Intrinsic organization of the suprachiasmatic nucleus in the capuchin monkey

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

Evolution of time-control mechanisms in subterranean organisms: cave fishes under light-dark cycles (Teleostei: Siluriformes, Characiformes)

Subterranean organisms are excellent models for chronobiological studies, yet relatively few taxa have been investigated with this focus. Former results were interpreted as a pattern of regression of circadian locomotor activity rhythms in troglobitic (exclusively subterranean) species. In this paper we report results of experiments with cave fishes showing variable degrees of troglomorphism (reduction of eyes, melanic pigmentation and other specializations related to the hypogean life) submitted to light-dark cycles, preceded and followed by several days in constant darkness. Samples from seven species have been monitored in our laboratory for the detection of significant circadian rhythms in locomotor activity: S. typhlops, an extremely troglomophic species, presented the lowest number of significant components in the circadian range (only one individual out of eight in DD1 and three other fish in LD), all weak (low values of spectral power). Higher incidence of circadian components was observed for P. kronei - only one among six studied catfish without significant circadian rhythms under DD1 and DD2; spectral powers were generally high. Intermediate situations were observed for the remaining species, however all of them presented relatively strong significant rhythms under LD. Residual oscillations (circadian rhythms in DD2) were detected in at least part of the studied individuals of all species but S. typhlops, without a correlation with spectral powers of LD rhythms, i.e., individuals exhibiting residual oscillations were not necessarily those with the strongest LD rhythms. In conclusion, the accumulated evidence for troglobitic fishes strongly supports the hypothesis of external, environmental selection for circadian locomotor rhythms.

Sleep length, working hours and socio-demographic variables are associated with time attending evening classes among working college students

It is the aim of the present study to assess factors associated with time spent in class among working college students. Eighty-two working students from 21 to 26 years old participated in this study. They were enrolled in an evening course of the University of Sao Paulo, Brazil. Participants answered a questionnaire on living and working conditions. During seven consecutive days, they wore an actigraph, filled out daily activity diaries (including time spent in classes) and the Karolinska Sleepiness Scale every three hours from waking until bedtime. Linear regression analyses were performed in order to assess the variables associated with time spent in classes. The results showed that gender, sleep length, excessive sleepiness, alcoholic beverage consumption (during workdays) and working hours were associated factors with time spent in class. Thus, those who spent less time in class were males, slept longer hours, reported excessive sleepiness on Saturdays, worked longer hours, and reported alcohol consumption. The combined effects of long work hours (>40 h/week) and reduced sleep length may affect lifestyles and academic performance. Future studies should aim to look at adverse health effects induced by reduced sleep duration, even among working students who spent more time attending evening classes.

Intrinsic organization of the suprachiasmatic nucleus in the capuchin monkey

The suprachiasmatic nucleus (SCN), which is the main circadian biological clock in mammals, is composed of multiple cells that function individually as independent oscillators to express the self-sustained mRNA and protein rhythms of the so-called clock genes. Knowledge regarding the presence and localization of the proteins and neuroactive substances of the SCN are essential for understanding this nucleus and for its successful manipulation. Although there have been advances in the investigation of the intrinsic organization of the SCN in rodents, little information is available in diurnal species, especially in primates. This study, which explores the pattern of expression and localization of PER2 protein in the SCN of capuchin monkey, evaluates aspects of the circadian system that are common to both primates and rodents. Here, we showed that PER2 protein immunoreactivity is higher during the light phase. Additionally, the complex organization of cells that express vasopressin, vasoactive intestinal polypeptide, neuron-specific nuclear protein, calbindin and calretinin in the SCN, as demonstrated by their immunoreactivity, reveals an intricate network that may be related to the similarities and differences reported between rodents and primates in the literature.

