinical setting.

The CaSm (LSm1) oncogene promotes transformation, chemoresistance and metastasis of pancreatic cancer cells.

The cancer-associated Sm-like (CaSm) oncogene is overexpressed in 87% of human pancreatic tumor samples and CaSm knockdown has demonstrated therapeutic efficacy in murine models of pancreatic cancer. Evidence indicates that CaSm modulates messenger RNA degradation; however, its target genes and the mechanisms by which CaSm promotes pancreatic cancer remain largely unknown. Here, we demonstrate that the CaSm overexpression alters several hallmarks of cancer-including transformation, proliferation, chemoresistance and metastasis. Doxycycline-induced CaSm expression enhanced proliferation and both anchorage-dependent and -independent growth of the human Panc-1 cells in vitro. CaSm induction decreased gemcitabine-induced cytotoxicity and altered the expression of apoptotic regulation genes, including Bad, E2F1 and Bcl-XL. CaSm-overexpressing Panc-1 cells were twofold more migratory and fourfold more invasive than the driver controls and demonstrated characteristics of epithelial-to-mesenchymal transition such as morphological changes and decreased E-cadherin expression. CaSm induction resulted in changes in RNA expression of metastasis-associated genes such as MMP1, SerpinB5, uPAR and Slug. Using a murine model of metastatic pancreatic cancer, injection of CaSm-induced Panc-1 cells resulted in a higher abundance of hepatic metastatic lesions. Overall, CaSm overexpression contributed to a more aggressive cancer phenotype in Panc-1 cells, further supporting the use of CaSm as a therapeutic target against pancreatic cancer.

PAX3 and ETS1 synergistically activate MET expression in melanoma cells.

Melanoma is a highly aggressive disease that is difficult to treat owing to rapid tumor growth, apoptotic resistance and high metastatic potential. The MET proto-oncogene (MET) tyrosine kinase receptor promotes many of these cellular processes, but while MET is often overexpressed in melanoma, the mechanism driving this overexpression is unknown. As the MET gene is rarely mutated or amplified in melanoma, MET overexpression may be driven to increased activation through promoter elements. In this report, we find that transcription factors PAX3 and ETS1 directly interact to synergistically activate MET expression. Inhibition of PAX3 and ETS1 expression in melanoma cells leads to a significant reduction of MET receptor levels. The 300-bp 5' proximal MET promoter contains a PAX3 response element and two ETS1 consensus motifs. Although ETS1 can moderately activate both of these sites without cofactors, robust MET promoter activation of the first site is PAX dependent and requires the presence of PAX3, whereas the second site is PAX independent. The induction of MET by ETS1 via this second site is enhanced by hepatocyte growth factor-dependent ETS1 activation, thereby MET indirectly promotes its own expression. We further find that expression of a dominant-negative ETS1 reduces the ability of melanoma cells to grow both in culture and in vivo. Thus, we discover a pathway where ETS1 advances melanoma through the expression of MET via PAX-dependent and -independent mechanisms.

Transferrin receptor targeting nanomedicine delivering wild-type p53 gene sensitizes pancreatic cancer to gemcitabine therapy.

To overcome gene therapy barriers such as low transfection efficiency and nonspecific delivery, liposomal nanoparticles targeted by a single-chain antibody fragment to the transferrin receptor (TfRscFv) delivering wild-type (wt) human p53 (SGT-53) were developed for tumor-specific targeting. We hypothesize that SGT-53 in combination with gemcitabine will demonstrate enhanced therapeutic benefit in an in vivo metastatic pancreatic cancer model. Intrasplenic injection of 1 × 10(6) Panc02 murine pancreatic cancer cells was used to generate in vivo hepatic metastatic tumors. Nanoparticle localization was assessed by tail vein injection of TfRscFv with fluorescently labeled oligonucleotides (6-carboxyfluorescein phosphoramidite (6FAM) ODN) imaged by Xenogen IVIS 200 scan. SGT-53 (equivalent to 30 µg of p53 intravenously) and gemcitabine (20 mg/kg intraperitoneally) alone and in combination were administered biweekly and compared with untreated mice. Survival was determined by blinded daily assessment of morbidity. Human wtp53 expression and transferrin levels in the tumors were assessed by western blot analysis. Tumor burden was quantified by liver weight. Xenogen imaging demonstrated tumor-specific uptake of TfRscFv-6FAM ODN. Exogenous human wtp53 protein was detected in the SGT-53-treated tumors compared with control. Compared with untreated mice with metastatic tumors demonstrating median survival of 20 days, SGT-53, gemcitabine and the combination demonstrated improved median survival of 29, 30 and 37 days, respectively. The combination treatment prolonged median survival when compared with single drug treatment and decreased tumor burden. The tumor targeting liposomal-based SGT-53 nanoparticle is capable of sensitizing pancreatic cancer to conventional chemotherapy in pancreatic cancer models. This approach has the potential to be translated into a new, more effective therapy for pancreatic cancer. Further optimization is ongoing, moving towards a Phase 1B/2 clinical trial.

