Neurobehavioral Research Research Articles

Disparate trajectories of cognitive aging among American Indian and Alaskan Native people with and without HIV.

This study describes trajectories of cognitive aging among American Indian/Alaskan Native (AI/AN) adults with and without HIV and the role of immunosenescence longitudinally. We characterized trajectories of cognitive aging in a sample of 333 AI/AN and 309 non-Hispanic White (NHW) adults who were followed longitudinally for up to 20 years by the HIV Neurobehavioral Research Program (HNRP) across six U.S. research sites. We used growth curve modeling with autoregressive Lag-1 structures and heterogeneous residual variances to assess the role of ethnoracial identity and HIV grouping upon decline in trajectories of cognitive aging. HIV- AI/AN adults demonstrated earlier and steeper decline in normative trajectories of cognitive aging on tasks of processing speed, timed tasks of attention/working memory, executive function, and psychomotor speed in comparison to HIV- NHW adults. Accentuated trajectories of cognitive aging were evident in both HIV+ and HIV+ immunosuppressed groups in comparison to HIV- peers and were primarily driven by the role of immunosenescence. AI/AN disparities in trajectories of cognitive aging are evident and are likely explained by the interplay of biopsychosociocultural factors, including immunosenescence. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Standardizing zebrafish laboratory husbandry to ensure replicability and reproducibility of data in neurobehavioral research.

Standardizing zebrafish laboratory husbandry to ensure replicability and reproducibility of data in neurobehavioral research.

Contextual fear conditioning in zebrafish: Influence of different shock frequencies, context, and pharmacological modulation on behavior

Contextual fear conditioning is a protocol used to assess associative learning across species, including fish. Here, our goal was to expand the analysis of behavioral parameters that may reflect aversive behaviors in a contextual fear conditioning protocol using adult zebrafish (Danio rerio) and to verify how such parameters can be modulated. First, we analyzed the influence of an aversive stimulus (3 mild electric shocks for 5 s each at frequencies of 10, 100 or 1000 Hz) on fish behavior, and their ability to elicit fear responses in the absence of shock during a test session. To confirm whether the aversive responses are context-dependent, behaviors were also measured in a different experimental environment in a test session. Furthermore, we investigated the effects of dizocilpine (MK-801, 2 mg/kg, i.p.) on fear-related responses. Zebrafish showed significant changes in baseline activity immediately after shock exposure in the training session, in which 100 Hz induced robust contextual fear responses during the test session. Importantly, when introduced to a different environment, animals exposed to the aversive stimulus did not show any differences in locomotion and immobility-related parameters. MK-801 administered after the training session reduced fear responses during the test, indicating that glutamate NMDA-receptors play a key role in the consolidation of contextual fear-related memory in zebrafish. In conclusion, by further exploring fear-related behaviors in a contextual fear conditioning task, we show the effects of different shock frequencies and confirm the importance of context on aversive responses for associative learning in zebrafish. Additionally, our data support the use of zebrafish in contextual fear conditioning tasks, as well as for advancing pharmacological studies related to associative learning in translational neurobehavioral research.

Behavioral neuroscience in zebrafish: unravelling the complexity of brain-behavior relationships.

This paper reviews the utility of zebrafish (Danio rerio) as a model system for exploring neurobehavioral phenomena in preclinical research, focusing on physiological processes, disorders, and neurotoxicity biomarkers. A comprehensive review of the current literature was conducted to summarize the various behavioral characteristics of zebrafish. The study examined the etiological agents used to induce neurotoxicity and the biomarkers involved, including Aβ42, tau, MMP-13, MAO, NF-Кβ, and GFAP. Additionally, the different zebrafish study models and their responses to neurobehavioral analysis were discussed. The review identified several key biomarkers of neurotoxicity in zebrafish, each impacting different aspects of neurogenesis, inflammation, and neurodegeneration. Aβ42 was found to alter neuronal growth and stem cell function. Tau's interaction with tubulin affected microtubule stability and led to tauopathies under pathological conditions. MMP-13 was linked to oxidative assault and sensory neuron degeneration. MAO plays a role in neurotransmitter metabolism and neurotoxicity conversion. NF-Кβ was involved in pro-inflammatory pathways, and GFAP was indicative of neuroinflammation and astroglial activation. Zebrafish provide a valuable model for neurobehavioral research, adhering to the "3Rs" philosophy. Their neurotoxicity biomarkers offer insights into the mechanisms of neurogenesis, inflammation, and neurodegeneration. This model system aids in evaluating physiological and pathological conditions, enhancing our understanding of neurobehavioral phenomena and potential therapeutic interventions.

