Speaking with mask in the COVID-19 era: Multiclass machine learning classification of acoustic and perceptual parameters.

The intensive use of personal protective equipment often requires increasing voice intensity, with possible development of voice disorders. This paper exploits machine learning approaches to investigate the impact of different types of masks on sustained vowels /a/, /i/, and /u/ and the sequence /a'jw/ inside a standardized sentence. Both objective acoustical parameters and subjective ratings were used for statistical analysis, multiple comparisons, and in multivariate machine learning classification experiments. Significant differences were found between mask+shield configuration and no-mask and between mask and mask+shield conditions. Power spectral density decreases with statistical significance above 1.5 kHz when wearing masks. Subjective ratings confirmed increasing discomfort from no-mask condition to protective masks and shield. Machine learning techniques proved that masks alter voice production: in a multiclass experiment, random forest (RF) models were able to distinguish amongst seven masks conditions with up to 94% validation accuracy, separating masked from unmasked conditions with up to 100% validation accuracy and detecting the shield presence with up to 86% validation accuracy. Moreover, an RF classifier allowed distinguishing male from female subject in masked conditions with 100% validation accuracy. Combining acoustic and perceptual analysis represents a robust approach to characterize masks configurations and quantify the corresponding level of discomfort.

Prospective memory: the combined impact of cognitive load and task focality.

Prospective Memory (PM) entails a set of executive processes primarily associated with the activation of frontal and parietal regions. Both the number of PM-targets to be monitored (i.e. task load) and the relationship between the type of PM-targets and the ongoing (ONG) task (i.e. task focality) can impact executive monitoring and PM performance. In the present imaging study, we manipulated load and focality of an event-based PM task to test the hypothesis that common resources engage in situations requiring high levels of cognitive control: that is, in high-load (i.e. monitor multiple PM-targets) and non-focal conditions (i.e. monitor at the same time letters' identity and color). We investigated monitoring-related and detection-related processes by assessing behavior and brain activity separately for ONG trials (monitoring) and PM-targets (detection). At the behavioral level, we found a significant interaction between load and focality during detection, with slowest reaction times for focal, high-load PM-targets. The imaging analyses of the detection phase revealed the activation of the left intraparietal sulcus in the high-load conditions. Both in the monitoring and the detection phases, we found overlapping effects of non-focality and low-load in the fusiform gyrus. Our results suggest that under low-load conditions, cognitive control operates via early selection mechanisms in the ventral occipito-temporal cortex. By contrast, high-load conditions entail control at later processing stages within the dorsal parietal cortex. We conclude that load and focality operate via different mechanisms, with the level of task load largely determining how cognitive control selects the most relevant information.

Endothelial activation and injury by cigarette smoke exposure.

Endothelial activation/injury following exposure to cigarette smoke may explain incidence of atherosclerosis and cardiovascular disease in smokers. We investigated cigarette smoke extract (CSE) effects relative to activation, injury, and survival of human umbilical vein endothelial cells (HUVEC) and compared circulating levels of specific endothelial activation markers between smokers and healthy non-smokers before and after smoking cessation. Viability and toxicity of HUVEC were tested by MTT and LDH assay. Release (by endothelial cells) and circulating levels (in smokers) of von Willebrand Factor (vWF), thrombomodulin (TM), was evaluated by ELISA. Incubation with increasing concentrations of CSE reduced the percentage of viable cells, being 33.9%, 23.9% after CSE 4%, 6% respectively. Dose- and time-dependent release of LDH was observed after incubation with CSE. vWF, TM release were assayed after CSE 2% HUVEC stimulation. Significant 42%, 61%, 76% increase in vWF concentration was detected respectively at 30', 60', 120'. Reduction in circulating levels of vWF, from a median value of 144.0% to 123.7%, was observed in the quitters group after smoking cessation. Exposure to cigarette smoke is cytotoxic and induces activation/injury of endothelium in vitro and in vivo. These findings may provide pathogenetic basis by which smoking can predispose to development of atherothrombosis and cardiovascular disease.

