Cocaine and Amphetamine-Regulated Transcript-Containing Neurons in the Nucleus Accumbens Project to the Ventral Pallidum in the Rat and May Inhibit Cocaine-Induced Locomotion

We have previously demonstrated that cocaine- and amphetamine-regulated transcript (CART) peptide colocalizes with GABA, dynorphin, D1 receptors, and substance P in some neurons in the nucleusaccumbens (NAcc). One of the main nuclei that receive accumbal efferents is the ventralpallidum (VP), and both dynorphin and substance P have been shown to be present in the cell bodies and terminals of this projection. Thus, we investigated whether CART peptide is also present in the VP in terminals that originate in the accumbens. The anterograde tracer Phaseolus vulgaris leukoagglutinin (PHA-L) colocalized with CART in neuronal processes in the VP when injected into the NAcc. Also, CART colocalized with the retrograde tracer r-BDA in accumbens cell bodies after the tracer was injected into the VP. Using electron microscopic immunocytochemistry, we examined CART terminals in the VP and found that CART-immunoreactive terminals formed symmetric synapses consistent with inhibitory GABAergic synapses. These synapses closely resemble GABAergic synapses in the substantia nigra pars reticulata (SNr), another nucleus that receives some CART-containing accumbal efferents. Lastly, we found that intra-pallidal injection of CART 55-102 inhibited cocaine-induced locomotion, indicating that CART peptide in the VP can have functional effects

Interleukin-8, Interleukin-1β, and Interferon-γ Levels Are Linked to PRRS Virus Clearance

Infection with porcine reproductive and respiratory syndrome virus (PRRSV) results in a weak antiviral immune response that leads to a persistent infection in a subset of pigs. We investigated the intensity and timing of the early cytokine responses to PRRSV infection to determine their utility as a predictor of persistence. As part of the “Big Pig” project, we evaluated cytokine gene expression in lymphoid tissues collected from pigs for up 202 days post-infection (dpi); serum samples were collected biweekly. Cytokine mRNA levels were compared between pigs that cleared the viral infection from serum and tissues (non-persistent [NP] pigs) to those of persistent (P) pigs, that had viral RNA in their serum for up to 126 dpi. The gene expression studies in the tracheobronchial lymph nodes (TBLN) of all the pigs showed upregulation of interferon-γ (IFN-γ)-associated T-helper 1 (Th-1) markers from 14–84 dpi, and of T-regulatory interleukin-10 (IL-10), but no upregulation of innate markers (IFN-A, IL-1B, and IL-8). At later time points (\u3e112 dpi) these genes were no longer differentially expressed and thus were uninformative for persistence studies. Statistical analyses of serum cytokine levels indicated that innate cytokine (IL-1β and IL-8) levels were upregulated early after infection. Interestingly, serum IL-8 levels in NP pigs were significantly higher than in P pigs at 14 dpi. When analyzed together, variations in all three of the serum cytokines tested (IL-8, IL-1β, and IFN-γ) was significantly correlated with virus level, accounting for ∼84% of the variations observed. These results indicate that while each cytokine individually has minor effects on the length of virus replication, the combination of cytokine activities should be considered when understanding the role of immunity in persistence

Global transcriptional response of porcine mesenteric lymph nodes to Salmonella enterica serovar Typhimurium

To elucidate the host transcriptional response to Salmonella enterica serovar Typhimurium, Affymetrix porcine GeneChip analysis of pig mesenteric lymph nodes was used to identify 848 genes showing differential expression across different times after inoculation or when compared to non-inoculated controls. Annotation analyses showed that a high proportion of these differentially expressed (DE) genes are involved in immune and inflammatory responses. T helper 1, innate/inflammatory, and antigen-processing pathways were induced at 24 h post-inoculation (hpi) and/or 48 hpi, while apoptosis and antigen presentation/dendritic cell function pathways were downregulated at 8 hpi. Cluster analyses revealed that most DE genes annotated as NFκB targets were grouped into a specific induced subcluster, while many translation-related DE genes were found in a repressed subcluster. Quantitative polymerase chain reaction analyses confirmed the Affymetrix results, revealing transcriptional induction of NFκB target genes at 24 hpi and suppression of the NFκB pathway from 24 to 48 hpi. We propose that such NFκB suppression in antigen-presenting cells may be the mechanism by which S. Typhimurium eludes a strong inflammatory response to establish a carrier status in pigs