Alzheimer-Type Cerebral Amyloidosis in the Context of HIV Infection: Implications for a Proposed New Treatment Approach

Reverse transcriptase inhibitors (RTIs) are currently broadly prescribed for the treatment of HIV infection but are also thought to prevent Alzheimer’s disease (AD) progression by protecting against amyloidosis. Our study evaluates the hypothesis that reverse transcriptase inhibitors protect against Alzheimer-type brain amyloidogenesis in the context of HIV infection. We compiled a case series of participants from a prospective study of the neurological consequences of HIV infection at the HIV Neurobehavioral Research Program (HNRP) who had serial neuropsychological and neurological assessments and were on RTIs. Two participants had gross and microscopic examination and immunohistochemistry of the brain at autopsy; one was assessed clinically for Alzheimer’s disease by cerebrospinal fluid (CSF) analysis of phosphorylated-Tau, Total-Tau and Aβ42. Additionally, a larger cohort of 250 autopsied individuals was evaluated for presence of amyloid plaques, Tau, and related pathologies. Three older, virally suppressed individuals with HIV who had long-term treatment with RTIs were included in analyses. Two cases demonstrated substantial cerebral amyloid deposition at autopsy. The third case met clinical criteria for AD based on a typical clinical course and CSF biomarker profile. In the larger cohort of autopsied individuals, the prevalence of cerebral amyloidosis among people with HIV (PWH) was greater for those on RTIs. Our study showed that long-term RTI therapy did not protect against Alzheimer-type brain amyloidogenesis in the context of HIV infection in these patients. Given the known toxicities of RTIs, it is premature to recommend them to individuals at risk or with Alzheimer’s disease who do not have HIV infection.

Towards Fish Welfare in the Presence of Robots: Zebrafish Case

Zebrafish (Danio rerio) have emerged as a valuable animal model for neurobehavioral research, particularly in the study of anxiety-related states. This article explores the use of conceptual models to investigate stress, fear, and anxiety in zebrafish induced by bio-inspired mini-robotic fish with different components and designs. The objective is to optimize robotic biomimicry and its impact on fish welfare. Previous studies have focused on externally controlled fish models, whereas this study introduces prototypes of freely actuated swimming robots to examine interactions between a bio-inspired robot and individual zebrafish. By means of analysis of behavioral responses, certain robotic components have been identified as potential causes of anxiety in fish, which have provided insights that may be applicable to other species and future aquacultural robot designs.

Elevated Biomarkers of Inflammation and Vascular Dysfunction Are Associated with Distal Sensory Polyneuropathy in People with HIV.

