Neurologic Disorders in Immunocompetent Patients with Autochthonous Acute Hepatitis E.

Neurologic disorders, mainly Guillain-Barré syndrome and Parsonage­Turner syndrome (PTS), have been described in patients with hepatitis E virus (HEV) infection in industrialized and developing countries. We report a wider range of neurologic disorders in nonimmunocompromised patients with acute HEV infection. Data from 15 French immunocompetent patients with acute HEV infection and neurologic disorders were retrospectively recorded from January 2006 through June 2013. The disorders could be divided into 4 main entities: mononeuritis multiplex, PTS, meningoradiculitis, and acute demyelinating neuropathy. HEV infection was treated with ribavirin in 3 patients (for PTS or mononeuritis multiplex). One patient was treated with corticosteroids (for mononeuropathy multiplex), and 5 others received intravenous immunoglobulin (for PTS, meningoradiculitis, Guillain-Barré syndrome, or Miller Fisher syndrome). We conclude that pleiotropic neurologic disorders are seen in HEV-infected immunocompetent patients. Patients with acute neurologic manifestations and aminotransferase abnormalities should be screened for HEV infection.

Neuromodulatory role of acetylcholine in visually-induced cortical activation: behavioral and neuroanatomical correlates.

Acetylcholine is released in the primary visual cortex during visual stimulation and may have a neuromodulatory role in visual processing. The present study uses both behavioral and functional neuroanatomy investigations to examine this role in the rat. In the first set of experiments the cholinergic system was lesioned with 192 immunoglobulin G (IgG) saporin and the visual acuity and performance in a visual water maze task were assessed. The cholinergic lesion did not affect the visual acuity measured pre- and post-lesion but it did reduce the efficiency to learn a novel orientation discrimination task measured post-lesion. In order to better understand the involvement of the cholinergic system in the neuronal activity in the visual cortex c-Fos expression induced by patterned visual stimulation was further investigated. Results obtained following lesion of the cholinergic fibers (192 IgG-saporin or quisqualic acid), muscarinic inhibition (scopolamine), or NMDA receptor inhibition (CPP) were compared with control conditions. Double and triple immunolabeling was used in order to determine the neurochemical nature of the activated cortical cells. The results demonstrated that patterned stimulation elicited a significant increase in c-Fos immunolabeled neurons in layer IV of the contralateral primary visual cortex to the stimulated eye which was completely abolished by cholinergic fibers lesion as well as scopolamine administration. This effect was independent of NMDA receptor transmission. The c-Fos activation was predominantly observed in the glutamatergic spiny stellate cells and less frequently in GABAergic interneurons. Altogether, these results demonstrate a strong involvement of the basal forebrain cholinergic system in the modulation of post-synaptic visual processing, which could be related to cognitive enhancement or attention during visual learning.

Acetylcholine release is elicited in the visual cortex, but not in the prefrontal cortex, by patterned visual stimulation: a dual in vivo microdialysis study with functional correlates in the rat brain.

By its projections to the primary visual and the prefrontal cortices, the basal forebrain cholinergic system is involved in cognitive processing of sensory stimuli. It has been suggested that visual stimulus-induced cholinergic activation of the visual cortex may exert a permissive role on thalamocortical inputs. However, it is not known if visual stimulation elicits cholinergic activation of high-order brain areas in the absence of attentional need. In the present study, we measured the effects of patterned visual stimulation (horizontal grating) on the release of acetylcholine with dual-probe in vivo microdialysis in the visual and the prefrontal cortices of anesthetized rats. We also used retrograde tracing to determine the anatomical relationships of cholinergic neurons with neurons of the visual system and the prefrontal cortex. Finally, we evaluated a functional correlate of this stimulation, namely c-fos immunolabeling. Patterned visual stimulation elicited significant increases in acetylcholine release in the visual cortex, accompanied by an increased number of c-fos immunoreactive neurons in this brain area. In contrast, in the prefrontal cortex, neither the level of acetylcholine release nor the number of c-fos immunoreactive neurons was significantly changed because of the stimulation. Cholinergic basal forebrain neurons projecting to the visual or the prefrontal cortices were both localized within the horizontal limb of the diagonal band of Broca but were not immunoreactive for c-fos during visual stimulation. No parts of the visual system were found to directly project to these basal forebrain neurons. These results suggest the differential involvement of cholinergic projections in the integration of sensory stimuli, depending on the level of activity of the targeted cortical area.

