A localizing nanocarrier formulation enables multi-target immune responses to multivalent replicating RNA with limited systemic inflammation.

RNA vaccines possess significant clinical promise in counteracting human diseases caused by infectious or cancerous threats. Self-amplifying replicon RNA (repRNA) has been thought to offer the potential for enhanced potency and dose sparing. However, repRNA is a potent trigger of innate immune responses in vivo, which can cause reduced transgene expression and dose-limiting reactogenicity, as highlighted by recent clinical trials. Here, we report that multivalent repRNA vaccination, necessitating higher doses of total RNA, could be safely achieved in mice by delivering multiple repRNAs with a localizing cationic nanocarrier formulation (LION). Intramuscular delivery of multivalent repRNA by LION resulted in localized biodistribution accompanied by significantly upregulated local innate immune responses and the induction of antigen-specific adaptive immune responses in the absence of systemic inflammatory responses. In contrast, repRNA delivered by lipid nanoparticles (LNPs) showed generalized biodistribution, a systemic inflammatory state, an increased body weight loss, and failed to induce neutralizing antibody responses in a multivalent composition. These findings suggest that in vivo delivery of repRNA by LION is a platform technology for safe and effective multivalent vaccination through mechanisms distinct from LNP-formulated repRNA vaccines.

Acoustic correlates of sexual orientation and gender-role self-concept in women's speech.

Compared to studies of male speakers, relatively few studies have investigated acoustic correlates of sexual orientation in women. The present investigation focuses on shedding more light on intra-group variability in lesbians and straight women by using a fine-grained analysis of sexual orientation and collecting data on psychological characteristics (e.g., gender-role self-concept). For a large-scale women's sample (overall n = 108), recordings of spontaneous and read speech were analyzed for median fundamental frequency and acoustic vowel space features. Two studies showed no acoustic differences between lesbians and straight women, but there was evidence of acoustic differences within sexual orientation groups. Intra-group variability in median f0 was found to depend on the exclusivity of sexual orientation; F1 and F2 in /iː/ (study 1) and median f0 (study 2) were acoustic correlates of gender-role self-concept, at least for lesbians. Other psychological characteristics (e.g., sexual orientation of female friends) were also reflected in lesbians' speech. Findings suggest that acoustic features indexicalizing sexual orientation can only be successfully interpreted in combination with a fine-grained analysis of psychological characteristics.

The Effect of Fundamental Frequency on Gender Perception in Prepubertal Children: Insights from the LoKiS Database.

This study examines the impact of fundamental frequency on gender perception in prepubertal children in the LoKiS database - a longitudinal project collecting and analyzing recordings of approximately 60 German primary school children aged 6 to 10years. Spontaneous and content-controlled audio recordings were collected in two German primary schools. Three distinct listening experiments with over 100 listeners were conducted. In the first experiment, listeners judged the gender of the voices on a seven-point scale. The second experiment explored the relationships between perceptual attribute ratings and corresponding acoustic parameters associated with fundamental frequency. The third experiment utilized voice morphing techniques to investigate the influence of fundamental frequency on gender perception while controlling for other acoustic parameters. About one-third of the children receive unambiguous gender attributions. The perceived gender difference between children assigned female at birth (AFAB) and assigned male at birth (AMAB) increases from first to third grade. The feminine-sounding children were perceived as significantly higher-pitched and more melodious. A strong correlation between perceived pitch and measured fundamental frequency was found. While the acoustic analysis revealed only a few significant differences between AFAB and AMAB children in general, the feminine-sounding children exhibited markedly higher values than the masculine-sounding ones. Stronger differences of fundamental frequency and semitone range occur as AFAB and AMAB children get older. Linear mixed models confirm a significant influence of fundamental frequency and semitone range on gender perception. Other interacting factors include the speech material used, as well as the gender of the listener. The influence of fundamental frequency was even more pronounced when controlling for other acoustic parameters.

Effects of testosterone on speech production and perception: Linking hormone levels in males to vocal cues and female voice attractiveness ratings.

