A systematic approach to the analysis of illicit drugs for DNA with an overview of the problems encountered.

Due to the restricted nature of illicit drugs, it is difficult to conduct research surrounding the analysis of this drug material for any potential DNA in sufficient quantities acceptable for high numbers of replicates. Therefore, the current research available in peer reviewed journals thus far regarding analysing illicit drugs for DNA has been performed under varying experimental conditions, often using surrogate chemicals in place of illicit drugs. The data presented within this study originated from the analysis of genuine illicit drugs prepared both in controlled environments and those seized at the Australian border (and therefore from an uncontrolled environment) to determine if DNA can be obtained from this type of material. This study has been separated into three main parts (total n=114 samples): firstly, methamphetamine synthesised within a controlled environment was spiked with both saliva and trace DNA to determine the yield following DNA extraction; secondly, methamphetamine also synthesised in a controlled environment but on a larger scale was tested for the amount of DNA added incidentally throughout the synthesis, including the additional steps of recrystallising, homogenising and "cutting" the drug material to simulate preparation for distribution; and thirdly, the detection of human DNA within samples of cocaine and heroin seized at the Australian border. The DNA Fast Flow Microcon Device was utilised to concentrate all replicates from the same source into one combined extract to improve the DNA profiles for the samples where no DNA spiking occurred. Full STR profiles were successfully obtained from drug samples spiked with both saliva and trace DNA. Methamphetamine was present in the final DNA extracts and caused incompatibilities with the quantification of DNA using Qubit. The yields of DNA from drugs not spiked with DNA sources were much lower, resulting in 36 % of samples yielding alleles where all others did not. These results were not unexpected given these were realistic drug samples where the history of the drug material was unknown. This is the first study to obtain DNA profiles from genuine illicit drug material in both controlled and uncontrolled environments and indicates that the analysis of illicit drugs for DNA is an avenue worth pursuing to provide information which can in turn assist with disrupting the supply of these drugs. Given that DNA profiling is carried out worldwide using essentially the same systems as described within this study, the potential for impact is on a national and international scale.

KCL TEST: an open-source inspired asymptomatic SARS-CoV-2 surveillance programme in an academic institution.

Rapid and accessible testing was paramount in the management of the COVID-19 pandemic. Our university established KCL TEST: a SARS-CoV-2 asymptomatic testing programme that enabled sensitive and accessible PCR testing of SARS-CoV-2 RNA in saliva. Here, we describe our learnings and provide our blueprint for launching diagnostic laboratories, particularly in low-resource settings. Between December 2020 and July 2022, we performed 158277 PCRs for our staff, students, and their household contacts, free of charge. Our average turnaround time was 16 h and 37 min from user registration to result delivery. KCL TEST combined open-source automation and in-house non-commercial reagents, which allows for rapid implementation and repurposing. Importantly, our data parallel those of the UK Office for National Statistics, though we detected a lower positive rate and virtually no delta wave. Our observations strongly support regular asymptomatic community testing as an important measure for decreasing outbreaks and providing safe working spaces. Universities can therefore provide agile, resilient, and accurate testing that reflects the infection rate and trend of the general population. Our findings call for the early integration of academic institutions in pandemic preparedness, with capabilities to rapidly deploy highly skilled staff, as well as develop, test, and accommodate efficient low-cost pipelines.

Comparison of three DNA extraction methods tested on illicit drug-related powders.