Distal sensory polyneuropathy (DSP) is a disabling, chronic condition in people with HIV (PWH), even those with viral suppression of antiretroviral therapy (ART), and with a wide range of complications, such as reduced quality of life. Previous studies demonstrated that DSP is associated with inflammatory cytokines in PWH. Adhesion molecules, essential for normal vascular function, are perturbed in HIV and other conditions linked to DSP, but the link between adhesion molecules and DSP in PWH is unknown. This study aimed to determine whether DSP signs and symptoms were associated with a panel of plasma biomarkers of inflammation (d-dimer, sTNFRII, MCP-1, IL-6, IL-8, IP-10, sCD14) and vascular I integrity (ICAM-1, VCAM-1, uPAR, MMP-2, VEGF, uPAR, TIMP-1, TIMP-2) and differed between PWH and people without HIV (PWoH). A cross-sectional study was conducted among 143 participants (69 PWH and 74 PWoH) assessed by studies at the UC San Diego HIV Neurobehavioral Research Program. DSP signs and symptoms were clinically assessed for all participants. DSP was defined as two or more DSP signs: bilateral symmetrically reduced distal vibration, sharp sensation, and ankle reflexes. Participant-reported symptoms were neuropathic pain, paresthesias, and loss of sensation. Factor analyses reduced the dimensionality of the 15 biomarkers among all participants, yielding six factors. Logistic regression was used to assess the associations between biomarkers and DSP signs and symptoms, controlling for relevant demographic and clinical covariates. The 143 participants were 48.3% PWH, 47 (32.9%) women, and 47 (33.6%) Hispanics, with a mean age of 44.3 ± 12.9 years. Among PWH, the median (IQR) nadir and current CD4+ T-cells were 300 (178-448) and 643 (502-839), respectively. Participants with DSP were older but had similar distributions of gender and ethnicity to those without DSP. Multiple logistic regression showed that Factor 2 (sTNFRII and VCAM-1) and Factor 4 (MMP-2) were independently associated with DSP signs in both PWH and PWoH (OR [95% CI]: 5.45 [1.42-21.00], and 15.16 [1.07-215.22]), respectively. These findings suggest that inflammation and vascular integrity alterations may contribute to DSP pathogenesis in PWH, but not PWoH, possibly through endothelial dysfunction and axonal degeneration.

Neurodevelopmental toxicity and molecular mechanism of environmental concentration of tetrabromobisphenol A bis (2- hydroxyethyl) ether exposure to sexually developing male SD rats

Tetrabromobisphenol A bis (2- hydroxyethyl) ether (TBBPA-DHEE), as one of the main derivatives of Tetrabromobisphenol A, been attracted attention for its health risks. In this study, the neurotoxicity, mechanism, and susceptivity of TBBPA-DHEE exposure to sexually developing male rats were systematically studied. Neurobehavioral research showed that TBBPA-DHEE exposure could significantly affect the behavior, learning,and memory abilities of male-developing rats, and aggravate their depression. TBBPA-DHEE exposure could inhibit the secretion of neurotransmitters. Transcriptomics studies show that TBBPA-DHEE can significantly affect gene expression, and a total of 334 differentially expressed genes are enriched. GO function enrichment analysis shows that TBBPA-DHEE exposure can significantly affect the expression of genes related to synapses and cell components. KEGG function enrichment analysis shows that TBBPA-DHEE exposure can significantly affect the expression of signal pathways related to nerves, nerve development, and signal transduction. Susceptibility analysis showed that female rats were more susceptible to TBBPA-DHEE exposure than male rats. Therefore, TBBPA-DHEE exposure has neurodevelopmental toxicity to male developmental rats, and female developmental rats are more susceptible than male developmental rats. Its possible molecular mechanism is that TBBPA-DHEE may inhibit the secretion of neurotransmitters and affect signal pathways related to neurodevelopment and signal transduction.

Identifying and distinguishing cognitive profiles among virally suppressed people with HIV.

Cognitive deficits are common among people with HIV (PWH), even when virally suppressed. We identified cognitive profiles among virally suppressed PWH and determined how sociodemographic, clinical/behavioral, and HIV disease characteristics distinguish profile membership. Participants included 704 virally suppressed PWH (Mage = 43.9 [SD = 10.2], 88% male, 58.9% non-Hispanic White) from the HIV Neurobehavioral Research Program. Demographically adjusted T scores were derived from a neuropsychological evaluation comprised of 13 tests. We implemented a pipeline involving dimension reduction and clustering to identify profiles of cognitive performance. Random forest models on a 70/30 training/testing set with internal cross-validation were used to identify sociodemographic, clinical/behavioral, and HIV disease correlates of profile membership. Six cognitive profiles were identified: (a) "unimpaired" (19.9%); (b) weakness in verbal learning and memory (15.5%); (c) weakness in executive function and learning (25.8%); (d) weakness in motor, processing speed, and executive function (8.1%); (e) impaired learning and recall with weak-to-impaired motor, processing speed, and executive function (13.1%); (f) global deficits (17.6%). The most discriminative sociodemographic, clinical/behavioral, and HIV disease characteristics varied by profile with self-reported mood symptoms and cognitive/functional difficulties (e.g., language/communication, memory, and overall everyday function complaints) most consistently associated with profile membership. Cognitive profiles and their associated factors among PWH are heterogeneous, but learning/memory deficits were most common and self-reported mood, and cognitive/functional difficulties were most consistently related to profile membership. This heterogeneity in cognitive profiles and their correlates in PWH suggests that differing mechanisms contribute to cognitive deficits and, thus, underscores the need for personalized risk reduction and therapeutic strategies among PWH. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

