Divergent Routes toward Wnt and R-spondin Niche Independency during Human Gastric Carcinogenesis.

Recent sequencing analyses have shed light on heterogeneous patterns of genomic aberrations in human gastric cancers (GCs). To explore how individual genetic events translate into cancer phenotypes, we established a biological library consisting of genetically engineered gastric organoids carrying various GC mutations and 37 patient-derived organoid lines, including rare genomically stable GCs. Phenotype analyses of GC organoids revealed divergent genetic and epigenetic routes to gain Wnt and R-spondin niche independency. An unbiased phenotype-based genetic screening identified a significant association between CDH1/TP53 compound mutations and the R-spondin independency that was functionally validated by CRISPR-based knockout. Xenografting of GC organoids further established the feasibility of Wnt-targeting therapy for Wnt-dependent GCs. Our results collectively demonstrate that multifaceted genetic abnormalities render human GCs independent of the stem cell niche and highlight the validity of the genotype-phenotype screening strategy in gaining deeper understanding of human cancers.

An organoid-based organ-repurposing approach to treat short bowel syndrome.

The small intestine is the main organ for nutrient absorption, and its extensive resection leads to malabsorption and wasting conditions referred to as short bowel syndrome (SBS). Organoid technology enables an efficient expansion of intestinal epithelium tissue in vitro1, but reconstruction of the whole small intestine, including the complex lymphovascular system, has remained challenging2. Here we generate a functional small intestinalized colon (SIC) by replacing the native colonic epithelium with ileum-derived organoids. We first find that xenotransplanted human ileum organoids maintain their regional identity and form nascent villus structures in the mouse colon. In vitro culture of an organoid monolayer further reveals an essential role for luminal mechanistic flow in the formation of villi. We then develop a rat SIC model by repositioning the SIC at the ileocaecal junction, where the epithelium is exposed to a constant luminal stream of intestinal juice. This anatomical relocation provides the SIC with organ structures of the small intestine, including intact vasculature and innervation, villous structures, and the lacteal (a fat-absorbing lymphatic structure specific to the small intestine). The SIC has absorptive functions and markedly ameliorates intestinal failure in a rat model of SBS, whereas transplantation of colon organoids instead of ileum organoids invariably leads to mortality. These data provide a proof of principle for the use of intestinal organoids for regenerative purposes, and offer a feasible strategy for SBS treatment.

Tracking age-related changes in voice and speech production with Landmark-based analysis of speech.

Voice and speech production change with age, which can lead to potential communication challenges. This study explored the use of Landmark-based analysis of speech (LMBAS), a knowledge-based speech analysis algorithm based on Stevens' Landmark Theory, to describe age-related changes in adult speakers. The speech samples analyzed were sourced from the University of Florida Aging Voice Database, which included recordings of 16 sentences from the Speech Perception in Noise test of Bilger, Rzcezkowski, Nuetzel, and Rabinowitz [J. Acoust. Soc. Am. 65, S98-S98 (1979)] and Bilger, Nuetzel, Rabinowitz, and Rzeczkowski [J. Speech. Lang. Hear. Res. 27, 32-84 (1984)]. These sentences were read in quiet environments by 50 young, 50 middle-aged, and 50 older American English speakers, with an equal distribution of sexes. Acoustic landmarks, specifically, glottal, bursts, and syllabicity landmarks, were extracted using SpeechMark®, MATLAB Toolbox version 1.1.2. The results showed significant age effect on glottal and burst landmarks. Furthermore, the sex effect was significant for burst and syllabicity landmarks. While the results of LMBAS suggest its potential in detecting age-related changes in speech, increase in syllabicity landmarks with age was unexpected. This finding may suggest the need for further refinement and adjustment of this analytical approach.

Identification of Quiescent LGR5+ Stem Cells in the Human Colon.

