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Social norms and legality surrounding the use of medical and recreational cannabis are changing rapidly. The prevalence of cannabis use in adolescence is increasing. The aim of this study was to assess any sex-based neurobiological effects of chronically inhaled, vaporised cannabis on adolescent female and male mice.Female and male mice were exposed daily to vaporised cannabis (10.3% Δ-9-tetrahydrocannabinol [THC] and 0.05% cannabidiol [CBD]) or placebo from postnatal day 23 to day 51. Following cessation of treatment, mice were examined for changes in brain structure and function using noninvasive multimodal magnetic resonance imaging (MRI). Data from voxel-based morphometry, diffusion weighted imaging and rest state functional connectivity were registered to and analysed with a 3D mouse atlas with 139 brain areas. Following imaging, mice were tested for their preference for a novel object.The effects were sexually dimorphic with females showing a unique distribution and inverse correlation between measures of fractional anisotropy and apparent diffusion coefficient localised to the forebrain and hindbrain. In contrast males displayed significant increased functional coupling with the thalamus, hypothalamus and brainstem reticular activating system as compared with controls. Cannabis males also presented with altered hippocampal coupling and deficits in cognitive function.Chronic exposure to inhaled vaporised cannabis had significant effects on brain structure and function in early adulthood corroborating much of the literature. Females presented with changes in grey matter microarchitecture, while males showed altered functional connectivity in hippocampal circuitry and deficits in object recognition.

Objective: The rapid increase of cannabis consumption reinforces the need to elucidate the health hazards of this practice. The presence of fine particulate matter in cannabis smoke and vapor poses...

Drivers claim to use cell phones for benefits such as getting work done and to increase productivity ( Sanbonmatsu, Strayer, Behrends, Medeiros-Ward, & Watson, in press ). However, individuals who use cell phones while driving may be more likely to rely on reconstructive processes in memory due to divided attention, making them more susceptible to errors, yielding an ironic effect of multitasking that, in fact, may diminish productivity rather than increase it. To test this possibility, the present study included three within-subject conditions: single-task driving in a high-fidelity simulator, single-task memory including encoding and retrieval using the Deese–Roediger–McDermott false memory paradigm ( Deese, 1959; Roediger & McDermott, 1995 ), and a dual-task combination of both the driving and memory tasks. The effects of divided attention in working memory were bidirectional, impairing both driving and episodic memory performance, likely due to competition for limited resources needed to successfully maintain task goals related to driving or memory alone. More specifically, under dual-task conditions, participants became increasingly reliant on reconstructive, error-prone processes in memory, with high levels of false recall. Taken together, these results indicate there is a productivity illusion associated with distracted driving in that individuals wrongly believe that combining cell phones with driving will make them more productive. Results are discussed in relation to theories of working memory and the domain-free ability to maintain task goals and to avoid distractions, whether this interference occurs in more traditional lab tasks or in more applied settings, highlighting the value of such converging evidence in sharpening theories of attention.

The current research sought to understand the sources of cognitive distraction stemming from voice-based in-vehicle infotainment systems (IVIS) to send and receive textual information. Three experiments each evaluated 1) a baseline single-task condition, 2) listening to e-mail/text messages read by a “natural” pre-recorded human voice, 3) listening to e-mail/text messages read by a “synthetic” computerized text-to-speech system, 4) listening and composing replies to e-mail/text messages read by a “natural” voice, and 5) listening and composing replies to e-mail/text messages read by a “synthetic” voice. Each task allowed the driver to keep their eyes on the road and their hands on the steering wheel, thus any impairment to driving was caused by the diversion of non-visual attention away from the task of operating the motor vehicle.

Objective: The cognitive workload of three Smartphone Digital Assistants (SDA) was manipulated in an on-off manner while participants drove an instrumented vehicle in order to measure the costs associated with intermittent dual tasking. Background: Previous research has shown costs in productivity when switching between two discrete tasks; however, similar costs have not yet been examined using intermittent, continuous dual tasks. Methods: Participating drivers completed 5 conditions: baseline driving, 3 SDA conditions, and a cognitively demanding math-memory operation span (OSPAN) task, each while responding to Detection Response Task (DRT) stimuli. Within the SDA conditions, on- and off-task DRT performance was compared to baseline driving and to the OSPAN task performance. Results: The on-task periods of the SDA conditions resulted in similar RTs as the OSPAN condition, while the off-task periods did not immediately return to baseline driving performance. Post hoc analyses of the on-off transitions within the SDA conditions revealed a gradual return to baseline driving performance over the course of 18 seconds. Conclusions: The delays in returning to baseline driving performance after completing a secondary task raise concerns about the usage of in-vehicle devices while driving as the effects of the delays last beyond the cessation of the SDA interaction.

Suicide, self-injury, and predisposing vulnerabilities aggregate in families. Those at greatest risk often show deficits in two biologically-mediated domains: behavioral control and emotion regulation. This pilot study explored electroencephalographic and cardiovascular indices of self-regulation among typical and suicidal adolescents (n = 30/group) and biological family members (mothers, fathers, and siblings). We measured event-related potentials during a flanker task designed to evoke impulsive responding and respiratory sinus arrhythmia (RSA) at rest and during social rejection. Multilevel models indicate control families’ RSA was unaffected by social rejection (slope = 0.136, p = .097, d = 0.09), whereas clinical families demonstrated RSA withdrawal (slope = −0.191, p = .036, d = −0.13). Clinical families displayed weaker positive voltage (Pe) deflections following behavioral errors relative to controls (coefficient = −2.723, p = .017, d = −0.45), indicating risk for compromised cognitive control. Thus, families with suicidal adolescents showed autonomic and central nervous system differences in biological markers associated with suicide risk.