Spinal cord injury: assessment of autonomic state-dependent control of cardiovascular system and body core temperature

Spinal cord injury (SCI) results not only in paralysis; but it is also associated with a range of autonomic dysregulation that can interfere with cardiovascular, bladder, bowel, temperature, and sexual function. The entity of the autonomic dysfunction is related to the level and severity of injury to descending autonomic (sympathetic) pathways. For many years there was limited awareness of these issues and the attention given to them by the scientific and medical community was scarce. Yet, even if a new system to document the impact of SCI on autonomic function has recently been proposed, the current standard of assessment of SCI (American Spinal Injury Association (ASIA) examination) evaluates motor and sensory pathways, but not severity of injury to autonomic pathways. Beside the severe impact on quality of life, autonomic dysfunction in persons with SCI is associated with increased risk of cardiovascular disease and mortality. Therefore, obtaining information regarding autonomic function in persons with SCI is pivotal and clinical examinations and laboratory evaluations to detect the presence of autonomic dysfunction and quantitate its severity are mandatory. Furthermore, previous studies demonstrated that there is an intimate relationship between the autonomic nervous system and sleep from anatomical, physiological, and neurochemical points of view. Although, even if previous epidemiological studies demonstrated that sleep problems are common in spinal cord injury (SCI), so far only limited polysomnographic (PSG) data are available. Finally, until now, circadian and state dependent autonomic regulation of blood pressure (BP), heart rate (HR) and body core temperature (BcT) were never assessed in SCI patients. Aim of the current study was to establish the association between the autonomic control of the cardiovascular function and thermoregulation, sleep parameters and increased cardiovascular risk in SCI patients.

Visual and quantitative electroencephalographic analysis of healthy young and adult cats under medetomidine sedation

A study was designed to investigate the effect of medetomidine sedation on quantitative electroencephalography (q-EEG) in healthy young and adult cats to determine objective guidelines for diagnostic EEG recordings and interpretation. Preliminary visual examination of EEG recordings revealed high-voltage low-frequency background activity. Spindles, k-complexes and vertex sharp transients characteristic of sleep or sedation were superimposed on a low background activity. Neither paroxysmal activity nor EEG burst-suppression were observed. The spectral analysis of q-EEG included four parameters, namely, relative power (%), and mean, median and peak frequency (Hz) of all four frequency bands (delta, theta, alpha and beta). The findings showed a prevalence of slow delta and theta rhythms as opposed to fast alpha and beta rhythms in both young (group A) and adult (group B) cats. A posterior gradient was reported for the theta band and an anterior gradient for the alpha and beta bands in both groups, respectively. The relative power value in group B compared to group A was significantly higher for theta, alpha and beta bands, and lower for the delta band. The mean and median frequency values in group B was significantly higher for delta, theta and beta bands and lower for the alpha band. The study has shown that a medetomidine sedation protocol for feline EEG may offer a method for investigating bio-electrical cortical activity. The use of q-EEG analysis showed a decrease in high frequency bands and increased activity of the low frequency band in healthy cats under medetomidine sedation.

Morningness-eveningness and anxiety among adults: A matter of sex/gender?

People’s health and well-being may be determined by the interaction of endogenous and external rhythms. Late chronotypes should become anxious because of the demand to perform tasks in a society oriented preferentially to morning work. In this study, we examined the relationship between morningness and anxiety in 559 adults (age range 40–63 years) from rural environments who completed morningness–eveningness and anxiety measures. Results indicated a negative correlation between morningness and anxiety in women, but not in men, suggesting the relevance of gender-related variables. When demographic characteristics were considered, women’s anxiety was predicted by low educational level and eveningness, whereas men’s anxiety was predicted by being single, having children, being unemployed, and eveningness. Some hypotheses of gender-related social roles are postulated as a possible explanation of the results.

Subjective and predicted sleepiness while driving in young adults

Sleepiness is a significant contributor to car crashes and sleepiness related crashes have higher mortality and morbidity than other crashes. Young adult drivers are at particular risk for sleepiness related car crashes. It has been suggested that this is because young adults are typically sleepier than older adults because of chronic sleep loss, and more often drive at times of increased risk of acute sleepiness. This prospective study aimed to determine the relationship between predicted and perceived sleepiness while driving in 47 young-adult drivers over a 4-week period. Sleepiness levels were predicted by a model incorporating known circadian and sleep factors influencing alertness, and compared to subjective ratings of sleepiness during 25 18 driving episodes. Results suggested that young drivers frequently drive while at risk of crashing, at times of predicted sleepiness (>7% of episodes) and at times they felt themselves to be sleepy (>23% of episodes). A significant relationship was found between perceived and predicted estimates of sleepiness. However, the participants nonetheless drove at these times. The results of this study may help preventative programs to specifically target factors leading to increased sleepiness when driving (particularly time of day), and to focus interventions to stop young adults from driving when they feel sleepy. (c) 2005 Elsevier Ltd. All rights reserved.

