The isatin-Schiff base copper(II) complex Cu(isaepy)(2) acts as delocalized lipophilic cation, yields widespread mitochondrial oxidative damage and induces AMP-activated protein kinase-dependent apoptosis

We previously demonstrated that Bis[(2-oxindol-3-ylimino)-2-(2-aminoethyl) pyridine-N, N`] copper(II) [Cu(isaepy)(2)] was an efficient inducer of the apoptotic mitochondrial pathway. Here, we deeply dissect the mechanisms underlying the ability of Cu(isaepy)(2) to cause mitochondriotoxicity. In particular, we demonstrate that Cu(isaepy)(2) increases NADH-dependent oxygen consumption of isolated mitochondria and that this phenomenon is associated with oxy-radical production and insensitive to adenosine diphosphate. These data indicate that Cu(isaepy)(2) behaves as an uncoupler and this property is also confirmed in cell systems. Particularly, SH-SY5Y cells show: (i) an early loss of mitochondrial transmembrane potential; (ii) a decrease in the expression levels of respiratory complex components and (iii) a significant adenosine triphosphate (ATP) decrement. The causative energetic impairment mediated by Cu(isaepy)(2) in apoptosis is confirmed by experiments carried out with rho(0) cells, or by glucose supplementation, where cell death is significantly inhibited. Moreover, gastric and cervix carcinoma AGS and HeLa cells, which rely most of their ATP production on oxidative phosphorylation, show a marked sensitivity toward Cu(isaepy)(2). Adenosine monophosphate-activated protein kinase (AMPK), which is activated by events increasing the adenosine monophosphate: ATP ratio, is deeply involved in the apoptotic process because the overexpression of its dominant/negative form completely abolishes cell death. Upon glucose supplementation, AMPK is not activated, confirming its role as fuel-sensing enzyme that positively responds to Cu(isaepy)(2)-mediated energetic impairment by committing cells to apoptosis. Overall, data obtained indicate that Cu(isaepy)(2) behaves as delocalized lipophilic cation and induces mitochondrial-sited reactive oxygen species production. This event results in mitochondrial dysfunction and ATP decrease, which in turn triggers AMPK-dependent apoptosis.

Pkc1 acts through Zds1 and Gic1 to suppress growth and cell polarity defects of a yeast eIF5A mutant

eIF5A is a highly conserved putative eukaryotic translation initiation factor that has been implicated in translation initiation, nucleocytoplasmic transport, mRNA decay, and cell proliferation, but with no precise function assigned so far. We have previously shown that high-copy PKCI suppresses the phenotype of tif51A-1, a temperature-sensitive mutant of eIF5A in S. cerevisiae. Here, in an attempt to further understand how Pkc1 functionally interacts with eIF-5A, it was determined that PKCI suppression of tif51A-1 is independent of the cell integrity MAP kinase cascade. Furthermore, two new suppressor genes, ZDS1 and GIC1, were identified. We demonstrated that ZDS1 and ZDS2 are necessary for PKC1, but not for GIC1 suppression. Moreover, high-copy GIC1 also suppresses the growth defect of a PKCI mutant (stt1), suggesting the existence of a Pkc1-Zds1-Gic1 pathway. Consistent with the function of Gic1 in actin organization, the tif51A-1 strain shows an actin polarity defect that is partially recovered by overexpression of Pkc1 and Zds1 as well as Gic1. Additionally, PCL1 and BNI1, important regulators of yeast cell polarity, also suppress tif51A-1 temperature sensitiviiy Taken together, these data strongly Support the correlated involvement of Pkc1 and eIF5A in establishing actin polarity, which is essential for bud formation and G1/S transition in S. cerevisiae.