BACKGROUND & AIMS: In the mouse intestinal epithelium, Lgr5+ stem cells are vulnerable to injury, owing to their predominantly cycling nature, and their progenies de-differentiate to replenish the stem cell pool. However, how human colonic stem cells behave in homeostasis and during regeneration remains unknown. METHODS: Transcriptional heterogeneity among colonic epithelial cells was analyzed by means of single-cell RNA sequencing analysis of human and mouse colonic epithelial cells. To trace the fate of human colonic stem or differentiated cells, we generated LGR5-tdTomato, LGR5-iCasase9-tdTomato, LGR5-split-Cre, and KRT20-ERCreER knock-in human colon organoids via genome engineering. p27+ dormant cells were further visualized with the p27-mVenus reporter. To analyze the dynamics of human colonic stem cells in vivo, we orthotopically xenotransplanted fluorescence-labeled human colon organoids into immune-deficient mice. The cell cycle dynamics in xenograft cells were evaluated using 5-ethynyl-2'-deoxyuridine pulse-chase analysis. The clonogenic capacity of slow-cycling human stem cells or differentiated cells was analyzed in the context of homeostasis, LGR5 ablation, and 5-fluorouracil-induced mucosal injury. RESULTS: Single-cell RNA sequencing analysis illuminated the presence of nondividing LGR5+ stem cells in the human colon. Visualization and lineage tracing of slow-cycling LGR5+p27+ cells and orthotopic xenotransplantation validated their homeostatic lineage-forming capability in vivo, which was augmented by 5-FU-induced mucosal damage. Transforming growth factor-ß signaling regulated the quiescent state of LGR5+ cells. Despite the plasticity of differentiated KRT20+ cells, they did not display clonal growth after 5-FU-induced injury, suggesting that occupation of the niche environment by LGR5+p27+ cells prevented neighboring differentiated cells from de-differentiating. CONCLUSIONS: Our results highlight the quiescent nature of human LGR5+ colonic stem cells and their contribution to post-injury regeneration.

A mechanism of melanogenesis mediated by E-cadherin downregulation and its involvement in solar lentigines.

OBJECTIVE: Intensive studies have revealed that pleiotropic melanocytic factors are associated with age-spot formation. Dysfunctional keratinocyte differentiation is thought to be an upstream cause of age-spot formation. Although it has been shown that keratinocyte differentiation is mediated by the cell-cell contact factor E-cadherin, its involvement in age-spot formation remains unknown. Thus, to determine the origin of age-spots and an integrated solution for the same, we focused on E-cadherin expression in the present study. METHODS: First, we assessed the solar lentigines in cutaneous and cultured cells by means of immunofluorescence staining. Following that, keratinocytes treated with siRNAs against E-cadherin were co-cultured with melanocytes, and the secreted factors were identified by means of proteomic analysis of the culture supernatants. We also performed quantitative PCR to assess melanogenesis activity and screen ingredients. For behavioural analysis of melanocytes, we performed time-lapse imaging using confocal laser scanning microscopy. RESULTS: E-cadherin expression was downregulated in the epidermis of the solar lentigines, suggesting its involvement in age-spot formation. E-cadherin knocked down keratinocytes not only promoted the secretion of melanocytic/inflammatory factors but also increased melanogenesis by upregulating the expression of melanogenesis factors. Furthermore, live-imaging showed that the downregulation of E-cadherin inhibited melanocyte dynamics and accelerated melanin uptake. Finally, we identified Rosa multiflora fruit extract as a solution that can upregulate E-cadherin expression in keratinocytes. CONCLUSION: Our findings showed that E-cadherin downregulation triggers various downstream melanocytic processes, such as the secretion of melanocytic factors and melanogenesis. Additionally, we showed that the Rosa multiflora fruit extract upregulated E-cadherin expression in keratinocytes.