The detection of human DNA on and within illicit drug preparations is novel and a focus of current research. Previous studies have indicated that certain drug-related powders present in illicit drug preparations can interfere with downstream DNA analysis when directly added to the PCR. Therefore, it is important to determine if these drug-related powders are effectively removed during the DNA extraction or whether traces of powder remain to interfere with DNA processing. Three extraction methods were selected to assess their efficiency at removing drug-related powders for downstream processes using DNA from both saliva and touch depositions. This is the first study to compare efficiencies of DNA extraction methods from drug-related powders. The extraction methods compared were the DNA IQ™ System, the QIAamp® DNA Investigator Kit and the combination of a simple lysis step followed by use of the Microcon® DNA Fast Flow device. Saliva was added to dimethylsulfone (DMS), nitrostyrene and PROSOLV® tablet mixture to determine the effect of powder type (based on solubility). Saliva was also added to 0, 50, 200 and 400 mg of DMS to determine the effect of an increase in DMS quantity. Trace DNA was deposited onto DMS using a worn glove approach. These samples were re-tested six months post-DNA deposition and profiled for further comparisons. Ten replicates were conducted for each condition with five replicates of saliva positive controls per method (n = 255 samples). A subset of samples was chemically analysed to determine if DMS was present in the final DNA eluant. The readily soluble DMS did not interfere with any of the extraction methods at lower amounts, however increasing the DMS to 400 mg reduced the relative DNA yields using the Microcon® and Investigator methods. The tablet mixture reduced the relative DNA yield of all three methods, however the nitrostyrene (which was relatively insoluble) only reduced the relative DNA yield of the DNA IQ™. The Investigator method performed the best with the trace samples, followed by the Microcon® method and then the DNA IQ™. DMS was detected in all extracts chemically analysed from the DNA IQ™ and Microcon®, whereas only one sample tested from the Investigator kit contained DMS in the extract and was in a relatively low amount compared to the other samples. Not one kit outperformed the others in all comparisons, however the Investigator kit was the most efficient overall at optimising the DNA yield whilst also removing the powders more effectively.

Pausing and reorienting behaviors enhance the performance of a spatial working memory task.

During spatial working memory tasks, animals need to retain information about a previous trial in order to successfully select their next trajectory. Specifically, the delayed non-match to position task requires rats to follow a cued sample trajectory, then select the opposite route after a delay period. When faced with this choice, rats will occasionally exhibit complex behaviors, such as pausing and sweeping their head back and forth. These behaviors, called vicarious trial and error (VTE), are thought to be a behavioral manifestation of deliberation. However, we identified similarly complex behaviors during sample-phase traversals, despite the fact that these laps do not require a decision. First, we identified that these behaviors occurred more often after incorrect trials than before them, indicating that rats are retaining information between trials. Next, we determined that these pause-and-reorient (PAR) behaviors increased the likelihood of the next choice being selected correctly, suggesting that these behaviors assist the rat in successful task performance. Finally, we identified similarities between PARs and choice-phase VTEs, suggesting that VTEs may not only be reflective of deliberation, but may also contribute to a strategy for successful performance of spatial working memory tasks.

Recovery of integrated and surface trace DNA from illicit drug tablets.

In many parts of the world, tablets are a commonly encountered form of illicit drug preparation. Whilst previous research has investigated the feasibility of detecting trace DNA on illicit drug capsules, this has not been performed for tablets. Tablets have a unique substrate surface and therefore the amount of DNA transferring to them and persisting on them may be different to capsules; there may also be differences in the collection efficiency and the outcome of downstream DNA processing and analysis steps. The ability to profile the DNA from individuals who handled tablets during their preparation and distribution would add another level of discrimination between various drug seizures or corroborate chemical profiling outcomes which may link various seizures to a common origin. DNA from two different individuals (male and female) was added to the tablets in two stages. Firstly, tablet powder was spiked with DNA from one individual to mimic the situation where DNA traces are incorporated during the drug synthesis or final drying stages. The powder was then pressed into tablets in a clean environment without intentional addition of DNA. Subsequently, a second individual counted out the tablets into bags of ten to mimic the preparation for distribution at a user level. The exterior of the tablet was swabbed and then the entire tablet and the swab were put through separate DNA extractions, yielding two DNA extracts for each tablet. Swabs of the exterior tablet surface yielded single source DNA profiles that identified the tablet handler in 100 % of samples. The tablet extract yielded the donor of the DNA intentionally added within the drug powder in 80% of samples with varying levels of support, however contributions of the exterior handler were detected in 60 % of samples. The identification of individuals potentially involved in the synthesis of the drugs compared to the distribution of the tablets will provide invaluable strategic intelligence related to illicit drug investigations and to law enforcement agencies.

