Structural recovery in lesioned adult mammalian spinal cord by x-irradiation of the lesion site.

Mechanical injury to the adult mammalian spinal cord results in permanent morphological disintegration including severance/laceration of brain-cord axons at the lesion site. We report here that some of the structural consequences of injury can be averted by altering the cellular components of the lesion site with x-irradiation. We observed that localized irradiation of the unilaterally transected adult rat spinal cord when delivered during a defined time-window (third week) postinjury prevented cavitation, enabled establishment of structural integrity, and resulted in regrowth of severed corticospinal axons through the lesion site and into the distal stump. In addition, we examined the natural course of degeneration and cavitation at the site of lesion with time after injury, noting that through the third week postinjury recovery processes are in progress and only at the fourth week do the destructive processes take over. Our data suggest that the adult mammalian spinal cord has innate mechanisms required for recovery from injury and that timed intervention in certain cellular events by x-irradiation prevents the onset of degeneration and thus enables structural regenerative processes to proceed unhindered. We postulate that a radiation-sensitive subgroup of cells triggers the delayed degenerative processes. The identity of these intrusive cells and the mechanisms for triggering tissue degeneration are still unknown.

Estrogen reduces atherosclerotic lesion development in apolipoprotein E-deficient mice.

We have studied the effects of endogenous and exogenous estrogen on atherosclerotic lesions in apolipoprotein E-deficient mice. Female mice ovariectomized (OVX) at weaning displayed increases (P < 0.01) in fatty streak lesions in the proximal aorta and aortic sinus compared with female mice with intact ovarian function. These differences between the OVX and sham controls were apparent in both chow- and "Western-type" diet-fed mice. Moreover, increases in lesion size following OVX occurred without changes in plasma cholesterol. Hormone replacement with subdermal 17-beta-estradiol pellets releasing either 6, 14, or 28 micrograms/day significantly decreased (P < 0.001) atherosclerotic lesion area in both male and OVX female mice. In contrast, neither 17-alpha-estradiol (28 micrograms/day) or tamoxifen (85 micrograms/day) affected lesion progression in OVX female mice. In the Western diet-fed group, exogenous estradiol markedly reduced plasma cholesterol and triglycerides, whereas, in animals fed the chow diet, exogenous estrogen and tamoxifen treatment only decreased plasma and very low density lipoprotein triglycerides. However, lesion area was only weakly correlated with plasma cholesterol and triglycerides, 0.35 and 0.44 tau values, respectively (P < 0.01). In summary, in the apolipoprotein E-deficient mouse 17-beta-estradiol protects against atherosclerotic lesion formation, and this can only be partially explained through effects on plasma lipoprotein levels.

Axotomized neonatal motoneurons overexpressing the bcl2 proto-oncogene retain functional electrophysiological properties.

Bcl2 overexpression prevents axotomy-induced neuronal death of neonatal facial motoneurons, as defined by morphological criteria. However, the functional properties of these surviving lesioned transgenic neurons are unknown. Using transgenic mice overexpressing the protein Bcl2, we have investigated the bioelectrical properties of transgenic facial motoneurons from 7 to 20 days after neonatal unilateral axotomy using brain-stem slices and whole cell patch-clamp recording. Nonaxotomized facial motoneurons from wild-type and transgenic mice had similar properties; they had an input resistance of 38 +/- 6 M omega and fired repetitively after injection of positive current pulses. When cells were voltage-clamped at or near their resting membrane potential, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), N-methyl-D-aspartic acid (NMDA), or vasopressin generated sustained inward currents. In transgenic axotomized mice, facial motoneurons could be found located ipsilaterally to the lesion; they had an input resistance of 150 +/- 30 M omega, indicating that they were smaller in size, fired repetitively, and were also responsive to AMPA, NMDA, and vasopressin. Morphological measurements achieved 1 week after the lesion have shown that application of brain-derived neurotrophic factor prevented the reduction in size of axotomized transgenic motoneurons. These data indicate that Bcl2 not only prevents morphological apoptotic death of axotomized neonatal transgenic motoneurons but also permits motoneurons to conserve functional electrophysiological properties.

Distinct mechanisms underlie activation of hypothalamic neurosecretory neurons and their medullary catecholaminergic afferents in categorically different stress paradigms.