OBJECTIF: Des études intensives ont révélé que les facteurs mélanocytaires pléiotropiques sont associés à la formation de taches de vieillesse. On pense que la différenciation des kératinocytes dysfonctionnels est une cause en amont de la formation des taches de vieillesse. Bien qu'il ait été démontré que la différenciation des kératinocytes est médiée par le facteur de contact cellule-cellule E-cadhérine, son implication dans la formation des taches de vieillesse reste inconnue. Ainsi, pour déterminer l'origine des taches de vieillesse et une solution intégrée pour celles-ci, nous nous sommes concentrés sur l'expression de la E-cadhérine dans la présente étude. MÉTHODES: Tout d'abord, nous avons évalué les lentigines solaires dans les cellules cutanées et cultivées au moyen d'une coloration par immunofluorescence. Par la suite, les kératinocytes traités avec des siRNA contre l'E-cadhérine ont été co-cultivés avec des mélanocytes, et les facteurs sécrétés ont été identifiés au moyen d'une analyse protéomique des surnageants de culture. Nous avons également effectué une PCR quantitative pour évaluer l'activité de la mélanogénèse et dépister les ingrédients. Pour l'analyse comportementale des mélanocytes, nous avons réalisé une imagerie accélérée à l'aide de la microscopie confocale à balayage laser. RÉSULTATS: L'expression de l'E-cadhérine a été régulée à la baisse dans l'épiderme des lentigines solaires, suggérant son implication dans la formation des taches de vieillesse. Les kératinocytes dans lesquels l'E-cadhérine a été réduite non seulement ont favorisé la sécrétion de facteurs mélanocytaires/inflammatoires, mais ont également accru la mélanogenèse en régulant à la hausse l'expression de facteurs de mélanogenèse. De plus, l'imagerie en direct a montré que la régulation négative de l'E-cadhérine inhibait la dynamique des mélanocytes et accélérait l'absorption de la mélanine. Enfin, nous avons identifié l'extrait de fruit de Rosa multiflora comme une solution capable de réguler positivement l'expression de l'E-cadhérine dans les kératinocytes. CONCLUSION: Nos résultats ont montré que la régulation négative de la E-cadhérine déclenche divers processus mélanocytaires en aval, tels que la sécrétion de facteurs mélanocytaires et la mélanogénèse. De plus, nous avons montré que l'extrait de fruit de Rosa multiflora régulait à la hausse l'expression de l'E-cadhérine dans les kératinocytes.

Simultaneous measurement of the size and methylation of chromosome 4qA-D4Z4 repeats in facioscapulohumeral muscular dystrophy by long-read sequencing.

BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant muscular disorder characterized by asymmetric muscle wasting and weakness. FSHD can be subdivided into two types: FSHD1, caused by contraction of the D4Z4 repeat on chromosome 4q35, and FSHD2, caused by mild contraction of the D4Z4 repeat plus aberrant hypomethylation mediated by genetic variants in SMCHD1, DNMT3B, or LRIF1. Genetic diagnosis of FSHD is challenging because of the complex procedures required. METHODS: We applied Nanopore CRISPR/Cas9-targeted resequencing for the diagnosis of FSHD by simultaneous detection of D4Z4 repeat length and methylation status at nucleotide level in genetically-confirmed and suspected patients. RESULTS: We found significant hypomethylation of contracted 4q-D4Z4 repeats in FSHD1, and both 4q- and 10q-D4Z4 repeats in FSHD2. We also found that the hypomethylation in the contracted D4Z4 in FSHD1 is moderately correlated with patient phenotypes. CONCLUSIONS: Our method contributes to the development for the diagnosis of FSHD using Nanopore long-read sequencing. This finding might give insight into the mechanisms by which repeat contraction causes disease pathogenesis.

Agreements between speech language pathologists and naïve listeners' judgements of intelligibility in children with cleft palate.

Speech-language pathologists (SLPs) working in interdisciplinary craniofacial teams need to know how much their judgments of speech intelligibility could predict the patient's communication difficulties with non-clinical communicative partners. This study examines the relationship between experienced SLPs and naïve listeners in judgments of speech intelligibility in speakers with cleft palate. A speech perception study was conducted using speech samples from 20 speakers with velopharyngeal insufficiency (VPI) following cleft palate. Speech samples were presented to 70 naïve listeners and 10 SLPs who were experts in cleft palate treatment. Speech intelligibility scores from naïve listeners' orthographic transcriptions were obtained as the percentage of correctly identified words. Speech intelligibility scores from SLPs were obtained using a five-point rating scale. Spearman rank correlation indicated a very high level of overall agreement between naïve listeners and SLPs at the speaker level scores (rs = -.94, p < .001). While the listeners' judgment seems highly related across highly intelligible speakers, the differences in agreements increase when the speaker is unintelligible. The high correlations between scores for naïve listeners and SLPs suggest that speech intelligibility in children with VPI could be predicted by ratings done in the clinic by expert SLPs.

