High temperature aggravates the effects of pesticides in goldfish.

In human-altered rivers, fish are often conjointly exposed to an increase in water temperature due to global warming and to a contamination by organic pollutants such as pesticides, but their combined effects are still elusive. Thermal and chemical stressors could potentially interact because high temperature increases metabolism and toxicant uptake, and can alter the ability of organisms to set up adequate stress responses and to maintain homeostasis. These combined stressors could thus potentially result in higher level of molecular and cellular damage, and stronger effects on behavior and physiology, but experimental evidence across biological levels is still scarce. In this study, goldfish Carassius auratus were experimentally exposed to an environmentally realistic cocktail of pesticides (S-metolachlor, isoproturon, linuron, atrazine-desethyl, aclonifen, pendimethalin and tebuconazol) commonly found in rivers of South-West of France at low or high dose in two different thermal conditions: a common summer temperature (22 °C) or a high temperature recorded during heat waves (32 °C). Results showed that high temperature alone caused behavioral and physiological changes (increased swimming activity, increased hepatosomatic index, decreased reproductive index) but limited cellular damage. However, high temperature aggravated the effects of pesticides at the molecular and cellular level. Indeed, pesticide exposure resulted in higher genotoxic effects (micronuclei rate) and irreversible cellular damage of the gills and liver (apoptosis, inflammation, necrosis) at 32 °C compared to 22 °C. This suggests potential synergistic effects of climate change and pollution, and highlights the need for multiple stress approaches to better predict the impacts of human activities on aquatic wildlife.

Quantification of Flavobacterium psychrophilum in rainbow trout, Oncorhynchus mykiss (Walbaum), tissues by qPCR.

A qPCR assay was developed for rapid and sensitive detection of Flavobacterium psychrophilum, the aetiological agent of bacterial cold-water disease and rainbow trout fry syndrome in salmonid fish worldwide. A set of F. psychrophilum-specific primers based on 16S rRNA gene sequences was designed and validated for specific detection and quantification of DNA isolated from representative strains of F. psychrophilum. The qPCR assay exhibited a high specificity for the 16S rRNA gene of F. psychrophilum (from 4 × 10(8) down to 11 copies per reaction) but not for other Flavobacterium species or other bacteria including fish pathogens. This qPCR-based method proved to be useful in the quantification of the F. psychrophilum titre present within organs dissected out from diseased fish. As the F. psychrophilum genome contains six copies of the 16S rRNA gene, we could infer a limit of detection corresponding to two bacteria per reaction, corresponding to 800 bacteria per fish tissue sample, and therefore 20 F. psychrophilum cells mg(-1) of tissue (for sample weighing 40 mg). The qPCR assay reported here could be a useful tool for veterinary diagnostic laboratories to monitor the F. psychrophilum infection level in fish farms.

Highly variable upper and abyssal overturning cells in the South Atlantic.

The Meridional Overturning Circulation (MOC) is a primary mechanism driving oceanic heat redistribution on Earth, thereby affecting Earth's climate and weather. However, the full-depth structure and variability of the MOC are still poorly understood, particularly in the South Atlantic. This study presents unique multiyear records of the oceanic volume transport of both the upper (<~3100 meters) and abyssal (>~3100 meters) overturning cells based on daily moored measurements in the South Atlantic at 34.5°S. The vertical structure of the time-mean flows is consistent with the limited historical observations. Both the upper and abyssal cells exhibit a high degree of variability relative to the temporal means at time scales, ranging from a few days to a few weeks. Observed variations in the abyssal flow appear to be largely independent of the flow in the overlying upper cell. No meaningful trends are detected in either cell.

Unilateral right parietal damage leads to bilateral deficit for high-level motion.

Patients with right parietal damage demonstrate a variety of attentional deficits in their left visual field contralateral to their lesion. We now report that patients with right lesions also show a severe loss in the perception of apparent motion in their "good" right visual field ipsilateral to their lesion. Three tests of attention were conducted, and losses were found only in the contralesional fields for a selective attention and a multiple object tracking task. Losses in apparent motion, however, were bilateral in all cases. The deficit in apparent motion in the parietal patients supports previous claims that this relatively effortless percept is mediated by attention. However, the bilateral deficit suggests that the disruption is due to a bilateral loss in the temporal resolution of attention to transient events that drive the apparent motion percept.

