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Purpose: Background noise and voice problems among teachers can degrade listening conditions in classrooms. The aim of this literature review is to understand how these acoustic degradations affect spoken language processing in 6- to 18-year-old children. Method: In a narrative report and meta-analysis, we systematically review studies that examined the effects of noise and/or impaired voice on children's response accuracy and response time (RT) in listening tasks. We propose the Speech Processing under Acoustic DEgradations (SPADE) framework to classify relevant findings according to three processing dimensions--speech perception, listening comprehension, and auditory working memory--and highlight potential moderators. Results: Thirty-one studies are included in this systematic review. Our meta-analysis shows that noise can impede children's accuracy in listening tasks across all processing dimensions (Cohen's d between -0.67 and -2.65, depending on signal-to-noise ratio) and that impaired voice lowers children's accuracy in listening comprehension tasks (d = -0.35). A handful of studies assessed RT, but results are inconclusive. The impact of noise and impaired voice can be moderated by listener, task, environmental, and exposure factors. The interaction between noise and impaired voice remains underinvestigated. Conclusions: Overall, this review suggests that children have more trouble perceiving speech, processing verbal messages, and recalling verbal information when listening to speech in noise or to a speaker with dysphonia. Impoverished speech input could impede pupils' motivation and academic performance at school.

Purpose: The aim of this study was to investigate children's processing of dysphonic speech in a realistic classroom setting, under the influence of added classroom noise. Method: Typically developing 6-year-old primary school children performed two listening tasks in their regular classrooms--a phoneme discrimination task to assess speech perception and a sentence-picture matching task to assess listening comprehension. Speech stimuli were played back in either a typical or an impaired voice quality. Children performed the tasks in the presence of induced classroom noise at signal-to-noise ratios between +2 and +9 dB. Results: Children's performance in the phoneme discrimination task decreased significantly when the speaker's voice was impaired. The effect of voice quality on sentence-picture matching depended on task demands: Easy sentences were processed more accurately in the impaired-voice condition than in the typical-voice condition. Signal-to-noise ratio effects are discussed in light of methodological constraints. Conclusions: Listening to a dysphonic teacher in a noisy classroom may impede children's perception of speech, particularly when phonological discrimination is needed to disambiguate the speech input. Future research regarding the interaction of voice quality and task demands is necessary.

Purpose: Our aim was to investigate isolated and combined effects of speech-shaped noise (SSN) and a speaker's impaired voice quality on spoken language processing in first-grade children. Method: In individual examinations, 53 typically developing children aged 5-6 years performed a speech perception task (phoneme discrimination) and a listening comprehension task (sentence-picture matching). Speech stimuli were randomly presented in a 2 × 2 factorial design with the factors "noise" (no added noise vs. SSN at 0- dB SNR) and "voice quality" (normal voice vs. impaired voice). Outcome measures were task performance and response time (RT). Results: SSN and impaired voice quality significantly lowered children's performance and increased RTs in the speech perception task, particularly when combined. Regarding listening comprehension, a significant interaction between noise and voice quality indicated that children's performance was hindered by SSN when the speaker's voice was impaired but not when it was normal. RTs in this task were unaffected by noise or voice quality. Conclusions: Results suggest that speech signal degradations caused by a speaker's impaired voice and background noise generate more processing errors and increased listening effort in young school-aged children. This finding is vital for classroom listening and highlights the importance of ensuring teachers' vocal health and adequate room acoustics.

Purpose: Although a global picture exists of teachers' voice demands in general, few studies have compared specific groups of teachers to determine whether some are more at risk than others. This study compared the vocal loadings of kindergarten and elementary school teachers; professional and nonprofessional vocal load were determined for both groups. Method: Twelve kindergarten and 20 elementary school female teachers without voice problems were monitored during 1 workweek using the Ambulatory Phonation Monitor. Vocal loading parameters analyzed were F0, SPL, time dose, distance dose, and cycle dose. Results: Comparisons between the groups showed significantly higher cycle dose and distance dose for kindergarten teachers than for elementary school teachers, in both professional and nonprofessional environments. Professional and nonprofessional voice use comparisons showed significant differences for all parameters, indicating that vocal load was higher in the professional environment for both groups. Conclusions: The higher vocal doses measured in kindergarten teachers suggest that particular attention should be paid to this specific group of teachers. Although nonprofessional vocal load is lower than professional vocal load, it is important to take both into account because of their cumulative effects.

This study measured the impact of articulation exercises using a cork and articulation exercises for lip spreading on the formant frequencies of vowels and listener perceptions of femininity in transgender women. Thirteen transgender women were recorded before and after the cork exercise and before and after the lip spreading exercise. Speech samples included continuous speech during reading and were analyzed using Praat software. Vowel formant frequencies (F1, F2, F3, F4, F5) and vowel space were determined. A listening experiment was organized using naïve cisgender women and cisgender men rating audio samples of continuous speech. Masculinity/femininity, vocal quality and age were rated, using a visual analogue scale (VAS). Concerning vowel formant frequencies, F2 /a/ and F5 /u/ significantly increased after the lip spreading exercise, as well as F3 /a/, F3 /u/ and F4 /a/ after the cork exercise. The lip spreading exercise had more impact on the F2 /a/ than the cork exercise. Vowel space did not change after the exercises. The fundamental frequency (f o) increased simultaneously during both exercises. Both articulation exercises were associated with significantly increased listener perceptions of femininity of the voice. Subtle changes in formant frequencies can be observed after performing articulation exercises, but not in every formant frequency or vowel. Cisgender listeners rated the speech of the transgender women more feminine after the exercises. Further research with a more extensive therapy program and listening experiment is needed to examine these preliminary findings. [ABSTRACT FROM AUTHOR]

Summary Objectives This study aimed (1) to investigate music theory teachers' professional and extra-professional vocal loading and background noise exposure, (2) to determine the correlation between vocal loading and background noise, and (3) to determine the correlation between vocal loading and self-evaluation data. Methods Using voice dosimetry, 13 music theory teachers were monitored for one workweek. The parameters analyzed were voice sound pressure level (SPL), fundamental frequency (F0), phonation time, vocal loading index (VLI), and noise SPL. Spearman correlation was used to correlate vocal loading parameters (voice SPL, F0, and phonation time) and noise SPL. Each day, the subjects self-assessed their voice using visual analog scales. VLI and self-evaluation data were correlated using Spearman correlation. Results Vocal loading parameters and noise SPL were significantly higher in the professional than in the extra-professional environment. Voice SPL, phonation time, and female subjects' F0 correlated positively with noise SPL. VLI correlated with self-assessed voice quality, vocal fatigue, and amount of singing and speaking voice produced. Conclusions Teaching music theory is a profession with high vocal demands. More background noise is associated with increased vocal loading and may indirectly increase the risk for voice disorders. Correlations between VLI and self-assessments suggest that these teachers are well aware of their vocal demands and feel their effect on voice quality and vocal fatigue. Visual analog scales seem to represent a useful tool for subjective vocal loading assessment and associated symptoms in these professional voice users. [ABSTRACT FROM AUTHOR]