Chromosome Engineering of Human Colon-Derived Organoids to Develop a Model of Traditional Serrated Adenoma.

BACKGROUND & AIMS: Traditional serrated adenomas (TSAs) are rare colorectal polyps with unique histologic features. Fusions in R-spondin genes have been found in TSAs, but it is not clear whether these are sufficient for TSA development, due to the lack of a chromosome engineering platform for human tissues. We studied the effects of fusions in R-spondin genes and other genetic alterations found in TSA using CRISPR-Cas9-mediated chromosome and genetic modification of human colonic organoids. METHODS: We introduced chromosome rearrangements that involve R-spondin genes into human colonic organoids, with or without disruption of TP53, using CRISPR-Cas9 (chromosome-engineered organoids). We then knocked a mutation into BRAF encoding the V600E substitution and overexpressed the GREM1 transgene; the organoids were transplanted into colons of NOG mice and growth of xenograft tumors was measured. Colon tissues were collected and analyzed by immunohistochemistry or in situ hybridization. We also established 2 patient-derived TSA organoid lines and characterized their genetic features and phenotypes. We inserted a bicistronic cassette expressing a dimerizer-inducible suicide gene and fluorescent marker downstream of the LGR5 gene in the chromosome-engineered organoids; addition of the dimerizer eradicates LGR5+ cells. Some tumor-bearing mice were given intraperitoneal injections of the dimerizer to remove LGR5-expressing cells. RESULTS: Chromosome engineering of organoids required disruption of TP53 or culture in medium containing IGF1 and FGF2. In colons of mice, organoids that expressed BRAFV600E and fusions in R-spondin genes formed flat serrated lesions. Patient-derived TSA organoids grew independent of exogenous R-spondin, and 1 line grew independent of Noggin. Organoids that overexpressed GREM1, in addition to BRAFV600E and fusions in R-spondin genes, formed polypoid tumors in mice that had histologic features similar to TSAs. Xenograft tumors persisted after loss of LGR5-expressing cells. CONCLUSIONS: We demonstrated efficient chromosomal engineering of human normal colon organoids. We introduced genetic and chromosome alterations into human colon organoids found in human TSAs; tumors grown from these organoids in mice had histopathology features of TSAs. This model might be used to study progression of human colorectal tumors with RSPO fusion gene and GREM1 overexpression.

Whole genome sequencing of 45 Japanese patients with intellectual disability.

Intellectual disability (ID) is characterized by significant limitations in both intellectual functioning and adaptive behaviors, originating before the age of 18 years. However, the genetic etiologies of ID are still incompletely elucidated due to the wide range of clinical and genetic heterogeneity. Whole genome sequencing (WGS) has been applied as a single-step clinical diagnostic tool for ID because it detects genetic variations with a wide range of resolution from single nucleotide variants (SNVs) to structural variants (SVs). To explore the causative genes for ID, we employed WGS in 45 patients from 44 unrelated Japanese families and performed a stepwise screening approach focusing on the coding variants in the genes. Here, we report 12 pathogenic and likely pathogenic variants: seven heterozygous variants of ADNP, SATB2, ANKRD11, PTEN, TCF4, SPAST, and KCNA2, three hemizygous variants of SMS, SLC6A8, and IQSEC2, and one homozygous variant in AGTPBP1. Of these, four were considered novel. Furthermore, a novel 76 kb deletion containing exons 1 and 2 in DYRK1A was identified. We confirmed the clinical and genetic heterogeneity and high frequency of de novo causative variants (8/12, 66.7%). This is the first report of WGS analysis in Japanese patients with ID. Our results would provide insight into the correlation between novel variants and expanded phenotypes of the disease.

Intelligibility of dysphonic speech in auralized classrooms.