Object recognition memory and cholinergic parameters in mice expressing human presenilin 1 transgenes.

Most autosomal dominant forms of Alzheimer disease (AD) are related to missense mutations in the human presenilin (PS) 1 gene. Although the underlying mechanisms associated with pathophysiology of AD have yet to be clearly established, pathogenic mutations in the PS1 gene influence the processing of beta-amyloid precursor protein, leading to increased production and deposition of highly fibrillogenic amyloid beta(1-42) peptide in the brains of AD patients. As cognitive dysfunction in AD is associated with a dramatic loss of cholinergic innervation particularly in the hippocampus and neocortex, we investigated learning and cholinergic neurochemistry in transgenic mice expressing pathogenic mutant L286V or wild-type(wt) human PS1 transgenes. Relative to wt, the L286V PS1 transgenic mice exhibited reduced sensorimotor activity and marked deterioration of object memory between 3 and 5 h after the first encounter. Activity of the biosynthetic enzyme choline acetyltransferase was not altered in the hippocampus, frontoparietal cortex, or striatum of mutant transgenic mice relative to wt transgenic or littermate nontransgenic controls. No differences in the densities of M1/[3H]pirenzepine, M2/[3H]AF-DX 384, or alpha(7) nicotinic/125I-alpha-bungarotoxin receptor binding sites were evident in any brain regions among L286V PS1 transgenic, wt PS1 transgenic, and littermate nontransgenic controls. These results suggest that overexpression of a mutated PS1 gene induces a subtle alteration in object memory without affecting cholinergic neurochemistry.

Amyloid beta peptide levels and its effects on hippocampal acetylcholine release in aged, cognitively-impaired and -unimpaired rats.

Excessive extracellular deposition of amyloid beta (Abeta) peptide in neuritic plaques and degeneration of forebrain cholinergic neurones, which innervate the hippocampus and the neocortex, are the invariant characteristic features of Alzheimer's disease (AD). Studies of the pathological changes that characterize AD, together with several other lines of evidence, indicate that Abeta accumulation in vivo may initiate and/or contribute to the process of neurodegeneration observed in the AD brain. However, the underlying mechanisms by which Abeta peptide influences/causes degeneration of the basal forebrain cholinergic neurones in AD brains remain obscure. We reported earlier that nM concentrations of Abeta-related peptides, under acute conditions, can potently inhibit K+-evoked endogenous acetylcholine (ACh) release from the hippocampus and the cortex but not from striatum in young adult rats (J. Neurosci. 16 (1996) 1034). In the present study, to determine whether the effects of Abeta peptides alter with normal aging and/or cognitive state, we have measured Abeta1-40 levels and the effects of exogenous Abeta1-40 on hippocampal ACh release in young adult as well as aged cognitively-unimpaired (AU) and -impaired (AI) rats. Endogenous levels of Abeta(1-40) in the hippocampus are significantly increased in aged rats. Additionally, 10 nM Abeta1-40 potently inhibited endogenous ACh release from the hippocampus of the three groups of rats, but the time-course of the effects clearly indicate that the cholinergic neurones of AI rats are more sensitive to Abeta peptides than either AU or young adult rats. These results, together with earlier reports, suggest that the processing of the precursor protein of Abeta peptide alters with normal aging and the response of the cholinergic neurones to the peptide possibly varies with the cognitive status of the animals.

Ovariectomy up-regulates neuronal neurofilament light chain mRNA expression with regional and temporal specificity.

