Astrocyte focal adhesion kinase reduces passive stress coping by inhibiting ciliary neurotrophic factor only in female mice.

Astrocytes have been implicated in stress responses and produce ciliary neurotrophic factor (CNTF), which we have shown in the mouse medial amygdala (MeA) to promote passive stress coping response only in females. Pharmacological inhibition of focal adhesion kinase (FAK) upregulates CNTF expression. Here, we found that inducible knockout of FAK in astrocytes or systemic treatment with an FAK inhibitor increased passive coping behavior, i.e., immobility, in an acute forced swim stress test in female, but not male, mice. Strikingly, four weeks of chronic unpredictable stress (CUS) did not further increase passive coping in female astrocytic FAK knockout mice, whereas it exacerbated it in female wildtype mice and male mice of both genotypes. These data suggest that astrocyte FAK inhibition is required for chronic stress-induced passive coping in females. Indeed, CUS reduced phospho-FAK and increased CNTF in the female MeA. Progesterone treatment after ovariectomy activated amygdala FAK and alleviated ovariectomy-induced passive coping in wildtype, but not astrocytic FAK knockout females. This suggests that progesterone-mediated activation of FAK in astrocytes reduces female stress responses. Finally, astrocytic FAK knockout or FAK inhibitor treatment increased CNTF expression in the MeA of both sexes, although not in the hippocampus. As mentioned, MeA CNTF promotes stress responses only in females, which may explain the female-specific role of astrocytic FAK inhibition. Together, this study reveals a novel female-specific progesterone-astrocytic FAK pathway that counteracts CNTF-mediated stress responses and points to opportunities for developing treatments for stress-related disorders in women.

Modulation of mGlu5 reduces rewarding associative properties of nicotine via changes in mesolimbic plasticity: Relevance to comorbid cigarette smoking in psychosis.

RATIONALE: Antipsychotic medications that are used to treat psychosis are often limited in their efficacy by high rates of severe side effects. Treatment success in schizophrenia is further complicated by high rates of comorbid nicotine use. Dopamine D2 heteroreceptor complexes have recently emerged as targets for the development of more efficacious pharmaceutical treatments for schizophrenia. OBJECTIVE: The current study sought to explore the use of the positive allosteric modulator of the mGlu5 receptor 3-Cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamide (CDPPB) as a treatment to reduce symptoms related to psychosis and comorbid nicotine use. METHODS: Neonatal treatment of animals with the dopamine D2-like receptor agonist quinpirole (NQ) from postnatal day (P)1-21 produces a lifelong increase in D2 receptor sensitivity, showing relevance to psychosis and comorbid tobacco use disorder. Following an 8-day conditioning paradigm, brain tissue in the mesolimbic pathway was analyzed for several plasticity markers, including brain derived neurotrophic factor (BDNF), phosphorylated p70 ribosomal S6 kinase (phospho-p70S6K), and cadherin-13 (Cdh13). RESULTS: Pretreatment with CDPPB was effective to block enhanced nicotine conditioned place preference observed in NQ-treated animals. Pretreatment was additionally effective to block the nicotine-induced increase in BDNF and sex-dependent increases in cadherin-13 in the ventral tegmental area (VTA), as well as increased phospho-p70S6K in the nucleus accumbens (NAcc) shell found in NQ-treated animals. CONCLUSION: In conjunction with prior work, the current study suggests positive allosteric modulation of the mGlu5 receptor, an emerging target for schizophrenia therapeutics, may be effective for the treatment of comorbid nicotine abuse in psychosis.

Reinstatement of nicotine conditioned place preference in a transgenerational model of drug abuse vulnerability in psychosis: Impact of BDNF on the saliency of drug associations.

