Exploring the relationship between plasma substance P and glottal incompetence in the elderly.

We speculated that increased blood-plasma levels of Substance P may serve as an indicator of glottal incompetence, which is usually indicated by reduced maximum phonation time. We performed an initial study to test the plausibility of this hypothesis. Patients with dysphonia caused by glottal incompetence were asked to perform vocal exercises for six months to reduce glottal incompetence and we compared the plasma concentration of Substance P before and after the vocal exercise to detect correlation between maximum phonation time and plasma concentration of Substance P. Based on the results, we further hypothesized that patients exhibiting dysphonia with maximum phonation time less than 14 s, in particular less than 10 sec, caused by glottal incompetence may have increased plasma concentration of Substance P with the results of elevated thresholds of cough reflex associated with subclinical aspiration in airways. Further study is needed on patients with decreased Substance P levels, with low scores on Activities of Daily Living and who are hospitalized with aspiration pneumonia.

Magnetic cortical oscillations associated with subjective auditory coolness during paired comparison of time-varying HVAC sounds.

The impressions of heating, ventilation, and air conditioning (HVAC) sounds are important for the comfort people experience in their living spaces. Revealing neural substrates of the impressions induced by HVAC sounds can help to develop neurophysiological indices of the comfort of HVAC sounds. There have been numerous studies on the brain activities associated with the pleasantness of sounds, but few on the brain activities associated with the thermal impressions of HVAC sounds. Seven time-varying HVAC sounds were synthesized as stimuli using amplitude modulation. Six participants took part in subjective evaluation tests and MEG measurements. Subjective coolness of the HVAC sounds was measured using the paired comparison method. Magnetoencephalographic (MEG) measurements were carried out while participants listened to and compared the time-varying HVAC sounds. Time-frequency analysis and cluster-based analysis were performed on the MEG data. The subjective evaluation tests showed that the subjective coolness of the amplitude-modulated HVAC sounds was affected by the modulation frequency, and that there was individual difference in subjective coolness. A cluster-based analysis of the MEG data revealed that the brain activities of two participants significantly differed when they listened to cooler or less cool HVAC sounds. The frontal low-theta (4-5 Hz) and the temporal alpha (8-13 Hz) activities were observed. The frontal low-theta and the temporal alpha activities may be associated with the coolness of HVAC sound. This result suggests that the comfort level of HVAC sound can be evaluated and individually designed using neurophysiological measurements.

Evaluation and Manipulation of Neural Activity using Two-Photon Holographic Microscopy.

Recent advances in optical bioimaging and optogenetics have enabled the visualization and manipulation of biological phenomena, including cellular activities, in living animals. In the field of neuroscience, detailed neural activity related to brain functions, such as learning and memory, has now been revealed, and it has become feasible to artificially manipulate this activity to express brain functions. However, the conventional evaluation of neural activity by two-photon Ca2+ imaging has the problem of low temporal resolution. In addition, manipulation of neural activity by conventional optogenetics through the optic fiber can only simultaneously regulate the activity of neurons with the same genetic background, making it difficult to control the activity of individual neurons. To solve this issue, we recently developed a microscope with a high spatiotemporal resolution for biological applications by combining optogenetics with digital holographic technology that can modify femtosecond infrared laser beams. Here, we describe protocols for the visualization, evaluation, and manipulation of neural activity, including the preparation of samples and operation of a two-photon holographic microscope (Figure 1). These protocols provide accurate spatiotemporal information on neural activity, which may be useful for elucidating the pathogenesis of neuropsychiatric disorders that lead to abnormalities in neural activity.

Whole exome analysis of patients in Japan with hearing loss reveals high heterogeneity among responsible and novel candidate genes.