Summary: Objective: The operatic singing technique is frequently used in classical music. Several acoustical parameters of this specific technique have been studied but how these parameters combine remains unclear. This study aims to further characterize the Western operatic singing technique by observing the effects of melody and technique on acoustical and musical parameters of the singing voice. Methods: Fifty professional singers performed two contrasting melodies (popular song and romantic melody) with two vocal techniques (with and without operatic singing technique). The common quality parameters (energy distribution, vibrato rate, and extent), perturbation parameters (standard deviation of the fundamental frequency, signal-to-noise ratio, jitter, and shimmer), and musical features (fundamental frequency of the starting note, average tempo, and sound pressure level) of the 200 sung performances were analyzed. Results: The results regarding the effect of melody and technique on the acoustical and musical parameters show that the choice of melody had a limited impact on the parameters observed, whereas a particular vocal profile appeared depending on the vocal technique used. Conclusions: This study confirms that vocal technique affects most of the parameters examined. In addition, the observation of quality, perturbation, and musical parameters contributes to a better understanding of the Western operatic singing technique. [Copyright &y& Elsevier]

Summary: Objective: The quality of a music performance can be lessened or enhanced if the performer experiences stressful conditions. In addition, the quality of a sung performance requires control of the fundamental frequency of the voice, which is particularly sensitive to stress. The present study aimed to clarify the effects of stress on singing voice accuracy. Methods: Thirty-one music students were recorded in a stressful condition (ie, a music examination) and a nonstressful condition. Two groups were defined according to the challenge level of the music examination (first and second music levels). Measurements were made by self-reported state anxiety (CSAI-2R questionnaire) and by observing heart rate activity (electrocardiogram) during each performance. In addition, the vocal accuracy of the sung performances was objectively analyzed. Results: As expected, state anxiety and heart rate were significantly higher on the day of the music examination than in the nonstressful condition for all the music students. However, the effect of stress was positive for the first-year students but negative for the second-year students, for whom the music examination was particularly challenging. In addition, highly significant correlations were found between the intensity of cognitive symptoms and the vocal accuracy criteria. Discussion: This study highlights the contrasting effects of stress on singing voice accuracy but also the need to consider the challenge level and perception of the symptoms in experimental and pedagogical settings. [Copyright &y& Elsevier]

Summary: Subjective evaluation of the voice by the patient is routinely assessed in the adult dysphonic population; this is, however, not the case in the pediatric population. There were three objectives of this study: the first goal was to study the ability of children aged 5–13 years to express themselves about physical, emotional, and sociofunctional aspects of their voice. The second goal was to explore if specific voice-related complaints were expressed by dysphonic children as compared with normophonic children. The third goal was to compare the dysphonic children''s voice-related complaints with those of their mothers. The overall objective was to set the grounds for the elaboration of a standardized questionnaire in French concerning subjective evaluation of voice in children. Twenty-five dysphonic children with vocal complaint (15 nodules, one polyp, one microweb, eight unspecified) and 55 normophonic children aged 5–13 years were interviewed. The interviews were semistructured based on a canvas of voice-related questions. The dysphonic children''s mothers were interviewed with the mean of a written questionnaire and were invited to discuss their answers orally with the examiner. The results were analyzed qualitatively and statistically. A Chi-square test and the Fisher''s test were used to analyze the differences between the complaints expressed by the dysphonic and the normophonic children, and a binomial test was used to compare the children''s answers with their mothers'' answers. The qualitative analysis of the interviews suggests that children are capable of reflecting over their own voice and of giving autonomous information about different aspects of their voice. It also appeared that voice is a complex phenomenon and that it needs to be clearly and cautiously defined to the children. We identified 27 different complaints related to the voice, out of which 17 were significantly more expressed by dysphonic than by normophonic children (P <0.05). Three of the 27 identified complaints show significant discordances between the mothers and the dysphonic children. The results suggest that children are capable of making a subjective and autonomous evaluation of their voice and that dysphonic children experience significantly more voice-related discomfort than nondysphonic children. The complaints expressed by the dysphonic children and their mothers are not all in concordance. The main conclusion is that a standardized subjective evaluation of the voice, not only by the parents but also by the child him/herself, would be relevant in the assessment of pediatric dysphonia. [Copyright &y& Elsevier]

Summary: Objectives: The aim of this study was to evaluate the objective voice results of Wendler''s glottoplasty in male-to-female transsexuals. Method: We retrospectively reviewed our patients treated with Wendler''s technique with minor modifications. Glottoplasty consisted in CO2-laser epithelial ablation of the anterior commissure and the two vocal folds in anterior third, suturing of the two vocal folds with two stitches of 3.0 resorbable thread, and application of fibrin sealant to strengthen the suture. Voice assessment was based mainly on fundamental frequency (F 0), frequency range, jitter, maximum phonation time, phonation quotient, estimated subglottic pressure (ESGP) grade of dysphonia (G), and voice handicap index (VHI). These measures were taken before surgery and on the last follow-up visit. Results: Our series included 15 patients with a mean age of 36 years. The mean follow-up period was 7.2 months. We did not observe any early complications related to the technique. The comparison between the preoperative and the postoperative measurements, using Wilcoxon signed rank test, showed a significant improvement of median F 0 from 139 to 191Hz (P =0.006) with an increase in the grade of dysphonia (G pre =0.2, G post =1, P =0.013) and ESGP (ESGPpre =8.1±3.2, ESGPpost =12.0±3.8, P =0.002). Other measurements, including VHI, did not show any significant differences pre- and postoperatively. Conclusion: Wendler''s glottoplasty can contribute to feminize the voice. [Copyright &y& Elsevier]

