The neurotoxic effects of enzyme inhibitors at the neuromuscular junction

Current knowledge of the long-term, low dose effects of carbamate (CB) anti-cholinesterases on skeletal muscle or on the metabolism and regulation of the molecular forms of acetylcholinesterase (AChE) is limited. This is largely due to the reversible nature of these inhibitors and the subtle effects they induce which has generally made their study difficult and preliminary investigations were conducted to determine suitable study methods. A sequential extraction technique was used to rapidly analyse AChE molecular form activity at the mouse neuromuscular junction and also in peripheral parts of muscle fibres. AChE in the synaptic cleft involved in the termination of cholinergic transmission was successfully assessed by the assay method and by an alternative method using a correlation equation which represented the relationship between synaptic AChE and the prolongation of extra-cellular miniature endplate potentials. It was found that inhibition after in vivo Carbamate (CB) dosing could not be maintained during tissue analysis because CB-inhibited enzyme complexes decarbamoylated vary rapidly and could not be prevented even when maintained on ice. The methods employed did not therefore give a measure of inhibition but presented a profile of metabolic responses to continual, low dose CB treatment. Repetitive and continual infusion with low doses of the CBs: pyridostigmine and physostigmine induced a variety of effects on mouse skeletal muscle. Both compounds induced a mild myopathy in the mouse diaphragm during continual infusion which was characterised by endplate deformation without necrosis; such deformation persisted on termination of treatment but had recovered slightly 14 days later. Endplate and non-endplate AChE molecular forms displayed selective responses to CB treatment. During treatment endplate AChE was reduced whereas non-endplate AChE was largely unaffected, and after treatment, endplate AChE recovered, whereas non-endplate AChE was up-regulated. The mechanisms by which these responses become manifest are unclear but may be due to CB-induced effects on nerve-mediated muscle activity, neurotrophic factors or morphological and physiological changes which arise at the neuromuscular junction. It was concluded that, as well as inhibiting AChE, CBs also influence the metabolism and regulation of the enzyme and induce persistent endplate deformation; possible detrimental effects of long-term, low-dose determination requires further investigation.

Cost-effective upgrade of WDM all-optical networks using overlay fibres and hop reduction links

Fibre overlay is a cost-effective technique to alleviate wavelength blocking in some links of a wavelength-routed optical network by increasing the number of wavelengths in those links. In this letter, we investigate the effects of overlaying fibre in an all-optical network (AON) based on GÉANT2 topology. The constraint-based routing and wavelength assignment (CB-RWA) algorithm locates where cost-efficient upgrades should be implemented. Through numerical examples, we demonstrate that the network capacity improves by 25 per cent by overlaying fibre on 10 per cent of the links, and by 12 per cent by providing hop reduction links comprising 2 per cent of the links. For the upgraded network, we also show the impact of dynamic traffic allocation on the blocking probability. Copyright © 2010 John Wiley & Sons, Ltd.

Towards agility in design in global component-based development

The potential benefits of implementing Component-Based Development (CBD) methodologies in a globally distributed environment are many. Lessons from the aeronautics, automotive, electronics and computer hardware industries, in which Component-Based (CB) architectures have been successfully employed for setting up globally distributed design and production activities, have consistently shown that firms have managed to increase the rate of reused components and sub-assemblies, and to speed up the design and production process of new products.

Cost-effective upgrade of WDM all-optical networks using overlay fibres and hop reduction links

Fibre overlay is a cost-effective technique to alleviate wavelength blocking in some links of a wavelength-routed optical network by increasing the number of wavelengths in those links. In this letter, we investigate the effects of overlaying fibre in an all-optical network (AON) based on GÉANT2 topology. The constraint-based routing and wavelength assignment (CB-RWA) algorithm locates where cost-efficient upgrades should be implemented. Through numerical examples, we demonstrate that the network capacity improves by 25 per cent by overlaying fibre on 10 per cent of the links, and by 12 per cent by providing hop reduction links comprising 2 per cent of the links. For the upgraded network, we also show the impact of dynamic traffic allocation on the blocking probability. Copyright © 2010 John Wiley & Sons, Ltd.