Analysis of Porcine Transcriptional Response to Salmonella enterica serovar Choleraesuis suggests novel targets of NFkappaB are activated in the Mesenteric Lymph Node

<p>Abstract</p> <p>Background</p> <p>Specific knowledge of the molecular pathways controlling host-pathogen interactions can increase our understanding of immune response biology as well as provide targets for drug development and genetic improvement of disease resistance. Toward this end, we have characterized the porcine transcriptional response to <it>Salmonella enterica </it>serovar Choleraesuis (<it>S</it>. Choleraesuis), a <it>Salmonella </it>serovar that predominately colonizes swine, yet can cause serious infections in human patients. Affymetrix technology was used to screen for differentially expressed genes in pig mesenteric lymph nodes (MLN) responding to infection with <it>S</it>. Choleraesuis at acute (8 hours (h), 24 h and 48 h post-inoculation (pi)) and chronic stages (21 days (d) pi).</p> <p>Results</p> <p>Analysis of variance with false discovery rate control identified 1,853 genes with significant changes in expression level (<it>p</it>-value < 0.01, <it>q</it>-value < 0.26, and fold change (FC) > 2) during infection as compared to un-inoculated control pigs. Down-regulation of translation-related genes at 8 hpi and 24 hpi implied that <it>S</it>. Choleraesuis repressed host protein translation. Genes involved in the Th1, innate immune/inflammation response and apoptosis pathways were induced significantly. However, antigen presentation/dendritic cell (DC) function pathways were not affected significantly during infection. A strong NF<it>κ</it>B-dependent response was observed, as 58 known NF<it>κ</it>B target genes were induced at 8, 24 and/or 48 hpi. Quantitative-PCR analyses confirmed the microarray data for 21 of 22 genes tested. Based on expression patterns, these target genes can be classified as an "Early" group (induced at either 8 or 24 hpi) and a "Late" group (induced only at 48 hpi). Cytokine activity or chemokine activity were enriched within the Early group genes GO annotations, while the Late group was predominantly composed of signal transduction and cell metabolism annotated genes. Regulatory motif analysis of the human orthologous promoters for both Early and Late genes revealed that 241 gene promoters were predicted to contain NF<it>κ</it>B binding sites, and that of these, 51 Early and 145 Late genes were previously not known to be NF<it>κ</it>B targets.</p> <p>Conclusion</p> <p>Our study provides novel genome-wide transcriptional profiling data on the porcine response to <it>S</it>. Choleraesuis and expands the understanding of NF<it>κ</it>B signaling in response to <it>Salmonella </it>infection. Comparison of the magnitude and timing of porcine MLN transcriptional response to different <it>Salmonella </it>serovars, <it>S</it>. Choleraesuis and <it>S</it>. Typhimurium, clearly showed a larger but later transcriptional response to <it>S</it>. Choleraesuis. Both microarray and QPCR data provided evidence of a strong NF<it>κ</it>B-dependent host transcriptional response during <it>S</it>. Choleraesuis infection. Our data indicate that a lack of strong DC-mediated antigen presentation in the MLN may cause <it>S</it>. Choleraesuis infected pigs to develop a systemic infection, and our analysis predicts nearly 200 novel NF<it>κ</it>B target genes which may be applicable across mammalian species.</p

First NuSTAR Limits on Quiet Sun Hard X-Ray Transient Events

We present the first results of a search for transient hard X-ray (HXR) emission in the quiet solar corona with the \textit{Nuclear Spectroscopic Telescope Array} (\textit{NuSTAR}) satellite. While \textit{NuSTAR} was designed as an astrophysics mission, it can observe the Sun above 2~keV with unprecedented sensitivity due to its pioneering use of focusing optics. \textit{NuSTAR} first observed quiet Sun regions on 2014 November 1, although out-of-view active regions contributed a notable amount of background in the form of single-bounce (unfocused) X-rays. We conducted a search for quiet Sun transient brightenings on time scales of 100 s and set upper limits on emission in two energy bands. We set 2.5--4~keV limits on brightenings with time scales of 100 s, expressed as the temperature T and emission measure EM of a thermal plasma. We also set 10--20~keV limits on brightenings with time scales of 30, 60, and 100 s, expressed as model-independent photon fluxes. The limits in both bands are well below previous HXR microflare detections, though not low enough to detect events of equivalent T and EM as quiet Sun brightenings seen in soft X-ray observations. We expect future observations during solar minimum to increase the \textit{NuSTAR} sensitivity by over two orders of magnitude due to higher instrument livetime and reduced solar background.Comment: 11 pages, 7 figures; accepted for publication in The Astrophysical Journa