Treatment outcomes in patients with laryngotracheal lesions and compromised airway during and prior to the coronovirus disease 2019 pandemic: a tertiary institution's experience.

OBJECTIVE: This study aimed to compare treatment outcomes in patients with laryngeal and tracheal stenosis treated during and prior to the coronavirus disease 2019 pandemic period. METHOD: Patients treated for laryngotracheal lesions with impending airway compromise during the active pandemic period were matched with those treated for similar lesions in the preceding years in a monocentric tertiary hospital setting. RESULTS: During the pandemic period of 55 days, 31 patients underwent 47 procedures. Seven patients (2 children, 5 adults) had open airway surgery, and one had an operation-specific complication. Twenty-four patients (10 children, 14 adults) underwent 40 endoscopic interventions without any complications. Operation specific results during and prior to the pandemic were comparable. CONCLUSION: The management strategy in patients with laryngotracheal lesions and impending airway compromise should not be altered during periods of risk from coronavirus disease 2019. Avoiding a tracheostomy by performing primary corrective surgery or proceeding with a definitive decannulation would be beneficial in these patients to reduce the risk of contagion.

Smartphones Offer New Opportunities in Clinical Voice Research.

Smartphone technology provides new opportunities for recording standardized voice samples of patients and sending the files by e-mail to the voice laboratory. This drastically improves the collection of baseline data, as used in research on efficiency of voice treatments. However, the basic requirement is the suitability of smartphones for recording and digitizing pathologic voices (mainly characterized by period perturbations and noise) without significant distortion. In this experiment, two smartphones (a very inexpensive one and a high-level one) were tested and compared with direct microphone recordings in a soundproof room. The voice stimuli consisted in synthesized deviant voice samples (median of fundamental frequency: 120 and 200 Hz) with three levels of jitter and three levels of added noise. All voice samples were analyzed using PRAAT software. The results show high correlations between jitter, shimmer, and noise-to-harmonics ratio measured on the recordings via both smartphones, the microphone, and measured directly on the sound files from the synthesizer. Smartphones thus appear adequate for reliable recording and digitizing of pathologic voices.

Tamoxifen inhibits nerve growth factor-induced proliferation of the human breast cancerous cell line MCF-7.

An array of polypeptide growth factors contribute to the development of breast cancer, the most common tumor-related cause of death in women of Western countries. Therefore, breast cancer therapy should be aimed at inhibition of growth factor-dependent breast cancerous cell proliferation. However, the relative contribution of each individual factor in the development and maintenance of the transformed phenotype is largely unknown. Here we report for the first time that the proliferative effects of nerve growth factor, (NGF) a typical neurotrophin, are similar to those of epidermal growth factor (EGF) and insulin-like growth factor II, and are enhanced by 17beta-estradiol in the human breast cancer cell line MCF-7. The effect of NGF appeared to be mediated by its trkA receptors (trkA(NGFR)), as suggested by the potent inhibition of both MCF-7 cell proliferation and trkA(NGFR) phosphorylation occurring upon treatment of cultures with the selective trkA(NGFR) inhibitor K252a. Surprisingly, the antiestrogen drug tamoxifen (TAM) inhibited NGF-induced MCF-7 cell proliferation and trkA(NGFR) phosphorylation in a concentration-related fashion. The effect of TAM seemed to be estrogen receptor-independent, because the pure estrogen receptor antagonist ICI 182.780 was unable to block NGF-induced trkA(NGFR) phosphorylation. Our data underline the new emerging role of trkA(NGFR) in breast tumor growth, and suggest a related novel therapeutic use of TAM in breast cancer.

Neutralization of TRAIL death pathway protects human neuronal cell line from beta-amyloid toxicity.