RATIONALE: Psychotic disorders such as schizophrenia are often accompanied by high rates of cigarette smoking, reduced quit success, and high relapse rates, negatively affecting patient outcomes. However, the mechanisms underlying altered relapse-like behaviors in psychosis are poorly understood. OBJECTIVES: The present study analyzed changes in extinction and reinstatement of nicotine conditioned place preference (CPP) and resulting changes in brain-derived neurotrophic factor (BDNF) in a novel heritable rodent model of psychosis, demonstrating increased dopamine D2 receptor sensitivity, to explore mechanisms contributing to changes in relapse-like behaviors. METHODS: Male and female offspring of two neonatal quinpirole-treated (1 mg/kg quinpirole from postnatal day (P)1-21; QQ) and two neonatal saline-treated (SS) Sprague-Dawley rats (F1 generation) were tested on an extended CPP paradigm to analyze extinction and nicotine-primed reinstatement. Brain tissue was analyzed 60 min after the last nicotine injection for BDNF response in the ventral tegmental area (VTA), the infralimbic (IfL) and prelimbic (PrL) cortices. RESULTS: F1 generation QQ offspring demonstrated delayed extinction and more robust reinstatement compared to SS control animals. In addition, QQ animals demonstrated an enhanced BDNF response to nicotine in the VTA, IfL and Prl cortices compared to SS offspring. CONCLUSIONS: This study is the first to demonstrate altered relapse-like behavior in a heritable rodent model with relevance to comorbid drug abuse and psychosis. This altered pattern of behavior is hypothesized to be related to elevated activity-dependent BDNF in brain areas associated with drug reinforcement during conditioning that persists through the extinction phase, rendering aberrantly salient drug associations resistant to extinction and enhancing relapse vulnerability.

An adenine/thymidine-rich region is integral to RepL-mediated DNA replication.

The lytic replication of bacteriophage P1 requires RepL expression and the lytic stage origin, oriL, which is postulated to be located within repL gene sequence. The exact sequence of P1 oriL and the mechanism(s) of RepL-mediated DNA replication, however, are not fully understood. By using repL gene expression to induce DNA replication of a gfp and a rfp reporter plasmids, we demonstrated that synonymous base substitution in an adenine/thymidine-rich region of repL gene sequence, termed AT2, significantly inhibited the RepL-mediated signal amplification. Contrastingly, mutations in an IHF and two DnaA binding sites did not affect the RepL-mediated signal amplification significantly. A truncated repL sequence with the AT2 region allowed RepL-mediated signal amplification in trans therefore verifying a significant role of the AT2 region in RepL-mediated DNA replication. A combination of repL gene expression and a non-protein-coding copy of repL gene sequence (termed nc-repL) was able to amplify the output of an arsenic biosensor. Furthermore, mutation(s) at single or multiple positions within the AT2 region produced varying levels of RepL-mediated signal amplification. Overall, our results provide novel insights into the identity and location of P1 oriL as well as demonstrating the potential of using repL constructs to amplify and modulate the output of genetic biosensors.

P1 Bacteriophage-Enabled Delivery of CRISPR-Cas9 Antimicrobial Activity Against Shigella flexneri.

The discovery of clustered, regularly interspaced, short palindromic repeats (CRISPR) and the Cas9 RNA-guided nuclease provides unprecedented opportunities to selectively kill specific populations or species of bacteria. However, the use of CRISPR-Cas9 to clear bacterial infections in vivo is hampered by the inefficient delivery of cas9 genetic constructs into bacterial cells. Here, we use a broad-host-range P1-derived phagemid to deliver the CRISPR-Cas9 chromosomal-targeting system into Escherichia coli and the dysentery-causing Shigella flexneri to achieve DNA sequence-specific killing of targeted bacterial cells. We show that genetic modification of the helper P1 phage DNA packaging site (pac) significantly enhances the purity of packaged phagemid and improves the Cas9-mediated killing of S. flexneri cells. We further demonstrate that P1 phage particles can deliver chromosomal-targeting cas9 phagemids into S. flexneri in vivo using a zebrafish larvae infection model, where they significantly reduce the bacterial load and promote host survival. Our study highlights the potential of combining P1 bacteriophage-based delivery with the CRISPR chromosomal-targeting system to achieve DNA sequence-specific cell lethality and efficient clearance of bacterial infection.

The Surgeon's Risk of SARS-CoV-2 Infection During the Initial Peak of the COVID-19 Pandemic in New Orleans.

Surgeons who care for patients with active SARS-CoV-2 infection represent a unique population of health care providers whose risk of infection has not been elucidated. The objective of this study was to examine SARS-CoV-2 seroprevalence among surgeons who cared for patients with active SARS-CoV-2 infection compared to other employees within our health care system and also the general public of New Orleans. 105 surgeons at our facilities provided direct surgical care to patients with active SARS-CoV-2 infection and underwent voluntary antibody testing. 2/105 (1.9% CI .2%-6.7%) tested positive for SARS-CoV-2 antibodies. 13 343 hospital employees underwent antibody testing and 1066/13 343 (8.0% CI 7.5%-8.5%) tested positive (1.9% vs. 8.0%; P = .03). We saw a significantly lower SARS-CoV-2 seroprevalence among surgeons who directly cared for infected patients versus other hospital employees. When compared to community seroprevalence (6.9% CI 6.0%-8.0%), seroprevalence among our surgeons is also significantly lower (1.9% vs. 6.9%; P = .04).

