Efeitos da senescência no núcleo supraquiasmático do sagui (Callithrix Jacchus): plasticidade morfológica e neuroquímica

The circadian timing system (CTS) is responsible for the generation and synchronization and the suprachiasmatic nucleus (SCN) of the hypothalamus has been described as the major circadian pacemaker in many mammalian species. The internal temporal organization managed by SCN is disturbed with aging bringing many pathological disorders that range from loss of complex cognitive performance to simple physiological functions. Therefore, our aim was perform a comparative study of the morphological aspects and neurochemical composition in the SCN of marmosets (Callithrix jacchus) adults and older using immunohistochemical techniques. We found morphometric and neurochemical changes in th SCN o folder animals in comparison to adults, among these a possible decreased in retinal projection to the SCN of older animals, found through a decline in CTB immunostaining, which can occur due atrophy and/or decreasing of fibers from the retinohypothalamic tract (RHT). The Klüver-Barrera histological technique strongly suggests a decrease in those fibers from RHT. Also, by means of a morphometric study, it is found a atrophy and numerical decline of neurons in SCN of aged animals, investigated by Nissl technique, and immunostaining with NeuN and calbindin. Relative optical density (ROD) analysis were used to evaluate the expression of some neurochemical components in SCN, such as GFAP expression, which was increased in older, result that indirectly reinforces that morphological changes occurs due the aging; the vasoactive intestinal polipeptide (VIP) showed no expression alteration in SCN of older animals; the serotonin (5-HT) was descreased in the dorsomedial portion of the SCN, and neurpeptide Y (NPY) apparently also decrease due to the increase of age. Many of these modifications were seen in other animals, such as rodents, human primates and non-human primates. These data about marmoset comes to add new information of the effect of aging on structures responsibles for the circadian rhytmicity, and that some behavioral changes controlled by th SCN, and founded in aged animals, may be caused by these morphological and neurochemical changes. Although some results have been quantitatively negative, qualitatively all analysis show significant change comparing adult and older animals, perhaps due to a low sampling number. In conclusion, the marmoset presents several morphological and neurochemical changes in the SCN of aged animals compared to adults, which may result in behavioral changes that favor pathology aging related

Plasticidade de células tronco mesenquimais de medula óssea de ratos na presença de meio condicionado do nervo facial e do fator de crescimento fibroblástico 2

A number of evidences show the influence of the growth of injured nerve fibers in Peripheral Nervous System (PNS) as well as potential implant stem cells (SCs) to make it more suitable for nerve regeneration medium. In this perspective, this study aimed to evaluate the plasticity of mesenchymal stem cells from bone marrow of mice in the presence of culture medium conditioned with facial nerve explants (D-10) and fibroblast growth factor-2 (FGF-2). In this perspective, the cells were cultivated only with DMEM (group 1), only with D-10(group 2), only with FGF-2(group 3) or with D-10 and FGF-2(group 4). The growth and morphology were assessed over 72 hours. Quantitative phenotypic analysis was taken from the immunocytochemistry for GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200 on the fourth day of cultivation. Cells cultured with conditioned medium alone or combined with FGF-2 showed distinct morphological features similar apparent at certain times with neurons and glial cells and a significant proliferative activity in groups 2 and 4 throughout the days. Cells cultived only with conditioned medium acquired a glial phenotype. Cells cultured with FGF-2 and conditioned medium expressed GFAP, OX-42, MAP-2, β-tubulin III, NeuN and NF-200. On average, area and perimeter fo the group of cells positive for GFAP and the área of the cells immunostained for OX-42 were higher than those of the group 4. This study enabled the plasticity of mesenchymal cells (MCs) in neuronal and glial nineage and opened prospects for the search with cell therapy and transdifferentiation

Centros rombencefálicos de processamento auditivo do sagui (Callithrix jacchus): uma análise citoarquitetônica e neuroquímica