Functional recombinant protein is present in the pre-induction phases of Pichia pastoris cultures when grown in bioreactors, but not shake-flasks

Background - Pichia pastoris is a widely-used host for recombinant protein production; expression is typically driven by methanol-inducible alcohol oxidase (AOX) promoters. Recently this system has become an important source of recombinant G protein-coupled receptors (GPCRs) for structural biology and drug discovery. The influence of diverse culture parameters (such as pH, dissolved oxygen concentration, medium composition, antifoam concentration and culture temperature) on productivity has been investigated for a wide range of recombinant proteins in P. pastoris. In contrast, the impact of the pre-induction phases on yield has not been as closely studied. In this study, we examined the pre-induction phases of P. pastoris bioreactor cultivations producing three different recombinant proteins: the GPCR, human A2a adenosine receptor (hA2aR), green fluorescent protein (GFP) and human calcitonin gene-related peptide receptor component protein (as a GFP fusion protein; hCGRP-RCP-GFP). Results - Functional hA2aR was detected in the pre-induction phases of a 1 L bioreactor cultivation of glycerol-grown P. pastoris. In a separate experiment, a glycerol-grown P. pastoris strain secreted soluble GFP prior to methanol addition. When glucose, which has been shown to repress AOX expression, was the pre-induction carbon source, hA2aR and GFP were still produced in the pre-induction phases. Both hA2aR and GFP were also produced in methanol-free cultivations; functional protein yields were maintained or increased after depletion of the carbon source. Analysis of the pre-induction phases of 10 L pilot scale cultivations also demonstrated that pre-induction yields were at least maintained after methanol induction, even in the presence of cytotoxic concentrations of methanol. Additional bioreactor data for hCGRP-RCP-GFP and shake-flask data for GFP, horseradish peroxidase (HRP), the human tetraspanins hCD81 and CD82, and the tight-junction protein human claudin-1, demonstrated that bioreactor but not shake flask cultivations exhibit recombinant protein production in the pre-induction phases of P. pastoris cultures. Conclusions - The production of recombinant hA2aR, GFP and hCGRP-RCP-GFP can be detected in bioreactor cultivations prior to methanol induction, while this is not the case for shake-flask cultivations of GFP, HRP, hCD81, hCD82 and human claudin-1. This confirms earlier suggestions of leaky expression from AOX promoters, which we report here for both glycerol- and glucose-grown cells in bioreactor cultivations. These findings suggest that the productivity of AOX-dependent bioprocesses is not solely dependent on induction by methanol. We conclude that in order to maximize total yields, pre-induction phase cultivation conditions should be optimized, and that increased specific productivity may result in decreased biomass yields.

Target/error overlap in jargonaphasia:the case for a one-source model, lexical and non-lexical summation, and the special status of correct responses

We present three jargonaphasic patients who made phonological errors in naming, repetition and reading. We analyse target/response overlap using statistical models to answer three questions: 1) Is there a single phonological source for errors or two sources, one for target-related errors and a separate source for abstruse errors? 2) Can correct responses be predicted by the same distribution used to predict errors or do they show a completion boost (CB)? 3) Is non-lexical and lexical information summed during reading and repetition? The answers were clear. 1) Abstruse errors did not require a separate distribution created by failure to access word forms. Abstruse and target-related errors were the endpoints of a single overlap distribution. 2) Correct responses required a special factor, e.g., a CB or lexical/phonological feedback, to preserve their integrity. 3) Reading and repetition required separate lexical and non-lexical contributions that were combined at output.