This study sets out to investigate the potential effect of males' testosterone level on speech production and speech perception. Regarding speech production, we investigate intra- and inter-individual variation in mean fundamental frequency (fo) and formant frequencies and highlight the potential interacting effect of another hormone, i.e. cortisol. In addition, we investigate the influence of different speech materials on the relationship between testosterone and speech production. Regarding speech perception, we investigate the potential effect of individual differences in males' testosterone level on ratings of attractiveness of female voices. In the production study, data is gathered from 30 healthy adult males ranging from 19 to 27 years (mean age: 22.4, SD: 2.2) who recorded their voices and provided saliva samples at 9 am, 12 noon and 3 pm on a single day. Speech material consists of sustained vowels, counting, read speech and a free description of pictures. Biological measures comprise speakers' height, grip strength, and hormone levels (testosterone and cortisol). In the perception study, participants were asked to rate the attractiveness of female voice stimuli (sentence stimulus, same-speaker pairs) that were manipulated in three steps regarding mean fo and formant frequencies. Regarding speech production, our results show that testosterone affected mean fo (but not formants) both within and between speakers. This relationship was weakened in speakers with high cortisol levels and depended on the speech material. Regarding speech perception, we found female stimuli with higher mean fo and formants to be rated as sounding more attractive than stimuli with lower mean fo and formants. Moreover, listeners with low testosterone showed an increased sensitivity to vocal cues of female attractiveness. While our results of the production study support earlier findings of a relationship between testosterone and mean fo in males (which is mediated by cortisol), they also highlight the relevance of the speech material: The effect of testosterone was strongest in sustained vowels, potentially due to a strengthened effect of hormones on physiologically strongly influenced tasks such as sustained vowels in contrast to more free speech tasks such as a picture description. The perception study is the first to show an effect of males' testosterone level on female attractiveness ratings using voice stimuli.

Accelerated prime-and-trap vaccine regimen in mice using repRNA-based CSP malaria vaccine.

Malaria, caused by Plasmodium parasites, remains one of the most devastating infectious diseases worldwide, despite control efforts to lower morbidity and mortality. Both advanced candidate vaccines, RTS,S and R21, are subunit (SU) vaccines that target a single Plasmodium falciparum (Pf) pre-erythrocytic (PE) sporozoite (spz) surface protein known as circumsporozoite (CS). These vaccines induce humoral immunity but fail to elicit CD8 + T-cell responses sufficient for long-term protection. In contrast, whole-organism (WO) vaccines, such as Radiation Attenuated Sporozoites (RAS), achieved sterile protection but require a series of intravenous doses administered in multiple clinic visits. Moreover, these WO vaccines must be produced in mosquitos, a burdensome process that severely limits their availability. To reduce reliance on WO while maintaining protection via both antibodies and Trm responses, we have developed an accelerated vaccination regimen that combines two distinct agents in a prime-and-trap strategy. The priming dose is a single dose of self-replicating RNA encoding the full-length P. yoelii CS protein, delivered via an advanced cationic nanocarrier (LIONTM). The trapping dose consists of one dose of WO RAS. Our vaccine induces a strong immune response when administered in an accelerated regimen, i.e., either 5-day or same-day immunization. Additionally, mice after same-day immunization showed a 2-day delay of blood patency with 90% sterile protection against a 3-week spz challenge. The same-day regimen also induced durable 70% sterile protection against a 2-month spz challenge. Our approach presents a clear path to late-stage preclinical and clinical testing of dose-sparing, same-day regimens that can confer sterilizing protection against malaria.

A self-amplifying RNA vaccine prevents enterovirus D68 infection and disease in preclinical models.

The recent emergence and rapid response to severe acute respiratory syndrome coronavirus 2 was enabled by prototype pathogen and vaccine platform approaches, driven by the preemptive application of RNA vaccine technology to the related Middle East respiratory syndrome coronavirus. Recently, the National Institutes of Allergy and Infectious Diseases identified nine virus families of concern, eight enveloped virus families and one nonenveloped virus family, for which vaccine generation is a priority. Although RNA vaccines have been described for a variety of enveloped viruses, a roadmap for their use against nonenveloped viruses is lacking. Enterovirus D68 was recently designated a prototype pathogen within the family Picornaviridae of nonenveloped viruses because of its rapid evolution and respiratory route of transmission, coupled with a lack of diverse anti-enterovirus vaccine approaches in development. Here, we describe a proof-of-concept approach using a clinical stage RNA vaccine platform that induced robust enterovirus D68-neutralizing antibody responses in mice and nonhuman primates and prevented upper and lower respiratory tract infections and neurological disease in mice. In addition, we used our platform to rapidly characterize the antigenic diversity within the six genotypes of enterovirus D68, providing the necessary data to inform multivalent vaccine compositions that can elicit optimal breadth of neutralizing responses. These results demonstrate that RNA vaccines can be used as tools in our pandemic-preparedness toolbox for nonenveloped viruses.

