Volabolomic Fingerprinting for Post-Mortem Interval Estimation: A Novel Physiological Approach.

Death is a multifaceted process wherein each individual cell and tissue has a metabolic homeostasis and a time of functional cessation defined by the dying process as well as by intrinsic and extrinsic factors. Decomposition is physiologically associated with the release of different types of volatile organic compounds (VOCs), and these form volaboloma mortis. The main purpose of this study was to record the volabolomic fingerprint produced by volatile molecules during the physiological decomposition process of human tissue and muscle cells. The volatile chemical signature has important implications for an open issue in forensics and pathology, namely the estimation of the postmortem interval (PMI), which decreases in accuracy with the passage of time. Volatile metabolites emitted from human tissues and muscle cells at 0, 24, 48, and 72 h were recorded in real time with an electronic nose sensor device. The key findings were the continuous sampling of VOCs emitted from tissues and cells. These showed a common behavior as time progressed; particularly, after 48 h the distributions became dispersed, and after 72 h they became more variable. Volabolomic fingerprinting associated with time progression relevant to the study of PMIs was reconstructed. Additionally, there may be broader applications, such as in dog training procedures for detecting human remains, and perhaps even for studying scavenger and insect attractants.

Aging and the carotid body: A scoping review.

The carotid body (CB) is a neuroepithelial tissue consisting of O2-sensitive glomus cells that constantly scan the arterial blood for O2 and generate a discharge as an inverse function of O2 content. Aging is a cumulative result of decreased O2 supply paralleled by a decreased O2 tissue demand and oxidative damage to cells derived from aerobic metabolism. Here we studied how CB affects the aging process. This is a study of CB ultrastructural morphometry and immunohistochemical expression of proteins underlying CB responsiveness. The study was based on human CBs obtained from cadavers of people who died due to traumatic events in young and old age. The study was supplemented by investigations of CBs obtained from young and old rats subjected to chronic normoxic and hypoxic conditions. We found changes in the old normoxic CBs akin to the effects of chronic hypoxia such as enhanced extracellular matrix, reduced synaptic contacts between glomus cells, fewer glomus cells, secretory vesicles, and mitochondria. These changes were accompanied by enhanced expressions of hypoxia-inducible factor one-alpha (HIF-1α), vascular endothelial growth factor (VEGF), and nitric oxide synthase (NOS2). We conclude that hypoxia and aging share a common background consisting of deficient O2 tissue supply, mitochondrial dysfunction, and a limited ability to deal with increased cellular oxidative stress. Aging leads to adaptative reductions in CB responsiveness to hypoxia shifting the chemosensory setpoint upward. We submit that the attenuated CB sensitivity at old age may be tantamount to "physiological denervation" leading to a gradual loss of the chemosensing role in the prevention of tissue hypoxia by increasing lung ventilation.

The Evanescent Bouquet of Individual Bear Fingerprint.

The evanescent and invisible communication carried by chemical signals, pheromones, or signature mixtures or, as we prefer, the pheromonal individual fingerprint, between members of the same species is poorly studied in mammals, mainly because of the lack of identification of the molecules. The difference between pheromones and the pheromonal individual fingerprint is that the former generate stereotyped innate responses while the latter requires learning, i.e., different receivers can learn different signature mixtures from the same individual. Furthermore, pheromones are usually produced by a particular gland, while the pheromonal individual fingerprint is the entire bouquet produced by the entire secreting gland of the body. In the present study, we aim to investigate the pheromonal individual fingerprint of brown bears in northern Italy. We collected the entire putative pheromone bouquet from all production sites in free-ranging bears and analyzed the entire crude extract to profile the individual fingerprint according to species-, sex- and subjective-specific characteristics. We were able to putatively characterize the brown bears' pheromonal individual fingerprints and compare them with the partial pheromone identifications published by other studies. This work is a step forward in the study of the complexity of chemical communication, particularly in a solitary endangered species.

Physiological discrimination and correlation between olfactory and gustatory dysfunction in long-term COVID-19.

