Patterns of ethanol intake in male rats with partial dopamine transporter deficiency.

Mesolimbic dopamine signaling plays a major role in alcohol and substance use disorders as well as comorbidities such as anxiety and depression. Growing evidence suggests that alcohol drinking is modulated by the function of the dopamine transporter (DAT), which tightly regulates extracellular dopamine concentrations. Adult male rats on a Wistar Han background (DAT+/+) and rats with a partial DAT deletion (DAT+/-) were used in this study. First, using fast-scan cyclic voltammetry in brain slices containing the nucleus accumbens core from ethanol-naïve subjects, we measured greater evoked dopamine concentrations and slower dopamine reuptake in DAT+/- rats, consistent with increased dopamine signaling. Next, we measured ethanol drinking using the intermittent access two-bottle choice paradigm (20% v/v ethanol vs. water) across 5 weeks. DAT+/- rats voluntarily consumed less ethanol during its initial availability (the first 30 min), especially after longer periods of deprivation. In addition, DAT+/- males consumed less ethanol that was adulterated with the bitter tastant quinine. These findings suggest that partial DAT blockade and concomitant increase in brain dopamine levels has potential to reduce drinking and ameliorate alcohol use disorder (AUD).

Teaching Hardy-Weinberg Equilibrium using Population-Level Punnett Squares: Facilitating Calculation for Students with Math Anxiety.

Hardy-Weinberg (HW) equilibrium and its accompanying equations are widely taught in introductory biology courses, but high math anxiety and low math proficiency have been suggested as two barriers to student success. Population-level Punnett squares have been presented as a potential tool for HW equilibrium, but actual data from classrooms have not yet validated their use. We used a quasi-experimental design to test the effectiveness of Punnett squares over 2 days of instruction in an introductory biology course. After 1 day of instruction, students who used Punnett squares outperformed those who learned the equations. After learning both methods, high math anxiety was predictive of Punnett square use, but only for students who learned equations first. Using Punnett squares also predicted increased calculation proficiency for high-anxiety students. Thus, teaching population Punnett squares as a calculation aid is likely to trigger less math anxiety and help level the playing field for students with high math anxiety. Learning Punnett squares before the equations was predictive of correct derivation of equations for a three-allele system. Thus, regardless of math anxiety, using Punnett squares before learning the equations seems to increase student understanding of equation derivation, enabling them to derive more complex equations on their own.

NeuroDAC: an open-source arbitrary biosignal waveform generator.

Objective.Researchers are developing biomedical devices with embedded closed-loop algorithms for providing advanced adaptive therapies. As these devices become more capable and algorithms become more complex, tasked with integrating and interpreting multi-channel, multi-modal electrophysiological signals, there is a need for flexible bench-top testing and prototyping. We present a methodology for leveraging off-the-shelf audio equipment to construct a biosignal waveform generator capable of streaming pre-recorded biosignals from a host computer. By re-playing known, well-characterized, but physiologically relevant real-world biosignals into a device under test, researchers can evaluate their systems without the need for expensivein vivoexperiments.Approach.An open-source design based on the proposed methodology is described and validated, the NeuroDAC. NeuroDAC allows for 8 independent channels of biosignal playback using a simple, custom designed attenuation and buffering circuit. Applications can communicate with the device over a USB interface using standard audio drivers. On-board analog amplitude adjustment is used to maximize the dynamic range for a given signal and can be independently tuned for each channel.Main results.Low noise component selection yields a no-signal noise floor of just 5.35 ± 0.063. NeuroDAC's frequency response is characterized with a high pass -3 dB rolloff at 0.57 Hz, and is capable of accurately reproducing a wide assortment of biosignals ranging from EMG, EEG, and ECG to extracellularly recorded neural activity. We also present an application example using the device to test embedded algorithms on a closed-loop neural modulation device, the Medtronic RC+S.Significance.By making the design of NeuroDAC open-source we aim to present an accessible tool for rapidly prototyping new biomedical devices and algorithms than can be easily modified based on individual testing needs.ClinicalTrials.gov Identifiers: NCT04281134, NCT03437928, NCT03582891.