The goal of this research was to examine the impact of voice-based interactions using 3 different intelligent personal assistants (Apple's Siri, Google's Google Now for Android phones, and Microsoft's Cortana) on the cognitive workload of the driver. In 2 experiments using an instrumented vehicle on suburban roadways, we measured the cognitive workload of drivers when they used the voice-based features of each smartphone to place a call, select music, or send text messages. Cognitive workload was derived from primary task performance through video analysis, secondary-task performance using the Detection Response Task (DRT), and subjective mental workload. We found that workload was significantly higher than that measured in the single-task drive. There were also systematic differences between the smartphones: The Google system placed lower cognitive demands on the driver than the Apple and Microsoft systems, which did not differ. Video analysis revealed that the difference in mental workload between the smartphones was associated with the number of system errors, the time to complete an action, and the complexity and intuitiveness of the devices. Finally, surprisingly high levels of cognitive workload were observed when drivers were interacting with the devices: "on-task" workload measures did not systematically differ from that associated with a mentally demanding Operation Span (OSPAN) task. The analysis also found residual costs associated using each of the smartphones that took a significant time to dissipate. The data suggest that caution is warranted in the use of smartphone voice-based technology in the vehicle because of the high levels of cognitive workload associated with these interactions. (PsycINFO Database Record

Technologies are needed to study gene expression at the level of individual cells within a population or microbial community. Fluorescent in situ hybridization (FISH) supplies high-resolution spatial information and has been widely applied to study microbial communities at the rRNA level. While mRNA-targeted FISH has been popular for studying gene expression in eukaryotic cells, very little success has been achieved with prokaryotes. At present, detection of specific mRNAs in individual prokaryotic cells requires the use of in situ RT-PCR or tyramide signal amplification (TSA). In this study we used DNA oligonucleotide probes labeled with a single near-infrared dye in FISH assays to detect multi-copy plasmid-based and endogenous mRNA molecules in Escherichia coli and Shewanella oneidensis MR-1. We took advantage of the fact that there is much less background signal produced by biological materials and support matrices in the near-infrared spectrum and thus long camera exposure times could be used. In addition, we demonstrate that a combination of probes targeting both rRNA and mRNA could be successfully employed within the same FISH assay. These results, as well as ongoing R&D improvements in NIR and infrared dyes, indicate that the FISH approach we demonstrated could be applied in certain environmental settings to monitor gene expression in mixed populations.

Herpes virus technology involving manipulation of GAD65 was used to study effects on audiogenic seizures (AGS). Audiogenic seizure behaviors were examined following injections of replication-defective herpes simplex virus (HSV-1) vectors incorporating sense or antisense toward GAD65 along with 10% lac -Z into the central nucleus of inferior colliculus (CNIC) of Long–Evans rats. In seizure-sensitive animals developmentally primed by intense sound exposure, injection of GAD65 in the sense orientation increased wild running latencies and reduced incidence of clonus compared with lac -Z only, unoperated, and vehicle seizure groups. In contrast, infection of CNIC with GAD65 antisense virus resulted in 100% incidence of wild running and clonus behaviors in AGS animals. Unprimed animals not operated continued to show uniform absence of seizure activity. Administration of GAD65 antisense virus into CNIC produced novel wild running and clonus behaviors in some unprimed animals. Staining for β-galactosidase in all vector animals revealed no differences in pattern or numbers of immunoreactive cells at injection sites. Qualitatively, typical small and medium multipolar/stellate and medium fusiform neurons appeared in the CNIC of vector animals. These results demonstrate that HSV-1 vector constructs implanted into the CNIC can predictably influence incidence and severity of AGS and suggest that viral vectors can be useful in studying GABA mechanisms with potential for therapeutic application in epilepsy. This article is part of a Special Issue entitled “Genetic and Reflex Epilepsies, Audiogenic Seizures and Strains: From Experimental Models to the Clinic”.

We examined the role of dual-task interference in working memory using a novel dual two-back task that requires a redundant-target response (i.e., a response that neither the auditory nor the visual stimulus occurred two back versus a response that one or both occurred two back) on every trial. Comparisons with performance on single two-back trials (i.e., with only auditory or only visual stimuli) showed that dual-task demands reduced both speed and accuracy. Our task design enabled a novel application of Townsend and Nozawa's (Journal of Mathematical Psychology 39: 321-359, 1995) workload capacity measure, which revealed that the decrement in dual two-back performance was mediated by the sharing of a limited amount of processing capacity. Relative to most other single and dual n-back tasks, performance measures for our task were more reliable, due to the use of a small stimulus set that induced a high and constant level of proactive interference. For a version of our dual two-back task that minimized response bias, accuracy was also more strongly correlated with complex span than has been found for most other single and dual n-back tasks.

To evade host resistance mechanisms, Salmonella enterica serovar Typhimurium (STM), a facultative intracellular pathogen, must alter its proteome following macrophage infection. To identify new colonization and virulence factors that mediate STM pathogenesis, we have isolated STM cells from RAW 264.7 macrophages at various time points following infection and used a liquid chromatography-mass spectrometry-based proteomic approach to detect the changes in STM protein abundance. Because host resistance to STM infection is strongly modulated by the expression of a functional host-resistant regulator, i.e. natural resistance-associated macrophage protein 1 (Nramp1, also called Slc11a1), we have also examined the effects of Nramp1 activity on the changes of STM protein abundances. A total of 315 STM proteins have been identified from isolated STM cells, which are largely housekeeping proteins whose abundances remain relatively constant during the time course of infection. However, 39 STM proteins are strongly induced after infection, suggesting their involvement in modulating colonization and infection. Of the 39 induced proteins, 6 proteins are specifically modulated by Nramp1 activity, including STM3117, as well as STM3118-3119 whose time-dependent abundance changes were confirmed using Western blot analysis. Deletion of the gene encoding STM3117 resulted in a dramatic reduction in the ability of STM to colonize wild-type RAW 264.7 macrophages, demonstrating a critical involvement of STM3117 in promoting the replication of STM inside macrophages. The predicted function common for STM3117-3119 is biosynthesis and modification of the peptidoglycan layer of the STM cell wall.

A nonimmune library of 10(9) human antibody scFv fragments has been cloned and expressed on the surface of yeast, and nanomolar-affinity scFvs routinely obtained by magnetic bead screening and flow-cytometric sorting. The yeast library can be amplified 10(10)-fold without measurable loss of clonal diversity, allowing its effectively indefinite expansion. The expression, stability, and antigen-binding properties of >50 isolated scFv clones were assessed directly on the yeast cell surface by immunofluorescent labeling and flow cytometry, obviating separate subcloning, expression, and purification steps and thereby expediting the isolation of novel affinity reagents. The ability to use multiplex library screening demonstrates the usefulness of this approach for high-throughput antibody isolation for proteomics applications.