Investigating the relationship between average speaker fundamental frequency and acoustic vowel space size.

The purpose of this study is to investigate the potential relationship between speaking fundamental frequency and acoustic vowel space size, thus testing a possible perceptual source of sex-specific differences in acoustic vowel space size based on the greater inter-harmonic spacing and a poorer definition of the spectral envelope of higher pitched voices. Average fundamental frequencies and acoustic vowel spaces of 56 female German speakers are analyzed. Several parameters are used to quantify the size and shape of the vowel space defined by /iː ε aː [symbol: see text] uː/ such as the area of the polygon spanned by the five vowels, the absolute difference in F1 or F2 between /iː/ and /uː/ or /aː/, and the Euclidian distance between /iː/ and /aː/. In addition, the potential impact of nasality on the vowel space size is examined. Results reveal no significant correlation between fundamental frequency and vowel space size suggesting other factors must be responsible for the larger female acoustic vowel space.

The Acoustic and Perceptual Correlates of Gender in Children's Voices.

PURPOSE: This study investigates the perceptual and acoustic correlates of gender in prepubertal voices. The study is part of a longitudinal project analyzing recordings of circa 60 German primary school children from the first to fourth grades (6- to 10-year-olds). METHOD: Spontaneous and content-controlled audio recordings were made of 62 first-grade children (29 girls, 33 boys; age: 6- to 7-year-olds) from two German primary schools. Information on gender conformity was also recorded. A total of 167 listeners judged the gender of the voices on a 7-point scale. The results of the listening experiments and gender conformity ratings were related to a range of typical acoustic parameters. RESULTS: Measures of self-reported gender conformity differ significantly between the boys and the girls. Sixteen of the 62 children show unambiguous gender attributions in the listening experiment. A hierarchical cluster analysis including gender perception, gender conformity, and acoustic parameters shows four different types of speakers. Two multiple regression models revealed a significant main effect of fundamental frequency on the gender perception ratings of the listening experiment across and within gender. Significant correlations were found between the center of gravity and skewness of the sibilants and gender conformity, especially for the male speakers. CONCLUSIONS: Fundamental frequency plays an important role in influencing perceptual judgments, whereas sibilant spectra are correlated with gender conformity. In further listening experiments, we will examine in more detail the role of individual acoustic parameters and analyze how the vocal expression of gender and gender conformity in individual children develops before reaching puberty.

Accelerated prime-and-trap vaccine regimen in mice using repRNA-based CSP malaria vaccine.

Malaria, caused by Plasmodium parasites, remains one of the most devastating infectious diseases worldwide, despite control efforts that have lowered morbidity and mortality. The only P. falciparum vaccine candidates to show field efficacy are those targeting the asymptomatic pre-erythrocytic (PE) stages of infection. The subunit (SU) RTS,S/AS01 vaccine, the only licensed malaria vaccine to date, is only modestly effective against clinical malaria. Both RTS,S/AS01 and the SU R21 vaccine candidate target the PE sporozoite (spz) circumsporozoite (CS) protein. These candidates elicit high-titer antibodies that provide short-term protection from disease, but do not induce the liver-resident memory CD8+ T cells (Trm) that confer strong PE immunity and long-term protection. In contrast, whole-organism (WO) vaccines, employing for example radiation-attenuated spz (RAS), elicit both high antibody titers and Trm, and have achieved high levels of sterilizing protection. However, they require multiple intravenous (IV) doses, which must be administered at intervals of several weeks, complicating mass administration in the field. Moreover, the quantities of spz required present production difficulties. To reduce reliance on WO while maintaining protection via both antibodies and Trm responses, we have developed an accelerated vaccination regimen that combines two distinct agents in a prime-and-trap strategy. While the priming dose is a self-replicating RNA encoding P. yoelii CS protein, delivered via an advanced cationic nanocarrier (LION™), the trapping dose consists of WO RAS. This accelerated regime confers sterile protection in the P. yoelii mouse model of malaria. Our approach presents a clear path to late-stage preclinical and clinical testing of dose-sparing, same-day regimens that can confer sterilizing protection against malaria.