Summary: The objective of the study was to measure the subjective long-term effects of vocal therapy (more than 6 months) in 29 patients suffering from dysfunctional dysphonia. Four subjective measurements were used: one visual analog scale ranging from 0 (not efficient at all) to 100 (very efficient); one question “Is the speech therapy still efficient?,” answered by yes or no; the GRB parameters of the GRBAS scale (evaluated by the patient); and the VHI-10. The results show that 76% of the patients consider that the vocal therapy they received is still efficient. On the vocal quality (G), roughness (R), and breathiness (B) parameters, we observe a high degree of satisfaction on the G parameter (median=77). The scores on the R and B parameters are lower (median R =12, S =5). Roughness is significantly correlated to the Grade (Spearman coefficient r =0.516, P =0.004), whereas breathiness is not (P =0.251, NS). The comparison of the results on the VHI-10 pre and post treatment shows a significant decrease in the grade of perceived handicap (medians 15 vs 11, P =0.017). The correlations between the judgment of the treatment's efficacy and the vocal quality is significant (r =0.623, P <0.001) as well as the correlation between the VHI-10 and the length of the treatment (r =0.416, P =0.035). The conclusion can be made that speech therapy plays an important role in long-term treatment of dysfunctional dysphonias. [Copyright &y& Elsevier]

The objective of the study was to measure the subjective long-term effects of vocal therapy (more than 6 months) in 29 patients suffering from dysfunctional dysphonia. Four subjective measurements were used: one visual analog scale ranging from 0 (not efficient at all) to 100 (very efficient); one question 'Is the speech therapy still efficient?,' answered by yes or no; the GRB parameters of the GRBAS scale (evaluated by the patient); and the VHI-10. The results show that 76% of the patients consider that the vocal therapy they received is still efficient. On the vocal quality (G), roughness (R), and breathiness (B) parameters, we observe a high degree of satisfaction on the G parameter (median=77). The scores on the R and B parameters are lower (median R=12, S=5). Roughness is significantly correlated to the Grade (Spearman coefficient r=0.516, P=0.004), whereas breathiness is not (P=0.251, NS). The comparison of the results on the VHI-10 pre and post treatment shows a significant decrease in the grade of perceived handicap (medians 15 vs 11, P=0.017). The correlations between the judgment of the treatment's efficacy and the vocal quality is significant (r=0.623, P<0.001) as well as the correlation between the VHI-10 and the length of the treatment (r=0.416, P=0.035). The conclusion can be made that speech therapy plays an important role in long-term treatment of dysfunctional dysphonias. [ABSTRACT FROM AUTHOR]

The Acoustic Voice Quality Index (AVQI), version 03.01, is a tool for quantitative assessment of the overall severity of dysphonia. It is based on the recordings of a sustained vowel and part of a text read aloud. For the Dutch sample, 34 syllables must be read aloud to balance the duration of the two tasks. The first part of this study thus aimed to determine how many syllables of a commonly used text in the French-speaking part of Belgium should be used to achieve the same balance. The psychometric qualities of the AVQI have been confirmed by numerous publications in various languages. However, its validation in French relies on a small cohort of patients, who were not native French speakers. Furthermore, version 03.01 of the AVQI has not yet been validated at all on French samples. Hence, the main aim of this study was to assess the criterion-related concurrent validity and diagnostic accuracy of the AVQI 03.01 applied to a sample of native French speakers. For the first part of this study, the optimal part of the text to be used for the AVQI was identified, taking into account both its phonemic contents and its time balance with the sustained vowel. For the validation study, 90 recordings from the University Hospital of Liège's ENT caseload database were used, as well as 30 new recordings of normophonic individuals, composing a control group. Four judges assessed the recordings using the G parameter of the GRBAS scale. Once the intra- and inter-rater reliability of the perceptual ratings was confirmed, the AVQI 03.01's criterion validity was assessed on the French sample. The diagnostic accuracy of the AVQI 03.01 in French was measured, and the cut-off score allowing for the greatest diagnostic precision determined. The most appropriate syllable number of the text to be read aloud was found to be 27, in order to balance the time analyzed for both the sustained vowel and the continuous speech. Regarding the validation study, intra-rater reliability was substantial for each of the four vocologists (κ mean =.778, P < 0.0001), and inter-rater reliability was high (W =.895, P < 0.0001). The Spearman correlation between the perceptual judgments and the AVQI 03.01 score was strong (r s =.84, P < 0.0001). The receiver operating characteristic-curve parameters indicated that the ideal cut-off score allowing for the highest diagnostic accuracy of the AVQI, version 03.01, applied to a French sample is 2.33, with a sensitivity of 59.8%, a specificity of 100%, an infinite positive likelihood ratio (LR+) and a negative likelihood ratio (LR−) of 0.4. This study confirms the external validity of the AVQI 03.01 when applied on a French 27-syllable sample. The AVQI 03.01 is a robust, ecologically valid objective measure of overall voice quality. The cut-off score to be used is 2.33. However, clinicians should be cautious when the AVQI score is lower than 2.33. The AVQI 03.01 does not yield a sufficiently low negative likelihood ratio to be sure that this score indeed indicates normophonia. Also, taking into account the limitations regarding the perceptual judgements used in this study, a replication study should be carried out in order to confirm the cut-off score. [ABSTRACT FROM AUTHOR]

The Acoustic Voice Quality Index (AVQI), version 03.01, is a tool for quantitative assessment of the overall severity of dysphonia. Its computation includes six acoustic parameters, which are all analyzed with Praat freeware. It is based on recordings of a sustained vowel and part of a text read aloud. The psychometric qualities of this tool in various languages, including French, have been confirmed by numerous publications. However, studies investigating the correlation between tools for objective vocal assessment and voice-related quality of life show inconsistent results. Hence, the aim of this study was to contribute to the debate on measuring the correlations between the AVQI 03.01 score computed on French samples and the Voice Handicap Index (VHI). Data from 78 patients were used; they were collected during initial vocal assessment and stored in the ear, nose, and throat caseload database of the University Hospital of Liège. The Spearman rank-order correlations (r s) between the VHI total score and subscores and the AVQI 03.01 scores for French samples were measured. The correlation between the diagnostic decisions ("normophonia" versus "dysphonia") of both tools was assessed using Cramer's phi. The Spearman correlation between AVQI 03.01 score and total VHI score, controlling for age, was moderate (r s = 0.62, P < 0.0001). The correlations between the AVQI 03.01 score and the functional, emotional, and physical subscores of the VHI were also moderate (r s = 0.643, 0.543, and 0.514, respectively, P < 0.0001). The correlation between the two instruments' diagnostic decisions ("normophonia" versus "dysphonia") was also moderate (φ = 0.52, P = 0.000). Although AVQI 03.01 scores were moderately correlated with the VHI total score and subscores, they measure two different things. The AVQI 03.01 assesses overall voice quality in terms of acoustic parameters, whereas the VHI assesses the multidetermined impact on the patient's everyday life. Both results should thus be taken into account, as part of a comprehensive vocal assessment. [ABSTRACT FROM AUTHOR]