The ventral midline thalamus coordinates prefrontal-hippocampal neural synchrony during vicarious trial and error.

When faced with difficult choices, the possible outcomes are considered through a process known as deliberation. In rats, deliberation is thought to be reflected by pause-and-reorienting behaviors, better known as vicarious trial and errors (VTEs). While VTEs are thought to require medial prefrontal cortex (mPFC) and dorsal hippocampal (dHPC) interactions, no empirical evidence has yet demonstrated such a dual requirement. The nucleus reuniens (Re) of the ventral midline thalamus is anatomically connected with both the mPFC and dHPC, is required for HPC-dependent spatial memory tasks, and is critical for mPFC-dHPC neural synchronization. Currently, it is unclear if, or how, the Re is involved in deliberation. Therefore, by examining the role of the Re on VTE behaviors, we can better understand the anatomical and physiological mechanisms supporting deliberation. Here, we examined the impact of Re suppression on VTE behaviors and mPFC-dHPC theta synchrony during asymptotic performance of a HPC-dependent delayed alternation (DA) task. Pharmacological suppression of the Re increased VTE behaviors that occurred with repetitive choice errors. These errors were best characterized as perseverative behaviors, in which some rats repeatedly selected a goal arm that previously yielded no reward. We then examined the impact of Re suppression on mPFC-dHPC theta synchrony during VTEs. We found that during VTEs, Re inactivation was associated with a reduction in mPFC-dHPC theta coherence and mPFC-to-dHPC theta directionality. Our findings suggest that the Re contributes to deliberation by coordinating mPFC-dHPC neural interactions.

DNA on drugs (part 2): An extended study into the transfer and persistence of DNA onto illicit drug capsules using realistic scenarios.

Capsules are now the main form of ecstasy rather than tablets in Australia and therefore their examination is of interest to forensic drug chemists in Australia and possibly elsewhere. Recently, we used controlled experimental conditions to show that capsules may be a source of DNA that can be used to identify those involved in production and distribution of illicit drugs. The question remains: in realistic scenarios where there are more unknowns, can we still detect DNA, and determine whose it is, on the exterior of capsules? The concept of comprehensive forensic intelligence and investigations - utilizing both biological and chemical signatures - relating to illicit drug preparations (i.e., the capsules and their contents) may be of great use to law enforcement. Experiments were conducted with both semi-realistic and realistic scenarios where two volunteers were asked to firstly use an encapsulator and mimic the loading of capsules, then Volunteer 1 would count out the capsules that Volunteer 2 prepared, and vice versa. This was to simulate the scenario where one person was involved in the assembly of the capsules which were then separated into smaller bags of 10 capsules by a second person for distribution. Gelatine and vegetable capsules were tested, with 10 replicates used per capsule type, scenario, and volunteer (total n = 80 capsules). Volunteer 2 was included as a contributor to the DNA profiles generated from 100% of samples handled by them within the semi-realistic scenario, whereas the other volunteer could be included as a contributor in 65% of samples. For the realistic scenario, profiles could be generated with the inclusion of both volunteers as profile contributors in 15% of samples and from just one of the volunteers in a further 50% of samples (therefore in total, either both or one of the volunteers were detected in 65% of realistic samples). Surprisingly, it was not necessarily the case that the last person to handle the capsule was the major or only contributor. The potential variability in the DNA quantities that could be deposited onto the capsules of genuine illicit drugs is high and would vary on a case-by-case basis. Nevertheless, this study has indicated that in realistic scenarios where two people are involved in the later stages of illicit drug capsule preparation, that either one or both individuals may be identified, potentially opening new investigative leads for law enforcement agencies as well as offering new information for intelligence-led policing.

A survey of the effects of common illicit drugs on forensic DNA analysis.