Voice disorders can reduce the speech intelligibility of affected speakers. This study evaluated the effect of noise, voice disorders, and room acoustics on vowel intelligibility, listening easiness, and the listener's reaction time. Three adult females with dysphonia and three adult females with normal voice quality recorded a series of nine vowels of American English in /h/-V-/d/ format (e.g., "had"). The recordings were convolved with two oral-binaural impulse responses acquired from measurements in two classrooms with 0.4 and 3.1 s of reverberation time, respectively. The stimuli were presented in a forced-choice format to 29 college students. The intelligibility and the listening easiness were significantly higher in quiet than in noisy conditions, when the speakers had normal voice quality compared to a dysphonic voice, and in low reverberated environments compared to high reverberated environments. The response time of the listener was significantly longer for speech presented in noisy conditions compared to quiet conditions and when the voice was dysphonic compared with healthy voice quality.

Agreement between Transcription- and Rating-Based Intelligibility Measurements for Evaluation of Dysphonic Speech in Noise.

Dysphonia negatively affects a speaker's intelligibility, especially in noisy environments. Previously, our study showed this effect of dysphonia with the transcription-based intelligibility measurement. While this finding indicates the importance of intelligibility assessment for this population, implementing the transcription-based measurement may be difficult in clinical settings due to its resource-demanding nature. Using the same speakers, this study examined the agreement between transcription- and rating-based intelligibility measurements. Six sentences from the Consensus of Auditory-Perceptual Evaluation of Voice (CAPE-V) were recorded from 18 individuals with dysphonia (6 adult females, 6 adult males, and 6 children). Their dysphonia severity was determined through auditory-perceptual evaluation by two speech-language pathologists. Cafeteria noise was added to these recordings at SNR0 and paired with a sample from a healthy speaker in their age and/or gender group. Forty-five listeners rated intelligibility of the dysphonic samples on a 7-point rating scale. Spearman's rank correlation tests were conducted to examine the correlations between rating-based intelligibility measurement and the transcription-based measurement from our previous study, as well as the voice quality ratings and the rating-based intelligibility measurements. There was a strong positive correlation between the transcription- and rating-based measurements at all noise levels. The correlation between rating-based intelligibility measurement and breathiness rating was also strong. Our findings suggest that the rating-based intelligibility measurement could potentially be used as a substitute for the transcription-based analysis. Furthermore, the intelligibility deficit may be particularly problematic to patients who present with breathy dysphonia.

Toward clinical application of landmark-based speech analysis: Landmark expression in normal adult speech.

The goal of clinical speech analysis is to describe abnormalities in speech production that affect a speaker's intelligibility. Landmark analysis identifies abrupt changes in a speech signal and classifies them according to their acoustic profiles. These acoustic markers, called landmarks, may help describe intelligibility deficits in disordered speech. As a first step toward clinical application of landmark analysis, the present study describes expression of landmarks in normal speech. Results of the study revealed that syllabic, glottal, and burst landmarks consist of 94% of all landmarks, and suggest the effect of gender needs to be considered for the analysis.

Cognitive load associated with speaking clearly in reverberant rooms.

Communication is a fundamental aspect of human interaction, yet many individuals must speak in less-than-ideal acoustic environments daily. Adapting their speech to ensure intelligibility in these varied settings can impose a significant cognitive burden. Understanding this burden on talkers has significant implications for the design of public spaces and workplace environments, as well as speaker training programs. The aim of this study was to examine how room acoustics and speaking style affect cognitive load through self-rating of mental demand and pupillometry. Nineteen adult native speakers of American English were instructed to read sentences in both casual and clear speech-a technique known to enhance intelligibility-across three levels of reverberation (0.05 s, 1.2 s, and 1.83 s at 500-1000 Hz). Our findings revealed that speaking style consistently affects the cognitive load on talkers more than room acoustics across the tested reverberation range. Specifically, pupillometry data suggested that speaking in clear speech elevates the cognitive load comparably to speaking in a room with long reverberation, challenging the conventional view of clear speech as an 'easy' strategy for improving intelligibility. These results underscore the importance of accounting for talkers' cognitive load when optimizing room acoustics and developing speech production training.

Vocal Effort and Acoustic Analysis of Gargle Phonation Versus Water Swallow in Patients With Muscle Tension Dysphonia: A Clinical Trial.