Accelerated prime-and-trap vaccine regimen in mice using repRNA-based CSP malaria vaccine.

Malaria, caused by Plasmodium parasites, remains one of the most devastating infectious diseases worldwide, despite control efforts that have lowered morbidity and mortality. The only P. falciparum vaccine candidates to show field efficacy are those targeting the asymptomatic pre-erythrocytic (PE) stages of infection. The subunit (SU) RTS,S/AS01 vaccine, the only licensed malaria vaccine to date, is only modestly effective against clinical malaria. Both RTS,S/AS01 and the SU R21 vaccine candidate target the PE sporozoite (spz) circumsporozoite (CS) protein. These candidates elicit high-titer antibodies that provide short-term protection from disease, but do not induce the liver-resident memory CD8+ T cells (Trm) that confer strong PE immunity and long-term protection. In contrast, whole-organism (WO) vaccines, employing for example radiation-attenuated spz (RAS), elicit both high antibody titers and Trm, and have achieved high levels of sterilizing protection. However, they require multiple intravenous (IV) doses, which must be administered at intervals of several weeks, complicating mass administration in the field. Moreover, the quantities of spz required present production difficulties. To reduce reliance on WO while maintaining protection via both antibodies and Trm responses, we have developed an accelerated vaccination regimen that combines two distinct agents in a prime-and-trap strategy. While the priming dose is a self-replicating RNA encoding P. yoelii CS protein, delivered via an advanced cationic nanocarrier (LION™), the trapping dose consists of WO RAS. This accelerated regime confers sterile protection in the P. yoelii mouse model of malaria. Our approach presents a clear path to late-stage preclinical and clinical testing of dose-sparing, same-day regimens that can confer sterilizing protection against malaria.

Auditory adaptation in voice perception.

Perceptual aftereffects following adaptation to simple stimulus attributes (e.g., motion, color) have been studied for hundreds of years. A striking recent discovery was that adaptation also elicits contrastive aftereffects in visual perception of complex stimuli and faces [1-6]. Here, we show for the first time that adaptation to nonlinguistic information in voices elicits systematic auditory aftereffects. Prior adaptation to male voices causes a voice to be perceived as more female (and vice versa), and these auditory aftereffects were measurable even minutes after adaptation. By contrast, crossmodal adaptation effects were absent, both when male or female first names and when silently articulating male or female faces were used as adaptors. When sinusoidal tones (with frequencies matched to male and female voice fundamental frequencies) were used as adaptors, no aftereffects on voice perception were observed. This excludes explanations for the voice aftereffect in terms of both pitch adaptation and postperceptual adaptation to gender concepts and suggests that contrastive voice-coding mechanisms may routinely influence voice perception. The role of adaptation in calibrating properties of high-level voice representations indicates that adaptation is not confined to vision but is a ubiquitous mechanism in the perception of nonlinguistic social information from both faces and voices.

Effect of storage conditions on SARS-CoV-2 RNA quantification in wastewater solids.

SARS-CoV-2 RNA in wastewater settled solids is associated with COVID-19 incidence in sewersheds and therefore, there is a strong interest in using these measurements to augment traditional disease surveillance methods. A wastewater surveillance program should provide rapid turn around for sample measurements (ideally within 24 hours), but storage of samples is necessary for a variety of reasons including biobanking. Here we investigate how storage of wastewater solids at 4 °C, -20 °C, and -80 °C affects measured concentrations of SARS-CoV-2 RNA. We find that short term (7 or 8 d) storage of raw solids at 4 °C has little effect on measured concentrations of SARS-CoV-2 RNA, whereas longer term storage at 4 °C (35-122 d) or freezing reduces measurements by 60%, on average. We show that normalizing SARS-CoV-2 RNA concentrations by concentrations of pepper mild mottle virus (PMMoV) RNA, an endogenous wastewater virus, can correct for changes during storage as storage can have a similar effect on PMMoV RNA as on SARS-CoV-2 RNA. The reductions in SARS-CoV-2 RNA in solids during freeze thaws is less than those reported for the same target in liquid influent by several authors.