This report examines the utility of using Event-Related Brain Potentials (ERPs) to evaluate cognitive distraction in the context of driving an automobile. Across two studies, ERPs (both P300 latency and P300 amplitude) were found to be effective in quantifying the cognitive workload experienced by drivers when they interact with in-vehicle voice-command systems.

This study assessed the impact of cognitive distraction on drivers’ anticipatory glances. Participants drove an instrumented vehicle and executed a number of secondary tasks associated with increasing levels of mental workload including: listening to the radio or audiobook, talking on a handheld or hands-free cellphone, interacting with a voice-based e-mail/text system, and executing a highly demanding task (Operational Span task; OSPAN). Drivers’ visual scanning behavior was recorded by four different high definition cameras and coded offline frame-by-frame. Visual scanning behavior at road intersections with crosswalks was targeted because distraction is one of the major causes of accidents at these locations (NHTSA, 2010a). Despite the familiarity of the locations, results showed that as the secondary-task became more cognitively demanding drivers reduced the amount of anticipatory glances to potential hazards locations. For example, while interacting with a high fidelity voice-based email/text system, the probability of executing a complete scan of the intersection was reduced by 11% compared to the no-distraction control condition. These results document the effects of cognitive distraction on drivers’ visual scanning for potential hazards and highlight the detrimental role of voice based systems on driving behavior.

The present study investigated individual differences in information processing following errant behavior. Participants were initially classified as high or as low working memory capacity using the Operation Span Task. In a subsequent session, they then performed a high congruency version of the flanker task under both speed and accuracy stress. We recorded ERPs and behavioral measures of accuracy and response time in the flanker task with a primary focus on processing following an error. The error-related negativity was larger for the high working memory capacity group than for the low working memory capacity group. The positivity following an error (Pe) was modulated to a greater extent by speed-accuracy instruction for the high working memory capacity group than for the low working memory capacity group. These data help to explicate the neural bases of individual differences in working memory capacity and cognitive control.

The effect of priming for audiogenic seizures (AGS) on the development of epileptiform activity in the hippocampus was studied using in vitro kindling (IVK) in Long-Evans rats. AGS priming consists of intense auditory stimulation during a critical period of auditory development, resulting in sound-induced clonic convulsions upon subsequent testing. Between postnatal day (PND) 28 and 50, slices from subjects primed and sham-primed for AGS on PND 18 were used for recording responses in area CA1 of hippocampus following Schaffer collateral stimulation from stratum radiatum of area CA2/CA3. The developmental priming procedure, which enhances auditory brainstem excitability, resulted in fewer afterdischarges in slices from primed subjects across initial IVK stimulation sequences. These results suggest that changes in excitability that occur with acoustic priming can initially diminish selective epileptiform response characteristics in forebrain areas such as the hippocampus.

The Long-Evans rat is a hybrid rodent strain with little innate susceptibility to audiogenic seizures (AGS). The present study examines parameters of acoustic priming (induced susceptibility) and testing for AGS during postnatal development subsequent to auditory function, and identifies the effects of stimulus intensity, repeated testing, and gender upon AGS activity. Rats were exposed to 125-dB SPL 10-kHz tone bursts at 14-36 days of age and tested with white noise at 14 or 19 days following sound exposure. All priming/testing combinations yielded AGS susceptibility; animals primed at 18 days showed the highest incidence of clonic seizures when tested 14 days later. All subjects displayed clonus at testing intensities of 120 dB, although some seizure behaviors could be elicited at 100 dB. Repeated testing at 120 dB increased latency to clonus and clonus duration, and total wild running activity. Gender differences for AGS expression were minimal. These results demonstrate the viability of the seizure-resistant Long-Evans rat for study of AGS.

Audiogenic seizure susceptibility in the normally seizure-resistant Long-Evans rat may result from altered processing in the auditory pathway. Representative waveform latencies of the auditory brainstem responses (ABR) were recorded to examine generator alterations at different levels of the auditory neuraxis. Male Long-Evans rats primed for audiogenic seizures (AGS) on PND 14 with a 10 kHz pure tone at 120 dB SPL for 8 min were tested for AGS on PND 28 with 120 dB SPL continuous white noise. Primed subjects displayed wild running culminating in clonic convulsions. Following behavioral testing at 4-6 months, vertex recordings of ABR waves Ia-VI were made in anesthetized subjects using pure tone stimulus bursts. AGS subjects showed marginally elevated ABR thresholds. Shorter ABR wave latencies were elicited in AGS subjects for peripheral and central auditory components with stimulus intensities above 50 dB PeSPL at 8 and 40 kHz. Interpeak intervals were reduced for waves Ia-V and III-V in AGS subjects. These results reveal that intense sound stimulation during a sensitive period of development later reduces processing time at higher intensity levels.

The ability of Long-Evans hooded rats (n = 10) to detect sounds presented from sources in the horizontal plane at 0 degrees elevation and the effects of bilateral lesions of the inferior colliculus on these abilities were examined. Rats were trained on a directional detection task which required animals to suppress licking responses in a conditioned avoidance paradigm when 100-ms noise bursts were presented at random from speakers at 45 degrees intervals beginning at azimuth (0 degrees). A task performance rate was determined by reducing the correct lick suppression rate for signal trials by the proportion of incorrect suppression responses on non-signal trials. Higher performance rates were observed for stimuli presented from 0-90 degrees than for stimuli presented in the caudal hemifield prior to surgical procedures. Bilateral lesions restricted to the inferior colliculus reduced detection performance (p0.05) and shifted the best performance rates from sounds presented at 0-45 degrees to stimuli emitted from a 90 degrees source (p0.05). These results demonstrate that pigmented rats show differential detection levels for noise bursts presented from different locations throughout the horizontal interaural plane, and suggest that the inferior colliculus is involved in this aspect of directional hearing.