BACKGROUND: Heterogeneous genetic loci contribute to hereditary hearing loss; more than 100 deafness genes have been identified, and the number is increasing. To detect pathogenic variants in multiple deafness genes, in addition to novel candidate genes associated with hearing loss, whole exome sequencing (WES), followed by analysis prioritizing genes categorized in four tiers, were applied. RESULTS: Trios from families with non-syndromic or syndromic hearing loss (n = 72) were subjected to WES. After segregation analysis and interpretation according to American College of Medical Genetics and Genomics guidelines, candidate pathogenic variants in 11 previously reported deafness genes (STRC, MYO15A, CDH23, PDZD7, PTPN11, SOX10, EYA1, MYO6, OTOF, OTOG, and ZNF335) were identified in 21 families. Discrepancy between pedigree inheritance and genetic inheritance was present in one family. In addition, eight genes (SLC12A2, BAIAP2L2, HKDC1, SVEP1, CACNG1, GTPBP4, PCNX2, and TBC1D8) were screened as single candidate genes in 10 families. CONCLUSIONS: Our findings demonstrate that four-tier assessment of WES data is efficient and can detect novel candidate genes associated with hearing loss, in addition to pathogenic variants of known deafness genes.

A High Risk of Missing Congenital Cytomegalovirus-Associated Hearing Loss through Newborn Hearing Screening in Japan.

It remains unclear to what extent newborn hearing screening (NHS) detects congenital cytomegalovirus (cCMV)-associated sensorineural hearing loss (SNHL) in Japan. This study aimed to clarify the NHS results and audiological characteristics of patients with cCMV-associated SNHL. A total of 541 individuals with unilateral or bilateral hearing loss of unknown etiology were examined for cCMV infection. cCMV infection was defined by the presence of CMV DNA in the dried umbilical cord detected using real-time quantitative PCR. NHS results and audiological data were retrospectively obtained from medical records. Forty-four cases (8.1%) were positive for cCMV infection. Of them, 33 cases underwent NHS and 13 cases (39.4%) passed NHS bilaterally. The pure-tone audiograms of 21 patients were obtained. There were seven cases of unilateral SNHL, five cases of asymmetric bilateral SNHL, and nine cases of symmetric bilateral SNHL. cCMV-related hearing loss is highly heterogeneous, and there is a high risk of missing this condition through NHS.

Pain induces stable, active microcircuits in the somatosensory cortex that provide a therapeutic target.

Sustained neuropathic pain from injury or inflammation remains a major burden for society. Rodent pain models have informed some cellular mechanisms increasing neuronal excitability within the spinal cord and primary somatosensory cortex (S1), but how activity patterns within these circuits change during pain remains unclear. We have applied multiphoton in vivo imaging and holographic stimulation to examine single S1 neuron activity patterns and connectivity during sustained pain. Following pain induction, there is an increase in synchronized neuronal activity and connectivity within S1, indicating the formation of pain circuits. Artificially increasing neuronal activity and synchrony using DREADDs reduced pain thresholds. The expression of N-type voltage-dependent Ca2+ channel subunits in S1 was increased after pain induction, and locally blocking these channels reduced both the synchrony and allodynia associated with inflammatory pain. Targeting these S1 pain circuits, via inhibiting N-type Ca2+ channels or other approaches, may provide ways to reduce inflammatory pain.

Quantitative Evaluation of Handwriting Skills during Childhood.

BACKGROUND: Handwriting skills are very important skills for school-aged children, and consist of the abilities to control writing pressure and to assess shapes visually. Currently, various kinds of research have been conducted to clarify the developmental process of handwriting to establish methods for evaluating handwriting skills. However, a gold standard method has not been established. PURPOSE: This study aimed to clarify the developmental process of handwriting and to develop a new method for evaluating handwriting skills. METHOD: One hundred ninety-nine children aged from 4 to 12 years old and 17 young adults participated in this study. They were asked to draw a line carefully between two parallel lines with an electronic pen. Pressure during drawing and distance from the center point of the parallel lines were monitored continually using originally developed computer software. RESULTS: Depending on age, the average pressure increased until seven years old, and afterward it was sustained at almost the same level as adults. The distance from the center points was similarly reduced until seven years old, and afterward it also remained at the same level. On the other hand, the time required for performance increased until the age of 7, whereupon it decreased with age. Considering the factor of speed in the second step, handwriting skills gradually improved depending on age even after seven years old. CONCLUSION: In the development of handwriting skills, our study suggests that accuracy develops in an earlier stage and then the ability of speed control develops in the next stage. The new method developed in this study is expected to be useful as a tool for evaluating handwriting skills. Our method might be useful in evaluating and supporting children with neurodevelopmental disorders, such as autism spectrum disorder, who are often affected by a lack of dexterity．.