The auditory system is composed by a set of relays from the outer ear to the cerebral cortex. In mammals, the central auditory system is composed by cochlear nuclei, superior olivary complex, inferior colliculus and medial geniculate body. In this study, the auditory rombencephalic centers, the cochlear nuclear complex and the superior olivary complex were evaluated from the cytoarchitecture and neurochemical aspects, thorough Nissl staining and immunohistochemical techniques to reveal specific neuron nuclear protein (NeuN), glutamate (Glu), glutamic acid decarboxilase (GAD), enkephalin (ENK), serotonin (5-HT), choline acetyltransferase (ChAT) and calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV). The common marmoset (Callithrix jacchus), a little native primate of the Brazilian atlantic forest was used as an experimental animal. As results, it was noted that the cochlear nuclear complex is composed by anteroventral, posteroventral and dorsal nuclei, and the superior olivary complex is constituted by the lateral and medial superior olivary nuclei and the trapezoid body nucleus. Glu, GAD, ENK, ChAT, CB, CR, PV-immunoreactive cells, fibers and terminals besides besides only 5-HT terminals were found unhomogeneously in all nuclei, of both complex. The emerging data are discussed in a comparative and functional context, and represent an important contribution to knowledge of the central auditory pathways in the common marmoset, and then in primates

O Núcleo genuíno lateral dorsal do tálamo do sagüi (callithrix jacchus): Pprojeção retiniana, caracterização citoarquitetônica e neuroquimica da principal estação visual primária.

The thalamus plays an important role in the sensorial processing information, in this particular case, the visual information. Several neuronal groups have been characterized as conductors and processors of important sensorial information to the cerebral cortex. The lateral geniculate complex is one to them, and appears as a group very studied once it is responsible, in almost all totality, for the processing of visual information. Among the nuclei that constitute the lateral geniculate complex we highlight the dorsal lateral geniculate nucleus of the thalamus (DLG), the main thalamic relay for the visual information. This nucleus is located rostral and lateral to medial geniculate nucleus and ventral to thalamic pulvinar nucleus in most of the mammals. In the primates humans and non-humans, it presents as a laminate structure, arranged in layers, when observed in coronal sections. The objective of this work was to do a mapping of the retinal projections and a citoarchictetonic and neurochemical characterization of DLG in the marmoset (Callithrix jacchus), a New World primate. The retinal projections were traced by anterograde transport of subunit b of cholera toxin (CTb), the citoarchicteture was described by Nissl method, and to neurochemical characterization immunohistochemicals technical were used to examine the main neurotransmitters and neuroatives substances present in this neural center. In DGL of marmoset thalamus, in coronal sections labeled by Nissl method, was possible to visualize the division of this nucleus in four layers divided in two portions: magnocellular and parvocellular. The retinal projections were present being visualized fibers and terminals immunorreactives to CTb (IR-CTb) in the DLG ipsilateral and contralateral. And through the immunohistochemicals techniques was observed that DLG contain cells, fibers and/or terminals immunoreactives against neuronal nuclear protein, subunits of AMPA 15 glutamate receptors (GluR1, GluR2/3, GluR4), choline acetyltransferase, serotonin, glutamic acid decarboxylase, binding calcium proteins (calbindin, parvalbumin and calretinin), vasopressin, vasoactive intestinal polypeptide, and an astrocyte protein, glial fibrillary acidic protein.

Caracterização citoarquitetônica, neuroquímica e de aferência óptica do complexo parabraquial do sagui (Callithrix jacchus)