Profiling of DNA associated with illicit drug packages and paraphernalia is a common investigative tool. In addition, research is being conducted regarding the analysis of trace DNA present within illicit drugs and on capsules. The application of trace DNA analysis to illicit drugs has the potential to identify individuals involved in their manufacture and distribution. However, the inhibitory effects of illicit drugs and related compounds on downstream DNA analysis has not yet been investigated. If drug-induced polymerase chain reaction (PCR) inhibition occurs, the quality or informativeness of the resultant DNA profile may be impacted. In this study, the effects of a range of drugs, diluents, adulterants, and synthetic precursors on both quantitative PCR (qPCR) data and short tandem repeat (STR) DNA profiling results were examined. Twenty-two compounds representative of drug compounds and adulterants which may be encountered in drug seizures were spiked with 1 ng/µL and 0.05 ng/µL of control DNA and underwent DNA quantification using Quantifiler™ Trio. A subset of 13 compounds, including the majority that indicated potential inhibition in Quantifiler™ Trio, underwent STR profiling with VeriFiler™ Plus to determine if inhibition also occurred at this stage. The effect of diluting the DNA extract on the extent of inhibition of STR profiling was also investigated. Internal PCR controls within the qPCR were not a reliable indicator of inhibition, although suppression of the short and long autosomal fragments was observed in the presence of many compounds, and four compounds gave inconclusive results. STR internal quality controls indicated inhibition in 5 of the 13 compounds, however, profiles were affected by the presence of 11 of the 13 compounds in various ways such as a decreased average relative fluorescence units (RFU), drop out of certain alleles (some based on allele size range of locus) leading to a decreased likelihood ratio (LR), an increase in the proportion of stutter peaks and the presence of split or shoulder peaks. All profiles improved following a dilution of the compound in the PCR and allowing the generation of LR values in excess of 1 × 1025, indicating inhibition occurred rather than DNA degradation. The data obtained show that removal of some of these compounds is required through an effective DNA extraction process for successful downstream trace DNA profiling. Upon successful PCR, the resultant DNA profiles provide the opportunity for opening new investigative avenues for law enforcement agencies.

Evaluating the Impact of Wellness Days on Enrollment of Underserved Women in the Connecticut Early Detection and Prevention Program.

Women in underserved communities are disproportionately affected by chronic diseases such as cardiovascular disease and cancer. The Connecticut Early Detection and Prevention Program (CEDPP) has taken a streamlined approach to improve access to comprehensive preventive health services for minority women and those with incomes below the federal poverty threshold. The CEDPP has implemented Wellness Days to improve outreach in the community and offer opportunities for health assessments, screenings, and education around chronic disease prevention and management. CEDPP contractors coordinated 47 Wellness Days in 2019, reaching 2,509 women and successfully enrolling 107 (4.3%) in the CEDPP. While the majority of Wellness Day events offered health education to participants, only 10.6% offered mammograms and 6.4% offered Papanicolaou (Pap) tests onsite. Through ongoing evaluation efforts, the CEDPP and its contractors have identified opportunities to enhance the success of Wellness Days to connect women with essential preventive services. By expanding its reach, the CEDPP will have a more widespread impact on women's health across Connecticut.

Optogenetic suppression of the medial septum impairs working memory maintenance.

Spatial working memory (SWM) is the ability to encode, maintain, and retrieve spatial information over a temporal gap, and relies on a network of structures including the medial septum (MS), which provides critical input to the hippocampus. Although the role of the MS in SWM is well-established, up until recently, we have been unable to use temporally precise circuit manipulation techniques to examine the specific role of the MS in SWM, particularly to distinguish between encoding, maintenance, and retrieval. Here, we test the hypothesis that the MS supports the maintenance of spatial information over a temporal gap using precisely timed optogenetic suppression delivered during specific portions of three different tasks, two of which rely on SWM and one that does not. In experiment 1, we found that MS optogenetic suppression impaired choice accuracy of a SWM dependent conditional discrimination task. Moreover, this deficit was only observed when MS suppression was delivered during the cue-sampling, but not the cue-retrieval, portion of the trial. There was also no deficit when MS neurons were optogenetically suppressed as rats performed a SWM-independent variant of the task. In experiment 2, we tested whether MS suppression affected choice accuracy on a delayed nonmatch to position (DNMP) task when suppression was limited to the sample, delay, and choice phases of the task. We found that MS suppression delivery during the delay phase of the DNMP task, but not during the sample or choice phases, impaired choice accuracy. Our results collectively suggest that the MS plays an important role in SWM by maintaining task-relevant information over a temporal delay.