An Analysis of Outcomes and Management Strategies for Patients With Cholecystostomy Tubes.

BACKGROUND: Percutaneous cholecystostomy tube (PCT) drainage is an effective management strategy for acute cholecystitis in patients medically unfit for surgery. However, little is known about the fate of patients managed by PCT. We conducted this study to determine tube management outcomes for patients with acute cholecystitis managed by PCT. METHODS: The electronic record was queried to identify patients with acute cholecystitis managed by PCT from 2012-2020. Patients were divided into three groups for analysis: 1) ultimately managed by cholecystectomy, 2) eventual confirmation of distal flow of bile from the gallbladder and tube removal, and 3) tubes left in place without further management. RESULTS: A total of 179 patients with acute cholecystitis treated by PCT were included. Sixty-six patients never fully recovered from the medical insult associated with their diagnosis of acute cholecystitis and had their tubes left in situ. Sixty-four of these 66 patients (97%) died during follow-up. The remaining 113 patients recovered from their illness and presented to clinic for evaluation for tube removal and/or cholecystectomy. When distal biliary flow was confirmed, tube removal was favored (n = 70). When cystic duct outflow occlusion persisted, cholecystectomy was planned for patients who became acceptable surgical candidates (n = 43). For patients managed by cholecystectomy, 8 were approached open and 35 laparoscopically, with 12 of 35 (34.3%) converted to open and 23 (65.7%) completed laparoscopically. CONCLUSION: Our study favors PCT removal for patients who recover from their acute illness when distal bile flow from the gallbladder is confirmed. We reserve cholecystectomy for patients who recover from their illness and demonstrate persistent cystic duct outflow obstruction.

Localization of NGF expression in mouse spleen and salivary gland: Relevance to pleotropic functions.

Our primary goal was to determine if leukocytes are a source of nerve growth factor (NGF) in mouse spleen. Noradrenergic nerves were localized to arteries and white pulp in normal spleens but only to arteries in ultra-immunodeficient R2G2 mice that lack leukocytes. NGF mRNA was detected in vascular cells and leukocytes of normal spleen, and several of the latter were T cells based on double labeling for NGF mRNA and CD3. Our findings indicate NGF is produced by vascular cells and to a lesser extent by leukocytes in spleen and provide support for pleiotropic actions in spleen and salivary glands.

Female-specific role of ciliary neurotrophic factor in the medial amygdala in promoting stress responses.

Ciliary neurotrophic factor (CNTF) is produced by astrocytes which have been implicated in regulating stress responses. We found that CNTF in the medial amygdala (MeA) promotes despair or passive coping, i.e., immobility in an acute forced swim stress, in female mice, while having no effect in males. Neutralizing CNTF antibody injected into the MeA of wildtype females reduced activation of downstream STAT3 (Y705) 24 and 48 h later. In concert, the antibody reduced immobility in the swim test in females and only after MeA injection, but not when injected in the central or basolateral amygdala. Antibody injected into the male MeA did not affect immobility. These data reveal a unique role of CNTF in female MeA in promoting despair or passive coping behavior. Moreover, 4 weeks of chronic unpredictable stress (CUS) increased immobility in the swim test and reduced sucrose preference in wildtype CNTF+/+, but not CNTF-/- littermate, females. Following CUS, 10 min of restraint stress increased plasma corticosterone levels only in CNTF+/+ females. In males, the CUS effects were present in both genotypes. Further, CUS increased CNTF expression in the MeA of female, but not male, mice. CUS did not alter CNTF in the female hippocampus, hypothalamus and bed nucleus of stria terminalis. This suggests that MeA CNTF has a female-specific role in promoting CUS-induced despair or passive coping, behavioral anhedonia and neuroendocrine responses. Compared to CNTF+/+ mice, CNTF-/- mice did not show differences in CUS-induced anxiety-like behavior and sensorimotor gating function as measured by elevated T-Maze, open field and pre-pulse inhibition of the acoustic startle response. Together, this study reveals a novel CNTF-mediated female-specific mechanism in stress responses and points to opportunities for developing treatments for stress-related disorders in women.

