PerBrain: a multimodal approach to personalized tracking of evolving state-of-consciousness in brain-injured patients: protocol of an international, multicentric, observational study.

BACKGROUND: Disorders of consciousness (DoC) are severe neurological conditions in which consciousness is impaired to various degrees. They are caused by injury or malfunction of neural systems regulating arousal and awareness. Over the last decades, major efforts in improving and individualizing diagnostic and prognostic accuracy for patients affected by DoC have been made, mainly focusing on introducing multimodal assessments to complement behavioral examination. The present EU-funded multicentric research project "PerBrain" is aimed at developing an individualized diagnostic hierarchical pathway guided by both behavior and multimodal neurodiagnostics for DoC patients. METHODS: In this project, each enrolled patient undergoes repetitive behavioral, clinical, and neurodiagnostic assessments according to a patient-tailored multi-layer workflow. Multimodal diagnostic acquisitions using state-of-the-art techniques at different stages of the patients' clinical evolution are performed. The techniques applied comprise well-established behavioral scales, innovative neurophysiological techniques (such as quantitative electroencephalography and transcranial magnetic stimulation combined with electroencephalography), structural and resting-state functional magnetic resonance imaging, and measurements of physiological activity (i.e. nasal airflow respiration). In addition, the well-being and treatment decision attitudes of patients' informal caregivers (primarily family members) are investigated. Patient and caregiver assessments are performed at multiple time points within one year after acquired brain injury, starting at the acute disease phase. DISCUSSION: Accurate classification and outcome prediction of DoC are of crucial importance for affected patients as well as their caregivers, as individual rehabilitation strategies and treatment decisions are critically dependent on the latter. The PerBrain project aims at optimizing individual DoC diagnosis and accuracy of outcome prediction by integrating data from the suggested multimodal examination methods into a personalized hierarchical diagnosis and prognosis procedure. Using the parallel tracking of both patients' neurological status and their caregivers' mental situation, well-being, and treatment decision attitudes from the acute to the chronic phase of the disease and across different countries, this project aims at significantly contributing to the current clinical routine of DoC patients and their family members. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04798456 . Registered 15 March 2021 - Retrospectively registered.

The scented brain: pheromonal responses in humans.

Using PET, Savic et al., in this issue of Neuron, found a sexually dimorphic neural response to two putative human pheromones. The specific regions activated combined with the pronounced sex difference depict a pheromonal-type brain response in humans. Here, we preview this finding and suggest that human pheromones exist.

Dissociated neural representations of intensity and valence in human olfaction.

Affective experience has been described in terms of two primary dimensions: intensity and valence. In the human brain, it is intrinsically difficult to dissociate the neural coding of these affective dimensions for visual and auditory stimuli, but such dissociation is more readily achieved in olfaction, where intensity and valence can be manipulated independently. Using event-related functional magnetic resonance imaging (fMRI), we found amygdala activation to be associated with intensity, and not valence, of odors. Activity in regions of orbitofrontal cortex, in contrast, were associated with valence independent of intensity. These findings show that distinct olfactory regions subserve the analysis of the degree and quality of olfactory stimulation, suggesting that the affective representations of intensity and valence draw upon dissociable neural substrates.

Pseudomonas aeruginosa activates the quorum sensing LuxR response regulator through secretion of 2-aminoacetophenone.

In this study we identify a volatile compound produced by Pseudomonas aeruginosa, which can specifically activate the LuxR quorum-sensing response regulator of Vibrio fischeri. Comparative gas-chromatography analysis between P. aeruginosa wild type and a ΔlasR mutant strain implied that the active volatile is 2-aminoacetophenone. The use of synthetic 2-aminoacetophenone and in silico docking analyses verified the activity of the molecule and provided putative interacting residues within the binding site.

Sniffing a human sex-steroid derived compound affects mood and autonomic arousal in a dose-dependent manner.