A clinical and genetic study of 16 Japanese families with Waardenburg syndrome.

The purpose of this study is to profile the clinical and genetic features of Japanese Waardenburg syndrome (WS) patients and validate the W index. Sixteen Japanese WS families with congenital sensorineural hearing loss were included in the study. The inner canthal, interpupillary, and outer canthal distances (ICD, IPD, and OCD) were measured for all patients, and patients were screened for presence of PAX3, MITF, SOX10, and EDNRB mutations. The WS patients were clinically classified under the current W index as follows: 13 families with WS1, 2 families with WS2, and 1 family with WS4. In the 13 WS1 families, genetic tests found PAX3 mutations in 5 families, MITF mutations in 4 families, SOX10 mutations in 3 families, and EDNRB mutations in 1 family. 61% of clinically classified WS1 patients under the current W index conflicted with the genetic classification, which implies W index is not appropriate for Japanese population. Resetting the threshold of W index or novel index formulated with ethnicity matched samples is necessary for clinical classification which is consistent with genetic classification for WS patients with distinct ethnicity.

Cortical Patterns for Prediction of Subjective Preference Induced by Chords.

To extract an effective feature in prediction of subjective impressions from single-trial neurophysiological recordings, the spatial filter that extracts brain activities related to impressions were constructed using the common spatial pattern (CSP). We focus on subjective preference induced by chords composed of 3 notes with different frequency ratio. Magnetic cortical activities while hearing chords and comparative judgment on pair of them were measured. The predictive model that predicts the scale value of preference was trained using the CSP-based feature for each participant. The result of the evaluation experiment shows that the CSP-based feature improved the mean prediction accuracy in all participants, compared with the other features without spatially filtering. Furthermore, the capability of construction of a spatial filter that extracts cortical activities varying with degree of preference using the comparative judgments was indicated.

Study on the Language Formation Process of Very-Low-Birth-Weight Infants in Infancy Using a Formant Analysis.

Expressive language development depends on anatomical factors, such as motor control of the tongue and oral cavity needed for vocalization, as well as cognitive aspects for comprehension and speech. The purpose of this study was to examine the differences in expressive language development between normal-birth-weight (NBW) infants and very-low-birth-weight (VLBW) infants in infancy using a formant analysis. We also examined the presence of differences between infants with a normal development and those with a high risk of autism spectrum disorder who were expected to exist among VLBW infants. The participants were 10 NBW infants and 10 VLBW infants 12-15 months of age whose speech had been recorded at intervals of approximately once every 3 months. The recorded speech signal was analyzed using a formant analysis, and changes due to age were observed. One NBW and 3 VLBW infants failed to pass the screening tests (CBCL and M-CHAT) at 24 months of age. The formant frequencies (F1 and F2) of the three groups of infants (NBW, VLBW and CBCL·M-CHAT non-passing infants) were scatter-plotted by age. For the NBW and VLBW infants, the area of the plot increased with age, but there was no significant expansion of the plot area for the CBCL·M-CHAT non-passing infants. The results showed no significant differences in expressive language development between NBW infants at 24 months old and VLBW infants at the corrected age. However, different language developmental patterns were observed in CBCL·M-CHAT non-passing infants, regardless of birth weight, suggesting the importance of screening by acoustic analyses.

Spatiotemporal Characteristics of Cortical Activities Associated with Articulation of Speech Perception.