High-Frequency, High-Throughput Quantification of SARS-CoV-2 RNA in Wastewater Settled Solids at Eight Publicly Owned Treatment Works in Northern California Shows Strong Association with COVID-19 Incidence.

A number of recent retrospective studies have demonstrated that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA concentrations in wastewater are associated with coronavirus disease 2019 (COVID-19) cases in the corresponding sewersheds. Implementing high-resolution, prospective efforts across multiple plants depends on sensitive measurements that are representative of COVID-19 cases, scalable for high-throughput analysis, and comparable across laboratories. We conducted a prospective study across eight publicly owned treatment works (POTWs). A focus on SARS-CoV-2 RNA in solids enabled us to scale up our measurements with a commercial lab partner. Samples were collected daily, and results were posted to a website within 24 h. SARS-CoV-2 RNA in daily samples correlated with the incidence of COVID-19 cases in the sewersheds; a 1 log10 increase in SARS-CoV-2 RNA in settled solids corresponds to a 0.58 log10 (4×) increase in sewershed incidence rate. SARS-CoV-2 RNA signals measured with the commercial laboratory partner were comparable across plants and comparable to measurements conducted in a university laboratory when normalized by pepper mild mottle virus (PMMoV) RNA. Results suggest that SARS-CoV-2 RNA should be detectable in settled solids for COVID-19 incidence rates of >1/100,000 (range, 0.8 to 2.3 cases per 100,000). These sensitive, representative, scalable, and comparable methods will be valuable for future efforts to scale up wastewater-based epidemiology. IMPORTANCE Access to reliable, rapid monitoring data is critical to guide response to an infectious disease outbreak. For pathogens that are shed in feces or urine, monitoring wastewater can provide a cost-effective snapshot of transmission in an entire community via a single sample. In order for a method to be useful for ongoing COVID-19 monitoring, it should be sensitive for detection of low concentrations of SARS-CoV-2, representative of incidence rates in the community, scalable to generate data quickly, and comparable across laboratories. This paper presents a method utilizing wastewater solids to meet these goals, producing measurements of SARS-CoV-2 RNA strongly associated with COVID-19 cases in the sewershed of a publicly owned treatment work. Results, provided within 24 h, can be used to detect incidence rates as low as approximately 1/100,000 cases and can be normalized for comparison across locations generating data using different methods.

Physicochemical structure of a polyacrylic acid stabilized nanoparticle alum (nanoalum) adjuvant governs TH1 differentiation of CD4+ T cells.

The growing shift to subunit antigen vaccines underscores the need for adjuvants that can enhance the magnitude and quality of immune response. Aluminum salts or alums are the first adjuvants with a long history of clinical use. Alum predominantly induces T helper 2 (TH2) type immunity in animal models, characterized by antibody production with little to no induction of antigen-specific T cells. The lack of cell-mediated or T helper 1 (TH1) immunity makes alum adjuvants ineffective in mounting durable responses against diseases like tuberculosis, malaria and HIV. Here we show that the clinically approved adjuvant, Alhydrogel, reformulated as a stable nanoparticle (nanoalum) with the anionic polymer polyacrylic acid (PAA) induces structure-dependent TH1 response against the recombinant tuberculosis antigen ID93. We found that PAA adsorption to Alhydrogel was a key parameter affecting nanoalum adjuvanticity. Adsorption depended on various factors, most notably formulation pH, and directly correlated with immunological response in mice, enhancing known hallmarks of a murine TH1 type response: induction of antigen-specific IFN-γ secreting CD4+ T cells and IgG2c subclass of antibodies. Our results demonstrate a correlation between a measurable nanoalum property and immunological response, providing a structural basis to derive a beneficial immunological outcome from a clinically approved adjuvant.

Squalene Emulsion Manufacturing Process Scale-Up for Enhanced Global Pandemic Response.