Protection issue discussion of DC network development:circuit breaker or fault-tolerant converter

This paper provides a discussion on future direct current (DC) network development in terms of system protection under DC-side fault scenarios. The argument between appropriate DC circuit breaker and new DC fault-tolerant converters is discussed after a review on DC technology development and bottleneck issues that require proper solutions. The overcurrent/cost curve of power-electronic DC circuit breakers (CB) superimposed to voltage-source converter (VSC) systems is derived and compared with other possible fault-tolerant power conversion options. This in-advance planning of protection capability is essential for the future development of DC networks.

g-C3N4/NaTaO3 organic–inorganic hybrid nanocomposite:high-performance and recyclable visible light driven photocatalyst

Novel g-C3N4/NaTaO3 hybrid nanocomposites have been prepared by a facile ultrasonic dispersion method. Our results clearly show the formation of interface between NaTaO3 and g-C3N4 and further loading of g-C3N4 did not affect the crystal structure and morphology of NaTaO3. The g-C3N4/NaTaO3 nanocomposites exhibited enhanced photocatalytic performance for the degradation of Rhodamine B under UV–visible and visible light irradiation compared to pure NaTaO3 and Degussa P25. Interestingly, the visible light photocatalytic activity is generated due to the loading of g-C3N4. A mechanism is proposed to discuss the enhanced photocatalytic activity based on trapping experiments of photoinduced radicals and holes. Under visible light irradiation, electron excited from the valance band (VB) to conduction band (CB) of g-C3N4 could directly inject into the CB of NaTaO3, making g-C3N4/NaTaO3 visible light driven photocatalyst. Since the as-prepared hybrid nanocomposites possess high reusability therefore it can be promising photocatalyst for environmental applications.

Pyrolysis and combustion of municipal solid wastes:evaluation of synergistic effects using TGA-MS

A thermogravimetric methodology was developed to investigate and semi-quantify the extent of synergistic effects during pyrolysis and combustion of municipal solid waste (MSW). Results from TGA-MS were used to compare the pyrolysis and combustion characteristics of single municipal solid waste components (polyvinyl chloride (PVC), polypropylene (PP), polystyrene (PS), branches (BR), leaves (LV), grass (GR), packaging paper (PK), hygienic paper (HP) and cardboard (CB)) and a mixture (MX) of PP, BR and CB. Samples were heated under dynamic conditions at 20°C/min from 25°C to 1000°C with the continuous record of their main evolved fragments. Synergistic effects were evaluated by comparing experimental and calculated weight losses and relative areas of MS peaks. Pyrolysis of the mixture happened in two stages, with the release of H2, CH4, H2O, CO and CO2 between 200 and 415°C and the release of CH4, CxHy, CO and CO2 between 415 and 525°C. Negative synergistic effect in the 1st stage was attributed to the presence of PP where the release of hydrocarbons and CO2 from BR and CB was inhibited, whereas positive synergistic effects were observed during the 2nd degradation stage. In a second part of the study, synergistic effects were related to the dependency of the effective activation energy (Eα) versus the conversion (α). Higher Eαs were obtained for MX during its 1st stage of pyrolysis and lower Eαs for the 2nd stage when compared to the individual components. On the other hand, mostly positive synergistic effects were observed during the combustion of the same mixture, for which lower Eαs were recorded.

The effectiveness of an enhanced cognitive behavioral intervention in improving behavioral outcomes related to HIV prevention

This study analyzed outcomes of an enhanced cognitive-behavioral intervention with dually diagnosed severely mentally ill adults. It specifically addressed the improvement of attitudes, skills, self-efficacy to use condoms and the heightening of condom use. The data were analyzed via a randomized three-group repeated measures design composed of the experimental (E-CB), standard care (SC) comparison or a no-treatment control condition as the between-subjects variable and pre-post measure as the within-subjects variable. The E-CB focused on cooperative, application, hands-on, skill-building and role-playing activities for sexual assertiveness, negotiation in risk-taking and proper condom use. The SC comparison, was didactic in its approach and addressed risk-taking and proper condom use in one session, but did not involve application approaches to problem-solving risky situations or condom use. Multiple assessments were conducted at pre-, post- and six months post-intervention. ^ The analysis indicated that the E-CB intervention led to more favorable attitudes toward condoms and to improved and maintained skills regarding their use by participants six months after the intervention compared to the standard care and control groups. No significant improvements in self-efficacy were found. A repeated measures ANOVA conducted on the transformed values of percentage of vaginal condom use indicated no significant differences between the experimental and standard care conditions but both had a significantly higher mean percentage vaginal condom use than the control group, averaged across pre- and six-month post-intervention. No gender differences were seen in attitudes, skills or self-efficacy to use condoms. ^ This study shed light upon the effectiveness of the instructional approach for the enhancement of attitudes, skills and self-efficacy outcomes related to HIV prevention. For heightened effectiveness, future approaches must address multiple factors impacting learning in this population. ^