Difference in severity of porcine circovirus type two-induced pathological lesions between Landrace and Pietrain pigs

Anecdotal information from the field suggests that there are host genetic differences in susceptibility to porcine circovirus type 2 (PCV2) associated disease among Landrace and Pietrain breeds. The objective of this study was to determine if a difference exists in PCV2 susceptibility between Landrace and Pi-etrain pigs under experimental conditions. Thirty-nine Landrace pigs and 39 Pietrain pigs were blocked by breed, sire, dam, and litter and randomly divided into the following 4 groups: Landrace noninoculated negative control (Landrace-NEG; n = 13), Pietrain noninoculated negative control (Pietrain-NEG; n = 13), Landrace-PCV2 (n = 26; Landrace), and Pietrain-PCV2 (n = 26; Pietrain). After waning of passively acquired anti-PCV2 antibodies, Landrace-PCV2 and Pietrain-PCV2 groups were inoculated with PCV2 isolate ISU-40895. The Landrace-NEG and Pietrain-NEG groups were housed in a separate room, remained noninoculated, and served as negative controls. All pigs in all groups were necropsied at 21 d post PCV2-inoculation. Onset of seroconversion and concentrations of anti-PCV2-IgM, anti-PCV2-IgG, and anti-PCV2 neutralizing antibodies were similar in Landrace-PCV2 and Pietrain-PCV2 groups. Furthermore, the amount of PCV2 DNA and cytokine concentrations in serum and plasma samples were not different between the 2 PCV2-inoculated groups. The severity of PCV2-associated microscopic lesions was different between Landrace and Pietrain pigs; Landrace-PCV2 pigs had significantly (P \u3c 0.05) more severe lymphoid lesions than the Pietrain-PCV2 pigs. Although the pigs originated from the same farm where their dams were commingled, passively acquired anti-PCV2-antibodies waned in Pietrain pigs by approximately 12 wk of age, whereas the majority of the Landrace pigs remained PCV2 seropositive until 18 wk of age and beyond. The results from this study indicate that a genetic difference exists between these 2 breeds of pigs in susceptibility to PCV2-associated lesions

The First Focused Hard X-ray Images of the Sun with NuSTAR

We present results from the the first campaign of dedicated solar observations undertaken by the \textit{Nuclear Spectroscopic Telescope ARray} ({\em NuSTAR}) hard X-ray telescope. Designed as an astrophysics mission, {\em NuSTAR} nonetheless has the capability of directly imaging the Sun at hard X-ray energies ($>$3~keV) with an increase in sensitivity of at least two magnitude compared to current non-focusing telescopes. In this paper we describe the scientific areas where \textit{NuSTAR} will make major improvements on existing solar measurements. We report on the techniques used to observe the Sun with \textit{NuSTAR}, their limitations and complications, and the procedures developed to optimize solar data quality derived from our experience with the initial solar observations. These first observations are briefly described, including the measurement of the Fe K-shell lines in a decaying X-class flare, hard X-ray emission from high in the solar corona, and full-disk hard X-ray images of the Sun.Comment: 11 pages, accepted to Ap

The Parasitic Wasp, Cotesia congregata (Say), Consists of Two Incipient Species Isolated by Asymmetric Reproductive Incompatibility and Hybrid Inability to Overcome Host Defenses

Parasitic wasps are highly diverse and play a major role in suppression of herbivorous insect pest populations. Several previously identified species of parasitic wasps have been found to be complexes of cryptic species resulting from adaptations to specific hosts or host foodplants. Cotesia congregata (Say) (Hymenoptera: Braconidae), which has long served as a model system for host-parasitoid interactions, can be used for investigating the process of diversification among sympatric populations that differ in host and host foodplant usage. Two incipient species of C. congregata have been identified in the USA mid-Atlantic region, “MsT wasps” originate from Manduca sexta (L.) (Lepidoptera: Sphingidae) on tobacco and “CcC wasps” originate from Ceratomia catalpae (Boisduval) (Lepidoptera: Sphingidae) on catalpa. Both wasp sources can develop in either host species. Hybrids resulting from MsT♂xCcC♀ crosses are fertile, whereas hybrids from CcC♂xMsT♀ crosses are typically sterile. In this study, we compared relative expression in vivo of seven C. congregata bracovirus (CcBV) genes among MsT and CcC parental and hybrid crosses. Also, we established hybrid crosses between MsT and CcC wasps and four additional host foodplant sources of C. congregata. Patterns of relative expression in vivo of MsT and CcC CcBV genes differed; a few were not expressed in hosts parasitized by CcC wasps. Overall, relative expression of CcBV genes from MsT and CcC wasps did not differ with respect to the host species parasitized. Low or absent expression of CcBV genes was found in hosts parasitized by sterile hybrids. For the most part, the other four host-foodplant wasp sources were reproductively compatible with either MsT or CcC wasps and hybrid crosses with the alternative wasp source were asymmetrically sterile. Crosses involving CcC males or MsT females produced sterile hybrids that lacked mature ovaries. Cumulatively, results indicate that C. congregata is composed of two sympatric incipient species that can utilize multiple host species rather than several host-associated races or cryptic species