AGE-VARYING COGNITIVE IMPAIRMENT AND SOCIAL SUPPORT AMONG PEOPLE AGING WITH AND WITHOUT HIV

Abstract There is compelling but limited evidence that the combination of HIV, methamphetamine (meth) use, and aging form a deleterious trio that undermines cognitive functioning. Currently, a gap exists in our understanding of the intertwining relationships among drug use, HIV, and cognitive function across biological ages and over time. Interventions to reduce meth use and support brain health among aging PWH are vital and improving social relationships is a promising area of intervention. Cross-sectional and limited longitudinal studies indicate that positive social relationships and social support are associated with better cognitive function among healthy older populations. This association has received limited attention among PWH who use drugs beyond adolescence and young adulthood. This study leverages NIH’s prior and ongoing investments in research to develop a new analytic cohort by combining participants from 23 studies at the HIV Neurobehavioral Research program. Enrollment in this cohort spans from 2009-2021. At baseline, our sample consists of 862 people with a mean age of 52 (range 18-81). Most participants are male (73%), White (74%), and over half (61%) are living with HIV. Using a life course perspective and time-varying estimation modeling, we examine age-varying cognitive impairment and variation in cognitive function across age by HIV status, meth use, and social relationships. First, we examine how cognitive impairment changes over time for the full sample then examine the differing transitions/turning points among those aging with and without HIV and meth use. Lastly, we show how positive and negative social relationships alter these trajectories among each group.

BARRIERS TO SUCCESSFUL AGING AMONG OLDER WOMEN AND MEN LIVING WITH HIV

Abstract This study identified and compared barriers to successful aging (SA) among older women and men living with HIV. Participants were recruited through 1) the Women’s Interagency HIV Study (WIHS), Atlanta and Brooklyn sites; and 2) the HIV Neurobehavioral Research Program at the University of California, San Diego. Our sample included 31 participants living with HIV: 17 women and 14 men, ages 52 to 73 years, 58% African American. We conducted semi-structured interviews, exploring SA with HIV. The interviews (~90 minutes long) were audio recorded, fully transcribed, and imported into MAXQDA software for analysis. Thematic and comparative analyses were conducted within a social-constructivist framework. We uncovered several themes related to SA barriers that were shared by women and men: challenging chronic conditions, stigmas, substance use, and lack of resources. We also found differences. While men stressed dreading old age and loneliness, women emphasized stress and ‘giving up’ as barriers to SA.

The clinical utility of three frailty measures in identifying HIV-associated neurocognitive disorders.