Campylobacter spp. recovered from the Upper Oconee River Watershed, Georgia in a 4-year study.

Waterways should be considered in the migration routes of Campylobacter, and the genus has been isolated from several water sources. Inferences on migration routes can be made from tracking genetic types in populations found in specific habitats and testing how they are linked to other types. Water samples were taken over a 4-year period from waterways in the Upper Oconee River Watershed, Georgia, to recover isolates of thermophilic Campylobacter. The isolates were typed by multilocus sequence typing (MLST) and analyzed to determine the overall diversity of Campylobacter in that environment. Forty-seven independent isolates were recovered from 560 samples (8.4 %). Two (~4 %) isolates were Campylobacter coli, three (~6 %) isolates were putatively identified as Campylobacter lari, and the remaining 42 (~90 %) were Campylobacter jejuni. The C. jejuni and C. coli isolates were typed by the Oxford MLST scheme. Thirty sequence types (STs) were identified including 13 STs that were not found before in the MLST database, including 24 novel alleles. Of the 17 previously described STs, 10 have been isolated from humans, 6 from environmental water, and 6 from wild birds (five types from multiple sources). Seven sites had multiple positive samples, and on two occasions, the same ST was isolated at the same site. The most common type was STST61 with four isolates, and the most common clonal complex was CC179 with nine isolates. CC179 has been commonly associated with environmental water. Although some Campylobacter STs that were found in the Oconee River engage in widespread migration, most are tightly associated with or unique to environmental water sources.

Toxicity of copper to early-life stage Kootenai River white sturgeon, Columbia River white sturgeon, and rainbow trout.

White sturgeon (Acipenser transmontanus) populations throughout western North America are in decline, likely as a result of overharvest, operation of dams, and agricultural and mineral extraction activities in their watersheds. Recruitment failure may reflect the loss of early-life stage fish in spawning areas of the upper Columbia River, which are contaminated with metals from effluents associated with mineral-extraction activities. Early-life stage white sturgeon (A. transmontanus) from the Columbia River and Kootenai River populations were exposed to copper during 96-h flow-through toxicity tests to determine their sensitivity to the metal. Similar tests were conducted with rainbow trout (RBT [Oncorhynchus mykiss]) to assess the comparative sensitivity of this species as a surrogate for white sturgeon. Exposures were conducted with a water quality pH 8.1-8.3, hardness 81-119 mg/L as CaCO(2), and dissolved organic carbon 0.2-0.4 mg/L. At approximately 30 days posthatch (dph), sturgeon were highly sensitive to copper with median lethal concentration (LC(50)) values ranging from 4.1 to 6.8 µg/L compared with 36.5 µg/L for 30 dph RBT. White sturgeon at 123-167 dph were less sensitive to copper with LC(50) values ranging from 103.7 to 268.9 µg/L. RBT trout, however, remained more sensitive to copper at 160 dph with an LC(50) value of 30.9 µg/L. The results indicate that high sensitivity to copper in early-life stage white sturgeon may be a factor in recruitment failure occurring in the upper Columbia and Kootenai rivers. When site-specific water-quality criteria were estimated using the biotic ligand model (BLM), derived values were not protective of early-life stage fish, nor were estimates derived by water-hardness adjustment.

Vibrio cholerae non-O1 infection in cirrhotics: case report and literature review.

Vibrio species are ubiquitous in the marine environment and can cause severe infections in cirrhotic patients. Patients with liver disease should be warned about the potential dangers of consuming raw or undercooked seafood, and avoiding exposure of wounds to seawater. We report a case of severe sepsis from Vibrio cholerae non-O1 in a patient with cirrhosis awaiting orthotopic liver transplant. This case is aimed to advise clinicians about the importance of V. cholerae subtypes, and non-cholera Vibrio species infections in cirrhotic patients, highlighting the need to educate these patients to stay away from undercooked seafood.

