[Biofeedback and drug-resistant epilepsy: back to an earlier treatment?]. / Biofeedback et épilepsie pharmacorésistante : le retour d'une thérapeutique ancienne ?

Biofeedback is a complementary non-pharmacological and non-surgical therapeutic developed over the last thirty years in the management of drug-resistant epilepsy. Biofeedback allows learning cognitive and behavioral strategies via a psychophysiological feedback loop. Firstly, this paper describes the different types of biofeedback protocols used for the treatment of drug-refractory epilepsy and their physiological justifications. Secondly, this paper analyzes the evidence of effectiveness, from a medical point of view, on reducing the numbers of seizures, and from a neurophysiological point of view, on the changing brain activity. Electroencephalography (EEG) biofeedback (neurofeedback) protocol on sensorimotor rhythms (SMR) has been investigated in many studies, the main limitation being small sample sizes and lack of control groups. The newer neurofeedback protocol on slow cortical potential (SCP) and galvanic skin response (GSR) biofeedback protocols have been used in a smaller number of studies. But, these studies are more rigorous with larger sized samples, matched control groups, and attempts to control the placebo effect. These protocols also open the way for innovative neurophysiological researches and may predict a renewal of biofeedback techniques. Biofeedback would have legitimacy in the field of clinical drug-resistant epilepsy at the interface between therapeutic and clinical neurophysiology.

[Interactive rTMS protocols in psychiatry]. / Protocoles de rTMS interactives en psychiatrie.

BACKGROUND: The efficiency of repetitive transcranial magnetic stimulation (rTMS) in the treatment of psychiatric disorders is robust for major depressive episode (MDE) while results are encouraging for schizophrenia. However, rTMS protocols need to be optimized. Basic researches in TMS led to the concept of "state dependency TMS". This concept suggests that the neural circuits' activation states, before and during the stimulation, influence the pulse effect. Indeed, TMS effect must be seen, not simply as a stimulus, but also as the result of an interaction between a stimulus and a level of brain activity. Those data suggest that rTMS efficiency could be increased in psychiatric disorders by triggering patients' neurocognitive activities during stimulation. Thus "interactive rTMS protocols" have been submitted. OBJECTIVES: This article provides a review and a classification of different interactive protocols implemented in the treatment of MDE and schizophrenia. Protocols' interactions with cognitive activities and brain electrical activities will be discussed. LITERATURE FINDINGS: Interactive rTMS protocols that manipulate cognitive activities have been developed for MDE treatments. They aim at regulating emotional states of depressed patients during the stimulation. The patients perform emotional tasks in order to activate cortical networks involving the left dorsolateral prefrontal cortex (DLPFC) into a state that may be more sensitive to the rTMS pulse effect. Simultaneous cognitive behavioral therapy ("CBT rTMS") and cognitive-emotional reactivation ("affective rTMS") have thus been tested during left DLPFC rTMS in MDE. Interactive rTMS protocols that manipulate brain electrical activities have been developed for MDE and schizophrenia treatments. Two categories of protocols should be identified. In the first set, personalized brain activity has been analyzed to determine the parameters of stimulation (i.e. frequency of stimulation) matching the patient ("personalized rTMS"). Personalized rTMS protocols can be made "online" or "offline" depending on whether the EEG activity is measured during or prior to rTMS. Online protocol is called "contingent rTMS": it consists in stimulating the brain only when a specific EEG pattern involving the intensity of alpha rhythm is recorded and recognized. Offline protocol is called "alpha rTMS", and relies on ascertaining frequency of stimulation in accordance with personalized alpha peak frequency prior to rTMS. In the second set, electrical brain activity is modulated before or during rTMS in order to stimulate the DLPFC in optimal conditions. Brain activity modulation may be obtained by transcranial direct current stimulation ("tDCS rTMS") or EEG-biofeedack ("EEG-biofeedback rTMS"). CONCLUSION: Interactive rTMS studies have various limitations, notably their exploratory character on a small sample of patients. Furthermore, their theoretical neurocognitive framework justification remains unclear. Nonetheless, interactive rTMS protocols allow us to consider a new field of rTMS, where cognitive and cerebral activities would no longer be considered as simple neural noise, leading to a kind of "first person rTMS", and certainly to innovative therapy in psychiatry.