Intermittent electrical footshock induces c-fos expression in parvocellular neurosecretory neurons expressing corticotropin-releasing factor and in other visceromotor cell types of the paraventricular hypothalamic nucleus (PVH). Since catecholaminergic neurons of the nucleus of the solitary tract and ventrolateral medulla make up the dominant loci of footshock-responsive cells that project to the PVH, these were evaluated as candidate afferent mediators of hypothalamic neuroendocrine responses. Rats bearing discrete unilateral transections of this projection system were exposed to a single 30-min footshock session and sacrificed 2 hr later. Despite depletion of the aminergic innervation on the ipsilateral side, shock-induced up-regulation of Fos protein and corticotropin-releasing factor mRNA were comparable in strength and distribution in the PVH on both sides of the brain. This lesion did, however, result in a substantial reduction of Fos expression in medullary aminergic neurons on the ipsilateral side. These results contrast diametrically with those obtained in a systemic cytokine (interleukin 1) challenge paradigm, where similar cuts ablated the Fos response in the ipsilateral PVH but left intact the induction seen in the ipsilateral medulla. We conclude that (i) footshock-induced activation of medullary aminergic neurons is a secondary consequence of stress, mediated via a descending projection transected by our ablation, (ii) stress-induced activation of medullary aminergic neurons is not necessarily predictive of an involvement of these cell groups in driving hypothalamic visceromotor responses to a given stressor, and (iii) despite striking similarities in the complement of hypothalamic effector neurons and their afferents that may be activated by stresses of different types, distinct mechanisms may underlie adaptive hypothalamic responses in each.

Differential replication of a single, UV-induced lesion in the leading or lagging strand by a human cell extract: fork uncoupling or gap formation.

We have constructed simian virus 40 minireplicons containing uniquely placed cis,syn-thymine dimers (T <> T) for the analysis of leading- and lagging-strand bypass replication. Assaying for replication in a human cell-free extract through the analysis of full-size labeled product molecules and restriction fragments spanning the T <> T site resulted in the following findings: (i) The primary site of synthesis blockage with T <> T in either the leading or lagging strand was one nucleotide before the lesion. (ii) Replicative bypass of T <> T was detected in both leading and lagging strands. The efficiency of synthesis past T <> T was 22% for leading-strand T <> T and 13% for lagging-strand T <> T. (iii) The lagging-strand T <> T resulted in blocked retrograde synthesis with the replication fork proceeding past the lesion, resulting in daughter molecules containing small gaps (form II' DNA). (iv) With T <> T in the leading-strand template, both the leading and lagging strands were blocked, representing a stalled replication fork. Uncoupling of the concerted synthesis of the two strands of the replication fork was observed, resulting in preferential elongation of the undamaged lagging strand. These data support a model of selective reinitiation downstream from the lesion on lagging strands due to Okazaki synthesis, with no reinitiation close to the damage site on leading strands [Meneghini, R. & Hanawalt, P.C. (1976) Biochim. Biophys. Acta 425, 428-437].

Glial cell line-derived neurotrophic factor but not transforming growth factor beta 3 prevents delayed degeneration of nigral dopaminergic neurons following striatal 6-hydroxydopamine lesion.

Glial cell line-derived neurotrophic factor (GDNF) and transforming growth factor beta 3 (TGF-beta 3) are members of the TGF-beta superfamily with high neurotrophic activity on cultured nigral dopamine neurons. We investigated the effects of intracerebral administration of GDNF and TGF-beta 3 on the delayed cell death of the dopamine neurons in the rat substantia nigra following 6-hydroxydopamine lesions of dopaminergic terminals in the striatum. Fluorescent retrograde tracer injections and tyrosine hydroxylase immunocytochemistry demonstrated nigral degeneration with an onset 1 week after lesion, leading to extensive death of nigral neurons 4 weeks postlesion. Administration of recombinant human GDNF for 4 weeks over the substantia nigra at a cumulative dose of 140 micrograms, starting on the day of lesion, completely prevented nigral cell death and atrophy, while a single injection of 10 micrograms 1 week postlesion had a partially protective effect. Continuous administration of TGF-beta 3, starting on the day of lesion surgery, did not affect nigral cell death or atrophy. These findings support the notion that GDNF, but not TGF-beta 3, is a potent neurotrophic factor for nigral dopamine neurons in vivo.

Localization of visual stimuli after striate cortex damage in monkeys: parallels with human blindsight.

Blindsight is a phenomenon in which human patients with damage to striate cortex deny any visual sensation in the resultant visual field defect but can nonetheless detect and localize stimuli when persuaded to guess. Although monkeys with striate lesions have also been shown to exhibit some residual vision, it is not yet clear to what extent the residual capacities in monkeys parallel the phenomenon of human blindsight. To clarify this issue, we trained two monkeys with unilateral lesions of striate cortex to make saccadic eye movements to visual targets in both hemifields under two conditions. In the condition analogous to clinical perimetry, they failed to initiate saccades to targets presented in the contralateral hemifield and thus appeared "blind." Only in the condition where the fixation point was turned off simultaneously with the onset of the target--signaling the animal to respond at the appropriate time--were monkeys able to localize targets contralateral to the striate lesion. These results indicate that the conditions under which residual vision is demonstrable are similar for monkeys with striate cortex damage and humans with blindsight.

Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation.