The spread of the SARS-CoV-2 virus produces a new disease termed COVID-19, the underlying physiological mechanisms of which are still being understood. Characteristic of the infection is the compromising of taste and smell. There is a persistent need to discriminate the dysfunctions and correlation between taste and smell, which are probably epiphenomena of other concealed conditions. Anosmic and ageusic long-term COVID-19 patients were re-evaluated after 1 year using a Volabolomic approach with an e-nose recording system coupled with olfactometric and gustometric tests. Here a range of sensory arrangements was found, from normal taste and smell to complete losses. The following patterns of olfactory threshold (OT)-taste threshold-olfactory uni- and cross-modal perception were found anosmia-severe hypogeusia-anosmia; hyposmia-hypogeusia-severe hyposmia; normosmia-ageusia-hyposmia; severe hyposmia -normogeusia-normosmia. There is a strong correlation between OT and olfactory uni- and cross-modal perception, a moderate correlation between olfactory and taste threshold and no correlation between OT and taste threshold. In conclusion, this study provides evidence for the feasibility of testing the chemical senses to directly objectify function in order to discriminate taste from olfactory impairment. Furthermore, it allows to hypothesize a long-term effect of the virus due to neuroinvasion through, probably, the olfactory system with injury in the related multisensory areas of taste and smell.

COVID-19 Smell Impairment and Crosstalk with Hypoxia Physiology.

Since its apomorphic appearance in 2019, severe acute respiratory syndrome Coronavirus type 2 (SARS-CoV-2) nowadays circulates as a plesiomorphic human virus in several synapomorphic variants. The respiratory tract is the most important site of infection, the viral effects in the lungs are well described, and more than half of the patients could develop shortness of breath and dyspnea and require ventilatory support. The physiological sign of this condition is the decrease in the partial pressure of oxygen in the blood, leading to acute hypoxia, which could be a factor in the disease. In severe patients, we recorded several physiological parameters: breath frequency (BF), partial pressure of oxygen in the blood (pO2), partial pressure of carbon dioxide in the blood (pCO2), hemoglobin (Hb), heart rate (HR), and blood pressure in correlation with the olfactory threshold. We found significant correlations between reduced olfactory threshold with pO2 and hemoglobin levels, changes in heart rate, and increased HR and pCO2. These results suggest that COVID-19 causes an impaired sense of smell that decreases in threshold corresponding to the disease severity.

Unmasking the 'Asymptomatic' COVID-19: A Nose Question.

The new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus has high infectivity, often masked by asymptomatic carriers, which allows it to spread rapidly and become a pandemic. Attempts to slow the pandemic at this stage depend on the ability to unmask asymptomatic carriers. The rapid diagnosis of active coronavirus disease 2019 (COVID-19) infection is one of the cornerstones of pandemic control, as the nasal cavity is the main gateway for SARS-CoV-2 entry and altered sense of smell is a feature of the current virus. In the present study, we therefore tested the olfactory threshold coupled with heart-lung parameters in subjects undergoing traditional molecular testing, resulting in a significantly different score between asymptomatic subjects and healthy controls. In total, 82% of asymptomatic positives showed olfactory impairment; of these, 46% had severe hyposmia and 7% had anosmia, while in the control 9% had severe hyposmia and 0% had anosmia, respectively, which agrees with heart rate, breathing rate, and blood pressure parameter variations. The olfactory test coupled with physiological parameters may help to identify asymptomatic people. In conclusion, our results suggest that most asymptomatic individuals could be unmasked by mass olfactory rapid threshold screening and then referred to traditional slower diagnostic tests.

Is Skull-Vibration-Induced Nystagmus Modified with Aging?

BACKGROUND: Despite clinical practice utilizing the Dumas test (SVINT), some questions remain unanswered, including the age-related changes in frequency (FN) and slow-phase angular velocity (SPAV). This study aims to retrospectively evaluate their variations in subjects affected by unilateral peripheral vestibular loss (UPVL). METHODS: We evaluated the selected samples based on the results of the SVINT, the results of the vestibular-evoked potentials (C-VEMP and O-VEMP), and the results of the head impulse test (HIT) and we compared the results against the age of the patients. We calculated the timing between the onset of UPVL and clinical evaluation in days. The presence or absence of VEMP indicated the UPVL severity. UPVL and BPPV patients with spontaneous or pseudo-spontaneous nystagmus were compared. RESULTS: Statistical analysis showed changes in the FN and SPAV depending on age and the side of the application of the stimulus. We also observed that, in the UPVL, the severity of the disease modifies the SPAV, but not the frequency. CONCLUSIONS: The SVINT is a simple, reliable, and straightforward test that, if evaluated instrumentally, can show significant differences with aging. Further studies need to be performed to refine the clinical significance of the test and clarify its physiological background.