[Mixed depressions: clinical and neurophysiological biomarkers]. / Épisodes dépressifs mixtes : clinique et biomarqueurs neurophysiologiques.

Epidemiological studies of major depressive episodes (MDE) highlighted the frequent association of symptoms or signs of mania or hypomania with depressive syndrome. Beyond the strict definition of DSM-IV, epidemiological recognition of a subset of MDE characterized by the presence of symptoms or signs of the opposite polarity is clinically important because it is associated with pejorative prognosis and therapeutic response compared to the subgroup of "typical MDE". The development of DSM-5 took into account the epidemiological data. DSM-5 opted for a more dimensional perspective in implementing the concept of "mixed features" from an "episode" to a "specification" of mood disorder. As outlined in the DSM-5: "Mixed features associated with a major depressive episode have been found to be a significant risk factor for the development of bipolar I and II disorder. As a result, it is clinically useful to note the presence of this specifier for treatment planning and monitoring of response to therapeutic". However, the mixed features are sometimes difficult to identify, and neurophysiological biomarkers would be useful to make a more specific diagnosis. Two neurophysiological models make it possible to better understand MDE with mixed features : i) the emotional regulation model that highlights a tendency to hyper-reactive and unstable emotion response, and ii) the vigilance regulation model that highlights, through EEG recording, a tendency to unstable vigilance. Further research is required to better understand relationships between these two models. These models provide the opportunity of a neurophysiological framework to better understand the mixed features associated with MDE and to identify potential neurophysiological biomarkers to guide therapeutic strategies.

[The measurement of electrodermal activity]. / La mesure de la réaction électrodermale.

INTRODUCTION: Electrodermal activity (EDA) is an early physiological index and the subject of constant interest, in spite of the bad reputation attached to "lie detectors". This interest is expected to increase in the future, following the development of research related to the neurobiological aspect of emotions of which it is an index. Recent data provided by functional cerebral imaging has added to the significance of this index and should result in further interest. AIM: The authors thus re-examined the various notions related to measuring EDA, and its practical aspect as well as its mechanisms. EDA should be useful both for authors wishing to use this variable and for readers wishing to form their own critical point of view. LITERATURE FINDINGS: The article first defines the various terms used to qualify EDA. Then, it analyses the mechanisms occurring at the sweat glands' level, showing that a distinct innervation of the sweat glands causes sweat to be released in the excretory channels, thereby allowing the recording of a negative surface potential in parallel to the lowering of skin conductance. Arguments are then pointed out to illustrate that the potential's positive phase following this first answer occurs in the case of high intensity stimulations. The study of the central command of sudation demonstrates that, several areas are involved and that different functions such as thermal regulation and motricity may interfere with emotive reactions. Difficulties regarding the mode of measurement of these answers as to their number and amplitude are also brought to light. DISCUSSION: A particular interest of measuring EDA is its ability to highlight individual characteristic and unconscious emotional reactivity. Subjects who constitutionally present many spontaneous and therefore habitual EDA can indeed be opposed to subjects whose EDA reflexes are very few and hardly habitual. A theory suggests that for the first category, whose subjects are named labiles, emotional control may be at the origin of EDA. This characteristic brings to mind the case of antisocial subjects whose rate of EDA is also reduced, although for the latter a primitive drop in behavioral inhibition is involved. The production of EDA in response to non-conscious emotive stimulations can be objectified in the rare cases of prosopagnosia. These subjects who are unable to recognize familiar faces can produce EDA when presented faces with an emotional load. These cases contrast with the delusional denial of the Capgras syndrome where subjects do not present EDA, suggesting that the dysfunction of visual analysis occurs at a different level. There are other rare cases represented by cortical blindness where EDA shows that an unconscious emotional analysis is preserved. These subjects are known however to be capable of unconscious visual discriminations, which are possibly accompanied by EDA. This possibility of a "blind vision" is experimentally studied via subliminal vision testing (backward masking tests). These demonstrate that a rudimentary visual analysis is carried out in the subcortical circuits while taking into account the affective aspect of stimulations. CONCLUSION: Present or future data should allow a greater comprehension of electrodermal signals, making it possible to overcome the difficulties related to their interpretation and facilitate their applications.