PURPOSE: To determine the effects of gargle phonation (GP) on self-perceived vocal improvement, vocal effort, acoustic parameters, and speech rate in patients with muscle tension dysphonia (MTD). We hypothesized that GP would improve voice, reduce phonatory effort, and alter acoustic and speech measures. STUDY DESIGN: Prospective randomized, single-blind cross-over clinical trial METHODS: Thirty-four participants (26 females, 8 males; average age 53 years) who were diagnosed with MTD completed the Voice Handicap Index-10 (VHI-10) and were assigned three study conditions: Baseline (B), GP, and Water Swallow (WS; sham), presented in one of two counterbalanced orders B-WS-GP (WS1st) or B-GP-WS (GP1st). Participants recorded stimuli from the Consensus Auditory-Perceptual Evaluation of Voice (CAPE-V) and rated their perceived vocal effort and vocal improvement. F0, vocal intensity, cepstral peak prominence (CPP), and speaking rate were measured. RESULTS: Average VHI-10 scores by group were 16 (min/max 2-29) for WS1st and 15 (min/max 3-40) for GP1st. About 73.5% reported more vocal improvement after GP, 17.65% after WS, and 8.8% noted no difference between conditions. Reduced effort was reported after GP, compared to B (P < 0.001) and WS (P = 0.005). Lower effort was also reported after the WS condition, compared to B (P = 0.011). Key acoustic findings included an increase in F0 after GP for sustained /i/ for females. CPP was significantly higher for females reading CAPE-V sentences after GP, when GP preceded WS, compared to B (P = 0.004) and WS (P = 0.003). Speech rate was faster for females after GP versus B (P = 0.029). CONCLUSIONS: GP may be beneficial in the treatment of MTD. CPP may be a useful marker for vocal improvement after GP for women with mild MTD. Further studies would benefit from having more male participants and those with moderate and severe MTD.

The Formant Bandwidth as a Measure of Vowel Intelligibility in Dysphonic Speech.

OBJECTIVE: The current paper examined the impact of dysphonia on the bandwidth of the first two formants of vowels, and the relationship between the formant bandwidth and vowel intelligibility. METHODS: Speaker participants of the study were 10 adult females with healthy voice and 10 adult females with dysphonic voice. Eleven vowels in American English were recorded in /h/-vowel-/d/ format. The vowels were presented to 10 native speakers of American English with normal hearing, who were asked to select a vowel they heard from a list of /h/-vowel-/d/ words. The vowels were acoustically analyzed to measure the bandwidth of the first and second formants (B1 and B2). Separate Wilcoxon rank sum tests were conducted for each vowel for normal and dysphonic speech because the differences in B1 and B2 were found to not be normally distributed. Spearman correlation tests were conducted to evaluate the association between the difference in formant bandwidths and vowel intelligibility between the healthy and dysphonic speakers. RESULTS: B1 was significantly greater in dysphonic vowels for seven of the eleven vowels, and lesser for only one of the vowels. There was no statistically significant difference in B2 between the normal and dysphonic vowels, except for the vowel /i/. The difference in B1 between normal and dysphonic vowels strongly predicted the intelligibility difference. CONCLUSION: Dysphonia significantly affects B1, and the difference in B1 may serve as an acoustic marker for the intelligibility reduction in dysphonic vowels. This acoustic-perceptual relationship should be confirmed by a larger-scale study in the future.

The Effect of Noise on Initiation and Maintenance of Clear Speech and Associated Mental Demand.

BACKGROUND/OBJECTIVES: The objectives of this study were to explore (a) the influence of different types of background noise and their informational content on the ability of speakers to initiate and maintain clear speech (CS), a widely utilized technique for enhancing speech intelligibility, and (b) the impact of background noise and CS usage on speakers' mental demand. METHOD: Five adult females were asked to read sentences using both habitual and CS under four distinct noise conditions: quiet, multitalker (MT) noise, reversed multitalker (RevMT) noise, and speech-shaped (SS) noise. Following this, speakers rated their perceived level of mental demand for each speaking condition using the modified NASA Task Load Index scale. A two-part listening experiment with 48 listeners was conducted to evaluate the speakers' effectiveness in initiating and maintaining CS. RESULTS: Speakers initiated CS more successfully in noise than in quiet, with better performance observed in the presence of RevMT noise as compared to SS noise. Regarding the maintenance of CS, none of the speakers were successful in a quiet environment. Furthermore, the ability to maintain CS was most adversely affected in MT noise, followed by RevMT noise and SS noise. CONCLUSIONS: Our findings suggest that the effect of background noise on speech production is complex and multifaceted. The noise type affected speakers' ability to initiate and maintain CS as well as the mental demand associated with the speech task. The results underscore the importance of considering the characteristics of background noise and cognitive aspects of speech production when training and evaluating speakers' performance.