Squalene emulsions are among the most widely employed vaccine adjuvant formulations. Among the demonstrated benefits of squalene emulsions is the ability to enable vaccine antigen dose sparing, an important consideration for pandemic response. In order to increase pandemic response capabilities, it is desirable to scale up adjuvant manufacturing processes. We describe innovative process enhancements that enabled the scale-up of bulk stable squalene emulsion (SE) manufacturing capacity from a 3000- to 5,000,000-dose batch size. Manufacture of concentrated bulk along with the accompanying viscosity change in the continuous phase resulted in a ≥25-fold process efficiency enhancement. Process streamlining and implementation of single-use biocontainers resulted in reduced space requirements, fewer unit operations, and minimization of cleaning requirements. Emulsion physicochemical characteristics were measured by dynamic light scattering, laser diffraction, and HPLC with charged aerosol detection. The newly developed full-scale process was demonstrated by producing two 5,000,000-dose batches of bulk concentrated SE. A scale-up of adjuvant manufacturing capacity through process innovation enables more efficient production capabilities for pandemic response.

Preparedness against pandemic influenza: Production of an oil-in-water emulsion adjuvant in Brazil.

Increasing pandemic influenza vaccine manufacturing capacity is considered strategic by WHO. Adjuvant use is key in this strategy in order to spare the vaccine doses and by increasing immune protection. We describe here the production and stability studies of a squalene based oil-in-water emulsion, adjuvant IB160, and the immune response of the H7N9 vaccine combined with IB160. To qualify the production of IB160 we produced 10 consistency lots of IB160 and the average results were: pH 6.4±0.05; squalene 48.8±.0.03 mg/ml; osmolality 47.6±6.9 mmol/kg; Z-average 157±2 nm, with polydispersity index (PDI) of 0.085±0.024 and endotoxin levels <0.5 EU/mL. The emulsion particle size was stable for at least six months at 25°C and 24 months at 4-8°C. Two doses of H7N9 vaccine formulated at 7.5 µg/dose or 15 µg/dose with adjuvant IB160 showed a significant increase of hemagglutination inhibition (HAI) titers in sera of immunized BALB/c mice when compared to control sera from animals immunized with the H7N9 antigens without adjuvant. Thus the antigen-sparing capacity of IB160 can potentially increase the production of the H7N9 pandemic vaccine and represents an important achievement for preparedness against pandemic influenza and a successful North (IDRI) to South (Butantan Institute) technology transfer for the production of the adjuvant emulsion IB160.

Effects of Gender, Parental Role, and Time on Infant- and Adult-Directed Read and Spontaneous Speech.

Purpose The study sets out to investigate inter- and intraspeaker variation in German infant-directed speech (IDS) and considers the potential impact that the factors gender, parental involvement, and speech material (read vs. spontaneous speech) may have. In addition, we analyze data from 3 time points prior to and after the birth of the child to examine potential changes in the features of IDS and, particularly also, of adult-directed speech (ADS). Here, the gender identity of a speaker is considered as an additional factor. Method IDS and ADS data from 34 participants (15 mothers, 19 fathers) is gathered by means of a reading and a picture description task. For IDS, 2 recordings were made when the baby was approximately 6 and 9 months old, respectively. For ADS, an additional recording was made before the baby was born. Phonetic analyses comprise mean fundamental frequency (f0), variation in f0, the 1st 2 formants measured in /i: ɛ a u:/, and the vowel space size. Moreover, social and behavioral data were gathered regarding parental involvement and gender identity. Results German IDS is characterized by an increase in mean f0, a larger variation in f0, vowel- and formant-specific differences, and a larger acoustic vowel space. No effect of gender or parental involvement was found. Also, the phonetic features of IDS were found in both spontaneous and read speech. Regarding ADS, changes in vowel space size in some of the fathers and in mean f0 in mothers were found. Conclusion Phonetic features of German IDS are robust with respect to the factors gender, parental involvement, speech material (read vs. spontaneous speech), and time. Some phonetic features of ADS changed within the child's first year depending on gender and parental involvement/gender identity. Thus, further research on IDS needs to address also potential changes in ADS.

Gender identity is indexed and perceived in speech.