Kappa Opioid receptor-induced aversion requires p38 MAPK activation in VTA dopamine neurons

The endogenous dynorphin-κ opioid receptor (KOR) system encodes the dysphoric component of the stress response and controls the risk of depression-like and addiction behaviors; however, the molecular and neural circuit mechanisms are not understood. In this study, we report that KOR activation of p38α MAPK in ventral tegmental (VTA) dopaminergic neurons was required for conditioned place aversion (CPA) in mice. Conditional genetic deletion of floxed KOR or floxed p38α MAPK by Cre recombinase expression in dopaminergic neurons blocked place aversion to the KOR agonist U50,488. Selective viral rescue by wild-type KOR expression in dopaminergic neurons of KOR(−/−) mice restored U50,488-CPA, whereas expression of a mutated form of KOR that could not initiate p38α MAPK activation did not. Surprisingly, while p38α MAPK inactivation blocked U50,488-CPA, p38α MAPK was not required for KOR inhibition of evoked dopamine release measured by fast scan cyclic voltammetry in the nucleus accumbens. In contrast, KOR activation acutely inhibited VTA dopaminergic neuron firing, and repeated exposure attenuated the opioid response. This adaptation to repeated exposure was blocked by conditional deletion of p38α MAPK, which also blocked KOR-induced tyrosine phosphorylation of the inwardly rectifying potassium channel (GIRK) subunit Kir3.1 in VTA dopaminergic neurons. Consistent with the reduced response, GIRK phosphorylation at this amino terminal tyrosine residue (Y12) enhances channel deactivation. Thus, contrary to prevailing expectations, these results suggest that κ opioid-induced aversion requires regulation of VTA dopaminergic neuron somatic excitability through a p38α MAPK effect on GIRK deactivation kinetics rather than by presynaptically inhibiting dopamine release. SIGNIFICANCE STATEMENT Kappa opioid receptor (KOR) agonists have the potential to be effective, nonaddictive analgesics, but their therapeutic utility is greatly limited by adverse effects on mood. Understanding how KOR activation produces dysphoria is key to the development of better analgesics and to defining how the endogenous dynorphin opioids produce their depression-like effects. Results in this study show that the aversive effects of κ receptor activation required arrestin-dependent p38α MAPK activation in dopamine neurons but did not require inhibition of dopamine release in the nucleus accumbens. Thus, contrary to the prevailing view, inhibition of mesolimbic dopamine release does not mediate the aversive effects of KOR activation and functionally selective κ opioids that do not activate arrestin signaling may be effective analgesics lacking dysphoric effects

Validation of a first-generation long-oligonucleotide microarray for transcriptional profiling in the pig

A first-generation porcine oligonucleotide set, representing 13,297 cDNAs and ESTs, has been designed by Qiagen–Operon for transcriptional profiling. To validate this set, microarrays containing each 70-mer oligonucleotide, referred to as the Qiagen–NRSP8 array, were hybridized with targets from porcine adult liver, lung, muscle, or small intestine. Transcriptome analyses showed that 11,328 of the oligonucleotides demonstrated expression in at least one tissue. Statistical analyses revealed that 1810 genes showed differential expression among tissues (Bonferroni adjusted p \u3c 0.05). Biological pathways identified by DAVID/EASE analysis using a list of 423 tissue-selective genes matched archetypal pathways in the corresponding human or mouse tissue. Real-time quantitative PCR confirmed expression patterns for 9 of 11 genes tested. Our results demonstrate that this first-generation porcine oligonucleotide array is informative and the specificity is high. This is essential validation for investigators using the Qiagen–NRSP8 array for porcine functional genomics and for using the pig in modeling important physiological problems