Synchronized oscillations at α and θ frequencies in the lateral geniculate nucleus

In relaxed wakefulness, the EEG exhibits robust rhythms in the alpha band (8-13 Hz), which decelerate to theta (approximately 2-7 Hz) frequencies during early sleep. In animal models, these rhythms occur coherently with synchronized activity in the thalamus. However, the mechanisms of this thalamic activity are unknown. Here we show that, in slices of the lateral geniculate nucleus maintained in vitro, activation of the metabotropic glutamate receptor (mGluR) mGluR1a induces synchronized oscillations at alpha and theta frequencies that share similarities with thalamic alpha and theta rhythms recorded in vivo. These in vitro oscillations are driven by an unusual form of burst firing that is present in a subset of thalamocortical neurons and are synchronized by gap junctions. We propose that mGluR1a-induced oscillations are a potential mechanism whereby the thalamus promotes EEG alpha and theta rhythms in the intact brain.

Nitrate metabolism in the dinoflagellate Lingulodinium polyedrum

Les dinoflagellés sont des eucaryotes unicellulaires retrouvés dans la plupart des écosystèmes aquatiques du globe. Ces organismes amènent une contribution substantielle à la production primaire des océans, soit en tant que membre du phytoplancton, soit en tant que symbiontes des anthozoaires formant les récifs coralliens. Malheureusement, ce rôle écologique majeur est souvent négligé face à la capacité de certaines espèces de dinoflagellés à former des fleurs d'eau, parfois d'étendue et de durée spectaculaires. Ces floraisons d'algues, communément appelées "marées rouges", peuvent avoir de graves conséquences sur les écosystèmes côtiers, sur les industries de la pêche et du tourisme, ainsi que sur la santé humaine. Un des facteurs souvent corrélé avec la formation des fleurs d'eau est une augmentation dans la concentration de nutriments, notamment l’azote et le phosphore. Le nitrate est un des composants principaux retrouvés dans les eaux de ruissellement agricoles, mais également la forme d'azote bioaccessible la plus abondante dans les écosystèmes marins. Ainsi, l'agriculture humaine a contribué à magnifier significativement les problèmes associés aux marées rouges au niveau mondial. Cependant, la pollution ne peut pas expliquer à elle seule la formation et la persistance des fleurs d'eau, qui impliquent plusieurs facteurs biotiques et abiotiques. Il est particulièrement difficile d'évaluer l'importance relative qu'ont les ajouts de nitrate par rapport à ces autres facteurs, parce que le métabolisme du nitrate chez les dinoflagellés est largement méconnu. Le but principal de cette thèse vise à remédier à cette lacune. J'ai choisi Lingulodinium polyedrum comme modèle pour l'étude du métabolisme du nitrate, parce que ce dinoflagellé est facilement cultivable en laboratoire et qu'une étude transcriptomique a récemment fourni une liste de gènes pratiquement complète pour cette espèce. Il est également intéressant que certaines composantes moléculaires de la voie du