Proton nuclear magnetic resonance investigation of the native and modified active site structure of heme proteins

Hemoproteins are a very important class of enzymes in nature sharing the essentially same prosthetic group, heme, and are good models for exploring the relationship between protein structure and function. Three important hemoproteins, chloroperoxidase (CPO), horseradish peroxidase (HRP), and cytochrome P450cam (P450cam), have been extensively studied as archetypes for the relationship between structure and function. In this study, a series of 1D and 2D NMR experiments were successfully conducted to contribute to the structural studies of these hemoproteins. ^ During the epoxidation of allylbenzene, CPO is converted to an inactive green species with the prosthetic heme modified by addition of the alkene plus an oxygen atom forming a five-membered chelate ring. Complete assignment of the NMR resonances of the modified porphyrin extracted and demetallated from green CPO unambiguously established the structure of this porphyrin as an NIII-alkylated product. A novel substrate binding motif of CPO was proposed from this concluded regiospecific N-alkylation structure. ^ Soybean peroxidase (SBP) is considered as a more stable, more abundant and less expensive substitute of HRP for industrial applications. A NMR study of SBP using 1D and 2D NOE methods successfully established the active site structure of SBP and consequently fills in the blank of the SBP NMR study. All of the hyperfine shifts of the SBP-CN- complex are unambiguously assigned together with most of the prosthetic heme and all proximal His170 resonances identified. The active site structure of SBP revealed by this NMR study is in complete agreement with the recombinant SBP crystal structure and is highly similar to that of the HRP with minor differences. ^ The NMR study of paramagnetic P450cam had been greatly restricted for a long time. A combination of 2D NMR methods was used in this study for P450cam-CN - complexes with and without camphor bound. The results lead to the first unequivocal assignments of all heme hyperfine-shifted signals, together with certain correlated diamagnetic resonances. The observed alternation of the assigned novel proximal cysteine β-CH2 resonances induced by camphor binding indicated a conformational change near the proximal side.^

The effectiveness of an enhanced congnitive behavioral intervention in improving behavioral outcomes related to HIV prevention

This study analyzed outcomes of an enhanced cognitive-behavioral intervention with dually diagnosed severely mentally ill adults. It specifically addressed the improvement of attitudes, skills, self-efficacy to use condoms and the heightening of condom use. The data were analyzed via a randomized three-group repeated measures design composed of the experimental (E-CB), standard care (SC) comparison or a no-treatment control condition as the between-subjects variable and pre-post measure as the within-subjects variable. The ECB focused on cooperative, application, hands-on, skill-building and role-playing activities for sexual assertiveness, negotiation in risk-taking and proper condom use. The SC comparison, was didactic in its approach and addressed risk- taking and proper condom use in one session, but did not involve application approaches to problem-solving risky situations or condom use. Multiple assessments were conducted at pre-, post- and six months post-intervention. The analysis indicated that the E-CB intervention led to more favorable attitudes toward condoms and to improved and maintained skills regarding their use by participants six months after the intervention compared to the standard care and control groups. No significant improvements in self-efficacy were found. A repeated measures ANOVA conducted on the transformed values of percentage of vaginal condom use indicated no significant differences between the experimental and standard care conditions but both had a significantly higher mean percentage vaginal condom use than the control group, averaged across pre- and six-month post-intervention. No gender differences were seen in attitudes, skills or self-efficacy to use condoms. This study shed light upon the effectiveness of the instructional approach for the enhancement of attitudes, skills and self-efficacy outcomes related to HIV prevention. For heightened effectiveness, future approaches must address multiple factors impacting learning in this population.

