Can the Brazilian Caffeine Expectancy Questionnaires Differentiate the CYP1A2 and ADORA2A Gene Polymorphisms?-An Exploratory Study with Brazilian Athletes.

This study investigated the ability of the Brazilian Caffeine Expectancy Questionnaire (CaffEQ-BR), full and brief versions, to differentiate genetic profiles regarding the polymorphisms of the CYP1A2 (rs 762551) and ADORA2A (rs 5751876) genes in a cohort of Brazilian athletes. One-hundred and fifty participants were genotyped for CYP1A2 and ADORA2A. After the recruitment and selection phase, 71 (90% male and 10% female, regular caffeine consumers) completed the CaffEQ-BR questionnaires and a self-report online questionnaire concerning sociodemographic data, general health status, and frequency of caffeine consumption. The order of completion of the CaffEQ-BR questionnaires was counterbalanced. The concordance between the full and brief versions of the CaffEQ-BR was analyzed using the intraclass correlation coefficient (ICC). To determine the discriminatory capacity of the questionnaires for genotype, the receiver operating characteristic (ROC) curve was applied for sensitivity and specificity (significance level of 5%). Mean caffeine intake was 244 ± 161 mg·day−1. The frequency of AA genotypes for CYP1A2 was 47.9% (n = 34) and 52.1% (n = 37) for C-allele carriers (AC and CC). The frequencies of TT genotypes for ADORA2A were 22.7% (n = 15) and 77.3% (n = 51) for C-allele carriers (TC and CC). All CaffEQ-BR factors, for the full and brief versions, were ICCs > 0.75, except for factor 6 (anxiety/negative effects; ICC = 0.60), and presented ROC curve values from 0.464 to 0.624 and 0.443 to 0.575 for CYP1A2 and ADORA2A. Overall, the CaffEQ-BR (full and brief versions) did not show discriminatory capacity for CYP1A2 and ADORA2A gene polymorphisms. In conclusion, the CaffEQ-BR was not able to differentiate genotypes for the CYP1A2 or ADORA2A genes in this group of Brazilian athletes.

Rapid fluctuations in brain oxygenation during glucose-drinking behavior in trained rats.

Proper inflow of oxygen into brain tissue is essential for maintaining normal neural functions. Although oxygen levels in the brain's extracellular space depend upon a balance between its delivery from arterial blood and its metabolic consumption, the use of high-speed electrochemical detection revealed rapid increases in brain oxygen levels elicited by various salient sensory stimuli. These stimuli also increase intrabrain heat production, an index of metabolic neural activation, but these changes are slower and more prolonged than changes in oxygen levels. Therefore, under physiological conditions, the oxygen inflow into brain tissue exceeds its loss due to consumption, thus preventing any metabolic deficit. Here, we used oxygen sensors coupled with amperometry to examine the pattern of real-time oxygen fluctuations in the nucleus accumbens during glucose-drinking behavior in trained rats. Following the exposure to a glucose-containing cup, oxygen levels rapidly increased, peaked when the rat initiated drinking, and relatively decreased during consumption. Similar oxygen changes but more episodic drinking occurred when Stevia, a calorie-free sweet substance, was substituted for glucose. When water was substituted for glucose, rats tested the water but refused to consume all of it. Although the basic pattern of oxygen changes during this water test was similar to that with glucose drinking, the increases were larger. Finally, oxygen increases were significantly larger when rats were exposed to concealed glucose and made multiple unsuccessful attempts to obtain and consume it. Based on these data, we discuss the mechanisms underlying behavior-related brain oxygen fluctuations and their functional significance.NEW & NOTEWORTHY Oxygen sensors coupled with high-speed amperometry were used to examine brain oxygen fluctuations during glucose-drinking behavior in trained rats. Oxygen levels rapidly increased following presentation of a glucose-contained cup, peaking at the initiation of glucose drinking, and relatively decreasing during drinking. Oxygen increases were larger when rats were exposed to concealed glucose and made multiple attempts to obtain it. We discuss the mechanisms underlying behavior-related brain oxygen fluctuations and their functional significance.