The medial geniculate body (MGB) is a thalamic structure that provides vital information flow to the forebrain for complex acoustic processing. The development of cytoarchitectural features of the MGB was examined in rat to identify age-related patterns of growth in major geniculate compartments that have been described previously (Clerici and Coleman [1990] J. Comp. Neurol. 297:14-31; Clerici et al. [1990] J. Comp. Neurol. 297:32-54): the ventral (MGv), dorsal (MGd), and medial (MGm) divisions. Results show that, on the day of parturition, all major nuclei of each division are characterized, including the ovoid (OV) and ventral (LV) nuclei of MGv; the dorsal, deep dorsal (DD), caudodorsal, limitans, and suprageniculate nuclei of MGd; and the MGm. The MGv and MGd, which display comparable areas at birth, show rapid growth to postnatal day 7 (PND7), which then slows until PND11, around the time of ear canal opening; subsequently, MGv accelerates growth to reach larger adult size. From PND11 to PND16, thionin facilitates parcellation by extensive staining of dendritic processes of MGd, MGm, and lateral posterior nucleus neurons but not neurons of the MGv or the dorsal lateral geniculate nucleus. Golgi stains after birth reveal restricted dendritic arborizations in MGv cells and dichotomous branching patterns of MGd neurons. Somal size in MGB increases dramatically subsequent to afferent innervation and again following onset of auditory function. Somal growth occurs between all postnatal age groups tested for OV, LV, and DD nuclei, although LV segments related to high and low frequencies do not differ. Cell packing density predicts the expanse of major MGB divisions better than somal size. These results demonstrate the integrity and growth patterns of MGB nuclei and divisions from nascence and provide a substrate for subsequent study of anatomical and physiological development of the MGB.

Previous work has suggested possible influences of ovarian hormones on evoked potentials in the auditory system. The aim of this project was to study the effects of ovariectomy and subsequent administration of estrogen replacement on the auditory brainstem response and the middle latency response. Groups of 90 day-old Long-Evans hooded rats were anesthetized for bilateral ovariectomies (ovex) and recordings made 3 weeks later. During the week prior to recordings some ovaricctomized groups received subcutaneous injections of 10, 100 or 500 μg/kg Premarin in peanut oil, and other unoperated animals received vehicle injections. Recordings from vertex/chin using needle electrodes and pure tone stimulus parameters were made under Rompun/Ketamine. The results using 40 kHz tone stimuli showed that mean latencies for ovex animals were longer than animals in the 100 μg/kg Premarin group for waves 1a, 1an, 1b, 11, 111, 111n, and 1V/V. Other posthoc comparisons at 40 kHz stimulation revealed differences between control and 100 μg/kg Premarin groups for latencies of waves 1b, 1bn, 11 and 111. Latency reduction appeared for waves 1b, 1bn, 11 and 111 for the 10 ovex group, but only at wave 11 for the 500 ovex group, compared to ovex-only animals. Data from 8 kHz stimulation also demonstrated significant differences between the ovex and ovex 100 groups at waves 1bn and Vn. Observations of interpeak latency differences, especially between waves 1a and 11, suggested central as well as cochlear involvement in hormone action. Examination of the middle-latency responses revealed considerable variability of latencies and amplitudes among subjects with latencies of ovariectomy animals significantly lengthened for the third component of the middle latency response at 40 kHz stimulation. These results suggest that changes in neurophysiological processing of auditory information followed by ovariectomy arc reversed by estrogen treatments which may alter activity in peripheral and central auditory structures. Modification of processing in the auditory system thus provides dependent measures for study in the surgical model of menopause.

This paper outlines the development of a new low-cost materials concept for polymer electrolyte membrane fuel cells (PEMFCs). Employing the roll bonding process to prepare a nickel clad steel laminate, the thin outer nickel layer is then passivated using a powder pack boridization technique. Results from energy dispersive X-ray (EDX) and X-ray diffraction (XRD) analyses and from scanning electron microscopy (SEM) indicated that a relatively homogeneous Ni 3 B layer grows on the exposed surfaces of the nickel and that the thickness of this layer can be readily controlled through the time and temperature over which boridization takes place. At high boridization temperatures, ≥ 700°C, and long periods of time, a Ni 2 B overlayer forms on top of the Ni 3 B. Preliminary exposure testing conducted at 80°C for 300hrs in 1M H 2 SO 4 containing 2ppm HF demonstrates a significant increase in the corrosion resistance that is attributable to the boridization treatment.

With semi-autonomous vehicles being pushed onto the market, driver monitoring systems play a relevant role in determining the user's level of engagement whenever an automated-to-manual transition of control is scheduled to occur. As current monitoring systems heavily rely on steering behaviour and ocular parameters as inputs for their algorithms, this study aims to investigate whether average heart rate represents a sensitive measure of driver mental workload. Participants from three different age groups (21-34, 35-53, 54-70 years old) were considered and the combined effects of mental workload (induced by interacting with the in-vehicle infotainment system via voice) and aging on average heart rate were investigated. Average heart rate was recorded using a portable heart rate monitor for commercial use. Results showed that average heart rate increased as the secondary task became more demanding. Furthermore, a significant task × age interaction suggested that as the secondary task became more demanding, younger drivers showed an increased heart rate compared to older drivers. These results are of the primary importance for the design of adaptive workload monitoring systems.