The effects of sniffing different concentrations of the human sex-steroid derived compound 4,16-androstadien-3-one (AND) on autonomic nervous system function and mood were measured in 60 subjects. The effects were sex-specific and concentration-dependent. Only high concentrations of AND (0.00625 M) increased positive mood (p < 0.03) and decreased negative mood (p < 0.05) in women compared to men, and had sympathetic-like effects in women (p < 0.003), and parasympathetic-like effects in men (p < 0.05). These findings further implicate AND in chemical communication between humans, but pose questions as to the path by which AND is transduced, whether through chemical sensing or transdermal diffusion.

Sex-steroid derived compounds induce sex-specific effects on autonomic nervous system function in humans.

The physiological and psychological effects of 2 human sex-steroid derived compounds, 4.16-androstadien-3-one (AND) and l,3,5(10),16-estratetraen-3-ol(EST) were measured in 24 subjects who participated in a within-subjects, double-blind experiment. A dissociation was evident in the physiological effects of AND, in that it increased physiological arousal in women but decreased it in men. EST did not significantly affect physiological arousal in women or men. Neither compound significantly affected mood. AND is an androgen derivative that is the most prevalent androstene in human male sweat, male axillary hair, and on the male axillary skin surface. The authors argue that AND's opposite effects on physiology in men and women further implicate this compound in chemical communication between humans.

A method for functional magnetic resonance imaging of olfaction.

A method for generating olfactory stimuli for humans within a functional magnetic resonance imaging (fMRI) experimental design is described. The system incorporates a nasal-mask in which the change from odorant to no-odorant conditions occurs in less than 500 ms and is not accompanied by visual, auditory, tactile, or thermal cues. The mask provides an ordorant-free environment following prolonged ordorant presence. Specific imaging parameters that are conducive to the study of the human olfactory system are described. In a pilot study performed using these methods, the specific patterns of activation observed converged with published experimental and clinical findings.

Rotational swimming tendencies in the dolphin (Tursiops truncatus).

Anecdotal evidence suggests that dolphins placed in a pool exhibit stereotypic swimming in circles. The present study confirmed these observations in a sample of thirteen dolphins. The majority of dolphins (84.6%) showed highly consistent directional swimming in counterclockwise circles. The latter directionality held throughout the circadian cycle and resisted environmental manipulations. Only social interaction was capable of altering the directionality of circumnavigation. The consistency of unidirectional swimming is considered paradoxical in view of the existing evidence regarding the alternating of hemispheric activity in sleeping dolphins.

Sniffing human sex-steroid derived compounds modulates mood, memory and autonomic nervous system function in specific behavioral contexts.

We asked whether the effects of exposure to two human sex-steroid derived compounds were context dependent. The effects of sniffing 4,16-androstadien-3-one (AND) and 1,3,5(10),16-estratetraen-3-ol (EST) on mood, memory, and autonomic nervous system responses were explored in 72 participants. Subjects were tested with AND, EST, or a Control compound within four mood contexts: neutral, sexually aroused, sad and happy. These moods were successfully induced using selected film segments (P < 0.0001). During the neutral context, none of the compounds affected mood or autonomic nervous system function. However, compound effects were significantly increased within arousing contexts. During the sexually arousing context, both compounds increased sexual arousal (P < 0.029). During the sad context, AND maintained positive mood in women (P< 0.050) and increased negative mood in men (P < 0.031). Memory for events during the sad context was impaired by AND in women (P < 0.047) but not in men. Finally, effects of AND on physiology were observed during the sexually arousing context whereby AND increased skin temperature in both sexes (P < 0.022) but reduced abdominal respiration rate in men only (P < 0.034). These results suggest that sex-steroidal compounds modulate mood, memory and autonomic nervous system responses and increase their significance within specific behavioral contexts. These findings lend support to a specific role for these compounds in chemical communication between humans.

Sister-chromatid exchanges and chromosomal aberrations in a population exposed to pesticides.