Estrogens can influence the survival, plasticity and function of many adult neurons. Many of these effects, such as neurite outgrowth and increased dendritic spine density, are mediated by changes in neuronal cytoskeletal architecture. Since neurofilament proteins play a key role in the maintenance and remodeling of the neuronal cytoskeleton, we postulated that changes in neurofilament light chain mRNA may parallel some of the alterations in neuronal architecture which follow bilateral ovariectomy. We measured neurofilament light chain mRNA levels using a ribonuclease protection assay at two time-points after ovariectomy in mature female rats. One week after ovariectomy, neurofilament light chain mRNA levels (corrected for glucose-6-phosphate dehydrogenase mRNA) did not differ from sham-operated animals in the five brain regions examined (hypothalamus, striatum, hippocampus, frontal cortex and occipital cortex). Four months after ovariectomy, neurofilament light chain mRNA levels were similarly unchanged in the hypothalamus and striatum. In contrast, statistically significant increases in neurofilament light chain mRNA expression were observed in the three regions receiving basal forebrain projections (hippocampus, frontal cortex and occipital cortex). In situ hybridization demonstrated increases in neurofilament light chain mRNA expression involving subpopulations of smaller medial septal neurons. There also appeared to be an increased number of larger septal neurons following long-term ovariectomy. We propose that atrophic changes involving basal forebrain projection fibers are followed by compensatory axonal growth by other 'intact' basal forebrain neurons. Increased neurofilament light chain mRNA expression and somatic hypertrophy in medial septal neurons may both be reflective of the need to sustain an axonal network which is larger and more complex. In contrast, increased neurofilament light chain mRNA expression observed in basal forebrain targets following long-term ovariectomy may be reflective of compensatory changes taking place in local neurons.

GABA neurons provide a rich input to microvessels but not nitric oxide neurons in the rat cerebral cortex: a means for direct regulation of local cerebral blood flow.

Basal forebrain neurons project to microvessels and the somata of nitric oxide (NO) synthase-containing neurons in the cerebral cortex, and their stimulation results in increases in cortical perfusion. gamma-Aminobutyric acid (GABA) is the second major neurotransmitter synthesized by these neurons and it has also been reported to modify cerebromicrovascular tone. We thus investigated by light and electron microscopy the association of GABA neurons (labeled for glutamic acid decarboxylase [GAD]) with cortical microvessels and/or NO neurons (identified by nicotinamide adenine dinucleotide [NADPH-D] histochemistry) within the frontoparietal and perirhinal cerebral cortex in the rat. On thick and semithin sections, a high density of GAD puncta was observed, several surrounded intracortical blood vessels and neuronal perikarya. In contrast, NADPH-D cell somata and proximal dendrites were only occasionally contacted by GAD nerve terminals. Perivascular and perisomatic GAD appositions were identified at the ultrastructural level as large (0.44-0.50 microm(2)) neuronal varicosities located in the immediate vicinity of, or being directly apposed to, vessels or unstained neuronal cell bodies. In both cortical areas, perivascular GAD terminals were located at about 1 microm from the vessels and were seen to frequently establish junctional contacts (synaptic frequency of 25-40% in single thin sections) with adjacent neuronal but not vascular elements. Ibotenic or quisqualic acid lesion of the substantia innominata did not significantly affect the density of cortical and perivascular GAD terminals, suggesting that they mostly originated locally in the cortex. These results suggest that GABA terminals can interact directly with the microvascular bed and that the somata and proximal dendrites of NO neurons are not a major target for cortical GABA neurotransmission. However, based on the colocalization of GABA and NADPH-D in a subset of cortical neurons, we suggest that these interneurons could be implicated in the cortical vascular response elicited by stimulation of basal forebrain neurons.

Reduced cortical vasodilatory response to stimulation of the nucleus basalis of Meynert in the aged rat and evidence for a control of the cerebral circulation.