The parabrachial complex (PB) is an area of the brainstem responsible for the processing and transmission of essential physiologic information for the survival of the organisms. This region is subdivided in approximately nine subregions, considering morphology, cytoarchitectural and functional characteristic. Its neurons have an extensive network of connections with other regions of the nervous system. The objective in this work was to map the retinal projection to the PB and make a citoarchitectonic and neurochemical characterization of this region in the common marmoset (Callithrix jacchus), a primate of the New World. The retinal projections were mapped by anterograde transport of the choleric toxin subunit b (CTb). The citoarchitecture was described through the Nissl method, and the neurochemical characterization was made through immunohistochemical technique to the some neurotransmitters and neuroactives substances present in this neural center. In marmoset PB, in the coronal sections labeled by Nissl method, we found a similar pattern to that evidenced in other animal species. The immunoreactivity against CTb was verified in the PBMv in fibers/terminal, characterizing such as retinal innervations in this area. The immunohistochemical technique reveled that the PB contain cells, fibers and/or terminals immunoreactives to the neuronal nuclear protein, Choline acetyl transferase, nitric oxide synthase, serotonin, enkephalin, substance P, Calcium-binding proteins (calbindin, calretinin e parvalbumin), and glial fibrillary acidic protein. The histochemical technique reveled cells and fibers NADPH-diaphorase reactive. Each one of those substances presented a characteristic pattern of demarcation in PB, and some serve as specific markers of subregions

Análise imunoistoquímica da distribuição de Serotonina, transportador de serotonina e receptores de serotonina no hipotálamo do sagui (callithrix jacchus)

The hypothalamus is a diencephalic portion located around the third ventricle below the hypothalamic sulcus, limited by the optic chiasm, and by the mammillary bodies, acting as a center that integrates behavioral and homeostatic functions. Serotonin is a neurotransmitter produced in limited sites in the midbrain and brain stem, but is distributed throughout the central nervous system and has many functions, acting through specific receptors that are also distributed throughout the nervous system. Using immunohistochemical techniques, the aim of this study was to delineate the hypothalamic nuclei of the marmoset (Callithrix jacchus) and study the distribution of serotonin transporter and serotonin receptors in the hypothalamus of this species. We used the Nissl method to determine the cytoarchitecture of the hypothalamic nuclei, and immunohistochemistry to reveal the presence of NeuN as a method to determine the contours of the hypothalamic nuclei. As a result, we found serotonin containing fibers and terminals throughout the rostrocaudal extent of the hypothalamus, more concentrated in some nuclei, and even absent in some. Like serotonin, serotonin transporter was observed between pre-optic area and tuberal region of the hypothalamus, in densities and distribution similar to serotonin. The 5-HT1A and 5-HT1B receptors were found with minor differences among itselves regarding the disposition and intensity of staining.

Efeito da degradação ambiental nas propriedades de cisalhamento de compósitos PPS/fibra de carbono

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

Avaliações térmica e reológica da matriz termoplástica PEKK utilizada em compósitos aeronáuticos

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

Projeção retiniana, caracterização citoarquitetônica e neuroquímica da zona incerta do moco (Kerodon rupestris)

The Zona Incerta (ZI) is embryologically derived from the ventral thalamus, in continuity with the reticular nucleus of the thalamus. Studies usingneural tracers technics have allowed identify a complex connectional map including the ZI. Futhermore, cytochemical, molecular and functional data have shown abundant variability in the neurochemical contend in the ZI, as well as,the involvement of the ZI in the modulation of nociception, attention, alertness, control and maintenance of posture and control of visceral activity. This work aims to characterize the cytoarchitecture, neurochemical content of the ZI in the rock cavy (Kerodon rupestris), and a direct retinal-ZI pathway present in this species. The Nissl staining is effective for the delineation and characterization of ZI citoarchitecture. ZIc receives a contralateral retinal projection showing varicosities, suggesting a modulatory character of photic information. The ZI in the rock cavy, as in others rodents and primates, is characterized by a complex neurochemical signature. The ZI neurochemistry presents great diversity, especially in the medial portion of ZIr, where we have found immunoreactivity of all neuroactive substances investigated, and that NOS-IR, GFAP and CR helped the delimitation of middle ZI in ZId and ZIv. Nevertheless, just 5-HT-IR fibers are present in all subdivisions of the ZI. These data demonstrate the great wealth of the neurochemistry of rock cavy s ZI and a direct retinal modulation in the ZI, helping to explain it s broad functional repertory