Atividades preventivas realizadas pelos cirurgiões-dentistas do PSF de Natal-RN

Family Health Program (FHP) presents itself as a restructured model on basic cares besides having prevention practices and promotion in health as essential parts. According to that, the purpose of this study is to perceive which prevention practices in oral health are accomplished by the dentists from the FHP of Natal-RN as well as the way they have been developed, the knowledge basis to execute the procedure and the use of evaluation instruments by the dentists and by the Oral Health Co-Ordinator. A list of dentists participating on the FHP of Natal in March, 2006 (n= 91) was obtained so it was possible to formulate a structured interview. After excluding those with less than six months attending to the program, and considering the loss, eighty dentists got to be interviewed. It had questions about individual and group preventive procedures, sources of basement to develop the activities and to verify the impact and evaluation instruments. Besides that, an interview was made with the Oral Health Co-Ordinator about the evaluating process and the existence of a protocol as well as about document analysis in the Ambulatory Informations System Unique Health System (AIS-UHS), Information of Basic Care System and Pact of Recorder. The main individual activities consist on Oral Hygiene Orientation (87,5%) and Fluoridization (95%). Group activities remain at scholar groups acts(91,25%) being fluoridization done by 91,25% and educative activities by 86,25% mainly through speeches (61,25%).Orientations about oral cancer were detached on the aged group (39,96%)and hypertensive and diabetic group (19,51%), as well as integration with the health team was respectively 21,93% and 39,02%. Speeches and talk rounds are the main activities for aged, hypertensive and diabetics, and are also expressive to pregnant women. For pregnant women and babies group predominate hygiene and diet orientation. About the evaluation instruments 73,75% of the dentists do not consider their existence, despite 73,75% consider that actions have impact specially due to a increase of hygiene and decrease of caries index on the children. The main sources of basement are clinical experience (42,5%) and preparative courses (33,75%), only 7,5% mentioned population need and local reality. The Oral Health Co-Ordinator related the Attention Pact Recorder and records from the AIS - UHS as evaluation source but also detached the lack of epidemiologic surveying. She mentioned the increase of supervised brushing as a positive result and detached the hardness of the professional to act inside the family and as a team. The AIS-UHS records showed that fluoridization is more used individually and the supervised brushing is predominant as a group action, besides that educative activities at the Health Establishment predominate due to actions at the community. Facing the results, it was verified the act in different groups, special attention given to scholars, as well as focus the activities on caries giving less emphasis to other oral problems. Despite that, there was no epidemiologic instruments or data of the local reality to guide the actions and to be part of the planning and evaluation process

MicroRNA-100 Acts as a Tumor Suppressor in Human Bladder Carcinoma 5637 Cells

Bladder carcinoma is one of the most common tumors in the world and, despite the therapy currently available, most of the patients relapse. Better understanding of the factors involved in disease pathogenesis would provide insights for the development of more effective strategies in treatment. Recently, differential miRNA expression profiles in bladder urothelial carcinomas identified miR-100 down-regulation and miR-708 up-regulation among the most common alterations, although the possible influence of these miRNAs in the control of basic mechanisms in bladder tumors has not been addressed. In this context, the present study aimed to evaluate the in vitro effects of miR-100 forced expression and miR-708 inhibition in the bladder carcinoma cell line 5637. Our results showed that overexpression of miR-100 significantly inhibited growth when compared to controls at both times tested (72 and 96 hours, p<0.01) with a maximum effect at 72 hours reducing proliferation in 29.6 %. Conversely, no effects on cell growth were observed after inhibition of miR-708. MiR-100 also reduced colony formation capacity of 5637 cells by 24.4%. No alterations in cell cycle progression or apoptosis induction were observed. The effects of miR-100 on growth and clonogenicity capacity in 5637 cells evince a possible role of this miRNA in bladder carcinoma pathogenesis. Further studies are necessary to corroborate our findings and examine the potential use of this microRNA in future therapeutic interventions.

Brazilian Medicinal Plant Acts on Prostaglandin Level and Helicobacter pylori

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

TNF-alpha acts in the hypothalamus inhibiting food intake and increasing the respiratory quotient - Effects on leptin and insulin signaling pathways

Acting in the hypothalamus, tumor necrosis factor-alpha (TNF-alpha) produces a potent anorexigenic effect. However, the molecular mechanisms involved in this phenomenon are poorly characterized. In this study, we investigate the capacity of TNF-alpha to activate signal transduction in the hypothalamus through elements of the pathways employed by the anorexigenic hormones insulin and leptin. High dose TNF-a promotes a reduction of 25% in 12 h food intake, which is an inhibitory effect that is marginally inferior to that produced by insulin and leptin. In addition, high dose TNF-a increases body temperature and respiratory quotient, effects not reproduced by insulin or leptin. TNF-alpha, predominantly at the high dose, is also capable of activating canonical pro-inflammatory signal transduction in the hypothalamus, inducing JNK, p38, and NF kappa B, which results in the transcription of early responsive genes and expression of proteins of the SOCS family. Also, TNF-a activates signal transduction through JAK-2 and STAT-3, but does not activate signal transduction. through early and intermediary elements of the insulin/leptin signaling pathways such as IRS-2, Akt, ERK and FOXO1. When co-injected with insulin or leptin, TNF-a, at both high and low doses, partially impairs signal transduction through IRS-2, Akt, ERK and FOXO1 but not through JAK-2 and STAT-3. This effect is accompanied by the partial inhibition of the anorexigenic effects of insulin and leptin, when the low, but not the high dose of TNF-alpha is employed. In conclusion, TNF-alpha, on a dose-dependent way, modulates insulin and leptin signaling and action in the hypothalamus. (c) Published by Elsevier B.V.