[Neurophysiological endophenotypes and schizophrenic disorder: emergence and evolution of a clinical concept]. / Endophénotypes neurophysiologiques et trouble schizophrénique : naissance et évolution d'un concept clinique.

It is proposed an historical approach to concepts leading to the development of operational paradigms for measuring objectives neurophysiological endophenotypes. It is hypothesized that psychiatric interest for paradigms measuring Event-Related Potential (ERP) come from Bleuler (1911) and McGhie and Chapman (1961) phenomenological and clinical descriptions. They noted, first that patients with schizophrenia generally feel as if they are being flooded by an overwhelming mass of sensory input combined with a heightened sensory perception, second that they were distractible to irrelevant sensory stimuli. These subjective abnormalities may be related, first to inability to filter incongruent information measured in a double click paradigm by a deficit in P50 amplitude gating, and second to an inability to select a stimulus of interest measured in the oddball paradigm by a deficit in P300 amplitude. The analysis of these P50 and P300 ERP in cohorts of patients with schizophrenia found most of Gottesman endophenotype criteria. P50 and P300 ERP are therefore relevant neurophysiological endophenotypes. However, from a clinical point of view, these endophenotypes lack specificity. The hypothesis of this article leads us to formulate ways of research. It is shown the value of combining objective neurophysiological measures with subjective measures using self-administered questionnaires ("offline") or psychophysiological tests ("online") to develop rigorous neurophysiological experimental paradigms especially as clinical observations of their origins are not forgotten.

[Electrophysiology and schizophrenic vulnerability: the P300 component as endophenotype candidate?]. / Électrophysiologie et vulnérabilité schizophrénique : la composante P300 comme endophénotype candidat?

INTRODUCTION: Studies on early stages of schizophrenia imply the observation of stable markers of vulnerability. Among other research fields, these early and objective markers, or potential endophenotypes, can be described in event-related potential (ERP) paradigms. LITERATURE FINDINGS: The P300 component, elicited during the allocation of attentional resources, is the most studied ERP among people with schizophrenia. In this review, we first develop the notion of endophenotypes in schizophrenia, notably in terms of stability, heritability and specificity. We also give a short account of the P300 component, its typical description, the classical paradigms which elicit it, and several interpretations of its significance. DISCUSSION: After reviewing the main features of the schizophrenic alterations of P300 (their topography, amplitude and latency), we discuss the relevance of P300 when described as a potential schizophrenic endophenotype. In spite of an important number of studies, results remain controversial and incomplete. First, P300 in schizophrenia shows complex patterns of temporal evolution, and thus can be described as either a stable trait or a state marker. Second, its heritability is still discussed among high-risk participants with genetic, schizotypal or clinical vulnerability. Third, the issue of its specificity is the less studied criteria. In line with the debate of its specificity, only little is known about specific alterations of P300 among unipolar or bipolar disorders. In the discussion, we describe a few possible origins of such controversial results in both empirical and conceptual perspectives, and we provide several experimental propositions in order to develop a more systematic exploration of P300 alterations.

[Single-voxel proton spectroscopy using a press sequence with a 135 ms echotime: normal values]. / Valeurs controles obtenues avec une sequence press 135 ms en neurospectroscopie monovoxel du proton.

The clinical value of MR spectroscopy is now well established and this technique has been added to the current French classification of medical acts (CCAM). This paper presents a set of normal control values for 3 metabolite ratios obtained using a PRESS sequence with a TE of 135 ms at 1.5T: NAA/Cho, NAA/Cr and Cho/CR. Spectroscopy data acquisition were obtained from the following 12 anatomical regions: parieto-occipital white matter, centrum semiovale, frontal white matter, thalamus, basal ganglia, cerebellum (hemisphere, including dentate nucleus), brain stem (including pons, medulla and midbrain), anterior and posterior temporal lobe, parietal, occipital and pre-frontal cortices. The presented data allow radiologists equipped with a similar MR system to implement a clinical spectroscopy program without undergoing research protocols in order to obtain control values.