The cytoarchitecture, myeloarchitecture, and neocortical connectivity of the rat medial geniculate body (MGB) were comprehensively studied in adult and immature rats to define major anatomical divisions and nuclei. The MGB is a highly intricate structure composed of the ventral (MGv), dorsal (MGd), and medial (MGm) divisions and component nuclei, each having reciprocal connections with auditory neocortex. The MGv lies inferior to the midgeniculate bundle and extends to the rostral, but not caudal MGB tip. The MGv is composed of ventral and ovoid nuclei bounded by a marginal zone, each region containing dark staining small and medium sized, densely packed neurons shown to have tufted dendritic morphology; in contrast to the MGd, but similar to the dorsal lateral geniculate nucleus, only the perikarya of MGv neurons stain for Nissl in early postnatal material. Ventral nucleus cells align with afferent brachial axons, which penetrate the nucleus in a dorsoventral direction, whereas rostrocaudal cellular arrays are retrogradely labeled after injections of horseradish peroxidase (HRP) into auditory cortex. The ovoid nucleus is a double spiraled structure encircled and penetrated by afferent fibers that determine the orientation of constituent perikarya. Neurons in the transition zone align with a spray of axons emanating from the juncture of the ovoid and midgeniculate bundles. Marginal zone neurons are oriented in parallel to the free geniculate wall. The MGd resides within and superior to the midgeniculate bundle, and is composed of several nuclei that stain palely for myelin. In immature material, both dendritic processes and somata in the MGd stain for Nissl with our protocol; many of these cells show a stellate arborization pattern that distinguishes this region from the MGv, but is similar to the staining pattern of immature neurons of the lateral posterior nucleus. The adult dorsal nucleus has medium-sized, loosely packed neurons. The deep dorsal nucleus is situated among the fibers of the midgeniculate bundle and contains loosely packed round and fusiform cells; the latter cell type constitutes a minor proportion of the adult neuronal population but the major cell type in immature animals. The caudodorsal nucleus, which occupies the caudal tip of the MGB and rostrally courses superior to the dorsal nucleus, contains small, dark staining multipolar cells; the ventrolateral nucleus courses inferior to the MGv. The suprageniculate and limitans nuclei are included in the auditory thalamus on the basis of connections with auditory neocortex; the former has medium to dark staining mixed-sized cells, and the latter has densely packed cells which form a vertical column. HRP-labeled cells of dorsal division nuclei typically show either stellate or tufted dendritic morphologies. The MGm is traversed by dense myelinated fiber bundles emanating from the brachium of the inferior colliculus and includes very large “magnocellular” neurons, which predominate caudally, as well as medium-sized and small cells. “Magnocellular” neurons are elongate, triangular, or round, have loose, nonuniform packing, and tend to orient vertically; many of these cells have sparse, stellate dendritic arbors.

The medial geniculate body (MGB) of the rat was studied with Golgi methods to determine the distribution of neurons identified by dendritic morphology. These findings were compared with major divisions and constituent nuclei established by somatic and fiber architectonics, and by connections with temporal neocortex (Clerici et al.: Society of Neuroscience Abstracts 12:1272, 1986; 13:325, 1987; Anatomical Record 218:23, 1987; Winer and Larue: Journal of Comparative Neurology 257:282–315, 1987; Clerici and Coleman: Journal of Comparative Neurology 297:14–31, 1990). It was found that an elaboration of the prototypical scheme proposed by Morest (Journal of Anatomy 98:611–630, 1964) for partitioning the mammalian MGB is valid for characterizing the rat MGB. Two predominant categories of principal neuron dendritic patterning were identified: a bushy cell having tufted dendritic fields and a stellate cell with a radiate dendritic domain. Tufted neurons have large caliber dendritic trunks that divide profusely into daughter branches close to the soma with intertwining higher order branches that maintain a relatively restricted dendritic field. Stellate neurons typically emit primary dendrites in all directions that then divide dichotomously at wide angles at subsequent orders of branching to produce a somewhat spheroidal dendritic field. In the present study, the rat MGB is found to be a tripartite structure composed of ventral (MGv), dorsal (MGd), and medial (MGm) divisions, each uniquely characterized by constituent dendritic morphology. The paramount neuronal class of the MGv is the tufted principal cell. In the ventral and ovoid nuclei of the MGv the neuronal orientation of highly oriented bitufted cells is in register with afferent brachial axons. In the ventral nucleus, this arrangement approximates vertical with a dorsomedial tilt most prominent rostrally; in the ovoid nucleus, tufted cells adhere to the double spiraled course of afferent axons. The transition zone between ventral and ovoid nuclei contains tufted neurons that align with radially oriented fibers issuing from the junction of the ovoid and midgeniculate bundles. Bitufted neurons of the marginal zone parallel fibers at the lateral margin of the geniculate. Within the MGd the dorsal and caudodorsal nuclei are characterized by stellate cells with extensive dendritic arbors and bushy neurons with dendritic branches less tufted than those observed in the MGv. The deep dorsal nucleus contains bitufted neurons that polarize with the long axis of the midgeniculate bundle and intermingle with stellate neurons. The suprageniculate nucleus includes neurons with large somata and long, sparsely branched and dorsoventrally oriented dendrites orthagonal to corticothalamic axons, as well as smaller neurons and classical stellate cells. Neurons of the limitans nucleus have elongated dendritic fields directed along the medial border of the auditory thalamus. Among the variety of cell types of the MGm are ones with the largest somata in the MGB, the prominent “magnocellular” neurons. These cells emit only a few gradually tapering dendrites, which are encrusted with appendages; dendrites branch at irregular intervals and lie oblique to incoming axons from the brachium of the inferior colliculus. The MGm also contains smaller neurons, both tufted and radiate in typology, along with more classical stellate cells. The distribution of cells characterized by morphological features of dendrites corresponds to anatomical regions of the MGB identified by cytoarchitecture, myeloarchitecture, and necortical connectivity. The structural of the rat MGB are mostly typical of mammalian forms, which suggests the rat provides a use for the auditory forebrain.

To evade host resistance mechanisms, Salmonella enterica serovar Typhimurium (STM), a facultative intracellular pathogen, must alter its proteome following macrophage infection. To identify new colonization and virulence factors that mediate STM pathogenesis, we have isolated STM cells from RAW 264.7 macrophages at various time points following infection and used a liquid chromatography-mass spectrometry-based proteomic approach to detect the changes in STM protein abundance. Because host resistance to STM infection is strongly modulated by the expression of a functional host-resistant regulator, i.e. natural resistance-associated macrophage protein 1 (Nramp1, also called Slc11a1), we have also examined the effects of Nramp1 activity on the changes of STM protein abundances. A total of 315 STM proteins have been identified from isolated STM cells, which are largely housekeeping proteins whose abundances remain relatively constant during the time course of infection. However, 39 STM proteins are strongly induced after infection, suggesting their involvement in modulating colonization and infection. Of the 39 induced proteins, 6 proteins are specifically modulated by Nramp1 activity, including STM3117, as well as STM3118-3119 whose time-dependent abundance changes were confirmed using Western blot analysis. Deletion of the gene encoding STM3117 resulted in a dramatic reduction in the ability of STM to colonize wild-type RAW 264.7 macrophages, demonstrating a critical involvement of STM3117 in promoting the replication of STM inside macrophages. The predicted function common for STM3117-3119 is biosynthesis and modification of the peptidoglycan layer of the STM cell wall.