Utilização de microemulsão na remoção da cor de efluentes têxteis contendo corantes dispersos

Polyester fibers are the most used fibers in the world and disperse dyes are used for dyeing these fibers. After dyeing, the colorful dyebath is discharged into effluent streams, which needs a special treatment for color removal. Surfactants interaction with dyes has been evaluated in several studies, including the textile area, specifically in the separation of dyes from textile wastewater. In this work a cationic surfactant was used in a microemulsion system for the extraction of anionic dyes (disperses dyes) from textile wastewater. These microemulsion system was composed by dodecylamonium chloride (surfactant), kerosene oil (organic phase), isoamyl alcohol (cosurfactant) and the wastewater (aqueous phase). The wastewater that results after the dyeing process is acid (pH 5). It was observed that changing the pH value to above 12.8 the extraction could be made, resulting in an aqueous phase with low color level. The Scheffé net experimental design was used for the extraction process optimization, and the obtained results were evaluated using the program "Statistica 7.0". The optimal microemulsion system was composed by 59.8wt.% of wastewater, 30.1wt.% of kerosene, 3.37wt.% of surfactant and 6.73wt.% of cosurfactant, providing extraction upper than 96%. A mix of reactive dyebath (50%) and disperse dyebath (50%) was used as aqueous phase and it presented extraction upper than 98%. The water phase after extraction process can be reused in a new dyeing, being obtained satisfactory results, according to the limits established by textile industry for a good dyeing. Tests were accomplished seeking to study the influence of salt addition and temperature. An experimental design was used for this purpose, which showed that the extraction doesn't depend on those factors. In this way, the removal of color from textile wastewater by microemulsion is a viable technique (that does not depend of external factors such as salinity and temperature), being obtained good extraction results even with in wastewater mixtures

Produção e composição bromatológica de forrageiras em sistema de integração lavoura-pecuária em diferentes épocas de semeadura

O objetivo deste trabalho foi avaliar a produção de matéria seca e a composição bromatológica de forrageiras em sistema de integração lavoura-pecuária, em diferentes datas de semeadura. Foi usado delineamento experimental de blocos ao acaso, em esquema fatorial 4x3, com quatro forrageiras - braquiária, Urochloa brizantha; capim-moa, Setaria italica; milheto, Pennisetum glaucum; e sorgo forrageiro, Sorghum bicolor - semeadas em três datas nas estações inverno/primavera e verão/outono, em sucessão à soja. O milheto e o sorgo forrageiro produziram maior quantidade de matéria seca em todas as datas de semeadura, nas duas estações de cultivo e, ao contrário da braquiária e do capim-moa, não diminuíram o teor de nutrientes digestíveis totais no inverno/primavera. Os teores de proteína bruta reduziram com o avanço das datas de semeadura no inverno/primavera e aumentaram no verão/outono, exceto para braquiária e milheto. O sorgo apresentou os menores teores de fibras em detergente neutro nas segunda e terceira datas de semeadura no inverno/primavera, e na primeira data de semeadura no verão/outono. Nas segunda e terceiras datas do inverno/primavera, o sorgo apresentou menor teor de fibras em detergente ácido. Quanto aos constituintes da parede celular, apenas os teores de hemicelulose não se alteram nas forrageiras em virtude das datas de semeadura, em ambas as estações.

Double muscle innervation using end-to-side neurorrhaphy in rats

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

Efeito do diclofenaco de sódio na cicatrização da parede abdominal de ratos. Estudo histopatológico, da força de ruptura e do colágeno tecidual