This is a case-control study investigating the personality profiles of children with vocal fold nodules (VFNs). Personality has been suggested as a contributing factor to VFN in adults and in children. However, previous studies in children have focused on psychopathologic traits and have reported contradictory findings. The aim of our study was to explore whether general, nonpathologic personality structure predicts the presence of VFN in children. Personality traits were measured in 34 children with VFN, aged 6.0–13.6, and 34 age- and sex-matched normophonic controls by means of a pediatric five-factor personality questionnaire. Children with VFN were found to be significantly more extroverted than controls (P = 0.022, d = 0.61), and a binary logistic regression showed that extraversion significantly predicted the presence of VFN in our population (P = 0.027). The clinical implications of the results are discussed. [ABSTRACT FROM AUTHOR]

Little is known about the impact of smoking on voice acoustics. Some studies have found that tobacco affects the fundamental frequency of the voice, whereas others have not. This study aimed to overcome the major methodological limitations observed in previous studies by strictly controlling several variables that could clarify the effect of smoking on the speaking voice. Lebanese men were chosen for this study. Thirty nonsmokers, 30 cigarette smokers, and 30 water-pipe smokers were matched on the basis of age, height, and weight. The 90 participants were asked to complete the Voice Handicap Index, sustain the vowel /a/, read 10 sentences in French and Arabic, and speak spontaneously in both languages. The mean fundamental frequency (F0), speaking fundamental frequency (SFF), jitter, and standard deviation of F0 were measured using Praat and Vocalab4 and were compared between the groups. The Voice Handicap Index scores differed significantly between nonsmokers and cigarette smokers and between nonsmokers and water-pipe smokers. Results also show that cigarette smokers' F0 and SFF were significantly lower than nonsmokers' results. No significant differences were found between water-pipe smokers and nonsmokers. The jitter and the standard deviation of F0 did not differ significantly between the two groups. Our findings clearly demonstrate the effect of smoking on the voice: smokers reported more voice complaints, and cigarette smokers presented lower F0 and SFF in French and in Arabic when age, height, and weight were controlled. Further investigations using similar strict controls over individual variables and additional measures are encouraged to better understand the effect of water-pipe smoking on the voice. [ABSTRACT FROM AUTHOR]

Transgender MtF people (trans women) consult otorhinolaryngologists and vocologists with the aim of feminizing their voice and being consistently perceived as women. Treatment of these trans women always begins with a vocal assessment that is relatively unspecific as it was originally constructed for individuals with dysphonia. This study examines the subjective portion of the assessment and specifically the self-assessment questionnaire. There is no French-language questionnaire designed to identify the issues facing people who want voice feminization and quantify the impact of their voice disorder on their daily lives. We present a translation and adaption into French of the questionnaire developed by Dacakis et al [6]. This work follows the World Health Organization recommendations [12] regarding translation. Thirty-six Belgian and French trans women took part in this study. The results show excellent repeatability and reliability, while the construct validity measures show that the items correlate with six areas of concern for trans women identified by Davies and Johnson [7] in a previous study. The domains are the following: effect of voice on ease of social interaction, effect of voice on emotions, relationship between voice and gender identity, effort and concentration required to produce voice, physical aspects of voice production, and pitch. Concurrent validity could not be measured owing to lack of sufficiently detailed stories. The psychometric properties of the French version of the questionnaire are acceptable. The questionnaire can be used as is in daily clinical practice. [ABSTRACT FROM AUTHOR]

Summary Background The original Dysphonia Severity Index (ie, DSI) weighs and combines four voice markers in a single number to size dysphonia gradation in the clinic: percent jitter (from Multi-Dimensional Voice Program; KayPENTAX Corp., Montvale, NJ), softest intensity and highest fundamental frequency (both from Voice Range Profile; KayPENTAX Corp.), and maximum phonation time. To be more generally applied, however, implementation of DSI in the program Praat (Paul Boersma and David Weenink, Institute for Phonetic Sciences, University of Amsterdam, The Netherlands) would be advantageous for all voice clinicians. The presented project was therefore designed (1) to develop such a Praat application and (b) to test its concurrent validity. Methods The four voice markers for the original DSI, as well as ten additional voice markers in Praat , were administered on a total of 49 subjects in three different clinical voice centers. A crossover research design was implemented to counterbalance for possible exercise effects. First, stepwise multiple linear regression was applied to build a statistical model with the best combination of Praat predictors for the original DSI. Second, correlation statistics were applied to compare Praat 's DSI with the original DSI. Results Both DSI versions correlated strongly. A custom script was therefore written for automated DSI determination in Praat . Conclusion With this script, every voice clinician can easily determine DSI in the Praat program. [ABSTRACT FROM AUTHOR]

Summary Objective The purposes of this study were to assess the long-term efficacy of medialization laryngoplasty via injection of autologous fat in patients with unilateral laryngeal paralysis, and to discuss the results based on the volume of fat injected and the size of the initial glottic gap. Study design This was a retrospective study with clinical reevaluation of 18 patients treated for unilateral laryngeal paralysis at Liège University Hospital between April 1, 2011, and December 1, 2014. Method The voice examination included subjective scales (GRBAS and the Voice Handicap Index), a report on acoustic and aerodynamic voice parameters, and laryngostroboscopic assessment of glottic closure and mucosal wave (T0: preoperative; T1: immediate postoperative; T2: >1 year). Results The median follow-up duration was 19 months. We observed a significant improvement in maximum phonation time; mean flow rate; jitter; the Voice Handicap Index; the G, R, B, and A subscales of the GRBAS-I; and glottic closure at T1. These parameters remained stable over time, without any significant difference except for shimmer and glottic closure between T1 and T2. Regardless of the volume of fat injected and the magnitude of the initial glottic gap, we did not see any significant difference from one assessment time to another. Conclusion Medialization laryngoplasty by means of the injection of autologous fat is a safe, effective surgical technique for patients with unilateral laryngeal paralysis regardless of the size of the initial glottic gap. The results are maintained longer than 1 year, and no evidence of significant resorption of the fat was found in our study over a period of 12–58 months. The volume of fat injected does not seem to affect the functional results. [ABSTRACT FROM AUTHOR]