Complete sparing of high-contrast color input to motion perception in cortical color blindness.

It is widely held that color and motion are processed by separate parallel pathways in the visual system, but this view is difficult to reconcile with the fact that motion can be detected in equiluminant stimuli that are defined by color alone. To examine the relationship between color and motion, we tested three patients who had lost their color vision following cortical damage (central achromatopsia). Despite their profound loss in the subjective experience of color and their inability to detect the motion of faint colors, all three subjects showed surprisingly strong responses to high-contrast, moving color stimuli--equal in all respects to the performance of subjects with normal color vision. The pathway from opponent-color detectors in the retina to the motion analysis areas must therefore be independent of the damaged color centers in the occipitotemporal area. It is probably also independent of the motion analysis area MT/V5, because the contribution of color to motion detection in these patients is much stronger than the color response of monkey area MT.

[Pulmonary embolism: the extent of computerized tomography]. / Embolie pulmonaire : faut-il tendre vers le << tout angioscanner >> ?

The use of computerised tomography in the diagnosis of pulmonary embolism has been the subject of clinical research while, at the same time, technical progress has provided the current multidetector-row spiral equipment. Computerised tomography has been assessed both with respect to reference strategies as well as in extensive pragmatic trials. The preliminary evaluation of the clinical probability and the assay of d-dimers has progressively become imperative. The value of the venous doppler ultrasound of the legs, in particular in the elderly, is limited by the variable accessibility according to the centre. In rare cases, uncertainty persists, for example with a good quality negative multidetector-row spiral computerised tomography associated with a high clinical probability, leaving room for complementary explorations. The confrontation between clinicians and radiologists is then all the more pertinent.

Impaired fronto-temporal processing of emotion in schizophrenia.

AIMS: Abnormal emotion processing in schizophrenia affects social and functional outcome. Spatiotemporal brain mechanisms underlying this deficit are unclear. MATERIALS AND METHODS: Event-related potential (ERP) responses to emotional and neutral face processing during an implicit (gender detection) and an explicit (expression detection) task were compared between a group of healthy volunteers (n=10) and a group of patients with schizophrenia (n=10). RESULTS: Whereas patients had normal primary visual cortex responses, the early modulation of occipital, temporal, and frontal responses by emotional expression observed in controls was absent in patients. The occipito-temporal N170 amplitude was reduced in patients relative to controls during expression detection, but not during gender detection. Frontal activity within 180-250ms was reduced in patients compared to controls. As opposed to controls, no significant difference was seen in patients at the right temporal electrode (T6) between amplitudes of long-latency ERPs elicited by distinct emotions during the expression detection task. CONCLUSION: In patients with schizophrenia, abnormal early extraction of expression-related information in the occipito-temporal cortex (before 170ms) impairs structural encoding of facial expressions (N170) and may disrupt motivation- and task-dependent context processing (180-250ms time window) of expression-related facial features. Moreover, top-down neuromodulation from frontal and limbic structures to visual occipito-temporal cortex may not be sufficient to optimize the extraction of expression-specific face features.

[Part I: Face recognition]. / Les visages et leurs émotions.

Faces represent a crucial vector of interhuman communication. The message transmitted by the face has multiple features. Recognition of each feature can be impaired independently or in combination with others. In order to understand the behavioral consequences of such impairments, which can be a major social handicap, we first must specify the neural networks involved in face recognition. We propose in this first part to present the systems involved in face recognition, in particular the question of identity and prosopagnosia. Different neural networks are indeed implicated in the recognition of invariant facial features such as identity, gender, ethnicity, and recognition of variant features like facial expression and eye gaze. This paper is illustrated by some of our scalp and intracranial electrophysiological studies performed in humans allowing us to describe some aspects of face recognition dynamics combining an excellent spatial and temporal resolution. Intracranial recordings were performed in drug refractory epileptical patients implanted with depth electrodes. These studies demonstrate that numerous deep brain and cortical structures participate early and sometimes in a sustained manner in face recognition.