Myxobolus arcticus and Parvicapsula minibicornis infections in sockeye salmon Oncorhynchus nerka following downstream migration in British Columbia.

Factors influencing the health of sockeye salmon Oncorhynchus nerka in British Columbia, Canada, are important for fisheries management and conservation. Juvenile salmon originating from the Fraser River were screened for 3 enzootic parasites (Myxobolus arcticus, Parvicapsula minibicornis, Ceratonova shasta) and the bacterium Renibacterium salmoninarum. Fish were collected from the Strait of Georgia in 2010, 2011 and 2012 and genotyped to stock of origin. Trends in infection status were estimated by year, spawning zone and catch area. The annual prevalences of P. minibicornis (n = 1448) were 23.3, 6.5 and 8.1%, and for M. arcticus (n = 1343), annual prevalences were 40.4, 66.3 and 27.4%, respectively. Logistic regression showed that P. minibicornis was most strongly associated with salmon from the lower Fraser River spawning zone and increased with distance caught from the mouth of the Fraser River. In contrast, infection with M. arcticus was most strongly associated with salmon from the middle Fraser River spawning zone, and there was no trend related to distance from the Fraser River. Neither R. salmoninarum nor C. shasta were detected. These observations are discussed in the context of salmon life history and pathogen biology.

Two Novel Microsporidia in Skeletal Muscle of Pike-Perch Sander lucioperca and Burbot Lota lota in Finland.

Two new species of Microsporidia were recognized in skeletal muscle of freshwater fishes from Finland. Myosporidium spraguei n. sp. from pike-perch Sander lucioperca occurred as mature spores within sporophorous vesicles (SPVs) within a xenoma. The ovoid spores were 3.8 µm long and 2.4 µm wide, based on transmission electron micrographs (TEM). The exospore and endospore were equally thick, the nucleus was monokaryotic and the polar filament was isofilar with 12 coils in a single rank, entirely adjacent to the prominent posterior vacuole. Small subunit (SSU) rDNA sequence confirmed the presence of M. spraguei n. sp. in burbot Lota lota . The second species, Microsporidium luciopercae n. sp., also from pike-perch, occurred within SPVs that occupied only a fraction of the volume of the otherwise intact myocyte; no xenoma was produced. Myocyte degeneration and necrosis occurred as mature spores dispersed into direct contact with the sarcoplasm. The ovoid spores were 4.6 µm long and 2.8 µm wide (based on TEM); they were monokaryotic and the polar filament was isofilar with 25 coils in a single rank in the posterior of the spore. The exospore was relatively thin with an irregular profile. Neither infection elicited an inflammatory response, although degenerate spores were observed within host cells, suggesting phagocytosis. Phylogenetic analysis of SSU sequences placed both organisms on distinct clades within the Marinosporidia.

MAOA expression predicts vulnerability for alcohol use.

The role of the monoamines dopamine (DA) and serotonin (5HT) and the monoamine-metabolizing enzyme monoamine oxidase A (MAOA) have been repeatedly implicated in studies of alcohol use and dependence. Genetic investigations of MAOA have yielded conflicting associations between a common polymorphism (MAOA-LPR) and risk for alcohol abuse. The present study provides direct comparison of tissue-specific MAOA expression and the level of alcohol consumption. We analyzed rhesus macaque MAOA (rhMAOA) expression in blood from males before and after 12 months of alcohol self-administration. In addition, nucleus accumbens core (NAc core) and cerebrospinal fluid (CSF) were collected from alcohol access and control (no alcohol access) subjects at the 12-month time point for comparison. The rhMAOA expression level in the blood of alcohol-naive subjects was negatively correlated with subsequent alcohol consumption level. The mRNA expression was independent of rhMAOA-LPR genotype and global promoter methylation. After 12 months of alcohol use, blood rhMAOA expression had decreased in an alcohol dose-dependent manner. Also after 12 months, rhMAOA expression in the NAc core was significantly lower in the heavy drinkers, as compared with control subjects. The CSF measured higher levels of DA and lower DOPAC/DA ratios among the heavy drinkers at the same time point. These results provide novel evidence that blood MAOA expression predicts alcohol consumption and that heavy alcohol use is linked to low MAOA expression in both the blood and NAc core. Together, the findings suggest a mechanistic link between dampened MAOA expression, elevated DA and alcohol abuse.