In earlier studies we showed that electrical stimulation of the rat nucleus basalis of Meynert (NBM) induces large increases in cerebral blood flow, mainly through cholinergic mechanisms. We then investigated the effect of aging on this influence by measuring cortical blood flow (CoBF) and tissue gas partial pressures (PtO2, PtCO2) in the conscious young adult and aged rat. NBM stimulation increased frontal (+101%) and parietal (+29%) CoBF in young rats. The effects were halved in aged rats. Moreover, PtO2 was significantly increased in young but not in aged rats. By contrast, the corticovascular reactivity to hypercapnia did not differ between young and aged rats, nor did the potentiating vasodilator effect of physostigmine. In combined autoradiographic measurements of cerebral blood flow and cerebral glucose utilization, we recently found that the cortical circulatory response to NBM stimulation was not accompanied by significant metabolic change. Thus, the blood flow changes observed in the cortex cannot be ascribed to increased metabolic activity. The distribution of this uncoupling coincides with that of cholinergic NBM projections directly impinging on cortical microvessels. These data support the cortical microcirculation and suggest the possible involvement of NBM dysfunction in the pathology of cortical microcirculation.

Cholinergic basal forebrain projections to nitric oxide synthase-containing neurons in the rat cerebral cortex.

Stimulation of basal forebrain neurons elicits regional cerebral blood flow increases which are reportedly mediated by acetylcholine and nitric oxide. However, the modality of interaction between these two mediators remains unclear. Particularly, little is known about the source, i.e. endothelial, glial and/or neuronal, of the potent gaseous vasodilator nitric oxide. In the present study, we examined, by double immunocytochemical labelling of nitric oxide synthase and choline acteyltransferase at the light and electron microscopic level, the existence of morphological relationships between cortical nitric oxide synthase-containing neurons and cholinergic cells or nerve fibres. Using anterograde tract tracing and selective basal forebrain lesions, we further investigated the origin of the cholinergic input to cortical nitric oxide synthase neurons. The results confirm that cortical nitric oxide synthase-immunoreactive neurons are often associated with the local microvascular bed, show that intracortical neurons immunostained for nitric oxide synthase and choline acetyltransferase belong to two distinct neuronal populations and, further, that a subset of nitric oxide synthase-containing cell bodies and their proximal dendrites receive a cholinergic input which originates primarily from basalocortical projections. Altogether, these findings suggest that cholinergic basal forebrain neurons could increase cortical blood flow partly via a local nitric oxide relay neuron whereby the freely diffusing gas would be the direct smooth muscle vasodilator agent. It is concluded that this interaction might contribute to the complex relationships between the basal forebrain and the cortical microcirculation, interactions which result in fine regulation of cortical perfusion.

Autoradiographic evidence for flow-metabolism uncoupling during stimulation of the nucleus basalis of Meynert in the conscious rat.

We earlier reported that electrical stimulation of the rat nucleus basalis of Meynert (NBM) induces large cerebral blood flow increases, particularly in frontal cortical areas but also in some subcortical regions. The present study was designed to address the issue of blood flow control exerted by NBM projections. To this aim, we have determined whether these flow increases were associated with proportionate changes in metabolic activity as evaluated by cerebral glucose utilization (CGU) strictly under the same experimental conditions in the conscious rat. An electrode was chronically implanted in a reactive site of the NBM as determined by laser-Doppler flowmetry (LDF) of the cortical circulation. One to two weeks later, while the cortical blood flow was monitored by LDF, we measured CGU using the [14C]2-deoxyglucose autoradiographic technique during unilateral electrical stimulation of the NBM, and analyzed the local flow-metabolism relationship. The large increases in cortical blood flow induced by NBM stimulation, exceeding 300% in various frontal areas, were associated with at most 24% increases in CGU (as compared with the control group) in one frontal area. By contrast, strong increases in CGU exceeding 150% were observed in subcortical regions ipsilateral to the stimulation, especially in extrapyramidal structures, associated with proportionate CBF changes. Thus, none of the blood flow changes observed in the cortex can be ascribed to an increased metabolic activity, whereas CBF and CGU were coupled in many subcortical areas. This result indicates that different mechanisms, which do not necessarily involve any metabolic factor, contribute to the regulation of the cerebral circulation at the cortical and subcortical level. Because the distribution of the uncoupling is coincident with that of cholinergic NBM projections directly reaching cortical microvessels, these data strongly support the hypothesis that NBM neurons are capable of exerting a neurogenic control of the cortical microcirculation.