Enteropathogenic Escherichia coli (EPEC) causes a characteristic histopathology in intestinal epithelial cells called the attaching and effacing lesion. Although the histopathological lesion is well described the bacterial factors responsible for it are poorly characterized. We have identified four EPEC chromosomal genes whose predicted protein sequences are similar to components of a recently described secretory pathway (type III) responsible for exporting proteins lacking a typical signal sequence. We have designated the genes sepA, sepB, sepC, and sepD (sep, for secretion of E. coli proteins). The predicted Sep polypeptides are similar to the Lcr (low calcium response) and Ysc (yersinia secretion) proteins of Yersinia species and the Mxi (membrane expression of invasion plasmid antigens) and Spa (surface presentation of antigens) regions of Shigella flexneri. Culture supernatants of EPEC strain E2348/69 contain several polypeptides ranging in size from 110 kDa to 19 kDa. Proteins of comparable size were recognized by human convalescent serum from a volunteer experimentally infected with strain E2348/69. A sepB mutant of EPEC secreted only the 110-kDa polypeptide and was defective in the formation of attaching and effacing lesions and protein-tyrosine phosphorylation in tissue culture cells. These phenotypes were restored upon complementation with a plasmid carrying an intact sepB gene. These data suggest that the EPEC Sep proteins are components of a type III secretory apparatus necessary for the export of virulence determinants.

Lesion-induced reorganization in the brainstem is not completely expressed in somatosensory cortex.

Electrophysiological and neuroanatomical methods were used to determine the extent to which neonatal forelimb removal altered the organization of the cuneate nucleus and representations of the fore- and hindlimbs in the primary somatosensory cortex of adult rats. Neonatal forelimb removal resulted in invasion of the cuneate nucleus by sciatic nerve primary afferents and development of cuneothalamic projection neurons with split receptive fields that included both the hindlimb and forelimb stump. Mapping in the primary somatosensory cortex of the neonatally manipulated adult rats demonstrated abnormalities, but the major change observed in the cuneate nucleus was demonstrable at only a few (5%) cortical recording sites in the remaining stump representation and there were none at all in the hindlimb representation. These results suggest that lesion-induced brainstem reorganization may be functionally suppressed at either the thalamic or cortical level.

Avaliação em 3D da cabeça da mandíbula em crianças portadoras de mordida cruzada posterior unilateral por meio de tomografia computadorizada de feixe cônico

Durante o desenvolvimento da oclusão, a instalação de maloclusões podem resultar em desarmonias dento faciais de natureza e severidade diversas, podendo provocar alterações no desenvolvimento crânio facial, dentre as estruturas envolvidas as Articulações Temporo Mandibulares (ATM), podem sofrer alguma influência, dessa forma a avaliação desta região, no aspecto morfológico e funcional, constituí tema de interesse, sempre que levados em conta os aspectos funcionais da oclusão. A relação entre a forma e a função, tanto das cabeças da mandíbula, bem como o contorno da fossa mandibular com as maloclusões ainda é controversa e não está compreendida por completo, porém a literatura sobre o assunto, demonstra correlação entre a instalação de maloclusões e modificações neste sistema, mesmo que algumas alterações não sejam de ordem estatística e em amostras de indivíduos em tenra idade, as mesmas podem comprometer o desenvolvimento adequado em indivíduos adultos ou mesmo adultos jovens. Tendo como propósito nesse estudo a avaliação das cabeças da mandíbula quanto ao volume e superficíe dos lados direito e esquerdo, cruzado e não cruzado, a amostra selecionada foi de 20 indivíduos com mordida cruzada posterior unilateral, com idades entre 06 e 09 anos de idade, utilizando imagens de tomografia computadorizada por feixe cônico, imagens obtidas por um equipamento modelo i- Cat, sendo utilizado na reformatação e manipulação das imagens o programa computacional - NemoCeph 3D® versão 11.5. Nas medições propostas para esse estudo, utilizou-se o teste t pareado de Student para amostras com distribuição normal. Na observação das tabelas e seus respectivos gráficos, podemos verificar que na comparação entre os lados direito e esquerdo, e cruzado e não cruzado das cabeças da mandíbula, com relação ao volume e superfície, existem diferenças numéricas entre elas, porém não pode ser observado diferenças estatísticas significantes, nessa amostra especifica com a metodologia empregada para esse estudo. Assim foi possível concluir que nas Mordidas Cruzadas Posteriores Unilaterais as cabeças da mandíbula tanto em seu volume como em sua superfície não apresentaram diferenças estatisticamente significantes na amostra estudada.