The brain gene expression profile of dopamine D2/D3 receptors and associated signaling proteins following amphetamine self-administration.

Persistent neuroadaptations following chronic psychostimulant exposure include reduced striatal dopamine D2 receptor (D2R) levels. The signaling of D2Rs is initiated by Gαi/o proteins and terminated by regulator of G protein signaling (RGS) proteins. The purpose of this study is to examine the association of the drug taking behavior and gene expression profile of D2/D3Rs, and their associated signaling proteins in the ventral tegmental area (VTA) and nucleus accumbens (NAc) using a rodent model of amphetamine (AMPH) self-administration. Rats were allowed to self-administer AMPH (0.187 mg/kg/infusion for a maximum of 40 injections in 6h daily sessions) for 5 days during which rats showed an escalated rate of AMPH intake across days. AMPH self-administration induced profound brain region-dependent alterations of the targeted genes. There was a positive correlation of the messenger ribonucleic acid (mRNA) levels of RGS10 between the VTA and the NAc in the control animals, which was abolished by AMPH self-administration. AMPH self-administration also produced a negative correlation of the mRNA levels of RGS7 and RGS19 between the two brain regions, which was not present in the control group. Furthermore, AMPH taking behavior was associated with changes in certain gene expression levels. The mRNA levels of RGS2 and RGS4 in both the VTA and NAc were positively correlated with the rate of AMPH intake. Additionally, the rate of AMPH intake was also positively correlated with RGS10 and negatively correlated with RGS17 and the short form of D2Rs mRNA level in the VTA. Although there were significant changes in the mRNA levels of RGS7 and RGS8 in the NAc, none of these measures were correlated with the rate of AMPH intake. The present study suggested that short-term AMPH self-administration produced pronounced changes in the VTA that were more associated with AMPH taking behavior than changes in the NAc.

A method for estimating (41)Ar, (85)(,88)Kr and (131m,133)Xe doses to non-human biota.

A method is presented for estimating (41)Ar, (85,88)Kr and (131m,133)Xe dose rates to terrestrial wildlife without having to resort to comparisons with analogue radionuclides. The approach can be used to calculate the dose rates arising from external exposures to given ambient air concentrations of these isotopes. Dose conversion coefficient (DCC) values for a range of representative organisms are calculated, using a Monte Carlo approach to generate absorbed fractions based on representing animals as reference ellipsoid geometries. Plume immersion is the main component of the total DCC. DCC values calculated for a human-sized organism are compared with human dose conversion factors from ICRP Publication 119, demonstrating the consistency of the biota approach with that for humans. An example of application is provided for hypothetical nuclear power plant atmospheric discharges with associated exposures to birds and insects. In this example, the dose rates appear to be dominated by (133)Xe and (88)Kr, respectively. The biota considered would be protected from the effects of noble gas radiation from a population protection perspective.

Greater ethanol inhibition of presynaptic dopamine release in C57BL/6J than DBA/2J mice: Role of nicotinic acetylcholine receptors.

The mesolimbic dopamine system, originating in the ventral tegmental area (VTA) and projecting to the nucleus accumbens (NAc), has been heavily implicated in the reinforcing effects of ethanol. Recent slice voltammetry studies have shown that ethanol inhibits dopamine release selectively during high-frequency activity that elicits phasic dopamine release shown to be important for learning and reinforcement. Presently, we examined ethanol inhibition of electrically evoked NAc dopamine in two mouse strains with divergent dopamine responses to ethanol, C57BL/6 (C57) and DBA/2J (DBA) mice. Previous electrophysiology and microdialysis studies have demonstrated greater ethanol-induced VTA dopaminergic firing and NAc dopamine elevations in DBA compared to C57 mice. Additionally, DBA mice have greater ethanol responses in dopamine-related behaviors, including hyperlocomotion and conditioned place preference. Currently, we demonstrate greater sensitivity of ethanol inhibition of NAc dopamine signaling in C57 compared to DBA mice. The reduced sensitivity to ethanol inhibition in DBA mice may contribute to the overall greater ethanol-induced dopamine signaling and related behaviors observed in this strain. NAc cholinergic activity is known to potently modulate terminal dopamine release. Additionally, ethanol is known to interact with multiple aspects of nicotinic acetylcholine receptor activity. Therefore, we examined ethanol-mediated inhibition of dopamine release at two ethanol concentrations (80 and 160 mM) during bath application of the non-selective nicotinic receptor antagonist mecamylamine, as well as compounds selective for the ß2-(dihydro-ß-erythroidine hydrobromide; DhßE) and α6-(α-conotoxin MII [H9A; L15A]) subunit-containing receptors. Mecamylamine and DhßE decreased dopamine release and reduced ethanol's inhibitory effects on dopamine in both DBA and C57 mice. Further, α-conotoxin also reduced the dopamine release and the dopamine-inhibiting effects of ethanol at the 80 mM, but not 160 mM, concentration. These data suggest that ethanol is acting in part through nicotinic acetylcholine receptors, or downstream effectors, to reduce dopamine release during high-frequency activity.