Here we report that a novel member of the TNF-alpha family, TNF-related apoptosis-inducing ligand (TRAIL), contributes substantially to amyloid-induced neurotoxicity in human SH-SY5Y neuronal cell line. Involvement of TRAIL in the amyloid-induced cell death is supported by cDNA array, Northern blot, and Western blot data, demonstrating increased TRAIL expression after treatment of the cells with a neurotoxic fragment of amyloid protein (betaAP). TRAIL was also found to be released in the culture media after betaAP treatment with a time-course overlapping to contents of the intracellular protein. Contribution of TRAIL to betaAP neurotoxicity is demonstrated by data showing that TRAIL-neutralizing monoclonal antibody protects neuronal SH-SY5Y cells from betaAP neurotoxicity. Moreover, exposure of neuronal SH-SY5Y cells to TRAIL leads to cell death, indicating that this substance per se is endowed with neurotoxic properties. We also found that, similarly to betaAP and TRAIL, activation of the death-domain adaptor protein FADD results in neuronal cell death. Lack of FADD function, by overexpression of its dominant negative, rescued cells from either TRAIL- or betaAP-induced neurotoxicity, supporting the hypothesis that these three molecules share common intracellular pathways. Finally, we found that betaAP strongly activated caspase-8, and the cell-permeable, selective caspase-8 inhibitor z-IETD-FMK prevents both betaAP- and TRAIL-induced neurotoxicity. In view of TRAIL's potency in inducing neuronal death, and its role as mediator of betaAP, it is plausible to hypothesize that TRAIL can be regarded as a molecule that provides substantial contribution to betaAP-dependent cell death, which takes part in the progression of the neurodegenerative process and related chronic inflammatory response.

Protective effects of estradiol on TRAIL-induced apoptosis in a human oligodendrocytic cell line: evidence for multiple sites of interactions.

Demyelinating diseases are high impact neurological disorders. Steroids are regarded as protective molecules in the susceptibility to these diseases. Here, we studied the interactions between tumour necrosis factor-related apoptosis-inducing ligand (TRAIL), a potent proapoptotic molecule toxic to oligodendrocytes, and 17-beta-estradiol (E-17-beta), in human oligodendrocytic MO3.13 cells. Exposure of cells to TRAIL resulted in the upregulation of both death receptors DR4 and DR5 and apoptosis, as well as the activation of caspase-8 and -3, increased phosphorylation of Jun-N-terminal kinase and p38 kinase, and the reduction of bcl-2 and bcl-xL proteins. TRAIL-mediated MO3.13 cell apoptosis was abrogated by the dominant-negative form of the adaptor protein FADD and by caspase inhibitors. Preincubation with E-17-beta completely prevented both TRAIL-induced DR4 and DR5 upregulation and apoptosis. Estrogen-induced cytoprotection was time and concentration dependent and reverted by antiestrogens. Estrogen treatment per se reduced kinase phosphorylation, and upregulated bcl-2 and bcl-xL proteins. In conclusion, our data show that the detrimental role of TRAIL on oligodendrocytes can be effectively counteracted by estrogens, thus suggesting that the underlying molecular interactions can be of potential relevance in characterizing novel targets for therapy of demyelinating disorders.

Divergent effects of corticotropin releasing hormone on endothelial cell nitric oxide synthase are associated with different expression of CRH type 1 and 2 receptors.

1. Endothelium is a target for an array of factors involved in inflammation. Endothelial cells express receptors for CRH, a neuropeptide produced during inflammation. We report both the concentration-dependent inhibitory effect of CRH upon cytokine-stimulated nitrite release by H5V murine endothelioma cells, and its stimulatory one in HUVEC cells. 2. Western blot analysis showed that CRH inhibits cytokine-stimulated iNOS protein in H5V cells, and, instead, potentiated it in HUVEC cells. 3. H5V cells expressed both CRH receptors (CRH-R1 and R2) mRNAs, whereas HUVEC cells expressed the CRH-R2 mRNA solely. 4. CRH increased medium nitrites and iNOS protein expression in H5V cells pretreated with the selective CRH-R1 antagonist CP 154,526. However, the selective CRH-R2 antagonist anti-Svg-30 failed to produce similar effects. In fact, anti-Svg-30 inhibited CRH-induced increase of nitrite release and iNOS expression in HUVEC cells. 5. Our results confirm the activating role of CRH on endothelial cells, although it suggests its possible inhibitory role in the late phase of the inflammatory response. NO-mediated effects of CRH on endothelial cells could be exploited in therapeutic strategies related to inflammatory and/or degenerative diseases.