Fluid intake, hydration status and body mass changes in U-15 judo athletes during a training day

Despite numerous studies related to dehydration there is still a lack of scientific literature presenting hydration status and fluid intake of judo athletes during different periods. Therefore, the aim of this study was to investigate, fluid intake, hydration status and body weight changes of young judo athletes during a typical day of training in preparation period. Twenty-two young judo athletes (age: 12 ± 0.7 y, experience: 3.5 ± 1.1) voluntarily participated in this study. Hydration status and weight were examined in the morning, before and immediately after the training. All athletes trained 90 min and they consumed fluids ad libitum during the exercise. According to morning urine specific gravity (USG) values, 81.2% of the athletes were dehydrated while only 18.8% of the athletes were euhydrated. Pre-training urine measurements showed that 63.64% of the athletes presented dehydration and 77.27% of the athletes completed the training in dehydrated condition despite fluid availability during the training. Mean body weight loss during training was -0.64 ± 0.66%. It can be concluded that young judo athletes presented high prevalence of dehydration as indicated by USG values. Most of the athletes were dehydrated during a typical training day and completed the training in more dehydrated conditions compared to pre training values despite ad libitum fluid intake. It is of great importance to evaluate hydration status of the athletes before training to refrain from common practice of fluid restriction for weight loss and adverse effects of a persistent state of fluid deficit on physical and health related state.

Habitual caffeine consumption moderates the antidepressant effect of dorsomedial intermittent theta-burst transcranial magnetic stimulation.

BACKGROUND: Potentiating current antidepressant treatment is much needed. Based on animal studies, caffeine may augment the effects of currently available antidepressants. OBJECTIVE: Here, we tested whether habitual caffeine consumption moderates the antidepressant effects of repetitive transcranial magnetic stimulation (rTMS) using intermittent theta-burst stimulation (iTBS). METHODS: Forty patients with current depressive episodes were randomized to active iTBS (n = 19) or sham treatment (n = 21; shielded side of the coil and weak transcutaneous electrical stimulation) delivered two times per day for 10-15 weekdays. Neuronavigated stimulation was applied to the dorsomedial prefrontal cortex. Symptom improvement was measured using change in self-reported Montgomery-Åsberg Depression Rating Scale (MADRS) scores. Pretreatment habitual caffeine consumption was quantified using self-reports of number of cups of coffee and energy drinks consumed the 2 days before the treatment starts. RESULTS: Habitual caffeine consumption was associated with symptom improvement following active iTBS (r = 0.51, 95% confidence interval (CI): 0.08-0.78, p = 0.025) but not following sham treatment (r = -0.02, 95% CI: -0.45 to 0.42, p = 0.938). A multiple regression analysis corroborated the findings by showing a significant caffeine consumption × treatment group interaction (ß = 0.62, p = 0.043), but no main effects of treatment group (ß = 0.22, p = 0.140) or caffeine consumption (ß = -0.01, p = 0.948). No group differences in pretreatment symptom scores or caffeine consumption were detected (p values > 0.86). CONCLUSION: Habitual caffeine consumption moderated the antidepressant effect of dorsomedial iTBS, consistent with caffeine improving antidepressant pharmacological treatments in animals. Caffeine is an antagonist of adenosine receptors and may enhance antidepressant effects through downstream dopaminergic targets.

Impact of Reduced Deuterium Intake on Thermoregulation.

The role of stable hydrogen isotopes in the thermoregulation and its regulation is poorly studied. We analyzed fluctuations in body temperature and changes in thermoregulation parameters in mice under conditions of reduced deuterium intake. The study was performed on male C57BL/6 mice that consumed water with a low (10 ppm) and normal (146 ppm) deuterium content. In 7 days, fluctuations of body temperature, locomotor activity, and oxygen uptake were assessed. Deuterium depletion in the body reduced the mean value of minute fluctuations of body temperature and the mean spectral density of minute fluctuations in body temperature in the 2-20-min periods. This attested to a stabilizing effect of deuterium depletion on the rhythms of body temperature fluctuations, without significant shifts in the thermogenesis parameters. Thus, drinking water with reduced deuterium content makes them less sensitive to external influences.

Habitual Tea and Coffee Consumption and Mean Reaction Time Among Qatari Adults.