Development of the salmon louse Lepeophtheirus salmonis and its effects on juvenile sockeye salmon Oncorhynchus nerka.

Responses of sockeye salmon Oncorhynchus nerka during infection with Lepeophtheirus salmonis were assessed in controlled laboratory trials. Juvenile salmon were exposed to 100 copepodids fish-1 (Trials 1 and 2) or 300 copepodids fish-1 (Trial 3) at mean weights of approximately 40, 80 and 135 g, respectively. Infections occurred on all salmon in all trials, and mean abundances (infection densities) ranged between 3.3 and 19.4 lice fish-1 (0.08 and 0.44 lice g-1 fish) in Trial 1, between 7.2 and 18.3 (0.09 and 0.22) in Trial 2 and between 19.5 and 60.7 (0.15 and 0.46) in Trial 3. A cumulative mortality of 24.4% occurred in Trial 3. At attachment sites on gills, we observed hyperplasia of basal epithelial cells and fusion of secondary lamellae occasionally associated with a cellular infiltrate. At attachment sites on fins, partial to complete skin erosion occurred, with limited evidence of hyperplasia or inflammation. Scale loss and abrasions coincided with pre-adult lice around 20 d post infection (dpi). Plasma osmolality was significantly elevated in exposed fish in Trials 1 (21 dpi), 2 (15 and 36 dpi) and 3 (20 dpi), whereas haematocrit was significantly depressed in exposed fish in Trials 1 (21 and 28 dpi) and 3 (20 dpi). Plasma cortisol was significantly elevated in exposed fish at 20 dpi (Trial 3). Physiological changes and mortality were related to the intensity of infection and became most prominent with pre-adult stages, suggesting patterns of infection and response in sockeye salmon similar to those reported for Atlantic and Chinook salmon.

Allometric methodology for the assessment of radon exposures to terrestrial wildlife.

A practical approach to calculate (222)Rn daughter dose rates to terrestrial wildlife is presented. The method scales allometrically the relevant parameters for respiration in different species of wildlife, allowing inter-species calculation of the dose per unit radon concentration in air as simple base-and-exponent power functions of the mass. For plants, passive gas exchange through the leaf surface is assumed, also leading to specific power relationships with mass. The model generates conservative predictions in which the main contributor to the dose rate of target tissues of the respiratory system is from α radiation arising from (222)Rn daughters. Tabulated (222)Rn DPURn values are given for 69 species used by the England & Wales Environment Agency for habitats assessments. The approach is then applied to assess the authorised discharges of (222)Rn from sites in England, demonstrating that, from a whole-body dose perspective, the biota considered are protected from effects at the population level.

Parasites and hepatic lesions among pink salmon, Oncorhynchus gorbuscha (Walbaum), during early seawater residence.