Determination of the mechanical properties of Bionate 80A and Bionate 75D for the stress analysis of cushion form bearings.

Two biomedical grade polyurethanes, currently under investigation as an alternative tibial bearing for a total knee replacement, are analysed. Extensive mechanical testing has been performed on these materials to determine the mechanical properties in terms of strain energy density functions for use in the finite element package MSC. Marc 2001 that do not rely on linear elasticity. A variation in Poisson's ratio in the fourth decimal place can lead to significant errors in the magnitude and distribution of stresses in a linear elastic analysis. As large variations in this parameter (up to the first decimal place) were determined experimentally, it is concluded that a linear elastic approach to the stress analysis of these materials is not valid.

Salmonella, Campylobacter and Enterococcus spp.: their antimicrobial resistance profiles and their spatial relationships in a synoptic study of the Upper Oconee River basin.

Rivers may serve as reservoirs for enteric organisms. Very little is known about the boundaries of microbial communities in moving bodies of water so this study was undertaken to find the limits of distribution of some bacteria, focusing on enteric organisms. The presence of Salmonella, Campylobacter, and Enterococcus spp. and the antimicrobial resistance phenotypes carried by these organisms was evaluated for the Upper Oconee River basin, a small river in the lower Piedmont of northeastern Georgia, USA. Samples were obtained from 83 sites during a 3-h period on a spring day (April 2005) in an approximately 30 x 20 km region. Campylobacter spp. was isolated at 12 sites. The Campylobacter isolates from three sites were resistant to tetracycline. Of the five short-variable region (SVR) subtypes of Campylobacter that were found, three were found at more than one site, two types were found twice, and one subtype was found three times. Enterococcus was isolated at 71 sites. E. casseliflavus was the most common species. Based on species identification and antimicrobial resistance patterns, 24 types of Enterococcus were found. Salmonella was isolated from 62 sites. Of the 19 Salmonella serovars that were isolated, serovar Muenchen accounted for about 20% of the isolates. The next three most common serovars isolated, Rubislaw, Hartford, and Give, accounted for about 44% of the river isolates. Antimicrobial resistance profiling offered limited differentiation of Salmonella isolates because only seven isolates were resistant to any antimicrobial. The sites at which Salmonella, Campylobacter, or Enterococcus were isolated did not correlate with each other or with the total coliform number or Escherichia coli count for the site. However, isolates of some of the same species and type occurred in clusters that were restricted to areas within 5 to 6 km.

Thermoelastic analysis of high-pressure angioplasty balloons.

Failures in angioplasty balloons are investigated using typical destructive techniques. The material properties of moulded balloons are derived from tensile tests and used to establish the reasons for failure of the balloons. Thermoelastic stress analysis is used to determine the stress distribution in the balloons, and a means of interpreting the data to derive actual stresses is described. The departure from linear elastic behaviour in the angioplasty balloons is identified using thermoelastic analysis. The results from the thermoelastic analysis are discussed and compared with those from the destructive tests, and the thermoelastic technique is shown to be a potential new means for non-destructive analysis of angioplasty balloons.

The potential for chromium to affect the fertilization process of Chinook salmon (Oncorhynchus tshawytscha) in the Hanford reach of the Columbia River, Washington, USA.

The Hanford Nuclear Reservation in south central Washington was claimed by the federal government as a site for the production of plutonium. During the course of production and operation of the facilities at Hanford, radionuclides and chromium were discharged directly into the river and also contaminated the groundwater. This study was designed to assess the effects of chromium (Cr) on Chinook salmon (Oncorhynchus tshawytscha) fertilization under exposure conditions similar to those of the Hanford Reach of the Columbia River. Chinook salmon gametes were exposed to aqueous Cr concentrations ranging from 0 to 266 microg Cr l(-1). The current ambient water-quality criteria (AWQC) established for the protection of aquatic life (United States Environmental Protection Agency [USEPA] 1986) is 11 microg Cr l(-1). Cr has been measured in pore water from bottom sediments of the Columbia River at concentrations >600 microg Cr l(-1). Under exposure conditions designed to closely mimic events that occur in the river, the fertilization of Chinook salmon eggs was not affected by concentrations of Cr ranging from 11 to 266 microg Cr l(-1). Data suggest that the instantaneous nature of fertilization likely limits the potential effects of Cr on fertilization success. As a result, the current AWQC of 11 mug Cr l(-1) is most likely protective of Chinook salmon fertilization.