BACKGROUND: Tea and coffee consumption is associated with cognitive function in some studies. OBJECTIVE: We aimed to identify tea and coffee drinking patterns and their association with mean reaction time among Qatari adults. METHOD: The study included 1,000 adults aged 20 years and above attending the Qatar Biobank Study (QBB). Habitual tea and coffee consumption during the previous year was assessed by questionnaire. Tea and coffee drinking patterns were identified using factor analysis. In a computer-based self-administered touch screens test, mean reaction time (MRT) was used as an indicator of cognitive function. RESULTS: The mean age of the participants was 35.8 (SD 10.3) years. Herbal tea and regular coffee consumption was inversely associated with MRT. In the multivariable model, compared with non-consumers, the regression coefficients for MRT were -34.3 (-65.4, -3.3) and -37.9 (-71.0, -4.7) for daily consumers of herbal tea and regular coffee, respectively. Of the two tea and coffee drinking patterns identified, pattern 1 (high consumption of tea, Arabic coffee, and herbal tea) was not associated with MRT but pattern 2 (high loadings of instant coffee, regular coffee, and Karak) was inversely associated with MRT in the unadjusted model. There was a significant interaction between pattern 2 and low-density lipoprotein (LDL) in relation to MRT. Pattern 2 was inversely associated with MRT among those with a low LDL. CONCLUSION: There was an inverse association between regular coffee and herbal tea consumption with mean reaction time. There was an interaction between Western coffee pattern and LDL.

Microstructural changes in human ingestive behavior after Roux-en-Y gastric bypass during liquid meals.

BACKGROUNDRoux-en-Y gastric bypass (RYGB) decreases energy intake and is, therefore, an effective treatment of obesity. The behavioral bases of the decreased calorie intake remain to be elucidated. We applied the methodology of microstructural analysis of meal intake to establish the behavioral features of ingestion in an effort to discern the various controls of feeding as a function of RYGB.METHODSThe ingestive microstructure of a standardized liquid meal in a cohort of 11 RYGB patients, in 10 patients with obesity, and in 10 healthy-weight adults was prospectively assessed from baseline to 1 year with a custom-designed drinkometer. Statistics were performed on log-transformed ratios of change from baseline so that each participant served as their own control, and proportional increases and decreases were numerically symmetrical. Data-driven (3 seconds) and additional burst pause criteria (1 and 5 seconds) were used.RESULTSAt baseline, the mean meal size (909.2 versus 557.6 kCal), burst size (28.8 versus 17.6 mL), and meal duration (433 versus 381 seconds) differed between RYGB patients and healthy-weight controls, whereas suck volume (5.2 versus 4.6 mL) and number of bursts (19.7 versus 20.1) were comparable. At 1 year, the ingestive differences between the RYGB and healthy-weight groups disappeared due to significantly decreased burst size (P = 0.008) and meal duration (P = 0.034) after RYGB. The first-minute intake also decreased after RYGB (P = 0.022).CONCLUSIONRYGB induced dynamic changes in ingestive behavior over the first postoperative year. While the eating pattern of controls remained stable, RYGB patients reduced their meal size by decreasing burst size and meal duration, suggesting that increased postingestive sensibility may mediate postbariatric ingestive behavior.TRIAL REGISTRATIONNCT03747445; https://clinicaltrials.gov/ct2/show/NCT03747445.FUNDINGThis work was supported by the University of Zurich, the Swiss National Fund (32003B_182309), and the Olga Mayenfisch Foundation. Bálint File was supported by the Hungarian Brain Research Program Grant (grant no. 2017-1.2.1-NKP-2017-00002).

Availability of a Flavored Beverage and Impact on Children's Hydration Status, Sleep, and Mood.