On the correlation between perceptual inundation caused by realistic immersive environmental auditory scenes and the sensory gating inventory in schizophrenia.

BACKGROUND: In schizophrenia, perceptual inundation related to sensory gating deficit can be evaluated "off-line" with the sensory gating inventory (SGI) and "on-line" during listening tests. However, no study investigated the relation between "off-line evaluation" and "on-line evaluation". The present study investigates this relationship. METHODS: A sound corpus of 36 realistic environmental auditory scenes was obtained from a 3D immersive synthesizer. Twenty schizophrenic patients and twenty healthy subjects completed the SGI and evaluated the feeling of "inundation" from 1 ("null") to 5 ("maximum") for each auditory scene. Sensory gating deficit was evaluated in half of each population group with P50 suppression electrophysiological measure. RESULTS: Evaluation of inundation during sound listening was significantly higher in schizophrenia (3.25) compared to the control group (2.40, P<.001). The evaluation of inundation during the listening test correlated significantly with the perceptual modulation (n=20, rho=.52, P=.029) and the over-inclusion dimensions (n=20, rho=.59, P=.01) of the SGI in schizophrenic patients and with the P50 suppression for the entire group of controls and patients who performed ERP recordings (n=20, rho=-.49, P=.027). CONCLUSION: An evaluation of the external validity of the SGI was obtained through listening tests. The ability to control acoustic parameters of each of the realistic immersive environmental auditory scenes might in future research make it possible to identify acoustic triggers related to perceptual inundation in schizophrenia.

Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy.

Aspects of energetic and intermediary metabolism were studied in a colon adenocarcinoma cell line (HT29) by multinuclear magnetic resonance spectroscopy. Experiments were carried out on the HT29-D4 clone, which was isolated by limit dilution techniques. This clone, usually undifferentiated (D4-UD), can be maintained in a differentiated state (D4-D) in a glucose-free medium. Metabolic data were obtained by NMR analysis of perchloric acid extracts from D4-UD and D4-D cells. Phosphorus-31 and proton NMR spectra showed the presence of a large amount of choline and phosphorylcholine in the differentiated state (400% and 200%, respectively, of the levels found in D4-UD cells). Other differences appeared in the content of phosphocreatine (absent in D4-D cells) and myoinositol (absent in D4-UD cells). Carbon-13 spectra were recorded from perchloric acid extracts of cells incubated with [1-13C]-labeled glucose or [2-13C]-labeled acetate. The data indicated that both types of cells metabolize glucose through the glycolytic pathway to give lactate, but only D4-D cells were able to store glucose as glycogen at a very high level. A mathematical analysis of fluxes through the tricarboxylic acid (TCA) cycle was developed on the basis of models derived from previous 14C tracer studies. The model was based on the steady-state labeling of glutamate carbons by the 13C isotope and gave the fraction of labeled acetyl-Coa entering the TCA cycle, and the activity y of anaplerotic reactions relative to the flux through the citrate synthetase reaction. The data indicated that y greater than 0.3 in all cases. Only 15% and 30% of labeled acetyl CoA entered the TCA cycle in D4-UD and D4-D cells, respectively, under labeled glucose incubation: these values were significantly different upon labeled acetate feeding, reaching 55% for D4-UD cells and 85% for D4-D cells. The main result of this study is that the process of differentiation of HT29 cells is correlated with a large increase in the activity of oxidative metabolism.

Two-dimensional 1H NMR spectroscopy of normal and pathological human plasma: complete water suppression and further assignment of resonances.

Two-dimensional J-resolved and correlated 1H NMR spectra with complete water suppression have been obtained to further characterize a metabolic pattern for normal and pathological human plasma samples. 1H COSY spectra have been recorded on plasma from 12 patients with cancer in order to check for the possible presence of fucose. Our results show that there is no evidence for the presence of fucosylated lipids in the plasma of these patients.

Energetic metabolism of glucose, mannose and galactose in glucose-starved rat insulinoma cells anchored on microcarrier beads. A phosphorus-31 NMR study.