This paper presents a logical approach to designing a safety test plan for a space nuclear system. It is pointed out that two important facts need to underlie the development of a test plan: first, that sequential insults and the accumulation of damage are the rule; and second that the response of the nuclear system is stochastic (i.e., for any given set of conditions a probabilistic range of outcomes will occur regardless of the state of our knowledge). Because of these facts a deterministic approach can only be a starting point. The substance of the approach consists of undertaking and documenting three basic efforts: (1) a description of the analysts view of the problem and how it fits into the safety analysis, (2) a formal documentation of the purpose and requirements of the test plan (or test), and (3) an assessment of the use or usefulness of existing test data.

In order to more productively utilize the rich source of antigen-specific reagents present in the previously described non-immune single chain fragment variable (scFv) yeast display library, one must be able to efficiently isolate and characterize clones within the library. To this end, we have developed and validated a magnetic bead sorting technique utilizing the Miltenyi Macs system to recover greater than 90% of the antigen-specific clones present in the library. In combination with flow cytometry, we rapidly reduced diversity and enriched for antigen-specific clones in three rounds of selection. Furthermore, we demonstrate the use of pre-existing monoclonal antibodies (mAbs) for antigen labeling and subsequent flow cytometric sorting and characterization of epitope-specific scFv. Combining these two improvements in library screening allowed isolation and characterization of three epitope-specific scFv, including a previously uncharacterized epitope to a 6-kDa protein, epidermal growth factor.

This paper presents an analysis model devised specifically for evaluating sequential impact damage to a space nuclear system (barrier failure and fuel release for isotopic systems and geometry changes for reactor systems) with a nested safety system design. The analysis model is a relatively simple deterministic approach using disruptive work as the measure of damage. This approach offers a logical basis for assessing the impact damage to nested safety systems from sequential impacts, for designing safety tests, for analyzing subsequent test data, and finally for use in a safety analysis. Because of the wide variety of conditions that must be considered in a complete safety analysis and the stochastic nature of the damage states, the deterministic approach is only the starting point for a complete safety analysis.

Introduction: M cells are located in the epithelial layer covering the gut-associated lymphoid tissue and are responsible for delivery of macromolecules and microorganisms to the underlying lymphoid cells. It has been shown that the human colonic cell line Caco-2 can be converted to M cells in vitro following coculture with isolated lymphocytes from murine Peyer's patches. Studies were undertaken to evaluate and characterize the transepithelial transport of select macromolecules across these in vitro derived M cells. Methods: Caco-2 cells were converted to M cells as reported previously. The morphology of Caco-2 cells and M cells was compared by transmission electron microscopy (TEM). The transport properties of macromolecules such as horseradish peroxidase, FITC-conjugated polystyrene beads, and radiolabeled dextrans were examined. The activation of murine antigen-specific T cells following transport of the antigen ovalbumin across the M-cell barrier was assessed by measuring cytokine production. Results: M cells were shown to be irregular in shape and have fewer and shorter microvilli compared to the Caco-2 cell progenitors. These cells were still able to form tight junctions and monolayers on polycarbonate membranes. Time-course studies demonstrated that the transport of polystyrene beads and large-molecular-weight dextrans at physiological temperature across M-cell-containing monolayers was size dependent and more rapid than across Caco-2 cell monolayers. The transport of dextrans was also shown to be temperature and concentration dependent. Befitting the role of the M cell in mucosal defense, protein antigen could be delivered by these cells in order to be processed and presented to antigen-specific CD4+ T lymphocytes. Discussion: The M-cell permeability model is a functional and practical system for evaluating the transport properties of macromolecules and assessing the potential for intestinal mucosal antigen sampling to elicit immunological responses.

The purpose of the present study was to examine the effects of GABA-producing cell transplants on audiogenic seizures (AGS). The M213-2O cell line was derived from fetal rat striatum and has GABAergic properties. This cell line was further modified to express human GAD 67 and produce elevated levels of GABA. The present study compares the effects of parent M213-2O cell transplants with those of GAD 67 -modified M213-2O cells in AGS-prone Long–Evans rats. Two weeks following implantation of engineered cells, latency to AGS-typical wild running was increased compared to nonimplanted subjects. Survival of the transplanted cells was confirmed by immunochemical labeling of GAD 67 and Epstein–Barr virus nuclear antigen. These findings support the use of GABA-producing cell lines to modify seizure activity.

Audiogenic seizure (AGS) activity can be induced in the seizure-resistant Long-Evans rat by postnatal priming. This study examined the effects of unilateral lesions of the inferior colliculus (IC) and implantation of tectal grafts on AGS components. Animals were primed with a 10-kHz tone burst at 120 dB on postnatal day 14 and tested for AGS susceptibility on day 28, and then two groups were unilaterally lesioned including animals receiving embryonic day 16-17 grafts of caudal tectum. Subsequently, animals were repeatedly tested for wild running and clonic-tonic convulsion components of AGS. The results demonstrate that unilaterally grafted animals with partial IC lesions showed significant reduction in the incidence of clonus expression with greater terminal uniphasic wild running behavior. These effects were stronger than in animals with comparable unilateral lesions alone. Many neurons in graft cases were in direct contact with host tissues to provide a substrate for tissue interactions previously demonstrated to promote neuron survival and remediate IC functions.

The auditory brainstem response (ABR) was recorded from 20-month-old Long Evans hooded female rats to determine if latency reductions occur from estrogen replacement. The ABR in these post-breeding age rats was also examined for reductions in response latencies as a function of adult age. Tone pip stimuli (8 and 40 kHz) were presented at 21, 51, or 81 s(-1). Aging control and ovariectomized animals showed slower response latencies for waves Ib-VI than young adults for 8 and 40 kHz stimulation at 21 s(-1). Increased stimulus rate resulted in longer latencies for all waves at 20 months. In contrast to hormone treatment effects in young adults, ABR latencies in post-breeding age estrogen-treated animals were not reduced, consistent with a general decrease in CNS responsiveness to estrogen steroids associated with age. The results also suggest that sensorineural modifications in the auditory system which prolong ABR latencies can occur early in the aging process of adult female subjects.