Euhydration remains a challenge in children due to lack of access and unpalatability of water and to other reasons. The purpose of this study was to determine if the availability/access to a beverage (Creative Roots®) influences hydration in children and, therefore, sleep quality and mood. Using a crossover investigation, 46 participants were randomly assigned to a control group (CON) or an intervention group and received Creative Roots® (INT) for two-week periods. We recorded daily first morning and afternoon urine color (Ucol), thirst perception, and bodyweight of the two groups. Participants reported to the lab once per week and provided first morning urine samples to assess Ucol, urine specific gravity (USG), and urine osmolality (Uosmo). Participants also completed the questionnaires Profile of Mood States-Adolescents (POMS-a) and Pittsburgh Sleep Quality Index (PSQI). Dependent t-tests were used to assess the effects of the intervention on hydration, mood, and sleep quality. Uosmo was greater and Ucol was darker in the control group (mean ± SD) [Uosmo: INT = 828 ± 177 mOsm·kg-1, CON = 879 ± 184 mOsm·kg-1, (p = 0.037], [Ucol:INT = 5 ± 1, CON = 5 ± 1, p = 0.024]. USG, POMS-a, and PSQI were not significant between the groups. At-home daily afternoon Ucol was darker in the control group [INT = 3 ± 1, CON = 3 ± 1, p = 0.022]. Access to Creative Roots® provides a small, potentially meaningful hydration benefit in children. However, children still demonstrated consistent mild dehydration based on Uosmo, despite consuming the beverage.

Expression of the hippocampal PTCH during early abstinence is associated with drinking patterns in a rat model of voluntary alcohol intake.

Perinatal alcohol exposure induces fetal alcohol syndrome partially through Sonic Hedgehog (SHH) impairment; however, the relationship between SHH signaling cascade and alcohol drinking pattern in adulthood remains obscure. We studied the expression of SHH and components of respective signaling cascade [PTCH receptor (Patched), SMO co-receptor (Smoothened) and downstream transcriptional factor Glioma-associated oncogene (GLI)] during early abstinence in brain regions of rats demonstrating different drinking patterns in intermittent access two-bottle choice paradigm (IA2BC). Male Wistar rats were subjected to twenty 24-h sessions of free access to two-bottle choice (water or 20% ethanol) with 24-h withdrawal periods (water only). Control animals had access to water only. Quantitative PCR and western blotting were used to assess transcript and protein levels in the brain, respectively. During the course of the IA2BC, one part of animals demonstrated gradual escalation from low to high alcohol intake and preference of alcohol over water (group I), while the other one consumed alcohol at stable high level (group II) (Peregud et al., 2021). Three days after the last drinking session, PTCH mRNA elevated in the hippocampus of group I rats as compared to the control group. However, SHH, SMO and GLI mRNA levels in the hippocampus did not change. The protein content of PTCH in the hippocampus of group I rats was higher as compared to both control and group II. PTCH elevation is a known marker of SHH cascade activity. Thus, activated hippocampal SHH signaling cascade is a hallmark of rats demonstrating gradual escalation of alcohol intake in the IA2BC procedure.

Peer influence in adolescent drinking behavior: A meta-analysis of stochastic actor-based modeling studies.

OBJECTIVES: To measure the effects of peer influence and peer selection on drinking behavior in adolescence through a rigorous statistical approach designed to unravel these interrelated processes. METHODS: We conducted systematic searches of electronic databases, thesis collections and conference proceedings to identify studies that used longitudinal network design and stochastic actor-oriented modeling to analyze drinking behavior in adolescents. Parameter estimates collected from individual studies were analyzed using multilevel random-effects models. RESULTS: We identified 26 articles eligible for meta-analysis. Meta-analyses for different specifications of the peer influence effect were conducted separately. The peer influence effect was positive for every specification: for average similarity (avSim) mean log odds ratio was 1.27 with 95% confidence interval [0.04; 2.49]; for total similarity (totSim) 0.46 (95% CI = [0.44; 0.48]), and for average alter (avAlt) 0.70 (95% CI = [-0.01; 1.41]). The peer selection effect (simX) was also positive: 0.46 (95% CI = [0.28; 0.63]). Conversion log odds ratio values to Cohen's d gives estimates from 0.25 to 0.70, which is considered as medium to large effect. CONCLUSIONS: Advances in methodology for social network analysis have made it possible to accurately estimate peer influence effects free from peer selection effects. More research is necessary to clarify the roles of age, gender, and individual susceptibility on the changing behavior of adolescents under the influence of their peers. Understanding the effects of peer influence should inform practitioners and policy makers to design and deliver more effective prevention programs.

Post-diagnostic coffee and tea consumption and breast cancer survival.