Low intensity laser therapy (LILT) in vivo acts on the neutrophils recruitment and chemokines/cytokines levels in a model of acute pulmonary inflammation induced by aerosol of lipopolysaccharide from Escherichia coli in rat

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da atividade antiulcerogênica dos extratos metanólico e clorofórmico e da fração butanólica de Vochysia tucanorum (Vochysiacea)

Pós-graduação em Ciências Biológicas (Farmacologia) - IBB

Levantamento, análise e descrição de elementos paralinguísticos do português espontâneo

A fala apresenta aspectos paralinguísticos que não pertencem ao código linguístico convencional, mas contribuem significativamente para a unidade temática do discurso, Essas realizações se constituem em enunciados não-lexicalizados que funcionam que funcionam como atos de fala completos nas interações comunicativas interpessoais. Sobre essas emissões não-verbais, Campbell (2002a, 2002b, 2003 e 2004), Maekawa (2004), Fujie et. al (2004), Hoult (2004), Key (1958) apud Steimberg (1988) postulam que elas constribuem para a manifestação da fala expressiva. Para os autores, é justamente o fenômeno da paralinguagem que sinaliza informações sobre atitudes, opiniões e emoções do falante em relação ao interlocutor ou ao tópico discursivo. Nesse sentido, investigamos, neste trabalho, as manifestações paralinguísticas recorrentes em conversas informais para demonstrarmos seu papel expressivo na linguagem falada. Para tanto, fizemos um levantamento de 450 ocorrências de elementos paralinguísticos no processo de transcrição de amostras de falas do Português Regional Paraense produzidas em situações reais de conversação. Pressupondo que essas realizações não-verbais são caracterizadas por variações prosódicas, nós as submetemos a uma análise fonética por meio do software PRAAT. A partir dessa análise, constatamos a contribuição de duas propriedades: a frequência fundamental (F0) e o tempo de emissão, para a manifestação expressiva dos elementos paralinguísticos no discurso falado. Além disso, identificamos também a silabação como uma propriedade comum às realizações sonoras focalizadas. Após o processo de análise, fizemos a descrição do uso e do funcionamento desses elementos nas conversas, bem como da contribuição deles para a manifestação da fala expressiva. Os resultados nos mostram que os elementos paralinguísticos, além de contribuírem para a fluência do discurso falado, desempenham a função de sinalizar compreensão, interesse e/ou atenção, gerenciar relações interpessoais e expressar emoções, atitudes e afeto.

Pré-tratamento oxidativo de bagaço de cana com persulfato de sódio para sacarificação enzimática e produção de etanol celulósico

Pós-graduação em Química - IBILCE

Pausas na interpretação teatral: delimitação de constituintes prosódicos

We intend to observe the function of a linguistic resource – the pause – in theatrical interpretation. Connected to the field of speech therapy, we search for theoretical support in the Linguistics field, mainly in prosodic phonology – specifically, we highlight intonational phrase and phonological utterance, prosodic constituents –, proposing a dialogue between these fields, regarding the work with actors. In speech therapy literature, the work with actors focuses, centrally, in organic issues involved in the vocal process, such as “misuse” or “voice abuse”. To a smaller extent, we find, in this literature, researches that emphasize issues regarding interpretation and expressive resources, besides a few emphasizing the importance of linguistic resources in interpretation. Differently, in linguistics literature, the pause is approached, to a larger extent, from the phonetic perspective, related to several language levels. In this research, we analyzed audio recordings of four actors from a same theatrical group, acting the theatrical text Brutas flores, focused on these aims: (1) detect the place where pauses happen in the interpretation of a single text by four actors; (2) survey physical characteristics of length of these pauses; (3) check to what extent the length of a pause is related to the place where it happens, regarding the prosodic limits of intonational phrases (I) and phonological utterance (U). We could observe that, although the interpretation is characterized by the subjectivity of the actor, the interpretation is constructed based in the possibilities offered by the prosodic organization of the text itself, being more or less flexible.We were also able to confirm, by considering the length of VVs units containing pauses, the prosodic hierarchy proposed by Nespor & Vogel, once the length of these units in U's limits was significantly higher than the length in I's limits. Thus, our results reinforce the premise that a linguistic structure overlaps the subjectivity of the actor, i. e., the premise that the strength of linguistic structure organization acts on the possible individual operation/style.