Sister-chromatid exchanges (SCE) and chromosomal aberrations were studied in a population of floriculturists occupationally exposed to organophosphorus, carbamate and organochlorine pesticides. Blood samples from 36 individuals from a community of 154 persons of asiatic origin were obtained. Among the group sampled, 21 individuals exhibited at least one symptom of chronic intoxication. SCE analysis was performed in 14 symptomatic and 13 asymptomatic persons. The asymptomatic group showed a SCE frequency of 5.47 +/- 1.03 and the symptomatic group a frequency of 6.45 +/- 1.19. Comparison between both groups with the Mann-Whitney 'U' test revealed a significant difference (p 0.0409). Case-control analysis of 9 pairs matched by sex and age also showed significant differences between both groups (p 0.0104). In contrast, the frequencies of chromosomal aberrations were not correlated with intoxication symptomatology, though a significant increment of exchange-type aberrations in relation to a group of non floriculturists was observed in the population studied.

Breast cancer in Argentina: case-control study with special reference to meat eating habits.

An exploratory case-control study of the role of diet in the etiology of breast cancer was conducted in Buenos Aires, Argentina, where the mortality rate for this disease is high and the consumption of meat, mainly beef, is unusually elevated (76.2 kilograms per head were reported for 1987). One hundred and ninety-six women with breast cancer admitted to the Institute of Oncology "Angel H. Roffo" and 205 controls were interviewed to obtain information on demographic, socio-economic and reproductive variables, on frequency of consumption of 40 food items, and on methods of cooking. Special emphasis was given to different kinds of meat. After controlling for other risk factors for breast cancer the major dietary associations observed were a statistically insignificant trend of increasing risk with amount of beef consumed, an increase in risk in women who ate more than 3 eggs per week, and an increase in risk in women who ate a variety of fried foods.

Progestins in preventive hormone therapy. Including pharmacology of the new progestins, desogestrel, norgestimate, and gestodene: are there advantages?

Progestins have played an important role in hormone replacement therapy (HRT). This article reviews the history, structure, and pharmacology of synthetic progestins, including the new progestins, norgestimate, gestodene, and desogestrel. The benefits and potential effect on carcinoma of the breast for each HRT are summarized. Current treatment alternatives are addressed.

Dissociated representations of irritation and valence in human primary olfactory cortex.

Irritation and negative valence are closely associated in perception. However, these perceptual aspects can be dissociated in olfaction where irritation can accompany both pleasant and unpleasant odorants. Whereas the sensation of odor reflects transduction at olfactory receptors, irritation reflects concurrent transduction of the odorant at trigeminal receptors. Thus a stimulus can be either a pure olfactant activating the olfactory receptors only or a bimodal odorant activating both types of receptors. Using event-related functional magnetic resonance imaging and a 2 x 2 experimental design contrasting odorant valence (pleasant/unpleasant) and odorant type (pure olfactant/bimodal) we found activity in piriform cortex to be associated with valence, and not type, of odors. In contrast, activity in the olfactory tubercle was associated with type, and not valence, of odors. Importantly, this was found when perceived intensity was held equal across odorants. These findings suggest that dissociable neural substrates subserve the encoding of irritation and valence in olfaction.

Odorant-specific patterns of sniffing during imagery distinguish 'bad' and 'good' olfactory imagers.

There are large individual differences in the self-reported ability to form vivid olfactory mental imagery. Based on such self-reports, subjects have been classified as 'bad' or 'good' imagers. The present study examined whether a differential strategy in re-enacting the olfactomotor response during imagery may explain the dissociation between 'bad' and 'good' olfactory imagers. As previously reported, odor imagery was accompanied by sniffing. Although 'bad' and 'good' olfactory imagers did not differ in their overall sniffing volume, they used different strategies when re-enacting the motor component of olfaction during imagery. Particularly, as in real perception, 'good' but not 'bad' imagers generated bigger sniffs when imagining a pleasant smell compared with an unpleasant smell (P<0.02). Furthermore, preventing sniffing significantly hampered mental imagery of pleasant odors in 'good' but not 'bad' imagers (P<0.03). Taken together, these results suggest (i) the validity of the dissociation between 'bad' and 'good' olfactory imagers as revealed by self-report; (ii) that sniffing may be a causal factor in the creation of olfactory imagery; and (iii) that sniff measurements may serve as a reliable non-verbal tool in exploring individual differences in odor imagery.