Summary: Objective: Vocal accuracy of a sung performance can be evaluated by two methods: acoustic analyses and subjective judgments. Acoustic analyses have been presented as a more reliable solution but both methods are still used for the evaluation of singing voice accuracy. This article presents a first time direct comparison of these methods. Methods: One hundred sixty-six untrained singers were asked to sing the popular song “Happy Birthday.” These recordings constituted the database analyzed. Acoustic analyses were performed to quantify the pitch interval deviation, number of contour errors, and number of tonality modulations for each recording. Additionally, 18 experts in singing voice or music rated the global pitch accuracy of these performances. Results: A high correlation occurred between acoustic measurements and subjective rating. The total model of acoustic analyses explained 81% of the variance of the judges'' scores. Their rating was influenced by both tonality modulations and pitch interval deviation. Conclusions: This study highlights the congruence between objective and subjective measurements of vocal accuracy within this first time comparison. Our results confirm the relevance of the pitch interval deviation criterion in vocal accuracy assessment. Furthermore, the number of tonality modulations is also a salient criterion in perceptive rating and should be taken into account in studies using acoustic analyses. [Copyright &y& Elsevier]

Summary: Objectives: This study evaluates the effect of a 2-hour reading task between 70 and 75dB(A) in 16 normophonic and 16 dysphonic female teachers with vocal nodules. Methods: Objective measurements (acoustic analysis, voice range measurements, and aerodynamic measurements) and subjective self-ratings were collected before and every 30 minutes during the reading to determine the voice evolution in both groups. Results: Fundamental frequency, lowest frequency, highest frequency (F-High), highest intensity, and intensity range increase through the reading, whereas shimmer decreases. Maximum phonation time decreases after 30 minutes. Estimated subglottal pressure (ESP) and sound pressure level increase during the first hour. Afterward, ESP decreases. Self-ratings worsen through time. When comparing the normophonic and the dysphonic teachers, self-ratings reveal more complaints in the dysphonic group. Few differences in objective measurements are found between both groups: normophonic teachers show lower ESP, higher F-High, and greater frequency range. Conclusions: Frequency modifications from acoustic analysis and voice range measurements suggest an increased laryngeal tension during vocal load, while subjects perceive a worsening of voice. Aerodynamic parameters depict first a deterioration of voice efficiency and then an adaptation to the prolonged reading. The comparison between both groups shows a discrepancy between objective measurements and self-ratings, suggesting that both approaches are necessary to have a complete view of vocal load effects. Surprisingly, both groups behave similarly through vocal load, without more or quicker deterioration of voice in the dysphonic group. [Copyright &y& Elsevier]

Summary: Objectives: The objectives of this study are to evaluate the impact on voice of both duration and intensity level of 2 hours of continuous oral reading. Voice modifications accompanying changes in intensity level during prolonged reading tasks are analyzed. Methods: Fifty normophonic women undergo two sessions of voice loading in which the required intensity level of voice varied between 60–65dB(A) for the first session and 70–75dB(A) for the second session. The effects of loading on objective data (average fundamental frequency [F0], jitter%, shimmer%, noise-to-harmonic ratio, maximum phonation time, lowest frequency [F-Low], highest frequency [F-High], frequency range [Range], lowest intensity [I-Low] level, and highest intensity level) and self-ratings (voice quality, phonation effort, vocal fatigue, and laryngeal discomfort) are assessed every 30 minutes during the loading tasks. Results: Results indicate that average F0, F-Low, I-Low, maximum phonation time, feeling of phonation effort, vocal fatigue, and laryngeal discomfort increase during prolonged reading, whereas shimmer% and self-rating of voice quality decrease. Average F0, F-High, and Range are the only parameters influenced by the required intensity of vocal load; they are significantly higher in the 70- to 75-dB session compared with the 60- to 65-dB session. Concerning the subjective self-ratings, similar results for the four ratings used suggest that only one would suffice in future studies. Conclusions: These results confirm the importance of both duration and intensity level as loading factors, even if intensity level affects fewer variables than duration. [Copyright &y& Elsevier]

Summary: The aim of our study was to validate a Pediatric Voice Symptom Questionnaire (PVSQ) presenting with a parallel form for children and their parents. The items of the questionnaire were elaborated from the results of structured interviews with dysphonic children (DP) and normophonic children (NP) and their mothers and were tested for feasibility in a pilot study involving 42 normophonic children aged 5–13 years. The items were then administered in a test-retest mode to 333 children and their parents (154 boys and 179 girls with a mean age of 9 years, standard deviation: 1.8); 45 consulting DP, 34 nonconsulting dysphonics (NcDP), 163 NP, and 91 others. Classical statistical analyses and an item response modeling approach were used to analyze the results. High internal consistency and good test-retest stability were found. Significant differences between total score of the NP, DP, and NcDP groups were observed both in the children and the parents and also between parental and child score for the NP and NcDP groups (P <0.001–P =0.014). Correlations between child and parental scores were found only in the DP groups (r =0.478; P <0.001). Based on our results, the PVSQ is a valid and reliable instrument for the autoevaluation of dysphonia in the child population. [Copyright &y& Elsevier]

In their environment, cells must cope with mechanical stresses constantly. Among these, nanoscale deformations of plasma membrane induced by substrate nanotopography are now largely accepted as a biophysical stimulus influencing cell behavior and function. However, the mechanotransduction cascades involved and their precise molecular effects on cellular physiology are still poorly understood. Here, using homemade fluorescent nanostructured cell culture surfaces, we explored the role of Bin/Amphiphysin/Rvs (BAR) domain proteins as mechanosensors of plasma membrane geometry. Our data reveal that distinct subsets of BAR proteins bind to plasma membrane deformations in a membrane curvature radius-dependent manner. Furthermore, we show that membrane curvature promotes the formation of dynamic actin structures mediated by the Rho GTPase CDC42, the F-BAR protein CIP4, and the presence of PI(4,5)P 2 . In addition, these actin-enriched nanodomains can serve as platforms to regulate receptor signaling as they appear to contain interferon-γ receptor (IFNγ-R) and to lead to the partial inhibition of IFNγ-induced JAK/STAT signaling.