[Part II: Recognising facial expressions]. / Les visages et leurs émotions.

In this second part, we address particularly the question of the neural mechanisms and structures involved in the recognition of facial emotional expressions that are crucial in social cognition. Emotion recognition in others can be critically impaired in some neurodegenerative and neurovascular diseases. That dysfunction sometimes correlated to disabling behavioural disorders and interpersonal communication impairment must be further understood. The results of a series of scalp and intracranial event related potential recordings, as well as recent advances in the literature, are reported. ERPs to facial emotional expressions were thus recorded in multiple subcortical and cortical areas in drug refractory epileptical patients implanted with depth electrodes. The roles of amygdala, insula and prefrontal cortex located at crossroads between perceptive analysis and emotional conceptual knowledge are particularly underlined. Altogether, these studies demonstrate that facial expressions are widely processed in space and time, some structures reacting very early and automatically, others providing a sustained reaction depending on the attention.

Doxorubicin induces slow ceramide accumulation and late apoptosis in cultured adult rat ventricular myocytes.

OBJECTIVES: Anthracyclines cause apoptotic death in many cell types through activation of the ceramide pathway. We tested the hypothesis that doxorubicin induces cardiac myocyte apoptosis through ceramide generation. METHODS: Adult rat ventricular myocytes were grown in the presence of 10% fetal calf serum, and exposed to 0.5 microM doxorubicin (Dox) for 1 h on the day of cell isolation (day 0). We used the membrane-permeant ceramide analog C2-ceramide (C2-cer) to mimic the effects of endogenous ceramide and PDMP to induce endogenous ceramide accumulation. Apoptosis was assessed upon morphological criteria and DNA fragmentation by the TUNEL method and agarose gel electrophoresis. Ceramide concentration was assessed using the DAG kinase assay. RESULTS: Myocyte exposure to Dox was associated with cellular and nuclear alterations typical of apoptosis on day 7 but not on day 3. At day 7, the percentage of TUNEL-positive myocytes was markedly increased in Dox-treated cultures compared to control (Cl) cultures (82 +/- 3 vs. 12 +/- 1%, n = 7; p < 0.001); internucleosomal DNA fragmentation was confirmed by the observation of DNA ladders. These alterations were associated with an increase in the intracellular ceramide concentration (1715 +/- 243 vs. 785 +/- 99 pmol/mg prot, n = 5; p < 0.01), a phenomenon also detected on day 3 (731 +/- 59 vs. 259 +/- 37 pmol/mg prot, n = 5; p < 0.001). Incubation of myocytes at day 0 with 50 microM C2-cer induced rapid cell shrinkage and DNA fragmentation (45 +/- 3 vs. 10 +/- 1% TUNEL-positive myocytes on day 1 in C2-cer-treated and Cl cultures, respectively; n = 6, p < 0.001). Myocyte exposure to 10 microM PDMP for 7 days (n = 5), caused ceramide accumulation (1.7-fold increase vs. Cl, p < 0.01), and a marked increase in the percentage of TUNEL-positive myocytes (62 +/- 6 vs. 11 +/- 3% in Cl cultures, p < 0.001). Ventricles of rats injected intraperitoneally with a cumulative dose of 14 mg/kg Dox over a period of 2 weeks also showed an increased ceramide concentration 2 weeks later (11.01 +/- 0.64 vs. 5.24 +/- 0.88 pmol/mg prot, n = 6; p < 0.001). CONCLUSION: Our study confirms the existence of a functional ceramide pathway related to apoptosis in cardiac myocytes, and points to its possible involvement in doxorubicin-induced cardiomyopathy.

Decision-making in a changing world: a study in autism spectrum disorders.

To learn to deal with the unexpected is essential to adaptation to a social, therefore often unpredictable environment. Fourteen adults with autism spectrum disorders (ASD) and 15 controls underwent a decision-making task aimed at investigating the influence of either a social or a non-social environment, and its interaction with either a stable (with constant probabilities) or an unstable (with changing probabilities) context on their performance. Participants with ASD presented with difficulties in accessing underlying statistical rules in an unstable context, a deficit especially enhanced in the social environment. These results point out that the difficulties people with ASD encounter in their social life might be caused by impaired social cues processing and by the unpredictability associated with the social world.