Frailty measures vary widely and the optimal measure for predicting HIV-associated neurocognitive disorders (HAND) is unclear. A study was conducted to examine the clinical utility of three widely used frailty measures in identifying HIV-associated neurocognitive disorders. The study involved 284 people with HIV (PWH) at least 50 years enrolled at UC San Diego's HIV Neurobehavioral Research Program. Frailty measurements included the Fried Phenotype, the Rockwood Frailty Index, and the Veterans Aging Cohort Study (VACS) Index. HAND was diagnosed according to Frascati criteria. ANOVAs examined differences in frailty severity across HAND conditions. ROC analyses evaluated sensitivity and specificity of each measure to detect symptomatic HAND [mild neurocognitive disorder (MND) and HIV-associated dementia (HAD)] from no HAND. Across all frailty measures, frailty was found to be higher in HAD compared with no HAND. For Fried and Rockwood (not VACS), frailty was significantly more severe in MND vs. no HAND and in HAD vs. ANI (asymptomatic neurocognitive impairment). For discriminating symptomatic HAND from no HAND, Fried was 37% sensitive and 92% specific, Rockwood was 85% sensitive and 43% specific, and VACS was 58% sensitive and 65% specific. These findings demonstrate that Fried and Rockwood outperform VACS in predicting HAND. However, ROC analyses suggest none of the indices had adequate predictive validity in detecting HAND. The results indicate that the combined use of the Rockwood and Fried indices may be an appropriate alternative.

Zebra fish behavioral assessment by using Calycophyllum Spruceanum bark methanolic extract

In neurobehavioral research, animal model has played a crucial role in yielding experimental data as well as, in the development of new insights. Researchers are always trying to develop novel animal models to understand fundamental features of physiological, behavioral and psychological disorders. Zebra fish (Danio rerio) is now considered globally as a new and useful model in biomedical research. The objective of the present study was to assess behavioral changes in adult zebra fish after acute exposure to different pharmacological and herbal compound. Adult zebra fish of 4-5-months-old were exposed to different concentrations of unknown test drug with the standard for 5 min. The test was conducted separately for each drug concentration as well as control. Behavioral activity parameters viz. novel tank test and light and dark transition test were recorded for each animal during the exposure period. Results: Zebra fish exposed to Test drug (Calycophyllum Spruceanum) showed change in behavioral activity with significant to the standard drug caffeine. In view of the above findings, these results suggested that exposure of adult zebra fish with drug produce the expected changes in the behavior like stress and anxiety; therefore, adult zebra fish can be used an alternative approach for the assessment of new chemical entities for their effect on behavioral activity.Dark Transition Test.

Longitudinal trajectories of childhood and adolescent attention deficit hyperactivity disorder diagnoses in three cohorts.

Attention deficit/hyperactivity disorder (ADHD) is usually conceptualized as a childhood-onset neurodevelopmental disorder, in which symptoms either decrease steadily into adulthood or remain stable. A recent study challenged this view, reporting that for most with ADHD, diagnostic status fluctuates with age. We ask if such a 'fluctuating' ADHD symptom trajectory subgroup is present in other population-based and clinic-based cohorts, centered on childhood and adolescence. Cohorts were the population-based Adolescent Brain Cognitive Development (ABCD: N=9735), Neurobehavioral Clinical Research (NCR: N=258), and the Nathan Kline Institute-Rockland (NKI-Rockland: N=149). All participants had three or more assessments spanning different age windows. Participants were categorized into developmental diagnostic subgroups: fluctuant ADHD (defined by two or more switches between meeting and not meeting ADHD criteria), remitting ADHD, persisting ADHD, emerging ADHD and never affected. Data were collected between 2011 and 2022. Analyses were performed between May 2022 and April2023. A subgroup with fluctuant child and adolescent ADHD diagnoses was found in all cohorts (29.3% of participants with ADHD in ABCD, 26.6% in NCR and 17% in NKI-Rockland). While the proportion of those with fluctuant ADHD increased with the number of assessments, it never constituted the dominant subgroup. We provide further evidence in three cohorts for the existence of a fluctuant ADHD diagnostic subgroup during childhood and adolescence, albeit in a minority of cases. Such fluctuant child and adolescent ADHD diagnoses may suggest a natural history more akin to relapsing-remitting mood disorders and/or a marked sensitivity to environmental shifts that occur across development. Intramural programs of the NHGRI and NIMH.