This study investigates a possible relationship between perceived and self-ascribed gender identity and the respective acoustic correlates in a group of young heterosexual adult speakers. For the production study, a sample of 37 German speaking subjects (20 males, 17 females) filled out a questionnaire to assess their self-ascribed masculinity/femininity on two scales. A range of acoustic parameters (acoustic vowel space size, fundamental frequency, sibilant spectral characteristics) were measured in speech collected from a picture describing task. Results show that male speakers judging themselves to be less masculine exhibited larger vowel spaces and higher average fundamental frequency.For the perception experiment, a group of 21 listeners (11 males, 10 females) judged masculinity of single word male stimuli drawn from the collected speech sample. A significant correlation between speakers' self-ascribed and listeners' attributed gender identity was found with a stronger relationship for female listeners. Acoustic parameters used by listeners to attribute gender identity include those used by speakers to index masculinity/femininity.The investigation demonstrates the importance of including self-ascribed gender identity as a potential source of inter-speaker variation in speech production and perception even in a sample of heterosexual adult speakers.

"Do I Sound Straight?": Acoustic Correlates of Actual and Perceived Sexual Orientation and Masculinity/Femininity in Men's Speech.

Purpose: This study aims to give an integrative answer on which speech stereotypes exist toward German gay and straight men, whether and how acoustic correlates of actual and perceived sexual orientation are connected, and how this relates to masculinity/femininity. Hence, it tests speech stereotype accuracy in the context of sexual orientation. Method: Twenty-five gay and 26 straight German speakers provided data for a fine-grained psychological self-assessment (e.g., masculinity/femininity) and explicit speech stereotypes. They were recorded for an extensive set of read and spontaneous speech samples using microphones and nasometry. Recordings were analyzed for a variety of acoustic parameters (e.g., fundamental frequency and nasalance). Seventy-four listeners categorized speakers as gay or straight on the basis of the same sentence. Results: Most relevant explicitly expressed speech stereotypes encompass voice pitch, nasality, chromaticity, and smoothness. Demonstrating implicit stereotypes, speakers were perceived as sounding straighter, the lower their median f0, center of gravity in /s/, and mean F2. However, based on actual sexual orientation, straight men only showed lower mean F1 than gay men. Additionally, we found evidence that actual masculinity/femininity and the degree of sexual orientation were reflected in gay and straight men's speech. Conclusion: Implicit and explicit speech stereotypes about gay and straight men do not contain a kernel of truth, and differences within groups are more important than differences between them. Supplemental Material: https://doi.org/10.23641/asha.6484001.

Investigating the common set of acoustic parameters in sexual orientation groups: A voice averaging approach.

While the perception of sexual orientation in voices often relies on stereotypes, it is unclear whether speech stereotypes and accurate perceptions of sexual orientation are each based on acoustic cues common to speakers of a given group. We ask if the stereotypical belief, that members of the same sexual orientation group share similar acoustic patterns, is accurate to some degree. To address this issue, we are the first to use a novel voice morphing technique to create voice averages from voices that represent extremes of a given sexual orientation group either in terms of actual or perceived sexual orientation. Importantly, averaging preserves only those acoustic cues shared by the original speakers. 144 German listeners judged the sexual orientation of twelve natural-sounding sentence stimuli, each representing an average of five original utterances. Half of the averages were based on targets' self-ratings of sexual orientation: On a 7-point Kinsey-like scale, we selected targets who were most typical for a certain sexual orientation group according to their self-identifications. The other half were based on extreme ratings by others (i.e., on speech-related sexual-orientation stereotypes). Listeners judged sexual orientation from the voice averages with above-chance accuracy suggesting 1) that the perception of actual and stereotypical sexual orientation, respectively, are based on acoustic cues shared by speakers of the same group, and 2) that the stereotypical belief that members of the same sexual orientation group share similar acoustic patterns is accurate to some degree. Mean fundamental frequency and other common acoustic parameters showed systematic variation depending on speaker gender and sexual orientation. Effects of sexual orientation were more pronounced for stereotypical voice averages than for those based on speakers' self-ratings, suggesting that sexual-orientation stereotypes exaggerate even those differences present in the most salient groups of speakers. Implications of our findings for stereotyping and discrimination are discussed.