BACKGROUND: We examined the role of post-diagnostic coffee and tea consumption in relation to breast cancer-specific and all-cause mortality among women with breast cancer in prospective cohort studies. METHODS: We identified 8900 women with stage I-III breast cancer from 1980 through 2010 in the Nurses' Health Study (NHS) and from 1991 through 2011 in the NHSII. Post-diagnostic coffee and tea consumption was assessed by a validated food frequency questionnaire every 4 years after diagnosis. RESULTS: During up to 30 years of follow-up, we documented 1054 breast cancer-specific deaths and 2501 total deaths. Higher post-diagnostic coffee consumption was associated with a lower breast cancer-specific mortality: compared with non-drinkers, >3 cups/day of coffee was associated with a 25% lower risk (hazard ratio (HR) = 0.75, 95% confidence interval (CI) = 0.59-0.96; Ptrend = 0.002). We also observed a lower all-cause mortality with coffee consumption: compared with non-drinkers, >2 to 3 cups/day was associated with a 24% lower risk (HR = 0.76, 95% CI = 0.66-0.87) and >3 cups/day was associated with a 26% lower risk (HR = 0.74, 95% CI = 0.63-0.87, Ptrend < 0.0001). Post-diagnostic tea consumption was associated with a lower all-cause mortality: compared with non-drinkers, >3 cups/day was associated with a 26% lower risk (HR = 0.74, 95% CI = 0.58-0.95; Ptrend = 0.04). CONCLUSIONS: Among breast cancer survivors, higher post-diagnostic coffee consumption was associated with better breast cancer and overall survival. Higher post-diagnostic tea consumption may be related to better overall survival.

Flower consumption, ambient temperature and rainfall modulate drinking behavior in a folivorous-frugivorous arboreal mammal.

Water is vital for the survival of any species because of its key role in most physiological processes. However, little is known about the non-food-related water sources exploited by arboreal mammals, the seasonality of their drinking behavior and its potential drivers, including diet composition, temperature, and rainfall. We investigated this subject in 14 wild groups of brown howler monkeys (Alouatta guariba clamitans) inhabiting small, medium, and large Atlantic Forest fragments in southern Brazil. We found a wide variation in the mean rate of drinking among groups (range = 0-16 records/day). Streams (44% of 1,258 records) and treeholes (26%) were the major types of water sources, followed by bromeliads in the canopy (16%), pools (11%), and rivers (3%). The type of source influenced whether howlers used a hand to access the water or not. Drinking tended to be evenly distributed throughout the year, except for a slightly lower number of records in the spring than in the other seasons, but it was unevenly distributed during the day. It increased in the afternoon in all groups, particularly during temperature peaks around 15:00 and 17:00. We found via generalized linear mixed modelling that the daily frequency of drinking was mainly influenced negatively by flower consumption and positively by weekly rainfall and ambient temperature, whereas fragment size and the consumption of fruit and leaves played negligible roles. Overall, we confirm the importance of preformed water in flowers to satisfy the howler's water needs, whereas the influence of the climatic variables is compatible with the 'thermoregulation/dehydration-avoiding hypothesis'. In sum, we found that irrespective of habitat characteristics, brown howlers seem to seek a positive water balance by complementing the water present in the diet with drinking water, even when it is associated with a high predation risk in terrestrial sources.

A Young man with Peri-ictal water drinking.

Neuro-vegetative features have been linked to epilepsy arising from the temporal lobe, which can be seen during ictal events and play an important role in determining the focal side of the lesion. Among the rare known features is peri-ictal water drinking (PIWD). Here, we present the case of a 31-year-old male with refractory temporal epilepsy, having episodes of PIWD. The patient was considered a candidate for resective surgery and was investigated accordingly, including long-term video electroencephalogram (EEG), brain magnetic resonance imaging (MRI), and neuropsychology assessment, which pointed towards a lesioned temporal lobe over the non-dominant hemisphere. The patient had an excellent outcome following right anterior temporal lobectomy. The lateralization significance of PIWD has not yet been established definitively in the literature, partly due to a limited number of published cases. This case highlights the previously observed association between PIWD and the involvement of the non-dominant hemisphere in epilepsy patients.

Ventral arkypallidal neurons inhibit accumbal firing to promote reward consumption.

