Levels of Perineuronal Nets in the Basolateral Amygdala Are Correlated with Sex Differences in Fear Learning

Thesis advisor: John P. ChristiansonTrauma and exposure to extreme stressors greatly increases a person’s vulnerability to developing mental illnesses like post-traumatic stress disorder (PTSD). Patients with PTSD often have impaired fear and safety learning, and despite the fact that women are more than twice as likely to develop PTSD, much of the research on this disorder has relied on the use of male subjects. This paper will review potential contributors to the sex differences seen in PTSD and fear-related learning. Our group has found that female rats show greater fear discrimination abilities than their male counterparts, but show no difference in levels of safety learning. Analysis of specialized extracellular matrix structures called perineuronal nets (PNNs) revealed that females displayed a much higher density of PNNs in the basolateral amygdala (BLA) than males, but not in the prefrontal cortex (PFC).Thesis (BS) — Boston College, 2017.Submitted to: Boston College. College of Arts and Sciences.Discipline: Departmental Honors.Discipline: Psychology

Die Effekte einer Ärzte-Kurzschulung zur Raucherentwöhnung in einer pneumologischen Akutklinik eines Universitätsklinikums

Objective: The objective was to evaluate the effect of a short physician training in smoking cessation on the physicians’ performance of smoking cessation interventions. The effects on patients’ cessation rates were analyzed as well. A further aim was to identify barriers for providing cessation interventions. The study was conducted in an acute care pulmonology department of a German university hospital. Methods: 24 physicians of the pulmonology department of a German university hospital received a two-hour training in smoking cessation. 109 pre- and 89 post-training group patients were compared with regard to the frequencies of received smoking cessation interventions (Ask, Advise, Assist) and three- and six-month abstinence rates. Physicians estimated their intervention frequencies and gave reasons for not providing cessation interventions. Results: In a multivariable analysis (p<0.05), the physicians’ application of “Ask” (OR 3.28, 95% CI 1.13–9.53) and the six-month abstinence rates (OR 2.70, 95% CI 1.24–5.84) were significantly higher in the post-training group. The univariate analysis also showed a significant effect on “Assist” (OR 2.05, 95% CI 1.09–3.87). No significant effect was seen on “Advise to quit”. Physicians overestimated their intervention frequencies and reported the patients’ low motivation to stop, an oncological disease and palliative care situation as barriers to performing smoking cessation. Conclusion: A short physician training in a hospital department of pulmonology increases the use of guideline-based cessation strategies and may improve cessation rates. The findings show that hospital-based strategies such as physician trainings could be useful in the improvement of smoking cessation. Strategies for overcoming barriers for providing smoking cessation interventions are needed.Ziel: Das Ziel war die Evaluation der Effekte einer Ärzte-Kurzschulung zur Raucherentwöhnung auf die Anwendungshäufigkeit der Entwöhnungsstrategien durch die Ärzte. Ebenso wurden die Effekte auf die Abstinenzraten der Patienten untersucht. Außerdem sollten die Barrieren für die Durchführung von Raucherentwöhnungsmaßnahmen ermittelt werden. Die Studie wurde in einer pneumologischen Akutklinik eines deutschen Universitätsklinikums durchgeführt. Methoden: 24 Ärzte erhielten eine zweistündige Schulung zur Raucherentwöhnung. 109 Kontrollgruppenpatienten, die vor der Schulung in der Klinik behandelt wurden, wurden mit 89 nach der Schulung behandelten Studiengruppenpatienten hinsichtlich der Häufigkeit der ärztlichen Raucherentwöhnungsinterventionen (Ask, Advise, Assist) und ihrer Abstinenzraten drei und sechs Monate nach Klinikaufenthalt verglichen. Die Ärzte schätzten ihre Interventionshäufigkeiten und gaben Gründe an, warum sie in manchen Fällen keine Entwöhnungsinterventionen durchführten. Ergebnisse: Die Anwendung von „Ask“ (OR 3.28, 95% KI 1.13–9.53) durch die Ärzte und die Sechsmonats-Abstinenzraten (OR 2.70, 95% KI 1.24–5.84) waren in der Studiengruppe im multivariablen Modell signifikant höher (p<0.05). Bei „Assist“ zeigte nur das univariable Modell einen signifikanten Einfluss der Schulung (OR 2.05, 95% KI 1.09–3.87). Es wurde kein signifikanter Effekt der Schulung auf die Anwendung von „Advise“ festgestellt. Die Ärzte überschätzten ihre Interventionshäufigkeiten und nannten als Gründe für die Nicht-Durchführung einer Raucherentwöhnung eine wahrgenommene geringe Aufhörmotivation der Patienten und das Vorliegen einer onkologischen oder palliativen Krankheitssituation. Fazit: Eine Ärztekurzschulung zur Raucherentwöhnung in einer pneumologischen Akutklinik erhöht den Einsatz von leitlinienbasierten Raucherentwöhnungsstrategien und könnte einen positiven Effekt auf die Abstinenzraten haben. Implikationen: Die Ergebnisse zeigen, dass Krankenhaus-basierte Strategien wie Ärzteschulungen zur Verbesserung der Raucherentwöhnung nützlich sein können. Strategien zur Überwindung von Hindernissen für die Bereitstellung von Maßnahmen zur Raucherentwöhnung sind erforderlich