Olfactory Response to Altitude Hypoxia: A Pilot Study During a Himalayan Trek.

The adaptation of olfaction to extreme environments is an area of limited understanding. This study aimed to get insights into the factors that constrain olfactory function at high altitudes. To this end, we compared the results of smell tests performed on the same subjects at low (665 m) and high altitude (4,780 m) during the "Kanchenjunga Exploration and Physiology" project in the Himalayas. The most distinct adaptive differences found at high altitude were reductions in the odor intensity, color-odor bimodal association, and memory, and increases in flavor perception. Physiological adaptations of the sense of smell resulting from exposure to hypoxia at altitude may have explanatory validity in unraveling the mechanisms that shape changes in olfactory function in the processes of aging and neurodegenerative diseases.

Italian Expert Consensus on Clinical and Therapeutic Management of Multiple Chemical Sensitivity (MCS).

Multiple chemical sensitivity (MCS) is a multisystem, recurrent, environmental disorder that flares in response to different exposures (i.e., pesticides, solvents, toxic metals and molds) under the threshold limit value (TLV) calculated for age and gender in the general population. MCS is a syndrome characterized by cutaneous, allergic, gastrointestinal, rheumatological, endocrinological, cardiological and neurological signs and symptoms. We performed a systematic review of the literature to summarize the current clinical and therapeutic evidence and then oriented an eDelphi consensus. Four main research domains were identified (diagnosis, treatment, hospitalization and emergency) and discussed by 10 experts and an MCS patient. Thus, the first Italian MCS consensus had the double aim: (a) to improve MCS knowledge among healthcare workers and patients by standardizing the clinical and therapeutic management to MCS patients; and (b) to improve and shed light on MCS misconceptions not supported by evidence-based medicine (EBM).

Postpartum Uterine Involution in Martina Franca Jennies.

This study investigated the postpartum (PP) uterine involution in nine multiparous Martina Franca jennies with at term, normal, and singleton foaling. Transrectal ultrasonography performed at Days 1, 3, 7, 14, 21, and 28 showed that the uterine tip diameters did not differ between the post-pregnant (PPH) and non-post-pregnant uterine horns (NPPH), whereas the diameter of the middle PPH was larger than the NPPH until Day 7 (p < 0.05). The diameter of the corpora-cornual junction resulted larger in the PPH than NPPH at Day 7 (p < 0.05). At Day 3, the microcaruncolae were not detected. Endometrial glands (GL) number increased, with the highest value on Day 28. Endometrial GL area and perimeter decreased (p < 0.001) from Day 1 to Day 28. Epithelial thickness increased from Days 1-3 to Day 7 (p < 0.001), and concurrently with the foal heat to Day 14 (p < 0.001), with a decrease at Days 21-28. A marked neutrophils reduction on Day 7 and eosinophil increase from the first three days to Days 7-14 was observed. The results suggest that, in Martina Franca jennies with normal foaling, the PP uterine involution can be considered complete on Day 14.

Volatile organic compounds (VOCs) in exhaled breath as a marker of hypoxia in multiple chemical sensitivity.