Insulin-secreting cells (RINm5F) have successfully been grown on a large scale on poly-L-lysine coated-polystyrene microcarriers, providing a high cell number in a restricted volume under conditions that respect the metabolic integrity of these anchorage-dependent cells. The energetic metabolism of the perfused cells has been followed non-invasively by phosphorus-31 nuclear magnetic resonance spectroscopy. Glucose starvation induced a rapid decrease in nucleoside triphosphates (mainly ATP) pools, correlated with an increase in Pi level. The initial ATP level was rapidly recovered when the cells were refed with glucose or with mannose, but not with galactose, even after 2 h of perfusion. These differential effects of hexoses on energetic metabolism might be related to their various insulin-release actions on tumor islet cells.

Variations of plasma sialic acid and N-acetylglucosamine levels in cancer, inflammatory diseases and bone marrow transplantation: a proton NMR spectroscopy study.

Proton NMR spectroscopy allows the detection in plasma of resonances arising from N-acetyl-glucosamine (NAG) and N-acetyl-neuraminic acid (NANA) which have been shown to be borne by acute phase glycoproteins. These resonances can be identified using 2 different protocols of spectrum acquisition detecting different physical states in the global pool of glycoproteins, ie mobile and less mobile moieties of glycosylated chains. In this study we demonstrate that NMR spectroscopy allows a precise monitoring of the variations of glycosylated residues in cancers, inflammatory processes and bone marrow transplantation. The most important findings are that: i), the distribution of glycosylated residues varies with the origin of the cancerous tissue; ii), the level of these residues is a function of tumor development; iii), the concentrations in NAG and NANA are well correlated with the standard biological parameters of acute phase and leucocyte activation. Proton NMR spectroscopy of glycosylated residues in plasma may offer a new means of monitoring sialic acid in cancer and other pathological conditions.

High resolution NMR spectroscopy of physiological fluids: from metabolism to physiology.

High resolution NMR spectroscopy of physiological fluids provides quantitative, qualitative and dynamic information on the metabolic status of the interstitial and plasma compartments under a variety of pathophysiological conditions. The simultaneous detection and quantitation by NMR spectroscopy of numerous compounds of the intermediary metabolism offers a new insight in the understanding of the milieu intérieur. NMR spectroscopy of physiological fluids offers a unique way to define and monitor the global metabolic homeostasis in humans. The development of this analytical approach is still limited by the scarcity of pluridisciplinary teams able to fully exploit the wealth of information present on the NMR spectrum of a fluid. While application in pharmacology and toxicology is already established, the main areas of current development are cancer, hereditary metabolic disorders, organ transplantation and neurological diseases.

Brain proton magnetic resonance spectroscopy in HIV-related encephalopathy: identification of evolving metabolic patterns in relation to dementia and therapy.

Proton MRS has proved useful in the early diagnosis of HIV-related encephalopathy. The modifications of brain metabolism in HIV-related encephalopathy can be classified according to different metabolic patterns (Vion-Dury J et al. CR Acad Sci 1994;317:833-840). The present study describes the relative occurrence of these patterns and evaluates their evolution under zidovudine treatment. We have examined 112 HIV patients--35 neuroasymptomatic patients and 77 patients with ADC (AIDS dementia complex)--with localized proton MRS, using the PRESS 135-msec sequence. We have found the same metabolic modifications in N-acetylaspartate and choline-containing compounds as described in the literature. In addition, 14% of HIV patients with normal MRI displayed abnormal MRS, whatever their neurological status. The MRS-added diagnostic value in neuroasymptomatic patients reaches 30 %. The occurrence of undifferentiated (modification of NAA/Cho ratio only) and Cho (mainly an increase in choline signal) patterns is not significantly different in neuroasymptomatic and ADC patients. The NAA pattern (mainly a significant loss of NAA) is more frequent in ADC patients. Only ADC patients display the double pattern (with a significant increase in choline signal and a significant loss of NAA). Quantitated cerebral atrophy (bifrontal ratio) is related to the occurrence of NAA loss (in NAA and double patterns). An MRS follow-up study of 11 HIV patients showed that the clinical outcome was favorable after a 1000-mg/day zidovudine treatment in patients displaying an NAA pattern whereas this treatment had no effect on the patients displaying the Cho pattern. Consequently, MRS appears to be of great interest in predicting responsiveness to antiretroviral drugs and detecting early any resistance to treatment.