Endoscopic Vacuum Therapy via Pharyngostomy: Novel Access for Management of Upper Gastrointestinal Defects.

BACKGROUND: Perforation and anastomotic leakage of the upper gastrointestinal tract (UGI) has a high mortality and morbidity rate. Recently, UGI leaks have been treated with endoscopic vacuum therapy (EVT). However, this technique traditionally requires multiple EVT changes and a prolonged and uncomfortable nasoenteric intubation. We describe our experience using EVT through a novel pharyngostomy access to manage UGI leaks. METHODS: We describe our development and implementation of EVT via a novel pharyngostomy access to treat a variety of UGI defects. Preoperative, perioperative, and postoperative data were analyzed. RESULTS: Six patients with UGI perforations or anastomotic leaks were treated with an EVT using a pharyngostomy access. The median age was 69 years (IQR 53-71). Four patients leaked after an Ivor Lewis esophagectomy, one after a robotic para-esophageal hernia repair, and another after a Roux en Y esophagojejunostomy. Defects were detected on a median of 11.5 days (IQR 3-21). Median values for the duration of the EVT therapy and the number of EVT changes were 19.5 days (IQR 14-31) and 7 (IQR 6.5-9), respectively. Four of the patients were discharged with an EVT in place and were successfully managed as outpatients. At a median follow-up of 8 months, two patients developed strictures. None of the patients required any surgical re-intervention, they tolerated oral intake, and all leakages were confirmed closed by imaging and endoscopy. DISCUSSION: Endoscopic vacuum therapy can be successfully managed through a pharyngostomy access, as described. This access is easy, comfortable, and reliable and allows for a transition to outpatient management.

Effects of Manipulation of Noradrenergic Activities on the Expression of Dopaminergic Phenotypes in Aged Rat Brains.

This study investigated the effects of the pharmacological manipulation of noradrenergic activities on dopaminergic phenotypes in aged rats. Results showed that the administration of L-threo-3,4-dihydroxyphenylserine (L-DOPS) for 21 days significantly increased the expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the striatum and substantia nigra (SN) of 23-month-old rats. Furthermore, this treatment significantly increased norepinephrine/DA concentrations in the striatum and caused a deficit of sensorimotor gating as measured by prepulse inhibition (PPI). Next, old rats were injected with the α2-adrenoceptor antagonist 2-methoxy idazoxan or ß2-adrenoceptor agonist salmeterol for 21 days. Both drugs produced similar changes of TH and DAT in the striatum and SN. Moreover, treatments with L-DOPS, 2-methoxy idazoxan, or salmeterol significantly increased the protein levels of phosphorylated Akt in rat striatum and SN. However, although a combination of 2-methoxy idazoxan and salmeterol resulted in a deficit of PPI in these rats, the administration of 2-methoxy idazoxan alone showed an opposite behavioral change. The in vitro experiments revealed that treatments with norepinephrine markedly increased mRNAs and proteins of ATF2 and CBP/p300 and reduced mRNA and proteins of HDAC2 and HDAC5 in MN9D cells. A ChIP assay showed that norepinephrine significantly increased CBP/p300 binding or reduced HDAC2 and HDAC5 binding on the TH promoter. The present results indicate that facilitating noradrenergic activity in the brain can improve the functions of dopaminergic neurons in aged animals. While this improvement may have biochemically therapeutic indication for the status involving the degeneration of dopaminergic neurons, it may not definitely include behavioral improvements, as indicated by using 2-methoxy idazoxan only.

Improved Accuracy for the Sturm-Pastyr Localizer in the Presence of Image Noise.

Image guidance for frame-based stereotaxis is facilitated by incorporating three to four Sturm-Pastyr (SP) localizers into a stereotactic frame. Typically, each SP localizer enables the calculation of one set of [Formula: see text] coordinates in the three-dimensional coordinate system of the stereotactic frame, given three sets of [Formula: see text] coordinates created by the SP localizer in the two-dimensional coordinate system of a computed tomography (CT) image. Bouza and Brown propose formulas to calculate three sets of [Formula: see text] coordinates for each SP localizer. Monte Carlo (MC) simulation compares the accuracy of the new formulation to the accuracy of the original SP formulation that calculates only one set of [Formula: see text] coordinates for each SP localizer. Monte Carlo simulation reveals that the calculation of three sets of [Formula: see text] coordinates instead of only one set improves the accuracy of image guidance.