Effects of multiple routes of cadmium exposure on the hibernation success of the American toad (Bufo americanus).

The effects of multiple routes of cadmium exposure on juvenile American toads (Bufo americanus) were evaluated using environmentally relevant concentrations. During or after exposure, toads were individually hibernated for 172 days at approximately 4degreesC. The following experiments were conducted: (1) dermal exposure (hibernation in soil contaminated with up to 120 microg Cd/g (dry weight)); (2) injection exposure (single injection with cadmium to achieve a maximum whole-body nominal concentration of 3 microg Cd/g (wet weight) 12 days before hibernation in uncontaminated soil); and, (3) oral exposure (feeding with mealworms containing < or =16 microg Cd/g (dry weight) for 50 days before hibernation in uncontaminated soil). We hypothesized that sublethal levels of cadmium would become lethal during hibernation because of combined chemical and cold stress. No prehibernation mortality occurred in the injection and oral exposure studies. There was a significant treatment effect on whole-body cadmium concentration in toads orally or dermally exposed and on percent of cadmium retention in toads orally exposed. There was also a trend of increased time-to-burrowing and more toads partially buried with greater cadmium concentration in the dermal study, which indicated avoidance. In all 3 experiments, no significant differences were found among cadmium treatments in hibernation survival, percent of mass loss, or locomotor performance. However, toads fed mealworms averaging 4.7 microg Cd/g (dry weight) had only 56% survival compared with 100% survival for controls. Although our results suggest that environmentally relevant levels of cadmium do not pose a great risk to American toads, factors such as soil type or prey species may increase cadmium bioavailability, and other amphibian species may be more sensitive to cadmium than B. americanus.

Effects of Rootstock and Budding Height on Bacterial Canker in French Prune.

Bacterial canker is one of the most economically important diseases of stone fruit trees, including 'French' prune (Prunus domestica). Field trials were conducted to evaluate the effect of rootstock selection and budding height on the incidence and severity of bacterial canker in four orchards with low to high disease pressure. Treatments included French prune scions low-grafted on 'Lovell' peach (Prunus persica) rootstocks as well as Myrobalan 29C (Prunus cerasifera) plum rootstocks grafted at 15, 50, and 90 cm above the rootstock crown. Another treatment consisted of growing Myrobalan 29C plum rootstocks in the field for one growing season, then field-grafting French prune buds onto rootstock scaffolds. Lovell peach rootstock provided the greatest protection from bacterial canker as measured by disease incidence and tree mortality in all orchards. Field-budded rootstocks and rootstocks grafted at the highest budding height provided moderate levels of resistance to bacterial canker. These treatments reduced the incidence but not the severity of disease.

Solution structure of the third immunoglobulin domain of the neural cell adhesion molecule N-CAM: can solution studies define the mechanism of homophilic binding?

Homophilic binding of the neural cell adhesion molecule (N-CAM) mediates the calcium-independent cell-cell adhesion that is involved in neuronal development. Two hypotheses have been advanced for the mechanism of homophilic binding. Cell-based experiments have implicated each of the five extracellular immunoglobulin (Ig) domains of N-CAM in the homophilic adhesion interaction, and have predicted that the third domain (Ig III) self-associates. The alternative hypothesis is based on solution observations, which implicate a specific antiparallel interaction between the first two Ig domains (Ig I and Ig II). In order to test these hypotheses, we have determined a high-resolution solution structure of recombinant Ig III (sequence derived from chicken N-CAM) and examined the aggregation behavior of isolated Ig domains in solution. The structure shows that Ig III adopts a canonical Ig fold, in which the beta strands ABED and A'GFCC' form two beta sheets that are linked by a disulfide bond. In contrast to the demonstrated aggregation of Ig III on solid supports, we were unable to demonstrate self-association of Ig III under any of a variety of solution conditions. The structure shows that the surface of Ig III is dominated by two large acidic patches, which may explain our failure to observe self-association in solution. To evaluate the involvement of the Ig I-Ig II interaction in cell-cell adhesion, we designed a point mutation in Ig I (F19S) that proved sufficient to abrogate the Ig I-Ig II interaction seen in solution. However, the introduction of this mutation into full-length N-CAM expressed in COS-7 cells failed to affect N-CAM-mediated cell-cell adhesion. The inability to observe Ig III self-association in solution, combined with the failure of the F19S mutation to affect N-CAM-mediated cell-cell adhesion, suggests that, although solution studies can give important insights into the structures of individual domains, the interactions observed in solution between the domains may not be representative of the interactions that occur on the cell surface.