Increased ACTH and cortisol secretion after interleukin-alpha injection in the common marmoset (Callithrix jacchus jacchus).

We have studied the effect of intravenous injection of interleukin-1 (dose range: from 0.25 to 4.5 microg/kg of body weight) on plasma ACTH and cortisol levels in the marmoset, a primate paradygm of peripheral glucocorticoid resistance. Blood sampling were collected and body temperature recorded 0, 15, 30, 60, 120, 180, 240 and 300 min after injection. Interleukin-1 stimulated secretion of ACTH in a dose-dependent fashion. Maximal secretion occurred 120 min after injection, and lasted up to 240 min. Plasma ACTH levels returned to baseline 300 min after interleukin-1 injection. Plasma cortisol levels were related to ACTH levels. Body temperature elevation, which occurred 10-15 min after injection was dose-dependent, and lasted 3 h. Results suggest that the pyrogenic effect of interleukin is associated, in the marmoset, with integrated activation of the hypothalamic-pituitary-adrenal axis. In light of the proneness of marmosets to hyperimmune disorders, our data are consistent with the hypothesized central biological role of IL-1, as well as the pathophysiological relevance of the neuro-endocrine-immune cross-talk during the acute phase response.

Restoring abduction of paralyzed vocal cords in the cat using selective laryngeal reinnervation by phrenic motoneurons.

The aim of this work was to reestablish the respiratory function of the paralyzed larynx through reinnervation of the posterior cricoarytenoid (PCA) muscle by phrenic motoneurons. In nine adult cats the adductor branch of the recurrent laryngeal nerve (RLN) of one side was cut and ligated, while the abductor branch was left intact. The whole RLN was then transected lower in the neck and its distal stump anastomosed to the upper branch of the phrenic nerve. Periodical laryngoscopies under ketamine anesthesia assessed that the inspiratory abduction of the paralyzed vocal cord recovered within 45 days to 60 days in all cats. Abduction was caused by reinnervation of the PCA muscle from phrenic motoneurons, as demonstrated by electrophysiological and anatomical (retrograde transport of horseradish peroxidase) testings.

Recovery of inspiratory abduction of the paralyzed vocal cords after bilateral reinnervation of the cricoarytenoid muscles by one single branch of the phrenic nerve.

The aim of this study was to provide the bilateral reinnervation of the posterior cricoarytenoid (PCA) muscles by the superior root of the right phrenic nerve. In six adult cats, the right phrenic root was anastomosed to the distal stump of the transected recurrent laryngeal nerve (RLN) on the same side. The RLN adductor branch was then cut and anastomosed to a nerve graft whose end was carried contralaterally and sutured to the left RLN or to the left PCA muscle. The phrenic fibers regrowing along the RLN abductor branch reinnervated the right PCA muscle and restored the inspiratory abduction of the right vocal cord in all the animals. In five of the six cats, the fibers regenerated through the RLN adductor branch and the graft reached the left PCA muscle and also restored the inspiratory opening of the larynx on the left side. Histological nerve examination revealed a fairly symmetrical distribution of the regenerated phrenic axons to the right and left PCA muscles.

Innervation of the paralyzed laryngeal muscles by phrenic motoneurons. A quantitative study by light and electron microscopy.

In the cat, inspiratory opening of the paralyzed glottis recovered after unilateral or bilateral reinnervation of the posterior cricoarytenoid (PCA) muscles by phrenic axons. The morphometric analysis of the regenerated recurrent laryngeal nerves (RLNs), showed that proliferation was abundant; 4 months after the nerve anastomosis, more than 500 myelinated axonal branches repopulated the RLNs. The mean diameter of motor axons (3.5 to 5.0 microns) was lower than in normal phrenic and RLN (8 to 10 microns), and the mean internode length was about half that of the normal RLN. Histochemical examination of the PCA muscle revealed that muscle fiber composition (44% type I and 56% type II muscle fiber) was fairly similar to that of normal PCA. The contraction time of the reinnervated muscles was as long as 60 msec at the time of movement recovery, but it shortened to 25 to 30 msec when the reinnervation time increased. These anatomical and functional results support the choice of the phrenic nerve for laryngeal reinnervation.