Recently, brain computer interface (BCI) technologies that control external devices with human brain signals have been developed. However, most of the BCI systems, such as P300-speller, can only discriminate among options that have been given in advance. Therefore, the ability to decode the state of a person's perception and recognition, as well as that person's fundamental intention and emotions, from cortical activity is needed to develop a more general-use BCI system. In this study, two experiments were conducted. First, articulations were measured for Japanese monosyllabic utterances masked by several levels of noise. Second, auditory brain magnetic fields evoked by the monosyllable stimuli used in the first experiment were recorded, and neuronal current sources were localized in regions associated with speech perception and recognition - the auditory cortex (BA41), the Wernicke's area (posterior part of BA22), Broca's area (BA22), motor (BA4), and premotor (BA6) areas. Although the source intensity did not systematically change with SNR, the peak latency changed along SNR in the posterior superior temporal gyrus in the right hemisphere. The results suggest that the information associated with articulation is processed in this area.

Vocal Hygiene Education Program Reduces Surgical Interventions for Benign Vocal Fold Lesions: A Randomized Controlled Trial.

OBJECTIVES/HYPOTHESIS: Vocal fold polyps and nodules are common benign laryngeal lesions. Currently, the Japanese health insurance system covers surgical interventions. However, the establishment of more cost-effective conservative methods is required, because healthcare costs are viewed as a major concern, and the government and taxpayers are demanding more economical, effective treatments. In this situation, more suitable vocal hygiene education may be important for the success of cost-effective conservative treatment. In this study, we developed a novel reinforced vocal hygiene education program and compared the results of this program with those of previous methods of teaching vocal hygiene. STUDY DESIGN: Multicenter randomized controlled trial. METHODS: Patients who visited a National Hospital Organization (NHO) hospital for the surgical indication of hoarseness were included in the study. Before undergoing surgery, 200 patients with benign vocal fold lesions (vocal fold polyps/nodules) were enrolled and randomly allocated to the NHO-style vocal hygiene educational program (intervention group) or control education program (control group). Two months after enrollment, the patients in both groups underwent laryngeal fiberscopic examinations to determine whether the benign lesions had resolved or whether surgery was indicated for the vocal fold polyps/nodules. RESULTS: After 2 months, in the intervention group, the proportion of lesion resolution (61.3%) was significantly greater than that in the control group (26.3%) (P < .001, Fisher exact test). CONCLUSIONS: Our results clearly indicate that the quality and features of the education program could affect the outcome of the intervention. We found that a reinforced vocal hygiene education program increased the rate of the resolution of benign vocal fold polyps and nodules in a multicenter randomized clinical trial. LEVEL OF EVIDENCE: 1b Laryngoscope, 2593-2599, 2018.

Spatiotemporal properties of magnetic fields induced by auditory speech sound imagery and perception.

Brain computer interface (BCI) technologies, which enable direct communication between the brain and external devices, have been developed. BCI technology can be utilized in neural prosthetics to restore impaired movement, including speech production. However, most of the BCI systems that have been developed are the "P300-speller" type, which can only detect objects that users direct his/her attention at. To develop more versatile BCI systems that can detect a user's intention or thoughts, the brain responses associated with verbal imagery need to be clarified. In this study, the brain magnetic fields associated with auditory verbal imagery and speech hearing were recorded using magnetoencephalography (MEG) carried out on 8 healthy adults. Although the magnetic fields lagged slightly and were long-lasting, significant deflections were observed even for verbal imagery, in the temporal regions, as well as for actual speech hearing. Also, sources for the deflections were localized in the association auditory cortices. Cross-correlations were calculated between envelopes of the imagined/presented speech sound and the evoked brain responses in the temporal areas. Measurable correlations were obtained for the presented speech sound; however, no significant correlations were observed for the imagined speech sound. These results indicate that auditory verbal imagery undoubtedly activates the auditory cortex, at least, and generates some observable neural responses.