DNA on drugs! A preliminary investigation of DNA deposition during the handling of illicit drug capsules.

DNA profiling from capsules and tablets offers a complementary tool to that of chemical profiling when investigating the manufacture and trade in illicit drugs. By sampling the outside of capsules, individuals who may have handled them during production, assembly or distribution may have deposited their DNA and can be identified if matched to a nominated profile or one on a relevant DNA database. The profiles can also be compared to those found on other capsules to potentially link various drug seizures. This study sampled the exterior of capsules after they had been handled in a controlled scenario to determine if informative DNA profiles could be generated from this brief contact. Two individuals of intermediate shedder status washed their hands and waited for 30 min before handling ten gelatine, vegetable, and enteric vegetable capsules each (n = 60). Contact was made for 15 s. Each capsule was swabbed and DNA isolated. The amount of recovered human DNA was quantified and profiled using the Verifiler Plus DNA profiling kit. Profiles were generated from 82% (49/60) of capsules tested with LR values above 1 × 103 for the inclusion of the volunteer as a contributor. Inhibition of the PCR was detected in 24 of the 60 samples, however 16 of these still produced informative profiles when sufficient template DNA was available and only mild inhibition was detected, or by overcoming inhibition by dilution of the DNA extract. This pilot study demonstrates the potential for forensic science laboratories to recover human DNA from the exterior surface of capsules which are commonly used to encase illicit drugs such as MDMA, thus enabling both biological and chemical profiling methods to contribute to the investigation of clandestine drug production and distribution.

The nucleus reuniens orchestrates prefrontal-hippocampal synchrony during spatial working memory.

Spatial working memory, the ability to temporarily maintain an internal representation of spatial information for use in guiding upcoming decisions, has been shown to be dependent upon a network of brain structures that includes the hippocampus, a region known to be critical for spatial navigation and episodic memory, and the prefrontal cortex (PFC), a region known to be critical for executive function and goal directed behavior. Oscillatory synchronization between the hippocampus and the prefrontal cortex (PFC) is known to increase in situations of high working memory demand. Most of our knowledge about the anatomical connectivity between the PFC and hippocampus comes from the rodent literature. Thus, most of the findings that will be discussed here model human working memory using spatial working memory-dependent maze navigation tasks in rodents. It has been demonstrated that the ventral midline thalamic nucleus reuniens (Re) is reciprocally connected to both the infralimbic and prelimbic subregions of the PFC, collectively referred to as the medial PFC (mPFC), and the hippocampus. Given that the Re serves as a major anatomical route between the mPFC and hippocampus, it is perhaps not surprising that Re has been shown to be critical for spatial working memory. This review will describe the latest findings and ideas on how the Re contributes to prefrontal-hippocampal synchronization and spatial working memory in rodents. The review will conclude with possible future directions that will advance the understanding of the mechanisms that enable the Re to orchestrate long range synchrony in the prefrontal-hippocampal network.

Representations of On-Going Behavior and Future Actions During a Spatial Working Memory Task by a High Firing-Rate Population of Medial Prefrontal Cortex Neurons.

Spatial working memory (SWM) requires the encoding, maintenance, and retrieval of spatially relevant information to guide decision-making. The medial prefrontal cortex (mPFC) has long been implicated in the ability of rodents to perform SWM tasks. While past studies have demonstrated that mPFC ensembles reflect past and future experiences, most findings are derived from tasks that have an experimental overlap between the encoding and retrieval of trajectory specific information. In this study, we recorded single units from the mPFC of rats as they performed a T-maze delayed non-match to position (DNMP) task. This task consists of an encoding dominant sample phase, a memory maintenance delay period, and retrieval dominant choice phase. Using a linear classifier, we investigated whether distinct ensembles collectively reflect various trajectory-dependent experiences. We find that a population of high-firing rate mPFC neurons both predict a future choice and reflect changes in trajectory-dependent behaviors. We then developed a modeling procedure that estimated the number of high and low-firing rate units required to dissociate between various experiences. We find that low firing rate ensembles weakly reflect the direction that rats were forced to turn on the sample phase. This was in contrast to the highly active population that could effectively predict both future decision-making on early stem traversals and trajectory-divergences at T-junction. Finally, we compared the ensemble size necessary to code a forced trajectory to the size required to predict a decision. We provide evidence to suggest that a larger number of highly active neurons are employed during decision-making processes when compared to rewarded forced behaviors. Together, our study provides important insight into how specific ensembles of mPFC units support upcoming choices and ongoing behavior during SWM.