Safety analyses and evaluations must be scrutable, defensible, and credible. This is particularly true when nuclear systems are involved, with their attendant potential for releases of radioactive materials (source terms) to the unrestricted environment. Analytical projections of General Purpose Heat Source Radioisotope Thermoelectric Generator (GPHS-RTG) source terms, for safety analyses conducted to date, have relied upon generic data correlations using a single parameter of cladding damage, termed “distortion.” However, distortion is not an unequivocal measure of cladding insult, failure, or release. Furthermore, the analytical foundation, applicability, and broad use of distortion are argumentative and, thus, somewhat troublesome. In an attempt to avoid the complications associated with the use of distortion, a new methodology, referred to as the Energy Interaction Model (EIM), has been preliminarily developed. This new methodology is based upon the physical principles of energy and energy exchange during mechanical interactions. Specifically, the EIM considers the energy imparted to GPHS-RTG components (bare fueled clads, GPHS modules, and full GPHS-RTGs) when exposed to mechanical threats (blast/overpressure, shrapnel and fragment impacts, and Earth surface impacts) posed by the full range of potential accidents. Expected forms are developed for equations intended to project cladding failure probabilities, the number of cladding failures expected, release amounts, and the fraction released as respirable particles. The coefficients of the equations developed are then set to fit the GPHS-RTG test data, ensuring good agreement with the experimental database. This assured, fitted agreement with the test database, along with the foundation of the EIM in first principles, provides confidence in the model’s projections beyond the available database. In summary, the newly developed EIM methodology is described and discussed. The conclusions reached are that the EIM holds great promise as a predictive analytical tool for future GPHS-RTG safety assessments, and this promise can become a reality in the near future—given a modest level of advancement and refinement.

The goals of this research were to examine the role of the inferior colliculus (IC) in mediating sound localization behavior and the ability of tectal grafts to restore function after IC ablation in the Long-Evans rat. Previous work has suggested that the IC is a major center for processing of information used in localizing sound sources in space. Adult rats were trained on a lick suppression paradigm to discriminate the location of the second pulse in a noise burst pair presented in the horizontal interaural plane. Following baseline testing, rats received bilateral IC lesions, bilateral lesions followed in 1 week by bilateral tectal grafts, or were sham operated. Sound localization ability was then tested 15 to 30 days and 40 to 50 days following surgical procedures. Performance across experimental groups was statistically the same during baseline testing. During the first operative test period lesion-only and grafted animals showed deficits in sound localization ability relative to controls. By the second postoperative test period control and grafted animals did not differ statistically in sound localization ability and performance of both groups was superior to that of lesion-only animals. Histology revealed a similar extent of IC damage in lesion-only and lesion-graft animals and revealed the presence of implanted tectal tissue in all grafted animals. There was significant neuron loss in the dorsal nucleus of lateral lemniscus (DNLL) in lesion-only animals relative to grafted rats and sham controls. Behavioral results suggest that the IC of pigmented rat is important for sound localization ability. The sparing of DNLL neurons in grafted animals suggests that the tectal grafts may directly integrate into or aid intrinsic recovery of the host auditory pathway.

SPE Members Abstract Co-precipitation of strontium sulfate with barium sulfate is often observed in oil fields. Some explanations have been given, but the mechanism is not fully understood. A numerical simulation approach gives a detailed explanation of its occurrence. This results in an improved tool for operators in dealing with prevention and treatment procedures. The computer model simulates reactive flow through porous media since injection of an incompatible fluid can induce numerous chemical reactions in a reservoir. The model uses the Gibbs free energy minimization technique to calculate chemical reactions that include precipitation from dissolved ionic species and dissolution of formation rocks. The model applies finite difference technique to solve mass transfer of the dissolved ionic species by convective fluid flow and diffusion. A case study of a North Sea seawater flood revealed the mechanism of strontium sulfate formation and its relationship to barium sulfate precipitation. Once seawater contacted barium (in the carbonate form) in the reservoir, barium sulfate dropped out immediately. This reaction also created excess carbonate ion (CO3-). Furthermore, although strontium carbonate is more stable than strontium sulfate, redistribution of the available CO3- forced strontium carbonate to dissolve and strontium sulfate to form. The results also suggest that the formation of the sulfate scales is not only controlled by their equilibrium constants but also dependent upon the concentrations of all other ionic species in the immediate vicinity. The simulation technique leads to a better prediction of the quantity and composition of sulfate scale formation. It also provides in-depth understanding of the interactions of the reservoir rocks with injected fluids. The application can also be extended to quantify the formation of other types of scales which occur during production operations. P. 447

Tone pip stimuli were used at different repetition rates to examine influences upon auditory brainstem response components. Rate increases to 80.1/s result in significant latency increases in waves IV and V at all test frequencies. Rate effects occur in rostral brainstem response components which show frequency related latencies different from wave I. Amplitude measures decline at higher repetition rates for waves I and IV at most test frequencies. Rate effects are most pronounced at 8 kHz stimulation which is around the optimal behavioral frequency in rat. At higher rates the second peak of wave I is enhanced at 8 kHz, whereas the third peak of wave I at 40 kHz disappears. These results demonstrate that tone-specific stimuli provide sensitive measures of latency, amplitude and wave morphology of components of the auditory brainstem response as a function of stimulus repetition.

The inferior colliculus was selected as a brain stem site for study of neural grafting and identification of calcium binding proteins. Unilateral ablation sites of eight midbrain inferior colliculus in adult Long-Evans rats were implanted with E17-18 caudal tectum. After 2 to 9 months animals were sacrificed and sections reacted using antibodies for calbindin and parvalbumin. The central nucleus of normal inferior colliculus shows high density of neuronal and fiber staining for parvalbumin. Typical graft cores had similar staining distributions including discoid and stellate neuron populations. Graft cores showed low densities of reactivity for calbindin comparable to central nucleus. In surrounding graft regions there was substantive-neuronal and fiber labeling for calbindin and parvalbumin including stellate neuron populations normally found in the dorsal and lateral nuclei of inferior colliculus. These results demonstrate that the expression of calcium binding proteins in tectal grafts resembles that of inferior colliculus.