The nucleus accumbens shell (NAcSh) and the ventral pallidum (VP) are critical for reward processing, although the question of how coordinated activity within these nuclei orchestrates reward valuation and consumption remains unclear. Inhibition of NAcSh firing is necessary for reward consumption, but the source of this inhibition remains unknown. Here, we report that a subpopulation of VP neurons, the ventral arkypallidal (vArky) neurons, project back to the NAcSh, where they inhibit NAcSh neurons in vivo in mice. Consistent with this pathway driving reward consumption via inhibition of the NAcSh, calcium activity of vArky neurons scaled with reward palatability (which was dissociable from reward seeking) and predicted the subsequent drinking behavior during a free-access paradigm. Activation of the VP-NAcSh pathway increased ongoing reward consumption while amplifying hedonic reactions to reward. These results establish a pivotal role for vArky neurons in the promotion of reward consumption through modulation of NAcSh firing in a value-dependent manner.

Live predator stress in adolescence results in distinct adult behavioral consequences and dorsal diencephalic brain activation patterns.

Exposure to traumatic events during childhood increases the risk of adult psychopathology, including anxiety, depression, alcohol use disorders and their co-morbidity. Early life trauma also results in increased symptom complexity, treatment resistance and poor treatment outcomes. The purpose of this study was to establish a novel rodent model of adolescent stress, based on an ethologically relevant life-threatening event, live predator exposure. Rats were exposed to a live predator for 10 min. at three different time points (postnatal day (PND)31, 46 and 61). Adult depression-, anxiety-like behaviors and ethanol consumption were characterized well past the last acute stress event (two weeks). Behavioral profiles across assessments were developed to characterize individual response to adolescent stress. CNS activation patterns in separate groups of subjects were characterized after the early (PND31) and last predator exposure (PND61). Subjects exposed to live-predator adolescent stress generally exhibited less exploratory behavior, less propensity to venture into open spaces, a decreased preference for sweet solutions and decreased ethanol consumption in a two-bottle preference test. Additional studies demonstrated blunted cortisol response and CNS activation patterns suggestive of habenula, rostromedial tegmental (RMTg), dorsal raphe and central amygdala involvement in mediating the adult consequences of adolescent stress. Thus, adolescent stress in the form of live-predator exposure results in significant adult behavioral and neurobiological disturbances. Childhood trauma, its impact on neurodevelopment and the subsequent development of mood disorders is a pervasive theme in mental illness. Improving animal models and our neurobiological understanding of the symptom domains impacted by trauma could significantly improve treatment strategies.

Beverage Intakes and Toothbrushing During Childhood Are Associated With Caries at Age 17 Years.

BACKGROUND: Sugar-sweetened beverages (SSBs) have been associated with childhood caries; however, associations among lifelong beverage intakes and adolescent caries have received less attention. OBJECTIVE: To investigate associations between beverage intakes during childhood and adolescence and caries experience at 17 years of age, while adjusting for fluoride intakes and toothbrushing. DESIGN: Descriptive model analyses were conducted on data collected from a longitudinal birth cohort study. PARTICIPANTS/SETTING: Participants included Iowa Fluoride Study members (n = 318) recruited at birth between 1992 and 1995 with at least 6 beverage questionnaires completed from ages 1 to 17 years and a caries examination at age 17. EXPOSURE: Predictors included mean daily milk, juice (100% juice and juice drinks before age 9), SSB (including juice drinks after age 9), and water/sugar-free beverage (SFB) intakes; daily fluoride intakes; and daily toothbrushing frequencies for ages 1 to 17. MAIN OUTCOME MEASURES: The outcome was dental caries count at age 17, adjusted for the number of scored tooth surfaces (decayed and filled surfaces attack rate [DFSAR]). STATISTICAL ANALYSES PERFORMED: Univariable generalized linear models were fit for each predictor and the outcome DFSAR. Multivariable models assessed combined effects of beverage types, fluoride variables, toothbrushing, sex, and baseline socioeconomic status. RESULTS: Based on multivariable models, each 8 oz of additional daily juice and water/SFB decreased expected DFSAR by 53% (95% confidence interval [CI]: 17%-73%) and 29% (95% CI: 7%-46%), respectively, and 8 additional oz SSBs increased expected DFSAR by 42% (95% CI: 5%-92%), after adjustment for other beverage intakes, toothbrushing, total fluoride intake excluding SSB fluoride (non-SSB total fluoride), sex, and baseline socioeconomic status. Each additional daily toothbrushing event decreased expected DFSAR by 43% (95% CI: 14%-62%) after adjustment for beverage intakes, non-SSB total fluoride intake, sex, and baseline SES. CONCLUSIONS: Higher juice and water/SFB intakes and more toothbrushing were associated with lower caries at age 17, while higher SSB intakes were associated with higher caries.