Com o objetivo de estudar os efeitos do diclofenaco sódico sobre a cicatrização da parede abdominal de ratos, foram utilizados 80 animais da linhagem Wistar divididos em dois grupos: Grupo 1: formado por 40 animais submetidos à laparotomia mediana e à injeção intramuscular de soro fisiológico durante quatro dias. Grupo 2: formado por 40 animais submetidos à laparotomia mediana e à injeção intramuscular de diclofenaco sódico durante quatro dias. Animais de ambos os grupos foram analisados no 5°, 7°, 14° e 21° dias de pós-operatório, correspondendo, respectivamente à M1, M2, M3 e M4. em cada momento foram estudados 10 animais e os parâmetros analisados foram a evolução clínica, a força de ruptura, estudo histológico e o conteúdo de colágeno tecidual da parede abdominal. Os animais do grupo tratado apresentaram como complicações, deiscência e/ou hérnia incisional, perda ponderal e taxa de mortalidade, complicações estas não evidenciadas no grupo controle. Observamos também neste grupo, diminuição da força de ruptura no 7° e 14° dia de pós-operatório e diminuição da concentração de colágeno tecidual no 5°, 7° e 14° dia de pós-operatório. Ambos os parâmetros retornaram a valores normais no 21° dia de pós-operatório. Quanto ao estudo histológico, concluímos que a cicatriz da parede abdominal dos animais tratados com D.S. apresentam retardo do processo cicatricial em relação aos seus controles, caracterizado por uma menor fibrogênese, menor densidade de fibras colágenas, além de um número maior de complicações, como microabscessos, em torno dos fios de sutura.

Muscle reinnervation in one or two stages?: experimental study in rats with end-to-side nerve graft

OBJETIVO: Comparar a reinervação muscular com enxerto de nervo em um e dois tempos operatórios, utilizando a neurorrafia término-lateral (NTL) sem lesão do nervo doador. MÉTODOS: Vinte ratos foram distribuídos em quatro grupos. O grupo 1 (G1), um estágio, recebeu o enxerto que foi suturado ao nervo tibial (NT), por meio de NTL, e seu coto livre foi suturado por NTL ao coto distal do nervo peroneal (NP), seccionado a um centímetro do NT, na mesma cirurgia. O grupo 2 (G2), dois estágios, recebeu o enxerto de nervo na primeira cirurgia, como já descrito. Dois meses depois, na segunda cirurgia, o NP foi seccionado e seu coto distal ligado ao coto distal do enxerto como em G1. O grupo controle de normalidade (Gn) recebeu o enxerto da mesma forma, apenas. E o grupo controle de denervação (Gd), além de receber o enxerto, teve o NP seccionado e seus cotos sepultados na musculatura adjacente, com a finalidade de denervar o músculo tibial cranial (MTC), alvo deste estudo. Os parâmetros utilizados para avaliar a reinervação do MTC foram massa muscular, diâmetro mínimo da fibra muscular e área. RESULTADOS: O grupo G2 apresentou superioridade (p<0,0001) em relação ao G1 na massa do MTC, no diâmetro mínimo e na área das fibras musculares. Na comparação entre os quatro grupos, estes mesmos parâmetros tiveram sua expressão máxima em Gn e mínima em Gd, como era esperado. CONCLUSÃO: A reinervação muscular em dois estágios apresenta melhor resultado quando comparada à técnica em um tempo.

Muscle graft as a substitute for peripheral nerve graft in rats

OBJETIVO: Avaliar a aplicabilidade do uso de músculo autógeno, tratado de diversas maneiras, em substituição aos enxertos de nervo. MÉTODOS: Os ratos foram separados em sete grupos que receberam, como tratamento a uma lesão nervosa padronizada, os seguintes tipos de enxertos: músculo fresco, músculo fixado com formol 10%, músculo congelado em freezer, músculo congelado em refrigerador, músculo denervado, nervo periférico e um grupo ficou sem qualquer tratamento. Foi avaliado o aspecto histológico das fibras nervosas no segmento reparado. RESULTADOS: A avaliação do segmento nervoso reparado mostrou que existiam axônios em quase todos os grupos, mas a metodologia empregada não possibilitou caracterizar adequadamente as diferenças entre os grupos. CONCLUSÃO: Este estudo mostrou a migração de axônios por meio de todos os enxertos utilizados.