CDKL5 expression is modulated during neuronal development and its subcellular distribution is tightly regulated by the C-terminal tail

Mutations in the human X-linked cyclin-dependent kinase-like 5 (CDKL5) gene have been identified in patients with Rett syndrome (RTT), West syndrome, and X-linked infantile spasms, sharing the common feature of mental retardation and early seizures. CDKL5 is a rather uncharacterized kinase, but its involvement in RTT seems to be explained by the fact that it works upstream of MeCP2, the main cause of Rett syndrome. To understand the role of this kinase for nervous system functions and to address if molecular mechanisms are involved in regulating its distribution and activity, we studied the ontogeny of CDKL5 expression in developing mouse brains by immunostaining and Western blotting. The expression profile of CDKL5 was compared with that of MeCP2. The two proteins share a general expression profile in the adult mouse brain, but CDKL5 levels appear to be highly modulated at the regional level. Its expression is strongly induced in early postnatal stages, and in the adult brain CDKL5 is present in mature neurons, but not in astroglia. Interestingly, the presence of CDKL5 in the cell nucleus varies at the regional level of the adult brain and is developmentally regulated. CDKL5 shuttles between the cytoplasm and the nucleus and the C-terminal tail is involved in localizing the protein to the cytoplasm in a mechanism depending on active nuclear export. Accordingly, Rett derivatives containing disease-causing truncations of the C terminus are constitutively nuclear, suggesting that they might act as gain of function mutations in this cellular compartment

Genome Sequence of Magnetospirillum magnetotacticum Strain MS-1

Here, we report the genome sequence of Magnetospirillum magnetotacticum strain MS-1, which consists of of 36 contigs and 4,136 protein-coding genes

Lying about intentions

This thesis aims to explore whether lying about intentions can be detected using the same methods that are used to detect other types of lie. Part I, the introduction, contains a literature review of research into future thinking and intentions. In Part II non-verbal cues and thermal imaging are investigated as cues to deception. In Experiment 1 participants of twelve different ethnicities were interviewed at an airport about their forthcoming trip. Half of the participants were instructed to answer truthfully, the other half to answer deceptively. The participants’ veracity affected only three of the 17 non-verbal cues: struggling, thinking hard and controlling behaviour. These cues were reduced to two factors, cognitive load and animation. These two factors could correctly classify 52.3% of truth tellers and 62.6% of liars. The thermal imaging showed that liars tended to get warmer during the interview, while the truth tellers’ temperatures remained constant. This temperature rise was a cue to deception at above chance rates. In Part III two experiments are described, investigating verbal cues to deceit in longer and shorter interviews. In Experiment 2 participants were interviewed, answering truthfully or deceptively, about a forthcoming trip. The interview contained general questions that were expected by the participants and three types of unexpected questions. Half of the participants had been to the travel destination previously, while the other half had not been there. The results showed that truth tellers generated more details on the unexpected questions than liars, while liars provided more details on expected questions than truth tellers. This pattern was more pronounced with participants who had been to their travel destination before than for those who had never been to that destination. A positive predictive value of .69 (truth) and a negative predictive value of .75 (lie) were obtained. Experiment 3 investigated verbal cues to deceit in a short one question interview about a forthcoming trip. The interview question was asked in two forms, one prompting for time detail and one control. The results showed that truth tellers and those answering the time prompting question were more likely to provide temporal information. The results showed that 47.6% of truth tellers and 81% of liars could be classified correctly on the basis of the time prompt question. Part IV investigated reaction times as a cue to deception. In Experiment 4, participants were asked to perform a mission, either immediately or two weeks later. Half of the participants was instructed to answer any questions asked by experimenters en route truthfully, the other half was asked to answer deceptively. Using a computer, participants classified adjectives as positive or negative and were primed by words related to the real intention of truth tellers (to buy a present), which was also the cover story for liars. Contrary to the hypothesis, in the immediate condition liars answered faster on positive than negative adjectives, while the truth tellers responded equally fast to both adjectives. In the delayed condition no difference was found between truth tellers and liars. Part V presents the general discussion. The main findings of the thesis are summarised, practical and theoretical implications are discussed, the methodological and ethical limitations are considered and ideas for future research are given. Overall, the conclusion of the thesis is that false intentions contain cues to deceit that observers should be able to spot.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Antibacterial effects of two synthetic peptides against Enterococcus faecalis biofilms: A preliminary in vitro study