Involvement of antibody-dependent apoptosis in graft rejection.

BACKGROUND: Both humoral factors and apoptosis have been recently suggested to play a role in chronic allograft rejection. However, a link between alloantibodies and grafted cell apoptosis has never been proposed. Using the aortic allograft model in the rat, we have previously demonstrated the presence of IgG associated with the disappearance of donor endothelial and medial smooth muscle cells. In the present study, we tested the interaction between recipient allosera, enriched with antibodies by presensitization, and primary culture of cardiovascular cells of donor origin. METHODS: For this purpose endothelial cells, smooth muscle cells, adventitial fibroblasts, and cardiac myocytes of donor origin were cultured. Binding of alloantisera to these cells was analyzed by flow cytometry. Apoptosis of donor cells was evaluated by Tdt-mediated d' UTP-FITC nick end labeling, 4',6-diamidino-2-phenylindole and DNA ladder techniques. The alloantisera were compared with anti-MHC class I monoclonal antibodies. Finally the colocalization of antibodies and apoptosis was investigated in vivo. RESULTS: In vitro, alloantisera bind to cardiovascular cells of donor origin. These cells expressed MHC class I but not MHC class II. There was a partial competition between anti-MHC I mouse monoclonal antibody and alloantisera mainly of the IgG isotype. Alloantisera bound to, but did not induce lysis of, donor RBC. Alloantisera induced apoptosis of donor cardiovascular cells as assessed by the typical morphological aspect of the donor cells after 24 hr of incubation. These data were confirmed by the Tdt-mediated d' UTP-FITC nick end labeling positivity of the cells and the fragmentation of the nucleus visualized by 4',6-diamidino-2-phenylindole and DNA ladder techniques. Similar apoptosis was induced by specific monoclonal antibodies directed against the MHC class I of donor cells. Primary culture of similar vascular cells of recipient origin was insensitive to alloantisera directed against donor alloantigens. Finally, in vivo, using allopresentization and aortic allografts, an association of alloantibody binding and endothelial cell apoptosis was observed at day 5, and a similar association with smooth muscle cell apoptosis on day 12 after grafting. CONCLUSION: These data demonstrate the role of humoral injury in chronic allograft rejection and suggest new therapeutical approaches focused on the induction of resistance to antibody-dependent apoptosis.

Acylation and immunological properties of Mycoplasma gallisepticum membrane proteins.

The acylation of Mycoplasma gallisepticum membrane proteins was studied by electrophoresis after in vivo labelling with different 14C-fatty acids and by chemical analysis. The immunological properties of these proteins were investigated by Western blotting and crossed immunoelectrophoresis. Among the ca. 200 membrane polypeptides resolved by two-dimensional electrophoresis, 35 components (including the major protein p67) were covalently modified with acyl chains. These acylated proteins displayed lower pls than average (5.0-7.4 vs. 5.0-9.0) and proved to be the major membrane protein antigens and immunogens of M. gallisepticum. The apparent selectivity of fatty acid incorporation into proteins was, as suggested by in vivo labelling: palmitic acid (16:0) > myristic acid (14:0) > oleic acid (18:1c) > stearic acid (18:0) > linoleic acid (18:2c). However, the true order of selectivity, as revealed by chemical analysis, proved to be 18:2c > 16:0 > 18:1c > 18:0 > 14:0. More specifically, palmitic acid was the major O-ester-bound fatty acid and linoleic acid the major amide-linked fatty acid. The observed average ratio [O-ester-bound + amide-linked acyl chains]/O-ester-bound chains approximately 1.4 and the presence of S-glycerylcysteine suggest that, in M. gallisepticum, membrane proteins are lipid-modified according to a mechanism identical to that depicted for lipoproteins of Gram-negative eubacteria.

Adult cardiac myocytes survive and remain excitable during long-term culture on synthetic supports.