In the history of diagnostics, breath analysis was one of the first method used until the breakthrough of biochemical testing technology. Today, breath analysis has made a comeback with the development of gas analyzers and e-noses, demonstrating its power in its applicability for diagnosing a wide range of diseases. The physical basis of multiple chemical sensitivity (MCS), an emerging environmental disease, is difficult to understand because it is based on the scenario of chronic hypoxia, with a complex of chemical compounds that trigger the syndrome and result in multiple symptoms. The aim of this study was to investigate MCS by analyzing exhaled volatile organic compounds (VOCs). The volatile, metabolic picture could be a putative gold standard for understanding and diagnosing the disease. The study was based on recording in resting condition using the noninvasive passive e-nose contactless breath test, the Olfactory Real-Time Volatile Organic Compounds (ORT-VOC) test in MCS, and control samples. The VOCs profile distinguished between disease and health. It also distinguished the gender-related volatile profile with significant robustness. The results trace a putative compensatory physiological pathway elicited by increased lactate, leading to acidosis, and hyperventilation, resulting in the production of specific VOCs. We conclude that breath testing is a valuable tool to investigate the hypoxia-related VOC profile, facilitating MCS diagnosis.

Electrophysiological and olfactometric evaluation of long-term COVID-19.

COVID-19 is a public health emergency with cases increasing globally. Its clinical manifestations range from asymptomatic and acute respiratory disease to multiple organ dysfunction syndromes and effects of COVID-19 in the long term. Interestingly, regardless of variant, all COVID-19 share impairment of the sense of smell and taste. We would like to report, as far as we know, the first comprehensive neurophysiological evaluation of the long-term effects of SARS-CoV-2 on the olfactory system with potential-related neurological damage. The case report concerns a military doctor, with a monitored health history, infected in April 2020 by the first wave of the epidemic expansion while on military duty in Codogno (Milan). In this subject, we find the electrophysiological signal in the periphery, while its correlate is absent in the olfactory bulb region than in whole brain recordings. In agreement with this result is the lack of metabolic signs of brain activation under olfactory stimulation. Consequently, quantitative and qualitative diagnoses of anosmia were made by means of olfactometric tests. We strongly suggest a comprehensive series of olfactometric tests from the first sign of COVID-19 and subsequent patient assessments. In conclusion, electrophysiological and metabolic tests of olfactory function have made it possible to study the long-term effects and the establishment of neurological consequences.

Maternal Phylogenetic Relationships and Genetic Variation among Rare, Phenotypically Similar Donkey Breeds.

The mitochondrial DNA (mtDNA) D-loop of endangered and critically endangered breeds has been studied to identify maternal lineages, characterize genetic inheritance, reconstruct phylogenetic relations among breeds, and develop biodiversity conservation and breeding programs. The aim of the study was to determine the variability remaining and the phylogenetic relationship of Martina Franca (MF, with total population of 160 females and 36 males), Ragusano (RG, 344 females and 30 males), Pantesco (PT, 47 females and 15 males), and Catalonian (CT) donkeys by collecting genetic data from maternal lineages. Genetic material was collected from saliva, and a 350 bp fragment of D-loop mtDNA was amplified and sequenced. Sequences were aligned and evaluated using standard bioinformatics software. A total of 56 haplotypes including 33 polymorphic sites were found in 77 samples (27 MF, 22 RG, 8 PT, 19 CT, 1 crossbred). The breed nucleotide diversity value (π) for all the breeds was 0.128 (MF: 0.162, RG: 0.132, PT: 0.025, CT: 0.038). Principal components analysis grouped most of the haplogroups into two different clusters, I (including all haplotypes from PT and CT, together with haplotypes from MF and RG) and II (including haplotypes from MF and RG only). In conclusion, we found that the primeval haplotypes, haplogroup variability, and a large number of maternal lineages were preserved in MF and RG; thus, these breeds play putative pivotal roles in the phyletic relationships of donkey breeds. Maternal inheritance is indispensable genetic information required to evaluate inheritance, variability, and breeding programs.

Perception of Social Odor and Gender-Related Differences Investigated Through the Use of Transfer Entropy and Embodied Medium.