Bidirectional control of infant rat social behavior via dopaminergic innervation of the basolateral amygdala.

Social interaction deficits seen in psychiatric disorders emerge in early-life and are most closely linked to aberrant neural circuit function. Due to technical limitations, we have limited understanding of how typical versus pathological social behavior circuits develop. Using a suite of invasive procedures in awake, behaving infant rats, including optogenetics, microdialysis, and microinfusions, we dissected the circuits controlling the gradual increase in social behavior deficits following two complementary procedures-naturalistic harsh maternal care and repeated shock alone or with an anesthetized mother. Whether the mother was the source of the adversity (naturalistic Scarcity-Adversity) or merely present during the adversity (repeated shock with mom), both conditions elevated basolateral amygdala (BLA) dopamine, which was necessary and sufficient in initiating social behavior pathology. This did not occur when pups experienced adversity alone. These data highlight the unique impact of social adversity as causal in producing mesolimbic dopamine circuit dysfunction and aberrant social behavior.

Modulation of mGlu5 improves sensorimotor gating deficits in rats neonatally treated with quinpirole through changes in dopamine D2 signaling.

This study analyzed whether the positive allosteric modulator of metabotropic glutamate receptor type 5 (mGlu5) 3-Cyano-N-(1,3-diphenyl-1H-pyrazol-5-yl) benzamide (CDPPB) would alleviate deficits in prepulse inhibition (PPI) and affect dopamine (DA) D2 signaling in the dorsal striatum and prefrontal cortex (PFC) in the neonatal quinpirole (NQ) model of schizophrenia (SZ). Male and female Sprague-Dawley rats were neonatally treated with either saline (NS) or quinpirole HCL (1 mg/kg; NQ), a DAD2 receptor agonist, from postnatal days (P) 1-21. Rats were raised to P44 and behaviorally tested on PPI from P44-P48. Before each trial, rats were subcutaneous (sc) administered saline or CDPPB (10 mg/kg or 30 mg/kg). On P50, rats were given a spontaneous locomotor activity test after CDPPB or saline administration. On P51, the dorsal striatum and PFC were evaluated for both arrestin-2 (ßA-2) and phospho-AKT protein levels. NQ-treated rats demonstrated a significant deficit in PPI, which was alleviated to control levels by the 30 mg/kg dose of CDPPB. There were no significant effects of CDPPB on locomotor activity. NQ treatment increased ßA-2 and decreased phospho-AKT in both the dorsal striatum and PFC, consistent with an increase DAD2 signaling. The 30 mg/kg dose of CDPPB significantly reversed changes in ßA-2 in the dorsal striatum and PFC and phospho-AKT in the PFC equivalent to controls. Both doses of CDPPB produced a decrease of phospho-AKT in the PFC compared to controls. This study revealed that a mGlu5 positive allosteric modulator was effective to alleviate PPI deficits and striatal DAD2 signaling in the NQ model of SZ.

Novel Geometries for Stereotactic Localizers.

INTRODUCTION: The N-localizer is generally utilized in a 3-panel or, more rarely, a 4-panel system for computing stereotactic positions. However, a stereotactic frame that incorporates a 2-panel (bipanel) N-localizer system with panels affixed to only the left and right sides of the frame offers several advantages: improved ergonomics to attach the panels, reduced claustrophobia for the patient, mitigation of posterior panel contact with imaging systems, and reduced complexity. A bipanel system that comprises two standard N-localizer panels yields only two three-dimensional (3D) coordinates, which are insufficient to solve for the stereotactic matrix without further information. While additional information to determine the stereotactic positions could include scalar distances from Digital Imaging and Communications in Medicine (DICOM) metadata or 3D regression across the imaging volume, both have risks related to noise and error propagation. Therefore, we sought to develop new stereotactic localizers that comprise only lateral fiducials (bipanel) that leave the front and back regions of the patient accessible but that contain enough information to solve for the stereotactic matrix using each image independently.  Methods: To solve the stereotactic matrix, we assumed the need to compute three or more 3D points from a single image. Several localizer options were studied using Monte Carlo simulations to understand the effect of errors on the computed target location. The simulations included millions of possible combinations for computing the stereotactic matrix in the presence of random errors of 1mm magnitude. The matrix then transformed coordinates for a target that was placed 50mm anterior, 50mm posterior, 50mm lateral, or 50mm anterior and 50mm lateral to the centre of the image. Simulated cross-sectional axial images of the novel localizer systems were created and converted into DICOM images representing computed tomography (CT) images.  Results: Three novel models include the M-localizer, F-localizer, and Z-localizer. For each of these localizer systems, optimized results were obtained using an overdetermined system of equations made possible by more than three diagonal bars. In each case, the diagonal bar position was computed using standard N-localizer mathematics. Additionally, the M-localizer allowed adding a computation using the Sturm-Pastyr method. Monte Carlo simulation demonstrated that the Z-localizer provided optimal results. CONCLUSION: The three proposed novel models meet our design objectives. Of the three, the Z-localizer produced the least propagation of error. The M-localizer was simpler and had slightly more error than the Z-localizer. The F-localizer produced more error than either the Z-localizer or M-localizer. Further study is needed to determine optimizations using these novel models.