OBJECTIVE: Cardiomyocytes can be transplanted successfully into skeletal and cardiac muscle. Our goal was to determine the feasibility of grafting cardiomyocytes onto various synthetic supports to create an excitable and viable tissue for implantation. METHODS: Adult rat cardiomyocytes were cultured over an 8-week period onto different substitutes, including human glutaraldehyde-treated pericardium (n = 3), equine glutaraldehyde-treated pericardium (n = 3), polytetrafluoroethylene (n = 8), Dacron polyester (n = 16), and Vicryl polyglactin (n = 8). RESULTS: Only the cells seeded on the Dacron survived, with the synthetic fibers colonized at 8 weeks. On the other supports, the number of myocytes progressively decreased from the first week, with their density (number of cells per square millimeter) being, after 20 days, 17 +/- 2 on the polytetrafluoroethylene and 5 +/- 1 on the human or equine pericardium compared with 45 +/- 3 on the Dacron. After 8 weeks of culture on Dacron, the sarcomeric protein (sarcomeric alpha-actinin) was detected in all cells. In addition, the staining was regularly arranged and well aligned in a striated pattern. Spontaneous beating activity was obtained. Moreover, electrical stimulation of the cell preparation resulted in the generation of calcium transients, the frequency of which followed the frequency of the electrical stimulation. CONCLUSIONS: These results suggest that adult cardiac myocytes remain viable and excitable during long-term culture on a 3-dimensional Dacron support, which might constitute a new synthetic cardiac tissue.

Able to name, unable to compare: the visual abilities of a posterior split-brain patient.

A patient with a lesion of the posterior half of the corpus callosum correctly named simple (e.g. colors) and complex (e.g. faces) stimuli whether presented to one or both hemifields. Although proficient at these tasks, and also able to make same/different judgements for stimuli within an hemifield, he failed dramatically when required to compare stimuli between the two hemifields. These results indicate that, while the posterior portion of the corpus callosum may not be essential for naming visual stimuli, it is essential for comparing stimuli between the two visual fields.

Unmasking of visual deficits following unilateral prestriate lesions in man.

The primate visual cortex is a mosaic of different areas which are roughly organized into two major pathways, a dorsal one along an occipito-parietal channel and a ventral one along an occipito-temporal channel. It is known that visual stimuli are processed differently within these two systems and one might expect therefore that behavioral deficits would differ according to the channel damaged by cerebral lesions. Two patients with left prestriate lesions presented perceptual deficits in the right visual field. These deficits involved reading, recognition of line drawings and colour perception and would be compatible with a dysfunction of the ventral system. Magnetic resonance imaging analysis confirmed that this was the case.

Two different readers in the same brain after a posterior callosal lesion.

Two different types of reading, one in each hemifield, were exhibited by a patient with a lesion of the posterior half of the corpus callosum. The patient read normally when words and non-words were presented to his right visual field. However, with left visual field presentations, the patient could not read non-words and vocalized real words very slowly, especially abstract words, inflected verbs and function words. He often replaced concrete words by semantic associates. Such an abnormal reading pattern is similar to that known as deep dyslexia. This unilateral deficit reveals the competence of the right hemisphere to initiate some semantic processing and its inability to manage phonological coding. The hypothesis that deep dyslexia is due to right hemisphere reading is reinforced by the present case.

Failure to identify the target does not prevent the attentional blink effect.

The attentional blink has been attributed to capacity limitations at a central level of processing. We tested whether failure to identify the target would eliminate the blink. Two agnostic patients were presented with streams of letters, which they were able to identify, and streams of pictures, which they were unable to identify. The dual-task involved identification of a target and detection of a probe. With letters the duration of the blink was equivalent to that of the control subjects. A prolonged blink was observed in both patients for pictures irrespective of whether the target was identified. This result indicates that failure to identify the target does nevertheless mobilize attentional resources sufficiently to prevent detection of a second target stimulus.

Dependence of color on context in a case of cortical color vision deficiency.

Color constancy depends on sensitivity to change in both illumination spectral properties and object position. We investigated this latter form of color constancy by asking a cerebral achromatopsic to name the colors of papers that were presented atop black, gray or white backgrounds under identical illumination. Comparison of color names across background conditions reveals poor constancy, characterized by a contrasting of foreground and background values that is not corrected by proper anchoring.