Sex-specific associations between plasma interleukin-6 and depression in persons with and without HIV

BackgroundPersons with HIV (PWH) have both more frequent depression and higher levels of plasma inflammatory biomarkers compared to persons without HIV (PWoH). Inflammation and depressive symptoms are linked, including in PWH; however, it is unclear whether these associations differ by HIV serostatus and biological sex. MethodsSix plasma inflammatory biomarkers were assessed using samples from PWH and PWoH who participated in six NIH-funded studies through the UCSD HIV Neurobehavioral Research Program (HNRP) from 2011 to 2019. Factor analysis was performed to identify intercorrelated groups of biomarkers. Factors and their components were then examined for relationships with Beck Depression Inventory-II (BDI-II) and modifying effects of sex or HIV serostatus using multivariable linear regression, adjusting for demographics, substance use diagnoses, and relevant co-morbidities. ResultsParticipants included 150 PWH (age = 48.3 ± 13.1 yr; 88% biologically male) and 138 PWoH (age = 46.3 ± 15.9; 56% male). Two inflammatory factors were identified: Factor 1 loaded on interleukin-6 (IL-6), C-reactive protein (CRP), and D-dimer; Factor 2 loaded on interleukin-8, chemokine C–C ligand 2 (CCL2), and chemokine C-X-C ligand 10 (CXCL10). Sex modified the effect of Factor 1 on BDI-II, with a more positive association for men than women (p = 0.04). No significant association between Factor 2 and BDI-II was found. Of the biomarkers in Factor 1, only IL-6 was significantly associated with BDI-II and was modified by sex (p = 0.003). In sex-stratified analysis, a positive association was found for men (β = 5.42; 95% confidence interval = [1.32, 9.52]) but not women (β = −3.88; 95% C.I. = [-11.02, 3.26]). No HIV-related interactions were detected. InterpretationWe identified a depression-associated inflammatory factor present in both PWH and PWoH, consistent with prior studies of PWH only. The association was driven by a correlation between IL-6 and depression exclusively in men, suggesting that the depression-inflammation link differs by sex. Future studies of depression etiology or treatment, including those on persons with HIV, should consider the impact of biological sex in both design and analysis.

Exenatide as an adjunct to nicotine patch for smoking cessation and prevention of postcessation weight gain among treatment-seeking smokers with pre-diabetes and/or overweight: study protocol for a randomised, placebo-controlled clinical trial

IntroductionObesity and smoking are the two leading causes of preventable death in the USA. Unfortunately, most smokers gain weight after quitting. Postcessation weight gain (PCWG) is frequently cited as one...

Neuropsychological Test Norms for the Assessment of HIV-Associated Neurocognitive Impairment Among South African Adults

Reliable and valid neurocognitive (NC) test batteries that assess multiple domains of cognitive functioning are vital tools in the early detection of HIV-associated NC impairment. The HIV Neurobehavioral Research Center’s International Neurobehavioral Battery (HNRC Battery) is one such diagnostic tool and has shown cultural validity in several international neuroHIV studies. However, no published norms are currently available for the full HNRC Battery in South Africa. To accurately interpret NC test results, appropriate reference norms are required. In light of this challenge, data were collected from 500 healthy, HIV-uninfected participants to develop demographically corrected South African norms. When demographically corrected United States of America (U.S.) norms were applied to the performance scores of our neurologically intact, HIV-negative sample, an impairment rate of 62.2% was observed compared to a 15.0% impairment rate when the newly generated South African norms were applied. These results reiterate the findings of other low- and middle-income countries, highlighting the need for localized, country-specific norms when interpreting NC performance.

Screening for HIV-associated neurocognitive impairment: Development and validation of an abbreviated neuropsychological test battery for use in South African clinical settings.