Sniffing longer rather than stronger to maintain olfactory detection threshold.

Air flow-rate is usually higher in one nostril in comparison to the other. Also, within bounds, higher nasal flow-rate improves odorant detection. It follows from the above that odorant detection should be better in the nostril with higher flow-rate in comparison to the nostril with lower flow-rate. Paradoxically, previous research has shown that odorant detection thresholds are equal for the high and low flow-rate nostrils. Here we resolve this apparent paradox by showing that when detecting through the nostril with lower air flow-rate, humans sniffed longer than when detecting through the nostril with higher air flow-rate, thus equalizing performance between the nostrils. When this compensatory mechanism was blocked, a pronounced advantage in odorant detection was seen for the nostril with higher air flow-rate over the nostril with lower air flow-rate. Finally, we show that normal birhinal sniff duration may enable only one nostril to reach optimal threshold. This finding implies that during each sniff, each nostril conveys to the brain a slightly different image of the olfactory world. It remains to be shown how the brain combines these images into a single olfactory percept.

Immunochemical studies of estrogen receptors.

Fusion of splenic lymphocytes from Lewis rats, immunized with affinity-purified estrogen receptor from the cytosol of MCF-7 human breast cancer cells, with two different mouse myeloma lines, has provided 13 monoclonal hybridoma lines secreting antiestrophilin antibodies, each of which (with one possible exception) recognizes a different antigenic determinant in the human receptor molecule. Of this library of monoclonal antibodies, some react with estrophilin from all sources tested, some react with mammalian but not avian receptors, whereas one preparation appears specific for estrophilin from primate sources. By proteolytic digestion under controlled conditions with mercury-deactivated papain, chymotrypsin, and trypsin, respectively, it is possible to remove sequentially the determinants recognized by one, two or three of the monoclonal antibodies, leaving the epitopes for the six remaining antibodies investigated on the steroid-binding portion of the receptor. The proteolytic fragment containing the epitope most readily removed (by mercuripapain) also contains the DNA-binding domain of the activated receptor molecule. Immunocytochemical staining, using the peroxidase procedure with various monoclonal antibody preparations, of frozen sections of human breast cancer tissue, fixed in ethanol or in picric acid-formaldehyde reagent, shows clearly that the majority of the native receptor, which appears in the cytosol after tissue homogenization, is actually localized within the nuclear compartment in the intact cell. The immunocytochemical technique also permits the identification of mixed populations of receptor-containing and non-containing cells in human breast cancers.

Time course of odorant-induced activation in the human primary olfactory cortex.

Paradoxically, attempts to visualize odorant-induced functional magnetic resonance imaging (fMRI) activation in the human have yielded activations in secondary olfactory regions but not in the primary olfactory cortex-piriform cortex. We show that odorant-induced activation in primary olfactory cortex was not previously made evident with fMRI because of the unique time course of activity in this region: in primary olfactory cortex, odorants induced a strong early transient increase in signal amplitude that then habituated within 30-40 s of odorant presence. This time course of activation seen here in the primary olfactory cortex of the human is almost identical to that recorded electrophysiologically in the piriform cortex of the rat. Mapping activation with analyses that are sensitive to both this transient increase in signal amplitude, and temporal-variance, enabled us to use fMRI to consistently visualize odorant-induced activation in the human primary olfactory cortex. The combination of continued accurate odorant detection at the behavioral level despite primary olfactory cortex habituation at the physiological level suggests that the functional neuroanatomy of the olfactory response may change throughout prolonged olfactory stimulation.