Cigarette smoke and electronic cigarettes differentially activate bronchial epithelial cells

Background: The use of electronic cigarettes (ECIGs) is increasing, but the impact of ECIG-vapor on cellular processes like inflammation or host defense are less understood. The aim of the present study was to compare the acute effects of traditional cigarettes (TCIGs) and ECIG-exposure on host defense, inflammation, and cellular activation of cell lines and primary differentiated human airway epithelial cells (pHBE). Methods: We exposed pHBEs and several cell lines to TCIG-smoke or ECIG-vapor. Epithelial host defense and barrier integrity were determined. The transcriptome of airway epithelial cells was compared by gene expression array analysis. Gene interaction networks were constructed and differential gene expression over all groups analyzed. The expression of several candidate genes was validated by qRT-PCR. Results: Bacterial killing, barrier integrity and the expression of antimicrobial peptides were not affected by ECIG-vapor compared to control samples. In contrast, TCIGs negatively affected host defense and reduced barrier integrity in a significant way. Furthermore ECIG-exposure significantly induced IL-8 secretion from Calu-3 cells but had no effect on NCI-H292 or primary cells. The gene expression based on array analysis distinguished TCIG-exposed cells from ECIG and room air-exposed samples. Conclusion: The transcriptome patterns of host defense and inflammatory genes are significantly distinct between ECIG-exposed and TCIG-treated cells. The overall effects of ECIGs on epithelial cells are less in comparison to TCIG, and ECIG-vapor does not affect host defense. Nevertheless, although acute exposure to ECIG-vapor induces inflammation, and the expression of S100 proteins, long term in vivo data is needed to evaluate the chronic effects of ECIG use

Calcifediol-loaded liposomes for local treatment of pulmonary bacterial infections.

The influence of vitamin D3 and its metabolites calcifediol (25(OH)D) and calcitriol on immune regulation and inflammation is well described, and raises the question of potential benefit against bacterial infections. In the current study, 25(OH)D was encapsulated in liposomes to enable aerosolisation, and tested for the ability to prevent pulmonary infection by Pseudomonas aeruginosa. Prepared 25(OH)D-loaded liposomes were nanosized and monodisperse, with a negative surface charge and a 25(OH)D entrapment efficiency of approximately 23%. Jet nebulisation of liposomes was seen to yield an aerosol suitable for tracheo-bronchial deposition. Interestingly, 25(OH)D in either liposomes or ethanolic solution had no effect on the release of the proinflammatory cytokine KC from Pseudomonas-infected murine epithelial cells (LA-4); treatment of infected, human bronchial 16-HBE cells with 25(OH)D liposomes however resulted in a significant reduction in bacterial survival. Together with the importance of selecting an application-appropriate in vitro model, the current study illustrates the feasibility and practicality of employing liposomes as a means to achieve 25(OH)D lung deposition. 25(OH)D-loaded liposomes further demonstrated promising effects regarding prevention of Pseudomonas infection in human bronchial epithelial cells

Sex-specific associations of comorbidome and pulmorbidome with mortality in chronic obstructive pulmonary disease : results from COSYCONET

In patients with COPD, it has not been comprehensively assessed whether the predictive value of comorbidities for mortality difers between men and women. We therefore aimed to examine sex diferences of COPD comorbidities in regard with prognosis by classifying comorbidities into a comorbidome related to extrapulmonary disorders and a pulmorbidome, referring to pulmonary disorders. The study population comprised 1044 women and 1531 men with the diagnosis of COPD from COSYCONET, among them 2175 of GOLD grades 1–4 and 400 at risk. Associations of comorbidities with mortality were studied using Cox regression analysis for men and women separately. During the follow-up (median 3.7 years) 59 women and 159 men died. In men, obesity, hypertension, coronary artery disease, liver cirrhosis, osteoporosis, kidney disease, anaemia and increased heart rate (HR) predict mortality, in women heart failure, hyperuricemia, mental disorders, kidney disease and increased HR (p< 0.05 each). Regarding the pulmorbidome, signifcant predictors in men were impairment in difusion capacity and hyperinfation, in women asthma and hyperinfation. Similar results were obtained when repeating the analyses in GOLD 1–4 patients only. Gender diferences should be considered in COPD risk assessment for a tailored approach towards the treatment of COPD

Large-scale validation of miRNAs by disease association, evolutionary conservation and pathway activity.