The HIV Neurobehavioral Research Center International Neurobehavioral Battery is a culturally valid battery sensitive to the neurocognitive (NC) effects of HIV-infection. However, its lengthy administration time makes the battery impractical in resource-limited settings, like South Africa, which are often faced with an overwhelming disease burden, a lack of neurological and neuropsychological (NP) expertise, and staff shortages. The present study therefore sought to develop an abbreviated version of the HNRC Battery and validate this battery in a sample of people with HIV (PWH) in South Africa. The present study therefore sought to develop an abbreviated version of the HNRC battery and validate this battery in a sample of people with HIV (PWH) in South Africa. Six measures were selected based on the NC test performances of 103 HIV-positive and 135 HIV-negative South African adults. For the validation, a subgroup of 103 PWH completed the full version of the battery, while the other subgroup of 52 PWH completed only the abbreviated version. Deficit scores of each participant were calculated. These scores were used as the gold standard against which the abbreviated battery was compared. There was a reduction of 81% in administration time when compared to the full version of the battery. The abbreviated battery demonstrated good sensitivity (75.0%) and excellent specificity (94.9%) when compared with the full version. The abbreviated battery showed good diagnostic accuracy in identifying NC impairment in an HIV-positive South African sample with a significant reduction in administration time, making it a more practical option in busy South African clinic settings. The results of this study may facilitate the growth of neuroAIDS research and aid initial identification of HIV-related NC impairment in resource-constrained settings. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Getting a handle on rat familiarization: The impact of handling protocols on classic tests of stress in Rattus norvegicus.

Experimenter familiarization with laboratory rodents through handling prior to experimentation is an important practice in neurobehavioral research and is implicated in stress, study variability, and replicability. Unfortunately, different handling protocols have not been thoroughly examined. Determining optimal experimenter familiarization protocols is expected to reduce animal stress and thus improve welfare and data consistency. The impact of different handling protocols was determined through behavioral assessments (i.e. elevated plus maze, light/dark box, open field) as well as via analysis of fecal boli counts, ultrasonic vocalizations, and blood corticosterone. Male and female Sprague Dawley rats were distributed among three groups: never handled, picked-up, and handled for 5 min once daily over five days. Handled and picked-up rats spent more time in open arms and less time in closed arms of the elevated plus maze and more time in the center and less time at the perimeter of the open field compared to rats that were never handled, indicating that handled and picked-up rats were less anxious than those that were never handled. Male rats consistently defecated more frequently throughout the handling process and throughout behavioral testing, whereas females showed greater concentrations of blood corticosterone. Female rats were found to emit more 50-kHz calls and fewer 22-kHz calls compared to males. The results observed suggest that picking animals up may suffice as a handling method compared to time-intensive handling procedures, and that there are significant sex differences in response to handling.

A novel open-source raspberry Pi-based behavioral testing in zebrafish.

The zebrafish (Danio rerio) is widely used as a promising high-throughput model organism in neurobehavioral research. The mobility of zebrafish can be dissected into multiple behavior endpoints to assess its neurobehavioral performance. However, such facilities on the market are expensive and clumsy to be used in laboratories. Here, we designed a low-cost, automatic zebrafish behavior assay apparatus, barely without unintentional human operational errors. The data acquisition part, composed of Raspberry Pi and HQ Camera, automatically performs video recording and data storage. Then, the data processing process is also on the Raspberry Pi. Water droplets and inner wall reflection of multi-well cell culture plates (used for placing zebrafish) will affect the accuracy of object recognition. And during the rapid movement of zebrafish, the probability of zebrafish tracking loss increased significantly. Thus, ROI region and related thresholds were set, and the Kalman filter algorithm was performed to estimate the best position of zebrafish in each frame. In addition, all functions of this device are realized by the custom-written behavior analysis algorithm, which makes the optimization of the setup more efficient. Furthermore, this setup was also used to analyze the behavioral changes of zebrafish under different concentrations of alcohol exposure to verify the reliability and accuracy. The alcohol exposure induced an inverted U-shape dose-dependent behavior change in zebrafish, which was consistent with previous studies, showcasing that the data obtained from the setup proposed in this study are accurate and reliable. Finally, the setup was comprehensively assessed by evaluating the accuracy of zebrafish detection (precision, recall, F-score), and predicting alcohol concentration by XGBoost. In conclusion, this study provides a simple, and low-cost package for the determination of multiple behavioral parameters of zebrafish with high accuracy, which could be easily adapted for various other fields.