The perception of putative pheromones or social odors (PPSO) in humans is a widely debated topic because the published results seem ambiguous. Our research aimed to evaluate how cross-modal processing of PPSO and gender voice can affect the behavioral and psychophysiological states of the subject during a listening task with a bodily contact medium, and how these effects could be gender related. Before the experimental session, three embodied media, were exposed to volatilized estratetraenol (Estr), 5α-androst-16-en-3 α-ol (Andr), and Vaseline oil. The experimental session consisted in listening to a story that were transmitted, with a male or female voice, by the communicative medium via a Bluetooth system during a listening task, recorded through 64-active channel electroencephalography (EEG). The sense of co-presence and social presence, elicited by the medium, showed how the established relationship with the medium was gender dependent and modulated by the PPSO. In particular, Andr induced greater responses related to co-presence. The gender of the participants was related to the co-presence desire, where women imagined higher medium co-presence than men. EEG findings seemed to be more responsive to the PPSO-gender voice interaction, than behavioral results. The mismatch between female PPSO and male voice elicited the greatest cortical flow of information. In the case of the Andr-male voice condition, the trained model appeared to assign more relevance to the flow of information to the right frontotemporal regions (involved in odor recognition memory and social behavior). The Estr-male voice condition showed activation of the bilateral frontoparietal network, which is linked to cognitive control, cognitive flexibility, and auditory consciousness. The model appears to distinguish the dissonance condition linked to Andr matched with a female voice: it highlights a flow of information to the right occipital lobe and to the frontal pole. The PPSO could influence the co-presence judgements and EEG response. The results seem suggest that could be an implicit pattern linked to PPSO-related gender differences and gender voice.

Smell and Taste in Severe CoViD-19: Self-Reported vs. Testing.

One of the most striking reported symptoms in CoViD-19 is loss of smell and taste. The frequency of these impairments and their specificity as a potential central nervous system function biomarker are of great interest as a diagnostic clue for CoViD-19 infection as opposed to other similar symptomatologic diseases and because of their implication in viral pathogenesis. Here severe CoViD-19 was investigated by comparing self-report vs. testing of smell and taste, thus the objective severity of olfactory impairment and their possible correlation with other symptoms. Because a significant discrepancy between smell and taste testing vs. self-report results (p < 0.001) emerges in our result, we performed a statistical analysis highlighting disagreement among normosmia (p < 0.05), hyposmia, severe hyposmia, and anosmia (p < 0.001) and, in hypogeusia and severe hypogeusia, while no differences are observed in normogeusia and ageusia. Therefore, we analyzed the olfactory threshold by an objective test revealing the distribution of hyposmic (34%), severe hyposmic (48%), and anosmic (13%) patients in severe CoViD-19. In severe CoViD-19 patients, taste is lost in 4.3% of normosmic individuals, 31.9% of hyposmic individuals, 46.8% of severe hyposmic individuals, and 17% of anosmic individuals. Moreover, 95% of 100 CoViD-19 patients objectively tested were affected by smell dysfunction, while 47% were affected by taste dysfunction. Furthermore, analysis by objective testing also highlighted that the severity of smell dysfunction in CoViD-19 subjects did not correlate with age and sex. In conclusion, we report by objective testing that the majority of CoViD-19 patients report severe anosmia, that most of the subjects have olfactory impairment rather than taste impairment, and, finally, that the olfactory impairment correlate with symptom onset and hospitalization (p < 0.05). Patients who exhibit severe olfactory impairment had been hospitalized for about a week from symptom onset; double time has taken place in subjects with normosmia. Our results may be limited by the relatively small number of study participants, but these suggest by objective testing that hyposmia, severe hyposmia, and anosmia may relate directly to infection severity and neurological damage. The smell test assessment could be a potential screening symptom that might contribute to the decision to test suspected cases or guide quarantine instructions, further therapeutic approach, and evaluation of neurological damage.

Olfactory Event-Related Potentials and Exhaled Organic Volatile Compounds: The Slow Link Between Olfactory Perception and Breath Metabolic Response. A Pilot Study on Phenylethyl Alcohol and Vaseline Oil.