Cholinergic and behavioral neurotoxicity of carbaryl and cadmium to larval rainbow trout (Oncorhynchus mykiss).

Pesticides and heavy metals are common environmental contaminants that can cause neurotoxicity to aquatic organisms, impairing reproduction and survival. Neurotoxic effects of cadmium and carbaryl exposures were estimated in larval rainbow trout (RBT; Oncorhynchus mykiss) using changes in physiological endpoints and correlations with behavioral responses. Following exposures, RBT were videotaped to assess swimming speed. Brain tissue was used to measure cholinesterase (ChE) activity, muscarinic cholinergic receptor (MChR) number, and MChR affinity. ChE activity decreased with increasing concentrations of carbaryl but not of cadmium. MChR were not affected by exposure to either carbaryl or cadmium. Swimming speed correlated with ChE activity in carbaryl-exposed RBT, but no correlation occurred in cadmium-exposed fish. Thus, carbaryl exposure resulted in neurotoxicity reflected by changes in physiological and behavioral parameters measured, while cadmium exposure did not. Correlations between behavior and physiology provide a useful assessment of neurotoxicity.

A short segment within the cytoplasmic domain of the neural cell adhesion molecule (N-CAM) is essential for N-CAM-induced NF-kappa B activity in astrocytes.

The neural cell adhesion molecule (N-CAM) is expressed on the surface of astrocytes, where its homophilic binding leads to the activation of the transcription factor NF-kappaB. Transfection of astrocytes with a construct encompassing the transmembrane region and the cytoplasmic domain of N-CAM (designated Tm-Cyto, amino acids 685-839 in the full-length molecule) inhibited this activation up to 40%, and inhibited N-CAM-induced translocation of NF-kappaB to the nucleus. N-CAM also activated NF-kappaB in astrocytes from N-CAM knockout mice, presumably through binding to a heterophile. This activation, however, was not blocked by Tm-Cyto expression, indicating that the inhibitory effect of the Tm-Cyto construct is specific for cell surface N-CAM. Deletions and point mutations of the cytoplasmic portion of the Tm-Cyto construct indicated that the region between amino acids 780 and 800 were essential for inhibitory activity. This region contains four threonines (788, 793, 794, and 797). Mutation to alanine of T788, T794, or T797, but not T793, abolished inhibitory activity, as did mutation of T788 or T797 to aspartic acid. A Tm-Cyto construct with T794 mutated to aspartic acid retained inhibitory activity but did not itself induce a constitutive NF-kappaB response. This result suggests that phosphorylation of T794 may be necessary but is not the triggering event. Overall, these findings define a short segment of the N-CAM cytoplasmic domain that is critical for N-CAM-induced activation of NF-kappaB and may be important in other N-CAM-mediated signaling.

Behavioral dysfunctions correlate to altered physiology in rainbow trout (Oncorynchus mykiss) exposed to cholinesterase-inhibiting chemicals.

We selected four metrics of swimming behavior (distance swam, speed, rate of turning, and tortuosity of path) and the commonly used biochemical marker, brain cholinesterase (ChE) activity, to assess (1) the sensitivity and reliability of behavior as a potential biomarker in monitoring work, (2) the potential for these endpoints to be used in automated monitoring, and (3) the linkage between behavior and its underlying biochemistry. Malathion-exposed fish exhibited large decreases in distance and speed and swam in a more linear path than control fish after 24 h exposure. By 96 h exposure, fish still swam slower and traveled less distance; fish fully recovered after 48 h in clean water. Diazinon-exposed fish exhibited decreases in distance, speed, and turning rate compared to controls. After 48 h recovery in clean water, fish exposed to diazinon had not recovered to control levels. The behavioral responses provided measures of neurotoxicity that were easily quantifiable by automated means, implying that the inclusion of behavior in monitoring programs can be successful. Furthermore, correlations between behavior and biochemical endpoints, such as ChE inhibition, suggest that this approach can provide a meaningful link between biochemistry and behavior and can provide useful information on toxicant impacts.