Landmark-based analysis of speech differentiates conversational from clear speech in speakers with muscle tension dysphonia.

This study evaluated the feasibility of differentiating conversational and clear speech produced by individuals with muscle tension dysphonia (MTD) using landmark-based analysis of speech (LMBAS). Thirty-four adult speakers with MTD recorded conversational and clear speech, with 27 of them able to produce clear speech. The recordings of these individuals were analyzed with the open-source LMBAS program, SpeechMark®, matlab Toolbox version 1.1.2. The results indicated that glottal landmarks, burst onset landmarks, and the duration between glottal landmarks differentiated conversational speech from clear speech. LMBAS shows potential as an approach for detecting the difference between conversational and clear speech in dysphonic individuals.

Diet-mediated constitutive induction of novel IL-4+ ILC2 cells maintains intestinal homeostasis in mice.

Group 2 innate lymphoid cells (ILC2s) expressing IL-5 and IL-13 are localized at various mucosal tissues and play critical roles in the induction of type 2 inflammation, response to helminth infection, and tissue repair. Here, we reveal a unique ILC2 subset in the mouse intestine that constitutively expresses IL-4 together with GATA3, ST2, KLRG1, IL-17RB, and IL-5. In this subset, IL-4 expression is regulated by mechanisms similar to but distinct from those observed in T cells and is partly affected by IL-25 signaling. Although the absence of the microbiota had marginal effects, feeding mice with a vitamin B1-deficient diet compromised the number of intestinal IL-4+ ILC2s. The decrease in the number of IL-4+ ILC2s caused by the vitamin B1 deficiency was accompanied by a reduction in IL-25-producing tuft cells. Our findings reveal that dietary vitamin B1 plays a critical role in maintaining interaction between tuft cells and IL-4+ ILC2s, a previously uncharacterized immune cell population that may contribute to maintaining intestinal homeostasis.

Prospective study of the evaluation of the usefulness of tumor typing by narrow band imaging for the differential diagnosis of gastric adenoma and well-differentiated adenocarcinoma.

BACKGROUND AND AIM: Presently, the differential diagnosis of gastric adenoma and well-differentiated adenocarcinoma by endoscopy is very difficult. We carried out magnifying endoscopy with narrow band imaging (NBI) in lesions that required discrimination between gastric adenoma and well-differentiated adenocarcinoma, and prospectively evaluated whether the tumor typing that we propose is useful for their differential diagnosis. METHODS: The materials were 93 lesions that required differential diagnosis between gastric adenoma and well-differentiated adenocarcinoma among the gastric epithelial tumors for which endoscopic treatment was planned at three facilities during the 14 months between November 2008 and December 2009. According to the typing method proposed by our facility based on images of the mucosal ultrastructure and microvessels obtained by magnified endoscopy combined with NBI, type I-II and type III-V lesions were diagnosed as gastric adenoma and well-differentiated adenocarcinoma, respectively, before endoscopic treatment, and the accuracy of the diagnoses were prospectively examined by comparing them with the postoperative pathological findings. RESULTS: Of the 93 lesions, 87 could be typed into the five types of our typing method, but six lesions could not be classified. The 87 lesions consisted of 16 type I, 12 type II, 29 type III, 27 type IV, and three type V lesions. The percentages of accurate preoperative diagnoses of types I-II as adenoma and types III-V as well-differentiated adenocarcinoma were 79% and 93%, respectively. CONCLUSIONS: The tumor typing based on NBI was useful for the endoscopic differentiation of gastric adenoma and well-differentiated adenocarcinoma.