Thymic factors influence on behavior in rodents.

We studied the effect of thymopentin, a synthetic thymic peptide, on spontaneous behavior and stress models in BALB/c mice in which a WEHI 164 clone 13 murine fibrosarcoma had been implanted, as well as in the intact Sprague-Dawley rat. In untreated animals with tumors, spontaneous behavior was significantly inhibited. Resistance to swimming in cold water was also decreased in untreated animals. Thymopentin (10, 100, 1,000, and 5,000 micrograms/kg body weight, IP, 20 min before the test) enhanced spontaneous behavior in tumor-implanted mice. In addition, thymopentin partially restored floating capability of tumor bearers in either freely moving or animals on which an additional weight had been applied. In the latter test, plasma corticotropin and corticosterone levels were relatedly modified according to treatment. Rats treated with thymopentin showed a decreased sensitivity to painful stimuli. The effect of thymopentin was comparable to acetylsalicylate. Finally, thymic factors appeared capable of restoring the diminished behavioral activity of animals bearing tumors, as well as of increasing resistance to stressful stimuli and pain.

A possible sequela of transoral approach to the upper cervical spine. Velopharyngeal incompetence.

The authors describe a case of velopharyngeal incompetence (VPI), as a consequence to the neurosurgical treatment for a complex malformation of the cranio-spinal junction. A 61-year-old woman underwent a transoral-transvelar surgical approach for odontoid resection. One month later surgical fixation of the posterior spine with autologous iliac bone graft was performed. Following these operations the patient presented a marked alteration of speech intellegibility due to hypernasal voice resonance and through incapability to articulate the oral phonemes correctly. She also complained of nasal regurgitation of fluids and solids while swallowing. She underwent a clinical phoniatric assessment of voice and speech. Videonasopharyngoscopy allowed us to inspect the velopharyngeal sphincter and to show clearly the type and morphology of its closure defect. Correction of VPI was achieved by means of a velopharyngoplasty (pharyngeal flap), in spite of technical difficulties due to local scarring and to a problematic exposure of the surgical field.

Bilateral laryngeal pseudoparalysis in xanthoma disseminatum treated by endoscopic laser medial arytenoidectomy.

Xanthoma disseminatum is a rare non-Langerhans' cell histiocytosis, characterized by papular cutaneous eruption, possible mucosal involvement, and frequent association with vasopressin-sensitive diabetes insipidus. Herein we report a case of xanthoma disseminatum with pharyngolaryngeal involvement. In this patient, mucosal xanthomas involving the arytenoid cartilages and the interarytenoid area resulted in laryngeal stenosis and severe impairment of both cricoarytenoid joints' motility. Endoscopic CO2 laser medial arytenoidectomy, according to the technique described by Crumley (1993), and vaporization of interarytenoid xanthomas were successfully performed, thus reestablishing bilateral cordal motility and the laryngeal airway. Four years later, a CO2 laser revision was necessary because of recurrence of xanthomas in the posterior larynx. Two years after the latter operation, the patient has no signs of laryngeal obstruction and has a normal voice quality. This case report suggests that endoscopic medial arytenoidectomy may be successfully used in the treatment of bilateral laryngeal pseudoparalysis secondary to xanthoma disseminatum.

[Evaluation of chronic nasal respiratory obstruction in children using optic fiber rhinopharyngoscopy and rhinomanometry]. / Valutazione dell'ostruzione respiratoria nasale cronica del bambino mediante rinofaringoscopia a fibre ottiche e rinomanometria.