[Cholestasis in pregnancy. A pure cytolytic form]. / Cholestase gravidique. Une forme cytolytique pure.

BACKGROUND: Intrahepatic cholestatic disease in pregnancy is the second most frequent cause of jaundice after viral hepatitis. Elevated serum alkaline phosphatase is always found but is of little aid for diagnosis. CASE REPORT: A 32-year-old woman (third gestation, second parturition) consulted at 31 weeks gestation after progressive development of pruritus then jaundice. Transaminase levels were elevated. The diagnosis of intrahepatic cholestasis was retained despite normal alkaline phosphatase levels throughout the disease course. DISCUSSION: Intrahepatic cholestasis of pregnancy is rare in France compared with its frequency in Latin America and in Scandinavia. The pathogenesis remains uncertain. Diagnosis is based on clinical and laboratory findings as well as morphological arguments. Nevertheless, the results of liver tests vary widely. Transaminase levels may rise greatly but no prior case with normal alkaline phosphatase levels has been reported. Most importantly, clinical course is spontaneously regressive; pruritus and jaundice disappear within a few days after delivery allowing the diagnosis of intrahepatic cholestasis of pregnancy.

Cholinergic basal forebrain neurons project to cortical microvessels in the rat: electron microscopic study with anterogradely transported Phaseolus vulgaris leucoagglutinin and choline acetyltransferase immunocytochemistry.

Physiological evidence indicates that central cholinergic pathways are involved in the regulation of cerebral cortical blood flow. We investigated the possible contribution of basal forebrain cholinergic neurons from the substantia innominata (SI) to the innervation of cortical microvessels. Basalo-cortical perivascular nerve terminals were detected by light and electron microscopic immunocytochemistry of anterogradely transported Phaseolus vulgaris leucoagglutinin (PHA-L) following its injection in the SI, and were compared to cortical perivascular cholinergic (immunoreactive for choline acetyltransferase (ChAT)) terminals. The basal forebrain origin of cholinergic terminals was evaluated after unilateral ibotenic acid lesion of the SI. PHA-L varicose fibers reached and surrounded microvessels in all cortical subdivisions examined. When studied at the ultrastructural level in the fronto-parietal and perirhinal cortices, perivascular PHA-L nerve terminals were located significantly closer to microvessels in the fronto-parietal than in the perirhinal cortex (respective average distance of 0.98 +/- 0.09 and 1.34 +/- 0.07 micron; p < 0.01). PHA-L and ChAT terminals in the fronto-parietal cortex compared very well in their perivascular distribution but not in the perirhinal cortex. In both cortices, perivascular PHA-L terminals were similar in size to, but engaged more frequently in synaptic contacts than their ChAT counterparts. Following SI lesion, the density of cortical ChAT terminals including those reaching microvessels decreased significantly (56 and 63%, respectively, p < 0.005) in the fronto-parietal cortex, while the cortical and perivascular denervations were much less pronounced (26%, not significant and 35%, p < 0.05, respectively) in the perirhinal cortex. These results indicate that basal forebrain neurons project preferentially to fronto-parietal cortical microvessels and further show that a significant proportion of these projections are cholinergic. In addition, the difference in distribution and/or synaptic incidence between perivascular PHA-L and ChAT terminals suggested that noncholinergic SI neurons also contribute to these neurovascular associations, and more so in the perirhinal cortex, as further indicated by the lesion studies. Such finding corroborates recent physiological evidence for a functional innervation of the cortical microvascular bed by SI neurons, a role that might be relevant to the overall pathology of Alzheimer's dementia.

Autoradiographic distribution of cerebral blood flow increases elicited by stimulation of the nucleus basalis magnocellularis in the unanesthetized rat.