Juvenile pink salmon, Oncorhynchus gorbuscha (Walbaum), in the Broughton Archipelago region of western Canada were surveyed over 2 years for sea lice (Lepeophtheirus salmonis and Caligus clemensi), gross and microscopic lesions and evidence of infections with viruses and bacteria. The 1071 fish examined had an approximate ocean residence time no longer than 3 months. A high prevalence of degenerative liver lesions, renal myxosporean parasites and a low prevalence of skin lesions and sea lice were observed. No indications of viral or bacterial diseases were detected in either year. The monthly prevalence of sea lice in 2007 (18-51%) was higher than in 2008 (1-26%), and the infestation density exceeded the lethal threshold in only two fish. Degenerative hepatic lesions and renal myxosporean parasites occurred in approximately 40% of the pink salmon examined in June of both years, and the peak monthly prevalence of hepatocellular hydropic degeneration was greater in 2007 (32%, in May) than in 2008 (12%, in June). Logistic regression analysis found skin lesions and hepatocellular hydropic degeneration significantly associated with sea lice. Most parasites and lesions occurred during both years, but the prevalence was often higher in 2007. Fish weight was 35% less in June 2007 than in June 2008, but condition factor was not different. Further research is required to monitor inter-annual variations and aetiology of the liver lesions and to assess their potential role on pink salmon survival.

A critical role for NMDA receptors in parvalbumin interneurons for gamma rhythm induction and behavior.

Synchronous recruitment of fast-spiking (FS) parvalbumin (PV) interneurons generates gamma oscillations, rhythms that emerge during performance of cognitive tasks. Administration of N-methyl-D-aspartate (NMDA) receptor antagonists alters gamma rhythms, and can induce cognitive as well as psychosis-like symptoms in humans. The disruption of NMDA receptor (NMDAR) signaling specifically in FS PV interneurons is therefore hypothesized to give rise to neural network dysfunction that could underlie these symptoms. To address the connection between NMDAR activity, FS PV interneurons, gamma oscillations and behavior, we generated mice lacking NMDAR neurotransmission only in PV cells (PV-Cre/NR1f/f mice). Here, we show that mutant mice exhibit enhanced baseline cortical gamma rhythms, impaired gamma rhythm induction after optogenetic drive of PV interneurons and reduced sensitivity to the effects of NMDAR antagonists on gamma oscillations and stereotypies. Mutant mice show largely normal behaviors except for selective cognitive impairments, including deficits in habituation, working memory and associative learning. Our results provide evidence for the critical role of NMDAR in PV interneurons for expression of normal gamma rhythms and specific cognitive behaviors.

Low and high affinity dopamine transporter inhibitors block dopamine uptake within 5 sec of intravenous injection.

Extensive evidence suggests that the reinforcing effects of cocaine involve inhibition of dopamine transporters (DAT) and subsequent increases in dopamine (DA) levels in the striatum. We have previously reported that cocaine inhibits the DAT within 4-5 s of i.v. injection, matching the temporal profile of the behavioral and subjective effects of cocaine. Intravenous injection of GBR-12909, a high affinity, long-acting DAT inhibitor, also inhibits DA uptake within 5 s. Given that high affinity, long-acting drugs are considered to have relatively low abuse potential, we found it intriguing that GBR-12909 had an onset profile similar to that of cocaine. To further explore the onset kinetics of both low and high affinity DAT inhibitors, we examined the effects of i.v. cocaine (1.5 mg/kg), methylphenidate (1.5 mg/kg), nomifensine (1.5 mg/kg), GBR-12909 (1.5 mg/kg), PTT (0.5 mg/kg), and WF23 (0.5 mg/kg) on electrically-evoked DA release and uptake in the nucleus accumbens core. Results indicate that all of the DAT inhibitors significantly inhibited DA uptake within 5 s of injection. However, the timing of peak uptake inhibition varied greatly between the low and high affinity uptake inhibitors. Uptake inhibition following cocaine, methylphenidate, and nomifensine peaked 30 s following injection. In contrast, peak effects for GBR-12909, PTT, and WF23 occurred between 20 and 60 min following injection. These observations suggest that the initial onset for i.v. DAT inhibitors is extremely rapid and does not appear to be dictated by a drug's affinity.

An approach to the assessment of risk from chronic radiation to populations of European lobster, Homarus gammarus (L.).