Aim: Current endodontic techniques are unable to fully eradicate intracanal bacteria. Thus, new agents that effectively eliminate endodontic pathogens are needed. The aim of this study was to assess the antibacterial properties of two synthetic peptides, namely KP and L18R, against planktonic cells and biofilms of the endodontic pathogen Enterococcus faecalis. Methodology: KP and L18R bactericidal activity against E. faecalis ATCC 29212 was evaluated by colony forming unit assays and the half maximal effective concentration (EC50) was calculated. The effect of peptides on E. faecalis biofilm formation onto polystyrene plates was also assessed by the crystal violet assay. Confocal laser scanning microscopy (CLSM) analysis was carried out to compare the effects of KP, L18R and a Ca(OH)2 saturated solution in an in vitro model of dental infection consisting in 2-day-old E. faecalis biofilms grown on hydroxyapatite disks. Results: Both KP and L18R showed strong bactericidal activity against planktonic E. faecalis. L18R proved to be 10-folds more ef fective than KP (KP and L18R EC50 values=4.520×10-6 M and 3.624×10-7 M, respectively). Peptides inhibited E. faecalis biofilm formation in a dose-dependent manner and L18R resulted more effectivethan KP. CLSM images showed that Ca(OH)2, KP and L18R remarkably impaired E. faecal is biof i lms pre -grown on hydroxyapatite. Conclusions: KP and L18R effectively inhibited E. faecalis, both in planktonic and biofilm form. L18R demonstrated a more potent antibacterial activity than KP. These preliminary results suggest that antimicrobial peptides may represent a promising new strategy for endodontic infection control

Activity of two antimicrobial peptides against Enterococcus faecalis in a model of biofilm-mediated endodontic infection

Enterococcus faecalis is a common cause of biofilm-associated opportunistic infections, which are often difficult to treat. The formation of E. faecalis biofilms on the dentinal walls of the root canal is frequently the cause of endodontic treatment failure and secondary apical periodontitis. In a preliminary work, two recognized antifungal peptides, KP and L18R, showed antibacterial activity against planktonic E. faecalis cells at micromolar concentrations. Moreover, L18R proved to reduce the biomass in the early stage of E. faecalis biofilm development on polystyrene plates, while a qualitative biofilm inhibition was demonstrated on hydroxyapatite disks by confocal laser scanning microscopy (CLSM). The aim of this study was to better characterize the effect of both peptides on E. faecalis biofilm. A reduction in metabolic activity after peptide treatment was detected by Al-amar Blue assay, while a remarkable impairment in the architecture of E. faecalis biofilms on hy-droxyapatite disks, along with a significant reduction in viable bacteria, was caused mostly by L18R, as assessed by CLSM and scanning electron microscopy. The lack of cytotoxicity of the investigated peptides against L929 murine fibroblasts was also determined. Obtained results suggest L18R as a promising candidate for the development of new strategies for endodontic infection control

Negative serology of Fasciola hepatica infection in patients with liver cancer in Peru: A preliminary report

Introduction: The etiology of several hepatocellular carcinoma (HCC) cases remains largely unknown. Although Fasciola hepatica has been associated with liver fibrosis in Latin America, it has not yet been associated with HCC. This study aimed to determine the existence of specific IgG antibodies against F. hepatica in the serum samples of HCC patients. Methods: In total, 13 serum samples from 13 HCC patients were screened using Fas2-ELISA. Results: Fas2-ELISA demonstrated negative results in all HCC patients included in this study. Conclusions: The pre-existence of F. hepatica infection in HCC patients needs to be further investigated in epidemiological and experimental studies

Development of reverse genetics systems for infectious bursal disease virus by yeast-based homologous recombination.

Projeto/Plano de Ação: 05.11.10.200-04