Detecting Abnormal Word Utterances in Children With Autism Spectrum Disorders: Machine-Learning-Based Voice Analysis Versus Speech Therapists.

Abnormal prosody is often evident in the voice intonations of individuals with autism spectrum disorders. We compared a machine-learning-based voice analysis with human hearing judgments made by 10 speech therapists for classifying children with autism spectrum disorders ( n = 30) and typical development ( n = 51). Using stimuli limited to single-word utterances, machine-learning-based voice analysis was superior to speech therapist judgments. There was a significantly higher true-positive than false-negative rate for machine-learning-based voice analysis but not for speech therapists. Results are discussed in terms of some artificiality of clinician judgments based on single-word utterances, and the objectivity machine-learning-based voice analysis adds to judging abnormal prosody.

Microbiomes of the normal middle ear and ears with chronic otitis media.

OBJECTIVE: The aim of this study was to profile and compare the middle ear microbiomes of human subjects with and without chronic otitis media. STUDY DESIGN: Prospective multicenter cohort study. METHODS: All consecutive patients undergoing tympanoplasty surgery for chronic otitis media or ear surgery for conditions other than otitis media were recruited. Sterile swab samples were collected from the middle ear mucosa during surgery. The variable region 4 of the 16S rRNA gene in each sample were amplified using region-specific primers adapted for the Illumina MiSeq sequencer (Illumina, CA, USA)). The sequences were subjected to local blast and classified using Metagenome@KIN (World Fusion, Tokyo, Japan). RESULTS: In total, 155 participants were recruited from seven medical centers. Of these, 88 and 67 had chronic otitis media and normal middle ears, respectively. The most abundant bacterial phyla on the mucosal surfaces of the normal middle ears were Proteobacteria, followed by Actinobacteria, Firmicutes, and Bacteroidetes. The children and adults with normal middle ears differed significantly in terms of middle ear microbiomes. Subjects with chronic otitis media without active inflammation (dry ear) had similar middle ear microbiomes as the normal middle ears group. Subjects with chronic otitis media with active inflammation (wet ear) had a lower prevalence of Proteobacteria and a higher prevalence of Firmicutes than the normal middle ears. CONCLUSION: The human middle ear is inhabited by more diverse microbial communities than was previously thought. Alteration of the middle ear microbiome may contribute to the pathogenesis of chronic otitis media with active inflammation. LEVEL OF EVIDENCE: 2b. Laryngoscope, 127:E371-E377, 2017.

Evaluation of positron emission tomography imaging to detect lymph node metastases in patients with high-risk cutaneous squamous cell carcinoma.

Ten to twenty percent of high-risk cutaneous squamous cell carcinoma (cSCC) can metastasize to regional lymph nodes. Detection of nodal metastasis is mandatory to manage high-risk cSCC. This study was aimed to evaluate the 18 F-fluorodeoxyglucose positron emission tomography (FDG-PET) imaging to assess lymph node (LN) metastasis of high-risk cSCC patients. Twenty-six patients with histologically proven primary cSCC were enrolled. All patients underwent whole-body PET prior to lymphoscintigraphy and subsequent sentinel LN biopsy. The maximum standardized uptake value (SUVmax) of more than 2.5 is generally evaluated as a positive PET finding indicative of malignancy. On the basis of the histopathological and PET findings, 30 LN from 26 patients were categorized into four groups: (i) histologically negative and PET negative (true-negative; n = 22); (ii) histologically positive and PET negative (false-negative; n = 0); (iii) histologically positive and PET positive (true-positive; n = 3); and (iv) histologically negative and PET positive (false-positive; n = 5). The mean SUVmax was significantly higher in the true-positive cases (11.0 ± 2.8) than in the false-positive cases (3.4 ± 0.6). In the false-positive cases, the number of tumor-infiltrating inflammatory cells at the primary skin site was highest among the four groups, suggesting that inflammation contributed to the false-positive uptake of FDG. Although we cannot negate the possibility of the presence of PET-undetectable micrometastasis, the SUVmax value may be useful for avoidance of excess performance of sentinel LN biopsy.