A multicenter proton magnetic resonance spectroscopy study of neurological complications of AIDS.

Human immunodeficiency virus (HIV) infection as seen in Europe and the United States has predominantly been contracted through male homosexual sex or intravenous drug abuse. In infected subjects, the brain is frequently affected both clinically and neuropathologically. The aim of this multicenter study has been to evaluate the value of single-voxel proton magnetic resonance spectroscopy (MRS) in the assessment of the neurological complications of acquired immunodeficiency syndrome (AIDS). MRS (voxel size = 8 ml, TR/TE = 1600/135 msec) was performed in 137 HIV-1-seropositive patients and 64 healthy controls without risk factors at three clinical MR sites operating at 1.5 T. The first result of this multicenter trial is that good reproducibility of results among participating sites was found. This demonstrates the reliability and robustness of MRS in the study of in vivo brain metabolism. In HIV patients, there was no significant correlation between metabolite ratios of brain detected by MRS and CDC grouping of patients or CD4 count. In contrast, the variations of brain metabolite ratios (NA/Cr, NA/Cho, and Cho/Cr) were related to the occurrence of encephalopathy, brain atrophy, or diffuse white matter lesions. There was no significant difference in brain metabolites between male homosexual AIDS patients and male intravenous drug user AIDS patients, whatever their neurological status (neurosymptomatic or neuroasymptomatic). Thus, the mode of transmission of HIV infection does not appear to affect the cerebral changes observed in the proton spectra from AIDS patients. Because of its ease of implementation and high information content, single-voxel proton MRS is likely to play a significant role in the evaluation of HIV-related encephalopathies.

Cerebral metabolic alterations in human immunodeficiency virus-related encephalopathy detected by proton magnetic resonance spectroscopy. Comparison between sequences using short and long echo times.

RATIONALE AND OBJECTIVES: The aim of this study is to evaluate comparatively the metabolic information afforded by proton magnetic resonance (MR) spectroscopy with stimulated-echo acquisition mode (STEAM) (echo time [TE], 20 mseconds) and point-resolved spectroscopy sequence (PRESS) (TE, 135 mseconds) spectra in HIV-related encephalopathy. METHODS: Sixty-three human immunodeficiency virus (HIV) patients and 8 controls were examined by single-voxel proton MR spectroscopy at 1.5 tesla, using both PRESS (TE, 135 mseconds) and STEAM (TE, 20 mseconds) sequences performed during the same MR examination, in the same volume of interest. Cerebral atrophy was quantitated using bicaudate ratio (BCR) and bifrontal ratio (BFR). RESULTS: With the STEAM (TE, 20 mseconds) spectra, mean N-acetylaspartate (NAA)/choline (Cho) and NAA/creatine and phosphocreatine (Cr-PCr) ratios are reduced in acquired immunodeficiency syndrome (AIDS) dementia complex (ADC) patients but not in neuroasymptomatics. The proportion of inositol signal is increased, that of NAA decreased in ADC patients. NAA/Cho and NAA/ Cr-PCr mean values measured with PRESS (TE, 135 mseconds) spectra are significantly reduced in ADC and neuroasymptomatic patients. Bifrontal ratio only correlates with NAA/Cr-PCr and NAA/Cho measured on the PRESS spectrum. PRESS (TE, 135 mseconds) spectra allow a definition of different metabolic patterns in HIV-related encephalopathy. At last, no correlation has been found between the NAA raw signals measured on the PRESS (TE, 135 mseconds) and STEAM (TE, 20 mseconds) spectra obtained in the same MR examination. CONCLUSIONS: STEAM (TE, 20 mseconds) spectra provide more metabolic information-namely an evaluation of glial-neuronal status-than PRESS (TE, 135 mseconds) spectra, which afford a metabolic classification of the HIV-related encephalopathy. Because both sequences afford a similar diagnostic gain, MR spectroscopy examination probably requires spectrum acquisition with both sequences.