The validation of microRNAs (miRNAs) identified by next generation sequencing involves amplification-free and hybridization-based detection of transcripts as criteria for confirming valid miRNAs. Since respective validation is frequently not performed, miRNA repositories likely still contain a substantial fraction of false positive candidates while true miRNAs are not stored in the repositories yet. Especially if downstream analyses are performed with these candidates (e.g. target or pathway prediction), the results may be misleading. In the present study, we evaluated 558 mature miRNAs from miRBase and 1,709 miRNA candidates from next generation sequencing experiments by amplification-free hybridization and investigated their distributions in patients with various disease conditions. Notably, the most significant miRNAs in diseases are often not contained in the miRBase. However, these candidates are evolutionary highly conserved. From the expression patterns, target gene and pathway analyses and evolutionary conservation analyses, we were able to shed light on the complexity of miRNAs in humans. Our data also highlight that a more thorough validation of miRNAs identified by next generation sequencing is required. The results are available in miRCarta ( https://mircarta.cs.uni-saarland.de )

miR449 Protects Airway Regeneration by Controlling AURKA/HDAC6-Mediated Ciliary Disassembly

Airway mucociliary regeneration and function are key players for airway defense and are impaired in chronic obstructive pulmonary disease (COPD). Using transcriptome analysis in COPD-derived bronchial biopsies, we observed a positive correlation between cilia-related genes and microRNA-449 (miR449). In vitro, miR449 was strongly increased during airway epithelial mucociliary differentiation. In vivo, miR449 was upregulated during recovery from chemical or infective insults. miR0449-/- mice (both alleles are deleted) showed impaired ciliated epithelial regeneration after naphthalene and Haemophilus influenzae exposure, accompanied by more intense inflammation and emphysematous manifestations of COPD. The latter occurred spontaneously in aged miR449-/- mice. We identified Aurora kinase A and its effector target HDAC6 as key mediators in miR449-regulated ciliary homeostasis and epithelial regeneration. Aurora kinase A is downregulated upon miR449 overexpression in vitro and upregulated in miR449-/- mouse lungs. Accordingly, imaging studies showed profoundly altered cilia length and morphology accompanied by reduced mucociliary clearance. Pharmacological inhibition of HDAC6 rescued cilia length and coverage in miR449-/- cells, consistent with its tubulin-deacetylating function. Altogether, our study establishes a link between miR449, ciliary dysfunction, and COPD pathogenesis

miR449 Protects Airway Regeneration by Controlling AURKA/HDAC6-Mediated Ciliary Disassembly

Airway mucociliary regeneration and function are key players for airway defense and are impaired in chronic obstructive pulmonary disease (COPD). Using transcriptome analysis in COPD-derived bronchial biopsies, we observed a positive correlation between cilia-related genes and microRNA-449 (miR449). In vitro, miR449 was strongly increased during airway epithelial mucociliary differentiation. In vivo, miR449 was upregulated during recovery from chemical or infective insults. miR0449−/− mice (both alleles are deleted) showed impaired ciliated epithelial regeneration after naphthalene and Haemophilus influenzae exposure, accompanied by more intense inflammation and emphysematous manifestations of COPD. The latter occurred spontaneously in aged miR449−/− mice. We identified Aurora kinase A and its effector target HDAC6 as key mediators in miR449-regulated ciliary homeostasis and epithelial regeneration. Aurora kinase A is downregulated upon miR449 overexpression in vitro and upregulated in miR449−/− mouse lungs. Accordingly, imaging studies showed profoundly altered cilia length and morphology accompanied by reduced mucociliary clearance. Pharmacological inhibition of HDAC6 rescued cilia length and coverage in miR449−/− cells, consistent with its tubulin-deacetylating function. Altogether, our study establishes a link between miR449, ciliary dysfunction, and COPD pathogenesis

Expression of human beta-defensins 1 and 2 in kidneys with chronic bacterial infection

BACKGROUND: Constitutive expression and localization of antimicrobial human β-defensin-1 (HBD-1) in human kidneys as a potential mechanism of antimicrobial defense has been previously reported. Inducible expression of human β-defensin-2 (HBD-2) has been described in various epithelial organs but not for the urogenital tract. METHODS: We investigated the gene- and protein expression of HBD-1 and HBD-2 by reverse transcriptase-polymerase chain reaction, and immunohistochemistry in 15 normal human kidney samples and 15 renal tissues with chronic bacterial infection. Additionally, cell culture experiments were performed to study HBD gene expression by real-time RT-PCR in response to inflammatory cytokines TNFα and IL-1β as well as lipopolysaccharide from Gram-negative bacteria. RESULTS: Constitutive HBD-1 gene- and protein expression was detected in normal renal tissue and kidneys with chronic infection. As a novel finding, inducible HBD-2 gene- and protein expression was demonstrated in tubulus epithelia with chronic infection but not in normal renal tissue. In pyelonephritic kidneys HBD-1 and HBD-2 expression showed a similar pattern of localizaton in distal tubules, loops of Henle and in collecting ducts of the kidney. Furthermore, real-time RT-PCR of kidney derived cell lines stimulated with inflammatory agents TNF-α, IL-1β and LPS revealed a strong increase in relative HBD-2 transcription level and also a slight increase in relative HBD-1 transcription level. CONCLUSIONS: Upregulated HBD-2 expression in renal tubulus epithelium indicates a role of a wider range of human defensins for antimicrobial host defense in the urogenital tract than previously recognized