The basic principles underlying a four-discrete age group, logistic, growth model for the European lobster Homarus gammarus are presented and discussed at proof-of-concept level. The model considers reproduction, removal by predation, natural death, fishing, radiation and migration. Non-stochastic effects of chronic low linear energy transfer (LET) radiation are modelled with emphasis on (99)Tc, using three endpoints: repairable radiation damage, impairment of reproductive ability and, at higher dose rates, mortality. An allometric approach for the calculation of LD(50/30) as a function of the mass of each life stage is used in model calibration. The model predicts that at a dose rate of 1 Gy day(-1), lobster population reproduction and survival become severely compromised, leading eventually to population extinction. At 0.01 Gy day(-1), the survival rate of an isolated population is reduced by 10%, mainly through loss of fecundity, comparable to natural migration losses. Fishing is the main ecological stress and only dose rates in the range 0.03-0.1 Gy day(-1) can achieve discernible effects above it. On the balance of radiation and other ecological stresses, a benchmark value of 0.01 Gy day(-1) is proposed for the protection of lobster populations. This value appears consistent with available information on radiation effects in wildlife.

An approach for the assessment of risk from chronic radiation to populations of phytoplankton and zooplankton.

A conceptual model of the effects of chronic radiation on a population of phytoplankton and zooplankton in an oceanic nutrient layer is presented. The model shows that there are distinct threshold dose rates at which the different plankton populations become unsustainable. These are 10,400 microGy h(-1) for phytoplankton and 125 microGy h(-1) for zooplankton. Both these values are considerably greater than the current screening values for protection of 10 microGy h(-1). The model highlights the effects of predator-prey dynamics in predicting that when the zooplankton is affected by the radiation dose, the phytoplankton population can increase. In addition, the model was altered to replicate the dose rates to the plankton of a previous ERICA Irish Sea assessment (24 microGy h(-1) for zooplankton and 430 microGy h(-1) to phytoplankton). The results showed only a 10% decrease in the zooplankton population and a 15% increase in the phytoplankton population. Therefore, at this level of dose, the model predicts that although the dose rate exceeds the guideline value, populations are not significantly affected. This result highlights the limitations of a single screening value for different groups of organisms.

Radionuclide transfer to invertebrates and small mammals in a coastal sand dune ecosystem.

International intercomparisons of models to assess the impact of ionising radiation on wildlife have identified radionuclide transfer assumptions as a significant source of uncertainty in the modelling process. There is a need to improve the underpinning data sets on radionuclide transfer to reduce this uncertainty, especially for poorly-studied ecosystems such as coastal sand dunes. This paper presents the results of the first published study of radionuclide transfer to invertebrates and small mammals in a coastal sand dune ecosystem. Activity concentrations of (137)Cs, (238)Pu, (239+240)Pu and (241)Am are reported for detritivorous, herbivorous, carnivorous and omnivorous biota. Differences in activity concentrations measured in the sand dune biota are related to the trophic level of the organisms and the influence of sea-to-land transfer is apparent in the food chain transfer observed at the site. There are notable differences in the concentration ratios (CRs) calculated for the sand dune biota compared to other terrestrial ecosystems, especially for the small mammals which have CRs that are two orders of magnitude lower than the generic terrestrial ecosystem CRs published by the recent EC EURATOM ERICA project. The lower CRs at the sand dunes may be due to the influence of other cations from the marine environment (e.g. K and Na) on the net radionuclide transfer observed, but further research is required to test this hypothesis.

The phenomenology of inner speech: comparison of schizophrenia patients with auditory verbal hallucinations and healthy controls.

BACKGROUND: Despite the popularity of inner-speech theories of auditory verbal hallucinations (AVHs), little is known about the phenomenological qualities of inner speech in patients with schizophrenia who experience AVHs (Sz-AVHs), or how this compares to inner speech in the non-voice-hearing general population. METHOD: We asked Sz-AVHs (n=29) and a non-voice-hearing general population sample (n=42) a series of questions about their experiences of hearing voices, if present, and their inner speech. RESULTS: The inner speech reported by patients and controls was found to be almost identical in all respects. Furthermore, phenomenological qualities of AVHs (e.g. second- or third-person voices) did not relate to corresponding qualities in inner speech. CONCLUSIONS: No discernable differences were found between the inner speech reported by Sz-AVHs and healthy controls. Implications for inner-speech theories of AVHs are discussed.