Olfactory processing starts with the breath and elicits neuronal, metabolic and cortical responses. This process can be investigated centrally via the Olfactory Event-Related Potentials (OERPs) and peripherally via exhaled Volatile Organic Compounds (VOCs). Despite this, the relationship between OERPs (i.e., N1 and Late Positive Component LPC) and exhaled VOCs has not been investigated enough. The aim of this research is to study OERPs and VOCs connection to two different stimuli: phenylethyl alcohol (PEA) and Vaseline Oil (VO). Fifteen healthy subjects performed a perceptual olfactory task with PEA as a smell target stimulus and VO as a neutral stimulus. The results suggest that OERPs and VOCs distributions follow the same amplitude trend and that PEA is highly arousing in both psychophysiological measures. PEA shows ampler and faster N1, a component related to the sensorial aspect of the stimulus. The N1 topographic localization is different between PEA and VO: PEA stimulus evokes greater N1 in the left centroparietal site. LPC, a component elicited by the perceptual characteristic of the stimulus, shows faster latency in the Frontal lobe and decreased amplitude in the Central and Parietal lobe elicited by the PEA smell. Moreover, the delayed time between the onset of N1-LPC and the onset of VOCs seems to be about 3 s. This delay could be identified as the internal metabolic time in which the odorous stimulus, once perceived at the cortical level, is metabolized and subsequently exhaled. Furthermore, the VO stimulus does not allocate the attentive, perceptive and metabolic resource as with PEA.

Antioxidant treatment for impaired hypoxic ventilatory responses in experimental diabetes in the rat.

Inflammation, tissue hypoxia, and impaired hypoxic ventilatory response (HVR) are the intricately entwined features of diabetes which perpetuate the disease and its sequelae. Hyperglycemia, notably, is an oxygen consuming process due to enhanced cellular metabolism. Oxidative stress underlies diabetic pathogenesis and also is a crucial modulator of the hypoxic chemoreflex. The present study seeks to determine if suppressed ventilation in diabetes could be improved by antioxidant treatment. The study was performed in streptozotocin-induced diabetes in awake rats. Two weeks into full-fledged diabetes, the rats were divided into mangiferin (potent natural antioxidant)-treated and untreated, with the observation continued for another two weeks. The HVR was investigated plethysmographically and compared with the pre-diabetic baseline in the same animal. Thiobarbituric acid reactive substances, superoxide dismutase, and tumor necrosis factor-α were assayed in the serum. We demonstrate that mangiferin reversed the suppressed HVR and acted toward normalization of oxidative and inflammatory stress. In conclusion, mangiferin holds a therapeutic promise in breaking the mesh of chronic tissue hypoxia, inflammation, and oxidative stress in diabetes by enhancing ventilation.

The companion dog as a unique translational model for aging.

The dog is a unique species due to its wide variation among breeds in terms of size, morphology, behaviour and lifespan, coupled with a genetic structure that facilitates the dissection of the genetic architecture that controls these traits. Dogs and humans co-evolved and share recent evolutionary selection processes, such as adaptation to digest starch-rich diets. Many diseases of the dog have a human counterpart, and notably Alzheimer's disease, which is otherwise difficult to model in other organisms. Unlike laboratory animals, companion dogs share the human environment and lifestyle, are exposed to the same pollutants, and are faced with pathogens and infections. Dogs represented a very useful model to understand the relationship between size, insulin-like growth factor-1 genetic variation and lifespan, and have been used to test the effects of dietary restriction and immunotherapy for Alzheimer's disease. Very recently, rapamycin was tested in companion dogs outside the laboratory, and this approach where citizens are involved in research aimed at the benefit of dog welfare might become a game changer in geroscience.

Olfactory phenotypic expression unveils human aging.

The mechanism of the natural aging of olfaction and its declinein the absence of any overt disease conditions remains unclear. Here, we investigated this mechanism through measurement of one of the parameters of olfactory function, the absolute threshold, in a healthy population from childhood to old age. The absolute olfactory threshold data were collected from an Italian observational study with 622 participants aged 5-105 years. A subjective testing procedure of constant stimuli was used, which was also compared to the 'staircase' method, with the calculation of the reliability. The n-butanol stimulus was used as an ascending series of nine molar concentrations that were monitored using an electronic nose. The data were analyzed using nonparametric statistics because of the multimodal distribution. We show that the age-related variations in the absolute olfactory threshold are not continuous; instead, there are multiple olfactory phenotypes. Three distinct age-related phenotypes were defined, termed as 'juvenile', 'mature' and 'elder'. The frequency of these three phenotypes depends on age. Our data suggest that the sense of smell does not decrease linearly with aging. Our findings provide the basis for further understanding of olfactory loss as an anticipatory sign of aging and neurodegenerative processes.