Crianças e investigação: aspectos teórico-metodológicos e cuidados éticos nas produções dos grupos de pesquisa da UFRN

The concepts of childhood are part of a cultural construction and vary throughout history politically, economically and socially. Nouns such as "childhood" and "child" did not exist as sense of unity in periods prior to modernity, reinforcing their historical character as concepts that have been socially constructed as profess the Historical Social Psychology, theoretical perspective which will support this research. The sociology of childhood distinguishes the terms and changes the approach toward the adult researcher approach with respect to children, aiming to give voice to the child. When it comes to defining the role of children in research, the researcher must take into consideration several relevant criteria such as age, gender, time, listening to the adults close to the children, the level of language and education and which children that will be heard in a group. It is from this discussion, considering the importance of children's participation in the research that are established the purpose of this work, namely, to analyze the theoretical and methodological aspects and ethical guidelines considered in the research process with children, by researchers at the UFRN, presenting as specific objectives: analyze and discuss, from research reports, the adopted ethical procedures and methods used in research with children. Set the goal, it was made a search of the UFRN Research Groups in SIGAA in order to select those which investigate children for participating in our survey. Among the centers of UFRN, we selected the Health Sciences Centre - CCSA, the Humanities Center, Letters and Arts - CCHLA, the Biosciences Center - CB and the Health Sciences Center - CCS, a total of 64 finalized reports to be analyzed. And here it is the observation that the foolish reports were not analyzed for ethical reasons as the guiding of this dissertation is the coordinator of the research group mentioned. In this study, we chose the documentary analysis of the finalized reports from UFRN research projects teachers / researchers as procedure to set up the corpus of the research. Data analysis was performed from the qualitative analysis in the following categories were established: the concept of childhood, concept of child, age of the subjects, the research context, theoretical and methodological care adopted and ethical care. It is hoped that this study will contribute with reflections on the ethical and theoretical and methodological care on research with children. The research showed how some of the results: significant number of reports excluded by repetition, most reports did not bring any closing remarks, no different procedures for children, with rare exceptions, the ethical issue was not mentioned in 50% of reports.

A objetivação da linguagem no (con)texto do vestibular : mo(vi)mentos na história e no político

In this study, we join up in the theoretical assumptions of the French Discourse Analysis in order to analyze effects of the demand of objectification of language in the context of vestibular essays. More specifically, we analyze the operation of said objectification via discourses constructed by the traditional vestibular exam due to the requirement to have, in the students’ essays, paraphrases of statements from the motivating texts (TM) of the test in question. From our perspective, the objectification mechanism of language, the paraphrase, in the vestibular, its logic of clarity and non-contradiction of ideas, is made by (in)determination of senses in the order of its speech and, also, in its practice: the correction of the vestibular essay. Therefore, in spite of what is assumed as guarantee to language in the moment of the vestibular essay, we suggest there are regularization-recognition conflicts of same senses— the constitutive senses of TM — in the evaluative speech of two vestibular-essay correctors(CA and CB). These correctors, with their history of reading (grammar and Linguistic Textual), stress the concept of paraphrase taken by the vestibular instance for the correction of students’ essays. Such stress creates a dispute of speeches: the speech of knowledge (university policy) versus the speech of produce (neoliberal policy); the latter as reading policy that favors literal meanings, consensus. Because of all this, we question: what are the effects of senses produced in (and about) vestibular essays by the demand of determining of the saying there instituted? To answer this question, we build analysis from clippings of documents that regulate the vestibular exam (institutional texts) in our country and, also, analysis of two vestibular essays in which at times appear, at times not, according to the judgment of CA and CB of essays, paraphrases of TM statements of the essay. The analysis, in theory, punctuates effects of sense of the objectification process of the saying in vestibular, and primarily the rarefaction of legal-position subject-of-knowing by the current institution of the subject-of-making. Moreover, our work comprises affiliations of sense that relates to the subject-speech relationship in evaluative exercise of vestibular essays, on the question of authorship.

Implication du système cannabinoïde dans un nouveau modèle de douleur neuropathique périphérique

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.