Perda auditiva unilateral: benefício da amplificação na ordenação e resolução temporal e localização sonora

A perda auditiva unilateral (PAUn) é caracterizada pela diminuição da audição em apenas uma orelha. Indivíduos com este tipo de perda auditiva podem apresentar comprometimento nas habilidades auditivas de localização sonora, processamento temporal, ordenação e resolução temporal. O objetivo deste estudo foi verificar as habilidades auditivas de ordenação temporal, resolução temporal e localização sonora, antes e após a adaptação do aparelho de amplificação sonora individual (AASI). Foram avaliados 22 indivíduos, com idades entre 18 e 60 anos, com diagnóstico de PAUn sensorioneural ou mista, de graus leve a severo. O estudo foi dividido em duas etapas: a pré e a pós-adaptação de AASI. Em ambas as etapas, os indivíduos foram submetidos a uma anamnese, aplicação do Questionário de Habilidade Auditiva da Localização da fonte sonora, avaliação simplificada do processamento auditivo (ASPA) e Random Gap Detection Test (RGDT). O presente estudo encontrou diferença estatisticamente significante na avaliação da ASPA, exceto no teste de memória para sons não verbais em sequência (TMSnV), no RGDT e no Questionário de Habilidade Auditiva da Localização Sonora. A conclusão do estudo foi que com o uso efetivo do AASI, indivíduos com PAUn apresentaram melhora nas habilidades auditivas de localização sonora, ordenação e resolução temporal.

Avaliação de linguagem por ressonância magnética funcional em pacientes com epilepsia associada à esclerose mesial temporal unilateral: correlação com avaliação clínica de linguagem

Introdução: A esclerose mesial temporal (EMT) é a principal causa de epilepsia resistente ao tratamento medicamentoso. Pacientes com EMT apresentam dificuldades no processamento semântico e fonológico de linguagem e maior incidência de reorganização cerebral da linguagem (bilateral ou à direita) em relação à população geral. A ressonância magnética funcional (RMf) permite avaliar a reorganização cerebral das redes de linguagem, comparando padrões de ativação cerebral entre diversas regiões cerebrais. Objetivo: Investigar o desempenho linguístico de pacientes com EMT unilateral esquerda e direita e a ocorrência de reorganização das redes de linguagem com RMf para avaliar se a reorganização foi benéfica para a linguagem nestes pacientes. Métodos: Utilizamos provas clínicas de linguagem e paradigmas de nomeação visual e responsiva para RMf, desenvolvidos para este estudo. Foram avaliados 24 pacientes com EMTe, 22 pacientes com EMTd e 24 controles saudáveis, submetidos a provas de linguagem (fluência semântica e fonológica, nomeação de objetos, verbos, nomes próprios e responsiva, e compreensão de palavras) e a três paradigmas de linguagem por RMf [nomeação por confrontação visual (NCV), nomeação responsiva à leitura (NRL) e geração de palavras (GP)]. Seis regiões cerebrais de interesse (ROI) foram selecionadas (giro frontal inferior, giro frontal médio, giro frontal superior, giro temporal inferior, giro temporal médio e giro temporal superior). Índices de Lateralidade (ILs) foram calculados com dois métodos: bootstrap, do programa LI-Toolbox, independe de limiar, e PSC, que indica a intensidade da ativação cerebral de cada voxel. Cada grupo de pacientes (EMTe e EMTd) foi dividido em dois subgrupos, de acordo com o desempenho em relação aos controles na avaliação clinica de linguagem. O <= -1,5 foi utilizado como nota de corte para dividir os grupos em pacientes com bom e com mau desempenho de linguagem. Em seguida, comparou-se o desempenho linguístico dos subgrupos ao índices IL-boot. Resultados: Pacientes com EMT esquerda e direita mostraram pior desempenho que controles nas provas clínicas de nomeação de verbos, nomeação de nomes próprios, nomeação responsiva e fluência verbal. Os mapas de ativação cerebral por RMf mostraram efeito BOLD em regiões frontais e temporoparietais de linguagem. Os mapas de comparação de ativação cerebral entre os grupos revelaram que pacientes com EMT esquerda e direita apresentam maior ativação em regiões homólogas do hemisfério direito em relação aos controles. Os ILs corroboraram estes resultados, mostrando valores médios menores para os pacientes em relação aos controles e, portanto, maior simetria na representação da linguagem. A comparação entre o IL-boot e o desempenho nas provas clínicas de linguagem indicou que, no paradigma de nomeação responsiva à leitura, a reorganização funcional no giro temporal médio, e possivelmente, nos giros temporal inferior e superior associou-se a desempenho preservado em provas de nomeação. Conclusão: Pacientes com EMT direita e esquerda apresentam comprometimento de nomeação e fluência verbal e reorganização da rede cerebral de linguagem. A reorganização funcional de linguagem em regiões temporais, especialmente o giro temporal médio associou-se a desempenho preservado em provas de nomeação em pacientes com EMT esquerda no paradigma de RMf de nomeação responsiva à leitura