The GDT1 family is broadly spread and highly conserved among living organisms. GDT1 members have functions in key processes like glycosylation in humans and yeasts and photosynthesis in plants. These functions are mediated by their ability to transport ions. While transport of Ca 2+ or Mn 2+ is well established for several GDT1 members, their transport mechanism is poorly understood. Here, we demonstrate that H + ions are transported in exchange for Ca 2+ and Mn 2+ cations by the Golgi-localized yeast Gdt1 protein. We performed direct transport measurement across a biological membrane by expressing Gdt1p in Lactococcus lactis bacterial cells and by recording either the extracellular pH or the intracellular pH during the application of Ca 2+ , Mn 2+ or H + gradients. Besides, in vivo cytosolic and Golgi pH measurements were performed in Saccharomyces cerevisiae with genetically encoded pH probes targeted to those subcellular compartments. These data point out that the flow of H + ions carried by Gdt1p could be reversed according to the physiological conditions. Together, our experiments unravel the influence of the relative concentration gradients for Gdt1p-mediated H + transport and pave the way to decipher the regulatory mechanisms driving the activity of GDT1 orthologs in various biological contexts., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Recent advances in the field demonstrate the high diversity and complexity of endocytic pathways. In the current study, we focus on the endocytosis of L1CAM. This glycoprotein plays a major role in the development of the nervous system, and is involved in cancer development and is associated with metastases and poor prognosis. Two L1CAM isoforms are subject to endocytosis: isoform 1, described as a clathrin-mediated cargo; isoform 2, whose endocytosis has never been studied. Deciphering the molecular machinery of isoform 2 internalisation should contribute to a better understanding of its pathophysiological role. First, we demonstrated in our cellular context that both isoforms of L1CAM are mainly a clathrin-independent cargo, which was not expected for isoform 1. Second, the mechanism of L1CAM endocytosis is specifically mediated by the N-BAR domain protein endophilin-A3. Third, we discovered PSTPIP1, an F-BAR domain protein, as a novel actor in this endocytic process. Finally, we identified galectins as endocytic partners and negative regulators of L1CAM endocytosis. In summary, the interplay of the BAR proteins endophilin-A3 and PSTPIP1, and galectins fine tune the clathrin-independent endocytosis of L1CAM., (© 2023 The Authors. Traffic published by John Wiley & Sons Ltd.)

Endocytic mechanisms actively regulate plasma membrane composition and sustain fundamental cellular functions. Recently, we identified a clathrin-independent endocytic (CIE) modality mediated by the BAR domain protein endophilin-A3 (endoA3, encoded by SH3GL3), which controls the cell surface homeostasis of the tumor marker CD166 (also known as ALCAM). Deciphering the molecular machinery of endoA3-dependent CIE should therefore contribute to a better understanding of its pathophysiological role, which remains so far unknown. Here, we investigate the role of actin, Rho GTPases and microtubules, which are major players in CIE processes, in this mechanism. We show that the actin cytoskeleton is dynamically associated with endoA3- and CD166-positive endocytic carriers, and that its perturbation strongly inhibits the process of CD166 uptake. We also reveal that the Rho GTPase Rac1, but not Cdc42, is a master regulator of this endocytic route. Finally, we provide evidence that microtubules and kinesin molecular motors are required to potentiate endoA3-dependent endocytosis. Of note, our study also highlights potential compensation phenomena between endoA3-dependent CIE and macropinocytosis. Altogether, our data deepen our understanding of this CIE modality and further differentiate it from other unconventional endocytic mechanisms. This article has an associated First Person interview with the first author of the paper., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Competence for DNA transformation is a major strategy for bacterial adaptation and survival. Yet, this successful tactic is energy-consuming, shifts dramatically the metabolism, and transitory impairs the regular cell-cycle. In streptococci, complex regulatory pathways control competence deactivation to narrow its development to a sharp window of time, a process known as competence shut-off. Although characterized in streptococci whose competence is activated by the ComCDE signaling pathway, it remains unclear for those controlled by the ComRS system. In this work, we investigate competence shut-off in the major human gut commensal Streptococcus salivarius. Using a deterministic mathematical model of the ComRS system, we predicted a negative player under the control of the central regulator ComX as involved in ComS/XIP pheromone degradation through a negative feedback loop. The individual inactivation of peptidase genes belonging to the ComX regulon allowed the identification of PepF as an essential oligoendopeptidase in S. salivarius. By combining conditional mutants, transcriptional analyses, and biochemical characterization of pheromone degradation, we validated the reciprocal role of PepF and XIP in ComRS shut-off. Notably, engineering cleavage site residues generated ultra-resistant peptides producing high and long-lasting competence activation. Altogether, this study reveals a proteolytic shut-off mechanism of competence in the salivarius group and suggests that this mechanism could be shared by other ComRS-containing streptococci., Competing Interests: The authors of this manuscript have the following competing interests: PH is a Research Director at FNRS.

Human peroxiredoxin-5 (PRDX5) is a unique redox-sensitive protein that plays a dual role in brain ischemia-reperfusion injury. While intracellular PRDX5 has been reported to act as a neuroprotective antioxidative enzyme by scavenging peroxides, once released extracellularly from necrotic brain cells, the protein aggravates neural cell death by inducing expression of proinflammatory cytokines in macrophages through activation of Toll-like receptor (TLR) 2 (TLR2) and 4 (TLR4). Although recent evidence showed that PRDX5 was able to interact directly with TLR4, little is known regarding the role of the cysteine redox state of PRDX5 on its DAMP function. To gain insights into the role of PRDX5 redox-active cysteine residues in the TLR4-dependent proinflammatory activity of the protein, we used a recombinant human PRDX5 in the disulfide (oxidized) form and a mutant version lacking the peroxidatic cysteine, as well as chemically reduced and hyperoxidized PRDX5 proteins. We first analyzed the oxidation state and oligomerization profile by Western blot, mass spectrometry, and SEC-MALS. Using ELISA, we demonstrate that the disulfide bridge between the enzymatic cysteines is required to allow improved TLR4-dependent IL-8 secretion. Moreover, single-molecule force spectroscopy experiments revealed that TLR4 alone is not sufficient to discriminate the different PRDX5 redox forms. Finally, flow cytometry binding assays show that disulfide PRDX5 has a higher propensity to bind to the surface of living TLR4-expressing cells than the mutant protein. Taken together, these results demonstrate the importance of the redox state of PRDX5 cysteine residues on TLR4-induced inflammation.