Traditional clinical examination is frequently inadequate to evaluate children affected by chronic nasal obstruction: radiology may be of some use but it exposes children to radiation and gives only a limited lateral view of the nasopharynx. Therefore other means need to be tested for the anatomical and functional examination of the nasal and nasopharyngeal cavities. Adenoid hypertrophy and chronic nasal congestion are the most common causes of nasal respiratory impairment in children. The clinical expressions of adenoid enlargement are influenced by the osteo-cartilaginous nasal conformation and by functional alterations of nasal mucosa. Therefore a correct clinical examination is necessary to determine the indications for adenoidectomy so as to prevent unsatisfactory results. The aim of this study was to test the efficacy of rhinopharyngoscopy and rhinomanometry in the quantitative and qualitative evaluation of nasal and nasopharyngeal airways in 50 children (age-range 5-12 years) affected by nasal obstruction. Based on the rhinopharyngoscopic evaluation of adenoid volume, the children were divided into 3 groups affected by choanal obstruction defined as: severe (28 cases); partial (14 cases); minimal (8 cases). In these 3 groups anterior rhinomanometry, performed before and after administration of a nasal decongestant, allowed to evidence the role played by functional and organic factors in determining nasal stenosis. Abnormal values of total nasal resistance were found in 42 of the 50 children in the study. In 12 of these cases there was a marked decrease in nasal resistance following decongestion, evidencing the prevalence of functional factors in determining nasal obstruction and contra-indicating adenoidectomy. Rhinopharyngoscopy and rhinomanometry were well tolerated by all examined patients; the combined use of the two techniques allowed a more accurate diagnostic assessment of functional and anatomical causes of nasal obstruction in order to predict the benefits which can be obtained through adenoidectomy.

Ischemic tolerance modulates TRAIL expression and its receptors and generates a neuroprotected phenotype.

TNF-related apoptosis inducing ligand (TRAIL), a member of the TNF superfamily released by microglia, appears to be involved in the induction of apoptosis following focal brain ischemia. Indeed, brain ischemia is associated with progressive enlargement of damaged areas and prominent inflammation. As ischemic preconditioning reduces inflammatory response to brain ischemia and ameliorates brain damage, the purpose of the present study was to evaluate the role of TRAIL and its receptors in stroke and ischemic preconditioning and to propose, by modulating TRAIL pathway, a new therapeutic strategy in stroke. In order to achieve this aim a rat model of harmful focal ischemia, obtained by subjecting animals to 100 min of transient occlusion of middle cerebral artery followed by 24 h of reperfusion and a rat model of ischemic preconditioning in which the harmful ischemia was preceded by 30 mins of tMCAO, which represents the preconditioning protective stimulus, were used. Results show that the neuroprotection elicited by ischemic preconditioning occurs through both upregulation of TRAIL decoy receptors and downregulation of TRAIL itself and of its death receptors. As a counterproof, immunoneutralization of TRAIL in tMCAO animals resulted in significant restraint of tissue damage and in a marked functional recovery. Our data shed new light on the mechanisms that propagate ongoing neuronal damage after ischemia in the adult mammalian brain and provide new molecular targets for therapeutic intervention. Strategies aimed to repress the death-inducing ligands TRAIL, to antagonize the death receptors, or to activate the decoy receptors open new perspectives for the treatment of stroke.

Autologous fat injection to face and neck: from soft tissue augmentation to regenerative medicine.

Minimally-invasive autologous fat injection of the head and neck region can be considered a valid alternative to major invasive surgical procedures both for aesthetic and functional purposes. The favourable outcomes of autologous fat injection in otolaryngological practice are due to the filling of soft tissue and, mainly, to the potential regenerative effect of adipose-derived mesenchymal stem cells. Herewith, some important biological preliminary remarks are described underlying the potential of autologous fat injection in regenerative medicine, and personal experience in using it for both consolidated clinical applications, such as fat grafting to the face and vocal fold augmentation in the treatment of glottic incompetence, and more recent applications including the treatment of post-parotidectomy Frey syndrome and velopharyngeal insufficiency.