Aversion-resistant fentanyl self-administration in mice.

RATIONALE: Animal models of compulsive drug use that continues despite negative consequences can be used to investigate the neural mechanisms of addiction. However, models of punished or aversion-resistant opioid self-administration are notably lacking. OBJECTIVES: We sought to develop an aversion-resistant, oral fentanyl self-administration paradigm. METHODS: In Experiment 1, C57BL/6J male and female, adult mice consumed fentanyl (10 µg/mL) in a two-bottle drinking in the dark task and escalating concentrations of quinine were added to the bottles. In Experiment 2, mice were trained to administer oral fentanyl (10 µg/mL) in an operant response task. Quinine was next added to the fentanyl solution in escalating concentrations. In Experiment 3, mice were trained to respond for oral fentanyl or fentanyl adulterated with 500 µM quinine on every session. In Experiment 4, mice were trained to respond for a 1% sucrose solution before introduction of quinine. RESULTS: Quinine reduced two-bottle choice consumption in males but not in females. Both sexes demonstrated the ability to detect the selected concentrations of quinine in fentanyl. In the operant chamber, mice responded robustly for oral fentanyl but introduction of quinine at any stage of training was insufficient to reduce responding. In contrast, quinine reduced responding for sucrose at concentrations above 250 µM. CONCLUSIONS: Mice will respond for and consume oral fentanyl in both a two-bottle choice and an operant response task. Quinine is detectable in fentanyl but mice will continue to respond for and consume fentanyl with quinine in both paradigms. These data support the use of these models in behavioral studies of compulsive-like opioid use.

Distinct CCK-positive SFO neurons are involved in persistent or transient suppression of water intake.

The control of water-intake behavior is critical for life because an excessive water intake induces pathological conditions, such as hyponatremia or water intoxication. However, the brain mechanisms controlling water intake currently remain unclear. We previously reported that thirst-driving neurons (water neurons) in the subfornical organ (SFO) are cholecystokinin (CCK)-dependently suppressed by GABAergic interneurons under Na-depleted conditions. We herein show that CCK-producing excitatory neurons in the SFO stimulate the activity of GABAergic interneurons via CCK-B receptors. Fluorescence-microscopic Ca2+ imaging demonstrates two distinct subpopulations in CCK-positive neurons in the SFO, which are persistently activated under hyponatremic conditions or transiently activated in response to water drinking, respectively. Optical and chemogenetic silencings of the respective types of CCK-positive neurons both significantly increase water intake under water-repleted conditions. The present study thus reveals CCK-mediated neural mechanisms in the central nervous system for the control of water-intake behaviors.

Automated recognition of postures and drinking behaviour for the detection of compromised health in pigs.

Changes in pig behaviours are a useful aid in detecting early signs of compromised health and welfare. In commercial settings, automatic detection of pig behaviours through visual imaging remains a challenge due to farm demanding conditions, e.g., occlusion of one pig from another. Here, two deep learning-based detector methods were developed to identify pig postures and drinking behaviours of group-housed pigs. We first tested the system ability to detect changes in these measures at group-level during routine management. We then demonstrated the ability of our automated methods to identify behaviours of individual animals with a mean average precision of [Formula: see text], under a variety of settings. When the pig feeding regime was disrupted, we automatically detected the expected deviations from the daily feeding routine in standing, lateral lying and drinking behaviours. These experiments demonstrate that the method is capable of robustly and accurately monitoring individual pig behaviours under commercial conditions, without the need for additional sensors or individual pig identification, hence providing a scalable technology to improve the health and well-being of farm animals. The method has the potential to transform how livestock are monitored and address issues in livestock farming, such as targeted treatment of individuals with medication.