Blind smell: brain activation induced by an undetected air-borne chemical.

EEG and behavioural evidence suggests that air-borne chemicals can affect the nervous system without being consciously detected. EEG and behaviour, however, do not specify which brain structures are involved in chemical sensing that occurs below a threshold of conscious detection. Here we used functional MRI to localize brain activation induced by high and low concentrations of the air-borne compound oestra-1,3,5(10),16-tetraen-3yl acetate. Following presentations of both concentrations, eight of eight subjects reported verbally that they could not detect any odour (P = 0.004). Forced choice detection performed during the presentations revealed above-chance detection of the high concentration, but no better than chance detection of the low concentration compound. Both concentrations induced significant brain activation, primarily in the anterior medial thalamus and inferior frontal gyrus. Activation in the inferior frontal gyrus during the high concentration condition was significantly greater in the right than in the left hemisphere (P = 0.03). A trend towards greater thalamic activation was observed for the high concentration than the low concentration compound (P = 0.08). These findings localize human brain activation that was induced by an undetectable air-borne chemical (the low concentration compound).

An impairment in sniffing contributes to the olfactory impairment in Parkinson's disease.

Although the presence of an olfactory impairment in Parkinson's disease (PD) has been recognized for 25 years, its cause remains unclear. Here we suggest a contributing factor to this impairment, namely, that PD impairs active sniffing of odorants. We tested 10 men and 10 women with clinically typical PD, and 20 age- and gender-matched healthy controls, in four olfactory tasks: (i) the University of Pennsylvania smell identification test; (ii and iii) detection threshold tests for the odorants vanillin and propionic acid; and (iv) a two-alternative forced-choice detection paradigm during which sniff parameters (airflow peak rate, mean rate, volume, and duration) were recorded with a pneomatotachograph-coupled spirometer. An additional experiment tested the effect of intentionally increasing sniff vigor on olfactory performance in 20 additional patients. PD patients were significantly impaired in olfactory identification (P < 0.0001) and detection (P < 0.007). As predicted, PD patients were also significantly impaired at sniffing, demonstrating significantly reduced sniff airflow rate (P < 0.01) and volume (P < 0.002). Furthermore, a patient's ability to sniff predicted his or her performance on olfactory tasks, i.e., the more poorly patients sniffed, the worse their performance on olfaction tests (P < 0.009). Finally, increasing sniff vigor improved olfactory performance in those patients whose baseline performance had been poorest (P < 0.05). These findings implicate a sniffing impairment as a component of the olfactory impairment in PD and further depict sniffing as an important component of human olfaction.

Sniffing and smelling: separate subsystems in the human olfactory cortex.

The sensation and perception of smell (olfaction) are largely dependent on sniffing, which is an active stage of stimulus transport and therefore an integral component of mammalian olfaction. Electrophysiological data obtained from study of the hedgehog, rat, rabbit, dog and monkey indicate that sniffing (whether or not an odorant is present) induces an oscillation of activity in the olfactory bulb, driving the piriform cortex in the temporal lobe, in other words, the piriform is driven by the olfactory bulb at the frequency of sniffing. Here we use functional magnetic resonance imaging (fMRI) that is dependent on the level of oxygen in the blood to determine whether sniffing can induce activation in the piriform of humans, and whether this activation can be differentiated from activation induced by an odorant. We find that sniffing, whether odorant is present or absent, induces activation primarily in the piriform cortex of the temporal lobe and in the medial and posterior orbito-frontal gyri of the frontal lobe. The source of the sniff-induced activation is the somatosensory stimulation that is induced by air flow through the nostrils. In contrast, a smell, regardless of sniffing, induces activation mainly in the lateral and anterior orbito-frontal gyri of the frontal lobe. The dissociation between regions activated by olfactory exploration (sniffing) and regions activated by olfactory content (smell) shows a distinction in brain organization in terms of human olfaction.