The coupling between mitochondrial respiration and photosynthesis plays an important role in the energetic physiology of green plants and some secondary-red photosynthetic eukaryotes (diatoms), allowing an efficient CO 2 assimilation and optimal growth. Using the flagellate Euglena gracilis, we first tested if photosynthesis-respiration coupling occurs in this species harbouring secondary green plastids (i.e. originated from an endosymbiosis between a green alga and a phagotrophic euglenozoan). Second, we tested how the trophic state (mixotrophy and photoautotrophy) of the cell alters the mechanisms involved in the photosynthesis-respiration coupling. Energetic coupling between photosynthesis and respiration was determined by testing the effect of respiratory inhibitors on photosynthesis, and measuring the simultaneous variation of photosynthesis and respiration rates as a function of temperature (i.e. thermal response curves). The mechanism involved in the photosynthesis-respiration coupling was assessed by combining proteomics, biophysical and cytological analyses. Our work shows that there is photosynthesis-respiration coupling and membrane contacts between mitochondria and chloroplasts in E. gracilis. However, whereas in mixotrophy adjustment of the chloroplast ATP/NADPH ratio drives the interaction, in photoautotrophy the coupling is conditioned by CO 2 limitation and photorespiration. This indicates that maintenance of photosynthesis-respiration coupling, through plastic metabolic responses, is key to E. gracilis functioning under changing environmental conditions., (© 2021 The Authors. New Phytologist © 2021 New Phytologist Foundation.)

The maintenance of K+ and Ca2+ homeostasis is crucial for many cellular functions. Potassium is accumulated in cells at high concentrations, while the cytosolic level of calcium, to ensure its signalling function, is kept at low levels and transiently increases in response to stresses. We examined Ca2+ homeostasis and Ca2+ signalling in Saccharomyces cerevisiae strains lacking plasma-membrane K+ influx (Trk1 and Trk2) or efflux (Tok1, Nha1 and Ena1-5) systems. The lack of K+ exporters slightly increased the cytosolic Ca2+, but did not alter the Ca2+ tolerance or Ca2+-stress response. In contrast, the K+-importers Trk1 and Trk2 play important and distinct roles in the maintenance of Ca2+ homeostasis. The presence of Trk1 was vital mainly for the growth of cells in the presence of high extracellular Ca2+, whilst the lack of Trk2 doubled steady-state intracellular Ca2+ levels. The absence of both K+ importers highly increased the Ca2+ response to osmotic or CaCl2 stresses and altered the balance between Ca2+ flux from external media and intracellular compartments. In addition, we found Trk2 to be important for the tolerance to high KCl and hygromycin B in cells growing on minimal media. All the data describe new interconnections between potassium and calcium homeostasis in S. cerevisiae., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS.)

Plasma membrane (PM) intrinsic proteins (PIPs) are aquaporins facilitating the diffusion of water and small solutes. The functional importance of the PM organisation of PIPs in the interaction with other cellular structures is not completely understood. We performed a pull-down assay using maize (Zea mays) suspension cells expressing YFP-ZmPIP2;5 and validated the protein interactions by yeast split-ubiquitin and bimolecular fluorescence complementation assays. We expressed interacting proteins tagged with fluorescent proteins in Nicotiana benthamiana leaves and performed water transport assays in oocytes. Finally, a phylogenetic analysis was conducted. The PM-located ZmPIP2;5 physically interacts with the endoplasmic reticulum (ER) resident ZmVAP27-1. This interaction requires the ZmVAP27-1 cytoplasmic major sperm domain. ZmPIP2;5 and ZmVAP27-1 localise in close vicinity in ER-PM contact sites (EPCSs) and endocytic structures upon exposure to salt stress conditions. This interaction enhances PM water permeability in oocytes. Similarly, the Arabidopsis ZmVAP27-1 paralogue, AtVAP27-1, interacts with the AtPIP2;7 aquaporin. Together, these data indicate that the PIP2-VAP27 interaction in EPCSs is evolutionarily conserved, and suggest that VAP27 might stabilise the aquaporins and guide their endocytosis in response to salt stress., (©2020 The Authors. New Phytologist ©2020 New Phytologist Trust.)

The yeast Saccharomyces cerevisiae has been perceived over decades as a highly valuable model organism for the investigation of ion homeostasis. Indeed, many of the genes and biological systems that function in yeast ion homeostasis are conserved throughout unicellular eukaryotes to humans. In this context, measurement of the yeast cellular ionic content provides information regarding yeast response to gene deletion or exposure to chemicals for instance. We propose here a protocol that we tested for the analysis of 12 elements (Ba 2+ , Ca 2+ , Cd 2+ , Co 2+ , Cu 2+ , Fe 2+ , K + , Mg 2+ , Mn 2+ , Na + , Ni 2+ , Zn 2+ ) in yeast using Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES). This technique enables determination of the cellular content of numerous ions from one biological sample., Competing Interests: Competing interestsThe authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.)

The Uncharacterized Protein Family 0016 (UPF0016) gathers poorly studied membrane proteins well conserved through evolution that possess one or two copies of the consensus motif Glu-x-Gly-Asp-(Arg/Lys)-(Ser/Thr). Members are found in many eukaryotes, bacteria and archaea. The interest for this protein family arose in 2012 when its human member TMEM165 was linked to the occurrence of Congenital Disorders of Glycosylation (CDGs) when harbouring specific mutations. Study of the UPF0016 family is undergone through the characterization of the bacterium Vibrio cholerae (MneA), cyanobacterium Synechocystis (SynPAM71), yeast Saccharomyces cerevisiae (Gdt1p), plant Arabidopsis thaliana (PAM71 and CMT1), and human (TMEM165) members. These proteins have all been identified as transporters of cations, more precisely of Mn 2+ , with an extra reported function in Ca 2+ and/or H + transport for some of them. Apart from glycosylation in humans, the UPF0016 members are required for lactation in humans, photosynthesis in plants and cyanobacteria, Ca 2+ signaling in yeast, and Mn 2+ homeostasis in the five aforementioned species. The requirement of the UPF0016 members for key physiological processes most likely derives from their transport activity at the Golgi membrane in human and yeast, the chloroplasts membranes in plants, the thylakoid and plasma membranes in cyanobacteria, and the cell membrane in bacteria. In the light of these studies on various UPF0016 members, this family is not considered as uncharacterized anymore and has been renamed the Gdt1 family according to the name of its S. cerevisiae member. This review aims at assembling and confronting the current knowledge in order to identify shared and distinct features in terms of transported molecules, mode of action, structure, etc., as well as to better understand their corresponding physiological roles., Competing Interests: Conflict of interest: The authors declare no conflicts of interest., (Copyright: © 2020 Thines et al.)