nitrate chez cet organisme soient sous contrôle circadien. Ainsi, dans ce projet, j'ai utilisé des analyses physiologiques, biochimiques, transcriptomiques et bioinformatiques pour enrichir nos connaissances sur le métabolisme du nitrate des dinoflagellés et nous permettre de mieux apprécier le rôle de l'horloge circadienne dans la régulation de cette importante voie métabolique primaire. Je me suis tout d'abord penché sur les cas particuliers où des floraisons de dinoflagellés sont observées dans des conditions de carence en azote. Cette idée peut sembler contreintuitive, parce que l'ajout de nitrate plutôt que son épuisement dans le milieu est généralement associé aux floraisons d'algues. Cependant, j’ai découvert que lorsque du nitrate était ajouté à des cultures initialement carencées ou enrichies en azote, celles qui s'étaient acclimatées au stress d'azote arrivaient à survivre près de deux mois à haute densité cellulaire, alors que les cellules qui n'étaient pas acclimatées mourraient après deux semaines. En condition de carence d'azote sévère, les cellules arrivaient à survivre un peu plus de deux semaines et ce, en arrêtant leur cycle cellulaire et en diminuant leur activité photosynthétique. L’incapacité pour ces cellules carencées à synthétiser de nouveaux acides aminés dans un contexte où la photosynthèse était toujours active a mené à l’accumulation de carbone réduit sous forme de granules d’amidon et corps lipidiques. Curieusement, ces deux réserves de carbone se trouvaient à des pôles opposés de la cellule, suggérant un rôle fonctionnel à cette polarisation. La deuxième contribution de ma thèse fut d’identifier et de caractériser les premiers transporteurs de nitrate chez les dinoflagellés. J'ai découvert que Lingulodinium ne possédait que très peu de transporteurs comparativement à ce qui est observé chez les plantes et j'ai suggéré que seuls les membres de la famille des transporteurs de nitrate de haute affinité 2 (NRT2) étaient réellement impliqués dans le transport du nitrate. Le principal transporteur chez Lingulodinium était exprimé constitutivement, suggérant que l’acquisition du nitrate chez ce dinoflagellé se fondait majoritairement sur un système constitutif plutôt qu’inductible. Enfin, j'ai démontré que l'acquisition du nitrate chez Lingulodinium était régulée par la lumière et non par l'horloge circadienne, tel qu'il avait été proposé dans une étude antérieure. Finalement, j’ai utilisé une approche RNA-seq pour vérifier si certains transcrits de composantes impliquées dans le métabolisme du nitrate de Lingulodinium étaient sous contrôle circadien. Non seulement ai-je découvert qu’il n’y avait aucune variation journalière dans les niveaux des transcrits impliqués dans le métabolisme du nitrate, j’ai aussi constaté qu’il n’y avait aucune variation journalière pour n’importe quel ARN du transcriptome de Lingulodinium. Cette découverte a démontré que l’horloge de ce dinoflagellé n'avait pas besoin de transcription rythmique pour générer des rythmes physiologiques comme observé chez les autres eukaryotes.