Lymphatic transit rate as a novel predictive parameter for nodal metastasis in primary truncal skin cancers.

Prediction of nodal metastasis in skin cancer before sentinel lymph node (SLN) biopsies is ideal to avoid unnecessary SLN biopsy performance. Primary truncal skin cancers are characterized by the lymphatic flow that drains from the primary lesion, occasionally to plural nodal basins. The scintigraphic appearance time (SAT), defined as the time between radionuclide injection and first SLN visualization, can potentially predict nodal metastasis, and a short SAT is a predictive parameter for metastasis. We recently introduced a novel method to measure the lymphatic flow rate using dynamic lymphoscintigraphy exhibiting a time-activity curve in the SLN. The time at which the count reaches a plateau in the SLN is termed the scintigraphic saturation time (SST) and can be a good alternative to the SAT. Moreover, the value obtained by division of the distance between the primary lesion and the SLN by the SST was termed the lymphatic transit rate (LTR), which represents the scintigraphic saturation velocity. In the present study, we evaluated LTR as a predictive parameter for nodal metastasis. Data for 22 lymph nodes from 18 patients with primary truncal skin cancers were used. Histopathologically, nodal metastasis was determined in nine nodes of eight patients. Because the mean LTR were 1.84 cm/min in non-metastatic SLN and 2.38 cm/min in metastatic SLN, the LTR was significantly higher in metastatic SLN than in non-metastatic SLN. All SLN with LTR of less than 1.8 cm/min were histopathologically evaluated as non-metastatic. The LTR may be a predictive indicator for nodal metastasis.

Dexamethasone but not tacrolimus suppresses TNF-α-induced thymic stromal lymphopoietin expression in lesional keratinocytes of atopic dermatitis model.

BACKGROUND: Thymic stromal lymphopoietin (TSLP) initiates the Th2-type allergic inflammation, and is thought to play an important role in the pathogenesis of atopic dermatitis (AD). TNF-α is a key cytokine which is involved in the pathophysiology of various inflammatory diseases, and the expression level is elevated in the sera and skin of patients with AD. In addition, TNF-α has been reported to induce TSLP expression in epidermal keratinocytes. Topical glucocorticoids and calcineurin inhibitors are safe and effective agents for AD, but the effects of these agents on TNF-α-induced TSLP expression are not fully understood. OBJECTIVE: To investigate whether the glucocorticosteroid dexamethasone and the calcineurin inhibitor tacrolimus could affect TSLP expression induced by TNF-α in lesional keratinocytes of AD. METHODS: The effects of topical dexamethasone and tacrolimus on TSLP expression were evaluated in an AD mouse model induced by repeated 2,4,6-trinitro-1-chlorobenzene application. Co-immunostaining for TSLP and TNF-α was performed using skin samples from AD patients and the mouse model. Normal human epidermal keratinocytes (NHEKs) were cultured with dexamethasone or tacrolimus in the presence of TNF-α to analyze TSLP expression. RESULTS: Topical application of dexamethasone but not tacrolimus repressed TSLP expression in the mouse model. TSLP and TNF-α showed similar distribution pattern in epidermal keratinocytes of AD lesions and the mouse model. TSLP expression was induced by TNF-α via NF-κB in a dose-dependent and an autocrine and/or paracrine manner in NHEKs, which was significantly suppressed by dexamethasone but not by tacrolimus. Similarly to TSLP expression, IL-6, TNF-α, IL-8, and IL-36γ expression induced by TNF-α were significantly suppressed by dexamethasone but not by tacrolimus in NHEKs. CONCLUSION: Dexamethasone but not tacrolimus suppresses the TSLP expression induced by TNF-α in lesional keratinocytes of AD model. Our observations uncover the unreported functional difference between topical glucocorticosteroids and calcineurin inhibitors in cutaneous inflammatory diseases.