The V-Localizer for Stereotactic Guidance.

Image-guidance for frame-based stereotaxis is facilitated by incorporating three to four N-localizers or Sturm-Pastyr localizers into a stereotactic frame. An extant frame that incorporates only two N-localizers violates the fundamental principle of the N-localizer, which requires three non-colinear points to define a plane in three-dimensional space. Hence, this two N-localizer configuration is susceptible to error. The present article proposes the V-localizer that comprises multiple diagonal bars to provide four or more non-colinear points to minimize error.

Transgenerational evidence of increases in dopamine D2 receptor sensitivity in rodents: Impact on sensorimotor gating, the behavioral response to nicotine and BDNF.

BACKGROUND/AIMS: Neonatal quinpirole (NQ) treatment to rats increases dopamine D2 (DAD2) receptor sensitivity in adult animals. We investigated if increased DAD2 sensitivity would be passed to the next (F1) generation, and if these animals demonstrated sensorimotor gating deficits and enhanced behavioral responses to nicotine. METHODS: Male and female rats were intraperitoneal (IP) administered quinpirole (1 mg/kg) or saline (NS) from postnatal day (P)1-21. Animals were either behaviorally tested (F0) or raised to P60 and mated, creating F1 offspring. RESULTS: Experiment 1 revealed that F1 generation animals that were the offspring of at least one NQ-treated founder increased yawning behavior, a DAD2-mediated behavioral event, in response to acute quinpirole (0.1 mg/kg). F1 generation rats also demonstrated increased striatal ß arrestin-2 and decreased phospho-AKT signaling, consistent with increased G-protein independent DAD2 signaling, which was equal to F0 NQ-treated founders, although this was not observed in all groups. RNA-Seq analysis revealed significant gene expression changes in the F1 generation that were offspring of both NQ-treated founders compared to F0 NQ founders and controls, with enrichment in sensitivity to stress hormones and cell signaling pathways. In Experiment 2, all F1 generation offspring demonstrated sensorimotor gating deficits compared to controls, which were equivalent to F0 NQ-treated founders. In Experiment 3, all F1 generation animals demonstrated enhanced nicotine behavioral sensitization and nucleus accumbens (NAcc) brain-derived neurotrophic factor (BDNF) protein. Further, F1 generation rats demonstrated enhanced adolescent nicotine conditioned place preference equivalent to NQ-treated founders conditioned with nicotine. CONCLUSIONS: This represents the first demonstration of transgenerational effects of increased DAD2 sensitivity in a rodent model.

Restoration of Noradrenergic Function in Parkinson's Disease Model Mice.

Dysfunction of the central noradrenergic and dopaminergic systems is the primary neurobiological characteristic of Parkinson's disease (PD). Importantly, neuronal loss in the locus coeruleus (LC) that occurs in early stages of PD may accelerate progressive loss of dopaminergic neurons. Therefore, restoring the activity and function of the deficient noradrenergic system may be an important therapeutic strategy for early PD. In the present study, the lentiviral constructions of transcription factors Phox2a/2b, Hand2 and Gata3, either alone or in combination, were microinjected into the LC region of the PD model VMAT2 Lo mice at 12 and 18 month age. Biochemical analysis showed that microinjection of lentiviral expression cassettes into the LC significantly increased mRNA levels of Phox2a, and Phox2b, which were accompanied by parallel increases of mRNA and proteins of dopamine ß-hydroxylase (DBH) and tyrosine hydroxylase (TH) in the LC. Furthermore, there was considerable enhancement of DBH protein levels in the frontal cortex and hippocampus, as well as enhanced TH protein levels in the striatum and substantia nigra. Moreover, these manipulations profoundly increased norepinephrine and dopamine concentrations in the striatum, which was followed by a remarkable improvement of the spatial memory and locomotor behavior. These results reveal that over-expression of these transcription factors in the LC improves noradrenergic and dopaminergic activities and functions in this rodent model of PD. It provides the necessary groundwork for the development of gene therapies of PD, and expands our understanding of the link between the LC-norepinephrine and dopamine systems during the progression of PD.