The nucleus basalis magnocellularis (NBM) of the rat, equivalent of Meynert's nucleus in the primate, is the origin of the main cholinergic innervation of the cerebral cortex. Stimulation of this area has been previously shown to induced marked, cholinergically mediated, blood flow increases in the frontal and parietal cortices. However, the complete distribution of the cerebrovascular effects of NBM stimulation within the whole brain has not been determined. In the present study, we used the [14C]iodoantipyrine autoradiographic method to measure local cerebral blood flow (CBF) in the unanesthetized rat, chronically implanted with a stimulation electrode. We performed unilateral electrical stimulation of the NBM in order to compare both the interhemispheric differences in blood flow and the differences with a group of sham-stimulated rats. Considerable blood flow increases were found in most neocortical areas, exceeding 400% in the frontal area, compared to the control group. Marked responses also appeared in discrete subcortical regions such as the zona incerta, some thalamic nuclei and structures of the extrapyramidal system. These responses were mostly ipsilateral to the stimulation. The significance and the distribution of these blood flow increases are related first, to anatomical and functional data on mainly the cholinergic projections from the NBM, but also non-cholinergic pathways connected with the NBM, second, to biochemical data on the basalocortical system, and third, to the limited ultrastructural data on the innervation of microvascular elements. This cerebrovascular study represents a step in the elucidation of the function of the basalocortical system and provides data which may be related to certain deficits of degenerative disorders such as Alzheimer's disease in which this system is consistently affected.

Autoradiographic study of the cerebrovascular effects of stimulation of the substantia innominata: convenient stimulation paradigm.

The aim of this study was to determine the distribution within the whole brain of the vascular effects of stimulation of the substantia innominata. This basal forebrain nucleus is the major cholinergic input in the neocortex in the rodent. The local cerebral blood flow was measured by the autoradiographic [14C]iodoantipyrine technique in a group of control and a group of stimulated unanesthetized rats. The substantia innominata was electrically stimulated through a chronically implanted electrode. The stimulation induced blood flow increases exceeding 200% in the hemisphere ipsilateral to the stimulation and 100% in the contralateral hemisphere compared to the control group. The ipsilateral vasodilations were observed not only in the cortical areas but also in some subcortical structures. Comparison with previous data suggests that part of the effects is due to cholinergic neurons of the substantia innominata and part to non-cholinergic neurons and indirect effects. However, only two out of eight stimulated rats displayed this response. The low reproducibility of the results is discussed, considering the stimulation paradigm which has been developed for future measurements of the cerebral glucose utilization which requires a long duration stimulation period.

The cerebrovascular effects of physostigmine are not mediated through the substantia innominata.

This study sought to determine whether the cortical cholinergic projections from Meynert's nucleus are actually the target of the cholinesterase inhibitor physostigmine, which presents the ability to increase cortical blood flow. To this aim, the multiregional cerebrovascular effects of physostigmine in rats with and without lesion of the substantia innominata (SI), the equivalent of Meynert's nucleus of primates, were investigated. Unilateral SI lesions were made using ibotenic acid in three groups of rats. Four to 11 days later, the cortical choline acetyltransferase (ChAT) activity was measured in one group to assess the efficacy of the lesion. In the two other groups, the regional cerebral blood flow was measured using the [14C]iodoantipyrine technique, under physostigmine (0.2 mg/kg/h iv) or control conditions. SI lesion induced 27-59% fall in cortical ChAT activity in the ipsilateral hemisphere with the frontal area most affected. Despite these large biochemical differences, the lesion had little cerebrovascular effects. Side-to-side blood flow differences did not exceed 11% and did not strictly overlap the ChAT depletion. Physostigmine increased flow (38-66%) in all cortical areas, with no frontal predominance. Despite these considerable vasodilations, there were no significant differences between the lesioned and the intact hemisphere, nor any significant interaction between physostigmine and SI lesion. Thus, physostigmine does not actually activate the SI neuron terminals. This result suggests that cholinesterase inhibitors cannot be used as presynaptic markers of the cholinergic activity of this nucleus and casts doubts on their specificity as enhancement therapeutic agents in Alzheimer's disease.