Addressing disparities in the health of persons with HIV attributable to unstable housing in the United States: The role of the Ryan White HIV/AIDS Program.

Amy Griffin and co-authors discuss unstable housing and the response to HIV/AIDS in the United States.

The nucleus reuniens of the thalamus sits at the nexus of a hippocampus and medial prefrontal cortex circuit enabling memory and behavior.

The nucleus reuniens of the thalamus (RE) is a key component of an extensive network of hippocampal and cortical structures and is a fundamental substrate for cognition. A common misconception is that RE is a simple relay structure. Instead, a better conceptualization is that RE is a critical component of a canonical higher-order cortico-thalamo-cortical circuit that supports communication between the medial prefrontal cortex (mPFC) and the hippocampus (HC). RE dysfunction is implicated in several clinical disorders including, but not limited to Alzheimer's disease, schizophrenia, and epilepsy. Here, we review key anatomical and physiological features of the RE based primarily on studies in rodents. We present a conceptual model of RE circuitry within the mPFC-RE-HC system and speculate on the computations RE enables. We review the rapidly growing literature demonstrating that RE is critical to, and its neurons represent, aspects of behavioral tasks that place demands on memory focusing on its role in navigation, spatial working memory, the temporal organization of memory, and executive functions.

Optogenetic suppression of the nucleus reuniens selectively impairs encoding during spatial working memory.

The nucleus reuniens (Re) of the ventral midline thalamus is known to be a critical anatomical link between the hippocampus (HPC) and the medial prefrontal cortex (mPFC). Consistent with this anatomical connectivity, the Re has been shown to be crucial for HPC-mPFC oscillatory synchrony. Moreover, Re inhibition consistently results in spatial working memory (SWM) deficits. Together, these results suggest that SWM requires HPC-mPFC synchrony via the Re. In spite of these findings, an understanding of how the Re contributes to the encoding, maintenance, and retrieval of spatial information during a SWM task is lacking. To address this issue, we trained rats to perform a SWM-dependent delayed-non-match-to-position (DNMP) task in a T-maze. Using optogenetic inhibition of Re activity, we demonstrated that Re suppression during the sample phase, but not the delay or choice phase, significantly decreased choice accuracy. We conclude that the Re contributes to the encoding of spatial information during working memory.

Racial/Ethnic Disparities in Cervical Cancer Screening Services Among Contractors of the Connecticut Breast and Cervical Cancer Early Detection Program.

Purpose: Racial/ethnic minority women are at increased risk for cervical cancer. The objective of this study is to use performance management data from the Connecticut Breast and Cervical Cancer Early Detection Program (CBCCEDP) to determine whether race/ethnicity disparities exist in human papillomavirus (HPV) co-testing uptake across CBCCEDP contractors. Methods: Secondary analysis of Connecticut's Minimum Data Elements data for 2013-2015 among 10 contractors participating in the CBCCEDP. Participants included women aged 30-64 years and eligible to receive routine cervical cancer screening services through the CBCCEDP (n=5,262). HPV co-testing uptake was compared across contractors and race/ethnicity groups within each contractor using chi-square and Fisher's exact tests as appropriate. Results: Overall, 62.9% of women received HPV co-testing services. Significant differences in co-testing rates were detected between racial/ethnic groups when data were examined across all contractors (p<0.001). Black women were least likely to receive co-testing (49.1%), while Hispanic women were most likely to receive co-testing (68.2%). When data were examined at the individual contractor level, significant differences between racial/ethnic groups were observed in 50% of the contractors. Conclusions: This study identified racial/ethnic disparities in uptake of HPV co-testing both overall and within individual contractors involved in the CBCCEDP. These findings will be used to guide program improvement with the goal of increasing quality and consistency of care for all women seeking screening services.