Monte Carlo Simulation of Errors for N-localizer Systems in Stereotactic Neurosurgery: Novel Proposals for Improvements.

INTRODUCTION:  Frame-based stereotaxis has been widely utilized for precise neurosurgical procedures throughout the world for nearly 40 years. The N-localizer is an integral component of most of the extant systems. Analysis of targeting errors related to the N-localizer has not been carried out in sufficient detail. We highlight these potential errors and develop methods to reduce them.  Methods: N-localizer systems comprising three and four N-localizers of various geometries were analyzed using Monte Carlo (MC) simulations. The simulations included native and altered geometric dimensions (Width [W] x Height [H]). Errors were computed using the MC simulations that included the x- and y-axes of vertically oriented rods, that altered the W/H ratio, and that added a fourth N-localizer to a three N-localizer system.  Results: The inclusion of an overdetermined system of equations and the geometries of the N-localizer systems had significant effects on target errors. Root Mean Square Errors (RMS-e) computed via millions of MC iterations for each study demonstrated that errors were reduced by (1) inclusion of the x- and y-coordinates of the vertically oriented rods, (2) a greater triangular area enclosed by the diagonal fiducials of the N-localizer system (stereotactic triangle), (3) a larger W/H ratio, and (4) an N-localizer system that comprised four N-localizers. CONCLUSION: Monte Carlo simulations of Root Mean Square error (RMS-e) is a useful technique to understand targeting while using N-localizer systems in stereotactic neurosurgery. The application of vertical rod positions enhances computational accuracy and can be performed on any N-localizer system. Keeping the target point within the stereotactic triangle enclosed by the diagonal rods can also reduce errors. Additional optimizations of N-localizer geometry may also reduce potential targeting errors. Further analysis is needed to confirm these findings which may have clinical importance.

The effects of a novel inhibitor of tumor necrosis factor (TNF) alpha on prepulse inhibition and microglial activation in two distinct rodent models of schizophrenia.

Increased neuroinflammation has been shown in individuals diagnosed with schizophrenia (SCHZ). This study evaluated a novel immune modulator (PD2024) that targets the pro-inflammatory cytokine tumor necrosis factor-alpha (TNFα) to alleviate sensorimotor gating deficits and microglial activation employing two different rodent models of SCHZ. In Experiment 1, rats were neonatally treated with saline or the dopamine D2-like agonist quinpirole (NQ; 1 mg/kg) from postnatal day (P) 1-21 which produces increases of dopamine D2 receptor sensitivity throughout the animal's lifetime. In Experiment 2, rats were neonatally treated with saline or the immune system stimulant polyinosinic:polycytidylic acid (Poly I:C) from P5-7. Neonatal Poly I:C treatment mimics immune system activation associated with SCHZ. In both experiments, rats were raised to P30 and administered a control diet or a novel TNFα inhibitor PD2024 (10 mg/kg) in the diet from P30 until P67. At P45-46 and from P60-67, animals were behaviorally tested on auditory sensorimotor gating as measured through prepulse inhibition (PPI). NQ or Poly I:C treatment resulted in PPI deficits, and PD2024 treatment alleviated PPI deficits in both models. Results also revealed that increased hippocampal and prefrontal cortex microglial activation produced by neonatal Poly I:C was significantly reduced to control levels by PD2024. In addition, a separate group of animals neonatally treated with saline or Poly I:C from P5-7 demonstrated increased TNFα protein levels in the hippocampus but not prefrontal cortex, verifying increased TNFα in the brain produced by Poly I:C. Results from this study suggests that that brain TNFα is a viable pharmacological target to treat the neuroinflammation known to be associated with SCHZ.