Nitrate metabolism in the dinoflagellate Lingulodinium polyedrum

Les dinoflagellés sont des eucaryotes unicellulaires retrouvés dans la plupart des écosystèmes aquatiques du globe. Ces organismes amènent une contribution substantielle à la production primaire des océans, soit en tant que membre du phytoplancton, soit en tant que symbiontes des anthozoaires formant les récifs coralliens. Malheureusement, ce rôle écologique majeur est souvent négligé face à la capacité de certaines espèces de dinoflagellés à former des fleurs d'eau, parfois d'étendue et de durée spectaculaires. Ces floraisons d'algues, communément appelées "marées rouges", peuvent avoir de graves conséquences sur les écosystèmes côtiers, sur les industries de la pêche et du tourisme, ainsi que sur la santé humaine. Un des facteurs souvent corrélé avec la formation des fleurs d'eau est une augmentation dans la concentration de nutriments, notamment l’azote et le phosphore. Le nitrate est un des composants principaux retrouvés dans les eaux de ruissellement agricoles, mais également la forme d'azote bioaccessible la plus abondante dans les écosystèmes marins. Ainsi, l'agriculture humaine a contribué à magnifier significativement les problèmes associés aux marées rouges au niveau mondial. Cependant, la pollution ne peut pas expliquer à elle seule la formation et la persistance des fleurs d'eau, qui impliquent plusieurs facteurs biotiques et abiotiques. Il est particulièrement difficile d'évaluer l'importance relative qu'ont les ajouts de nitrate par rapport à ces autres facteurs, parce que le métabolisme du nitrate chez les dinoflagellés est largement méconnu. Le but principal de cette thèse vise à remédier à cette lacune. J'ai choisi Lingulodinium polyedrum comme modèle pour l'étude du métabolisme du nitrate, parce que ce dinoflagellé est facilement cultivable en laboratoire et qu'une étude transcriptomique a récemment fourni une liste de gènes pratiquement complète pour cette espèce. Il est également intéressant que certaines composantes moléculaires de la voie du nitrate chez cet organisme soient sous contrôle circadien. Ainsi, dans ce projet, j'ai utilisé des analyses physiologiques, biochimiques, transcriptomiques et bioinformatiques pour enrichir nos connaissances sur le métabolisme du nitrate des dinoflagellés et nous permettre de mieux apprécier le rôle de l'horloge circadienne dans la régulation de cette importante voie métabolique primaire. Je me suis tout d'abord penché sur les cas particuliers où des floraisons de dinoflagellés sont observées dans des conditions de carence en azote. Cette idée peut sembler contreintuitive, parce que l'ajout de nitrate plutôt que son épuisement dans le milieu est généralement associé aux floraisons d'algues. Cependant, j’ai découvert que lorsque du nitrate était ajouté à des cultures initialement carencées ou enrichies en azote, celles qui s'étaient acclimatées au stress d'azote arrivaient à survivre près de deux mois à haute densité cellulaire, alors que les cellules qui n'étaient pas acclimatées mourraient après deux semaines. En condition de carence d'azote sévère, les cellules arrivaient à survivre un peu plus de deux semaines et ce, en arrêtant leur cycle cellulaire et en diminuant leur activité photosynthétique. L’incapacité pour ces cellules carencées à synthétiser de nouveaux acides aminés dans un contexte où la photosynthèse était toujours active a mené à l’accumulation de carbone réduit sous forme de granules d’amidon et corps lipidiques. Curieusement, ces deux réserves de carbone se trouvaient à des pôles opposés de la cellule, suggérant un rôle fonctionnel à cette polarisation. La deuxième contribution de ma thèse fut d’identifier et de caractériser les premiers transporteurs de nitrate chez les dinoflagellés. J'ai découvert que Lingulodinium ne possédait que très peu de transporteurs comparativement à ce qui est observé chez les plantes et j'ai suggéré que seuls les membres de la famille des transporteurs de nitrate de haute affinité 2 (NRT2) étaient réellement impliqués dans le transport du nitrate. Le principal transporteur chez Lingulodinium était exprimé constitutivement, suggérant que l’acquisition du nitrate chez ce dinoflagellé se fondait majoritairement sur un système constitutif plutôt qu’inductible. Enfin, j'ai démontré que l'acquisition du nitrate chez Lingulodinium était régulée par la lumière et non par l'horloge circadienne, tel qu'il avait été proposé dans une étude antérieure. Finalement, j’ai utilisé une approche RNA-seq pour vérifier si certains transcrits de composantes impliquées dans le métabolisme du nitrate de Lingulodinium étaient sous contrôle circadien. Non seulement ai-je découvert qu’il n’y avait aucune variation journalière dans les niveaux des transcrits impliqués dans le métabolisme du nitrate, j’ai aussi constaté qu’il n’y avait aucune variation journalière pour n’importe quel ARN du transcriptome de Lingulodinium. Cette découverte a démontré que l’horloge de ce dinoflagellé n'avait pas besoin de transcription rythmique pour générer des rythmes physiologiques comme observé chez les autres eukaryotes.

Adult attachment style and cortisol responses in women in late pregnancy

Suporte financeiro e não financeiro da Maternidade Alfredo da Costa, Lisboa, Portugal

Uma contribuição ao estudo de sinais de EEG para avaliar estados emocionais e mentais de crianças com autismo na interação com robô móvel