A Tactile-visual Conditional Discrimination Task for Testing Spatial Working Memory in Rats.

This protocol describes a novel dual task comparison across two variants of a tactile-visual conditional discrimination (CD) T-maze task, one is dependent upon spatial working memory (SWM; CDWM) and the other one (CDSTANDARD) is not. The task variants are equivalent in their sensory and motor requirements and overt behavior of the rat. Therefore, differences between the two task variants in the dependent variables such as choice accuracy, neural firing patterns, and the effects of pharmacological or optogenetic inactivation in brain regions of interest can be attributed to SWM, ruling out confounding sensorimotor variables, such as tactile, visual and self-motion cues. The CDWM task protocol is published in Hallock et al., 2013b and Urban et al., 2014.

Ventral Midline Thalamus Is Critical for Hippocampal-Prefrontal Synchrony and Spatial Working Memory.

UNLABELLED: Maintaining behaviorally relevant information in spatial working memory (SWM) requires functional synchrony between the dorsal hippocampus and medial prefrontal cortex (mPFC). However, the mechanism that regulates synchrony between these structures remains unknown. Here, we used a unique dual-task approach to compare hippocampal-prefrontal synchrony while rats switched between an SWM-dependent task and an SWM-independent task within a single behavioral session. We show that task-specific representations in mPFC neuronal populations are accompanied by SWM-specific oscillatory synchrony and directionality between the dorsal hippocampus and mPFC. We then demonstrate that transient inactivation of the reuniens and rhomboid (Re/Rh) nuclei of the ventral midline thalamus abolished only the SWM-specific activity patterns that were seen during dual-task sessions within the hippocampal-prefrontal circuit. These findings demonstrate that Re/Rh facilitate bidirectional communication between the dorsal hippocampus and mPFC during SWM, providing evidence for a causal role of Re/Rh in regulating hippocampal-prefrontal synchrony and SWM-directed behavior. SIGNIFICANCE STATEMENT: Hippocampal-prefrontal synchrony has long been thought to be critical for spatial working memory (SWM) and the ventral midline thalamic reuniens and rhomboid nuclei (Re/Rh) have long been considered a potential site for synchronizing the hippocampus and medial prefrontal cortex. However, the hypothesis that Re/Rh are critical for hippocampal-prefrontal synchrony and SWM has not been tested. We first used a dual-task approach to identify SWM-specific patterns of hippocampal-prefrontal synchrony. We then demonstrated that Re/Rh inactivation concurrently disrupted SWM-specific behavior and the SWM-specific patterns of hippocampal-prefrontal synchrony seen during dual-task performance. These results provide the first direct evidence that Re/Rh contribute to SWM by modulating hippocampal-prefrontal synchrony.

Comparing Color and Leader Line Highlighting Strategies in Coordinated View Geovisualizations.

In most coordinated view geovisualization tools, a transient visual effect is used to highlight observations across views when brushed with a mouse or other input device. Most current geovisualization and information visualization systems use colored outlines or fills to highlight observations, but there remain a wide range of alternative visual strategies that can also be implemented and compared to color highlighting to evaluate user performance. This paper describes the results of an experiment designed to compare user performance with two highlighting methods; color and leader lines. Our study methodology uses eye-tracking to capture participant eye fixations while they answer questions that require attention to highlighted observations in multiple views. Our results show that participants extract information as efficiently from coordinated view displays that use leader line highlighting to link information as they do from those that use a specific color to highlight items. We also found no significant differences when changing the color of the highlighting effect from red to black. We conclude that leader lines show significant potential for use as an alternative highlighting method in coordinated multiple view visualizations, allowing color to be reserved for representing thematic attributes of data.