The inferior colliculus (IC) is a midbrain structure that receives ascending auditory input from brainstem nuclei via the lateral lemniscus, sends efferent fibers to the medial geniculate body of thalamus and receives descending projections from auditory cortex. In the rat, the IC consists of dorsal and external cortices surrounding the central nucleus of IC (CNIC) which is populated by discoid and stellate neurons; the CNIC has a laminar appearance arising from organization of lemniscal fibers and processes of discoid cells. The IC of adult rats was chosen for implantation of whole grafts of E16-17 caudal tectum into unilateral lesion sites. Dendritic and somal architecture of graft neurons was examined 1 to 4.5 months following implantation using rapid Golgi, HRP and Nissl methods. The CNIC of rat is dominated by principal neurons with relatively flattened dendritic fields. In grafts of caudal tectum the most common neuron class observed possesses flattened dendritic arbors which often parallel one another. These neurons also resemble CNIC neurons of host tissue adjacent to the graft border. Spine formations appear on both proximal and distal dendrites of this neural type in both normal and implanted tissues. In addition, comparable somal features of graft neurons include ovoid or fusiform shapes with regular nuclear membranes as found in the normal colliculus. In Golgi stained material fewer stellate class neurons appear as in the normal CNIC, although stellate cell classes are more abundant in the pericentral areas of normal tissue. Both neuron populations are retrogradely labelled in graft and normal IC after HRP injection into the medial geniculate body. These features suggest that the graft core typically consists of prototypic CNIC cells. Other features of neuron and glial cell density vary in graft material which also shows a complex network of vasculature. These results demonstrate that whole grafts of caudal tectum placed into the inferior colliculus can form organized neural architecture similar to the normal CNIC. The somal, dendritic and spine features of these neurons form a potential substrate for connectional and functional properties which establish this preparation as suitable for further investigation as a model for development and recovery of function in the central auditory system.

The known structural and functional features of the inferior colliculus provided an advantageous substrate for examining the outcome of neural transplantation. In the present study, tissue from the midbrain tectum of Sprague-Dawley rat fetuses was removed at 15, 16 and 19 days of gestation (E15, E16 and E19), incubated as a dissociated cell suspension for 12 to 15 h in a medium containing True Blue for prelabeling, and injected into the central nucleus of the inferior colliculus (CNIC) of 28 normal male adult conspecifics. Viable tectal grafts were found in 57.5% of host animals with 65.2% located in CNIC. Grafts of E16 tissue had a survival rate of 84.2%, which was twice that of E15 or E19 donors, and there was no difference in graft survival rate in hosts sacrificed up to 6 months after the implantation procedure. Neurons prelabeled with True Blue and counterstained with thionin were identified as uniquely green stained cells in brightfield illumination and brilliant opaque cells in darkfield microscopy. Grafts were organized in tightly packed clusters of cells including large rounded polygonal and smaller cells similar in size to normal CNIC large and small multipolar neurons. Cell morphology of the larger ovoid and fusiform grafted neurons most closely resembled that of typical adult CNIC cell populations; other implanted cell types appeared immature even when host post-implant survival was extended. At longer post-implant intervals graft and host cells further intermingled, and both cellular and fibrous bridges were observed at the interface between host and graft tissue. The present work demonstrates the viability and development of homotypic cells grafted into a central auditory structure utilizing a simple method of prelabeling donor tissue with a fluorescent dye.

Bilateral dorsal and ventral hippocampal and anterior and posterior neocortical electrical activity was recorded in 11 cats during free behavior situations and during stimulation of medial and lateral hypothalamic systems which influence hippocampal electrical activity and behavior in distinct and contrasting ways. Behavioral states characterized by alertness, attentiveness, scanning, or investigatory activity were accompanied by hippocampal theta rhythm and by desynchronization of neocortical electrical activity. Behavioral states characterized by relaxed wakefulness, inattentiveness, or drowsiness were accompanied by high voltage irregular patterns of electrical activity in the hippocampus with no organized theta rhythms. Specific behaviors (alerting, orienting, scanning, or investigatory) emerging from relaxed behavioral states occurred concomitantly with hippocampal theta rhythm. Electrical stimulation of the medial hypothalamic system at 100 Hz elicited hippocampal theta rhythm and specific behaviors of alerting, orienting, scanning, and investigatory activity. In contrast, lateral hypothalamic stimulation at 100 Hz caused low voltage desynchronization of hippocampal electrical activity accompanied by postural stability and fixation of gaze. The relation of hippocampal theta rhythm to behavior is discussed in terms of brain stem-hypothalamic-hippocampal systems.

Metabolic responses from three sectors of the inferior colliculus (IC) divided according to frequency representation were examined bilaterally in young (YA) and aged adult (AA) rats using the 2-deoxyglucose method under quiescent or pure tone stimulus conditions. Resting IC metabolism in YA rats is characterized by elevated glucose uptake in the ventromedial (high frequency) relative to intermediate and dorsolateral sectors. AA resting incorporation is not reduced in any sector and is more uniform than YA uptake. Monaural high frequency stimulation (50 kHz) in YA animals evokes discrete contralateral banding along the ventromedial IC border, while stimulation in the optimal auditory sensitivity range of the rat at 8 kHz activates the intermediate sector of the contralateral IC. Although 50 kHz evoked ventromedial uptake is slightly reduced in AA rats contralaterally, the overall uptake within the three sectors does not differ significantly between ages under either of the monaural stimulus conditions; therefore senescent stability is exhibited. However, in AA rats the focus to 8 kHz stimulation has less clearly defined borders even though uptake within the intermediate sector is undiminished compared to YA animals, while 50 kHz elicited activity is typically reduced to a discontinuous band in the ventromedial sector of AA animals. These senescent changes may represent compensation in IC metabolism to changes in cochlear processing which are reflected in the activity of ascending auditory pathways.