O Transtorno do Espectro do Autismo (TEA) caracteriza-se por uma série de distúrbios cognitivos e neurocomportamentais e sua prevalência mundial é estimada em 1 criança com TEA a cada 160 crianças com típico desenvolvimento (TD). Indivíduos com TEA apresentam dificuldade em interpretar as emoções alheias e em expressar sentimentos. As emoções podem ser associadas à manifestação de sinais fisiológicos, e, dentre eles, os sinais cerebrais têm sido muito abordados. A detecção dos sinais cerebrais de crianças com TEA pode ser benéfica para o esclarecimento de suas emoções e expressões. Atualmente, muitas pesquisas integram a robótica ao tratamento pedagógico do TEA, através da interação com crianças com esse transtorno, estimulando habilidades sociais, como a imitação e a comunicação. A avaliação dos estados mentais de crianças com TEA durante a sua interação com um robô móvel é promissora e assume um aspecto inovador. Assim, os objetivos deste trabalho foram captar sinais cerebrais de crianças com TEA e de crianças com TD, como grupo controle, para o estudo de seus estados emocionais e para avaliar seus estados mentais durante a interação com um robô móvel, e avaliar também a interação dessas crianças com o robô, através de escalas quantitativas. A técnica de registro dos sinais cerebrais escolhida foi a eletroencefalografia (EEG), a qual utiliza eletrodos colocados de forma não invasiva e não dolorosa sobre o couro cabeludo da criança. Os métodos para avaliar a eficiência do uso da robótica nessa interação foram baseados em duas escalas internacionais quantitativas: Escala de Alcance de Metas (do inglês Goal Attainment Scaling - GAS) e Escala de Usabilidade de Sistemas (do inglês System Usability Scale - SUS). Os resultados obtidos mostraram que, pela técnica de EEG, foi possível classificar os estados emocionais de crianças com TD e com TEA e analisar a atividade cerebral durante o início da interação com o robô, através dos ritmos alfa e beta. Com as avaliações GAS e SUS, verificou-se que o robô móvel pode ser considerado uma potencial ferramenta terapêutica para crianças com TEA.

Análise do nível de estresse baseada em sinais de eletroencefalografia e de condutância da pele

O estresse pode afetar qualquer pessoa, independente de idade, sexo ou etnia. O organismo humano o utiliza como uma resposta adaptativa frente a situações diversas, as quais requeiram alguma adaptação do organismo para que possa enfrentar tal situação. Dependendo do estímulo estressor, pode ser gerado no indivíduo desgastes físico, mental ou emocional, no entanto, o estresse não representa necessariamente algo ruim ou patológico; este é um mecanismo de adaptação vital para a sobrevivência da espécie humana. Porém, o número de pessoas que são afetadas de forma negativa pelo estresse tem crescido imensamente nas últimas décadas. Pesquisas destacam que nos Estados Unidos cerca de 60% a 90% dos atendimentos médicos estão relacionados de alguma maneira com o estresse, enquanto que no Brasil aproximadamente 80% da população sofre de estresse, sendo que desses, 30% encontram-se na fase mais crítica, a chamada fase de exaustão. Tendo em vista que a principal forma de identificação de estresse ainda é realizada através do uso de questionário de autorrelato. O presente estudo apresenta como contribuição uma metodologia de análise do nível de estresse baseada na variação da condutância galvânica da pele e de sinais de eletroencefalografia, sendo utilizados como parâmetros a assimetria do ritmo alfa, assim como a razão entre os ritmos beta e alfa no córtex frontal e pré-frontal. Para a gravação dos sinais de EEG foi utilizado um dispositivo portátil, com eletrodos especificamente situados nas posições aF3, F3, F4 e aF4, de acordo com o Sistema Internacional 10/20 de posicionamento de eletrodos. Os participantes deste estudo são Bombeiros Militares da 1ª Cia de Vitória-ES. Foram utilizadas três classes de estímulos emocionais positivos, calmos e negativos, através da utilização de imagens pertencentes ao banco de dados IAPS (International Affective Picture System). Os resultados de acurácia obtidos através de um classificador SVM (Support Vector Machine) chegam a 88,24% para classe de estímulos positivos, 84,09% para classe calma e de 92,86% para os estímulos negativos. Deste modo, esta pesquisa apresenta uma combinação de parâmetros que podem ser aferidos com equipamentos de baixo custo, e fornecem condições de diferenciar estímulos estressantes, podendo assim, ser utilizada para auxiliar no treinamento de profissionais da área de urgência e emergência.

Cognitive intervention protocol for age-related memory impairments

With the number of elderly people increasing tremendously worldwide, comes the need for effective methods to maintain or improve older adults' cognitive performance. Using continuous neurofeedback, through the use of EEG techniques, people can learn how to train and alter their brain electrical activity. A software platform that puts together the proposed rehabilitation methodology has been developed: a digital game protocol that supports neurofeedback training of alpha and theta rhythms, by reading the EEG activity and presenting it back to the subject, interleaved with neurocognitive tasks such as n-Back and Corsi Block-Tapping. This tool will be used as a potential rehabilitative platform for age-related memory impairments.