Cases of congenital disorders of glycosylation (CDG) have been associated with specific mutations within the gene encoding the human Golgi TMEM165 (transmembrane protein 165), belonging to UPF0016 (uncharacterized protein family 0016), a family of secondary ion transporters. To date, members of this family have been reported to be involved in calcium, manganese, and pH homeostases. Although it has been suggested that TMEM165 has cation transport activity, direct evidence for its Ca 2+ - and Mn 2+ -transporting activities is still lacking. Here, we functionally characterized human TMEM165 by heterologously expressing it in budding yeast ( Saccharomyces cerevisiae ) and in the bacterium Lactococcus lactis Protein production in these two microbial hosts was enhanced by codon optimization and truncation of the putatively autoregulatory N terminus of TMEM165. We show that TMEM165 expression in a yeast strain devoid of Golgi Ca 2+ and Mn 2+ transporters abrogates Ca 2+ - and Mn 2+ -induced growth defects, excessive Mn 2+ accumulation in the cell, and glycosylation defects. Using bacterial cells loaded with the fluorescent Fura-2 probe, we further obtained direct biochemical evidence that TMEM165 mediates Ca 2+ and Mn 2+ influxes. We also used the yeast and bacterial systems to evaluate the impact of four disease-causing missense mutations identified in individuals with TMEM165-associated CDG. We found that a mutation leading to a E108G substitution within the conserved UPF0016 family motif significantly reduces TMEM165 activity. These results indicate that TMEM165 can transport Ca 2+ and Mn 2+ , which are both required for proper protein glycosylation in cells. Our work also provides tools to better understand the pathogenicity of CDG-associated TMEM165 mutations., (© 2020 Stribny et al.)

Superoxide dismutases (SODs) act as a primary defence against reactive oxygen species (ROS) by converting superoxide anion radicals (O 2 - ) into molecular oxygen (O 2 ) and hydrogen peroxide (H 2 O 2 ). Members of this enzyme family include CuZnSODs, MnSODs, FeSODs, and NiSODs, depending on the nature of the cofactor that is required for proper activity. Most eukaryotes, including yeast, possess CuZnSOD and MnSOD. This protocol aims at assessing the activity of the yeast Saccharomyces cerevisiae MnSOD Sod2p from cellular extracts using nitroblue tetrazolium staining. This method can be used to estimate the cellular bioavailability of Mn 2+ as well as to evaluate the redox state of the cell., Competing Interests: Competing interestsThe authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.)

6-NADH and 6-NADPH are strong inhibitors of several dehydrogenases that may form spontaneously from NAD(P)H. They are known to be oxidized to NAD(P)+ by mammalian renalase, an FAD-linked enzyme mainly present in heart and kidney, and by related bacterial enzymes. We partially purified an enzyme oxidizing 6-NADPH from rat liver, and, surprisingly, identified it as pyridoxamine-phosphate oxidase (PNPO). This was confirmed by the finding that recombinant mouse PNPO oxidized 6-NADH and 6-NADPH with catalytic efficiencies comparable to those observed with pyridoxine- and pyridoxamine-5'-phosphate. PNPOs from Escherichia coli, Saccharomyces cerevisiae and Arabidopsis thaliana also displayed 6-NAD(P)H oxidase activity, indicating that this 'side-activity' is conserved. Remarkably, 'pyridoxamine-phosphate oxidase-related proteins' (PNPO-RP) from Nostoc punctiforme, A. thaliana and the yeast S. cerevisiae (Ygr017w) were not detectably active on pyridox(am)ine-5'-P, but oxidized 6-NADH, 6-NADPH and 2-NADH suggesting that this may be their main catalytic function. Their specificity profiles were therefore similar to that of renalase. Inactivation of renalase and of PNPO in mammalian cells and of Ygr017w in yeasts led to the accumulation of a reduced form of 6-NADH, tentatively identified as 4,5,6-NADH3, which can also be produced in vitro by reduction of 6-NADH by glyceraldehyde-3-phosphate dehydrogenase or glucose-6-phosphate dehydrogenase. As 4,5,6-NADH3 is not a substrate for renalase, PNPO or PNPO-RP, its accumulation presumably reflects the block in the oxidation of 6-NADH. These findings indicate that two different classes of enzymes using either FAD (renalase) or FMN (PNPOs and PNPO-RPs) as a cofactor play an as yet unsuspected role in removing damaged forms of NAD(P)., (© 2019 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

Endocytosis of membrane proteins in yeast requires α-arrestin-mediated ubiquitylation by the ubiquitin ligase Rsp5. Yet, the diversity of α-arrestin targets studied is restricted to a small subset of plasma membrane (PM) proteins. Here, we performed quantitative proteomics to identify new targets of 12 α-arrestins and gained insight into the diversity of pathways affected by α-arrestins, including the cell wall integrity pathway and PM-endoplasmic reticulum contact sites. We found that Art2 is the main regulator of substrate- and stress-induced ubiquitylation and endocytosis of the thiamine (vitamin B1) transporters: Thi7, nicotinamide riboside transporter 1 (Nrt1), and Thi72. Genetic screening allowed for the isolation of transport-defective Thi7 mutants, which impaired thiamine-induced endocytosis. Coexpression of inactive mutants with wild-type Thi7 revealed that both transporter conformation and transport activity are important to induce endocytosis. Finally, we provide evidence that Art2 mediated Thi7 endocytosis is regulated by the target of rapamycin complex 1 (TORC1) and requires the Sit4 phosphatase but is not inhibited by the Npr1 kinase., Competing Interests: The authors have declared that no competing interests exist.

The biological importance of manganese lies in its function as a key cofactor for numerous metalloenzymes and as non-enzymatic antioxidant. Due to these two essential roles, it appears evident that disturbed manganese homeostasis may trigger the development of pathologies in humans. In this context, yeast has been extensively used over the last decades to gain insight into how cells regulate intra-organellar manganese concentrations and how human pathologies may be related to disturbed cellular manganese homeostasis. This review first summarizes how manganese homeostasis is controlled in yeast cells and how this knowledge can be extrapolated to human cells. Several manganese-related pathologies whose molecular mechanisms have been studied in yeast are then presented in the light of the function of this cation as a non-enzymatic antioxidant or as a key cofactor of metalloenzymes. In this line, we first describe the Transmembrane protein 165-Congenital Disorder of Glycosylation (TMEM165-CDG) and Friedreich ataxia pathologies. Then, due to the established connection between manganese cations and neurodegeneration, the Kufor-Rakeb syndrome and prion-related diseases are finally presented., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.