Entrapment of gadolinium-DTPA in liposomes. Characterization of vesicles by P-31 NMR spectroscopy.

The use of paramagnetic ion chelates to enhance contrast between pathologic and surrounding parenchyma is extensively documented in the magnetic resonance imaging (MRI) literature. Liposomes can be used to increase chelate concentration in the pathologic area, thereby enhancing the efficiency of paramagnetic compounds as contrast agents. Liposomes (50 +/- 20 nm) were prepared by sonicating a solution of dipalmitoylphosphatidylcholine and cholesterol containing 16.5 mM Gadolinium-DTPA (Gd-DTPA) in pharmaceutical formulation (Schering Laboratories, France) and 25 mM inorganic phosphate (Pi). The solutions were dialyzed against 0.9% NaCl before analysis by phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy. Spectra of liposomes displayed a sharp resonance ascribed to Pi and a broad signal arising from the phosphate groups of the phospholipid bilayer. The content of Gd-DTPA in liposomes was directly estimated, based on specific modifications of the longitudinal relaxation rate of intraliposomal Pi. Entrapment ratio was estimated by P-31 NMR spectroscopy and atomic absorption spectroscopy to represent 2.5% to 5% of the initial Gd-DTPA content in the solution. This work illustrates the usefulness of NMR spectroscopy in the characterization of liposomes to be used for MRI applications.

A new immunization procedure for the obtention of anti-leucine enkephalin antibodies. Part I. Immunization procedure and physicochemical characteristics of antibodies.

We described here a new immunization procedure to obtain high titre and high specific antibodies against Leu-enkephalin (LE). The immunogen form is composed of one part of LE conjugate and one part of LE-Arg6 conjugate. We have observed an increase of titre, affinity and specificity of the antibodies in the coimmunization procedure compared to those obtained by conventional immunization involving only the LE conjugate. The Leu-enkephalin antibodies exhibit a high affinity (KD 8 X 10(-12) M) and we are able to detect the Leu-enkephalin at the 10(-15) mole level. These LE antibodies are highly specific of the C part of LE peptide and cross-react weakly with Met-enkephalin (1%).

A new immunization procedure for obtention of anti-leucine-enkephalin antibodies. Part II. Effects of olfactory bulb removal on pro-enkephalin related peptides in rat brain.

Bilateral Olfactory Bulb Removal (OBR) induced both complex behavioral alterations and a decrease of many neurotransmitter levels. We studied brain levels of the pro-enkephalin related peptides 45 days after OBR. Opioid levels were studied using three different highly specific antisera exhibiting very high affinities in radioimmunoassays in striatum, hypothalamus, hypophysis, brain stem and cortex. Methionine enkephalin levels increase significantly in striatum (42%), hypophysis (94%) and hypothalamus (25%) and non-significantly in the other areas. Leucine-enkephalin levels tended to increase in all dissected structures but a significant increase only occurred in striatum (42%). Octapeptide levels (Methionine-enkephalin-Arg-Gly-Leu) significantly increase in striatum (22%) and decrease in hypophysis (97%) and in brain stem (76%). All these results are partially consistent with the decrease of opiate binding described previously after OBR and suggest a complex imbalance in neurotransmitters after such a sensorial deprivation. It is suggested that the modifications of enkephalinergic neurotransmission might be related to the stressful state induced by OBR.

Analgesic properties of valproic acid might be related to activation of pro-enkephalin system in rat brain.

Recent in vivo and in vitro studies have shown the involvement of gamma-aminobutyric acid (GABA) in analgesia. We investigated if the analgesia induced by an acute valproic acid (VPA) administration might be related to the activation of the enkephalinergic system. VPA administration (i.p.) induces 30 min after treatment a significant and dose-dependent increase of Leu-enkephalin and Met-enkephalin in the striatum, hypothalamus, cortex and brainstem. In the hypophysis no modification was observed for these two neuropeptides. The Met-enkephalin-Arg-Gly-Leu levels are affected by VPA administration in a more complex pattern. Such results suggest the implication of an enkephalinergic system in GABAergic analgesia.