Otitis media susceptibility and shifts in the head and neck microbiome due to SPINK5 variants.

BACKGROUND: Otitis media (OM) susceptibility has significant heritability; however, the role of rare variants in OM is mostly unknown. Our goal is to identify novel rare variants that confer OM susceptibility. METHODS: We performed exome and Sanger sequencing of >1000 DNA samples from 551 multiethnic families with OM and unrelated individuals, RNA-sequencing and microbiome sequencing and analyses of swabs from the outer ear, middle ear, nasopharynx and oral cavity. We also examined protein localisation and gene expression in infected and healthy middle ear tissues. RESULTS: A large, intermarried pedigree that includes 81 OM-affected and 53 unaffected individuals cosegregates two known rare A2ML1 variants, a common FUT2 variant and a rare, novel pathogenic variant c.1682A>G (p.Glu561Gly) within SPINK5 (LOD=4.09). Carriage of the SPINK5 missense variant resulted in increased relative abundance of Microbacteriaceae in the middle ear, along with occurrence of Microbacteriaceae in the outer ear and oral cavity but not the nasopharynx. Eight additional novel SPINK5 variants were identified in 12 families and individuals with OM. A role for SPINK5 in OM susceptibility is further supported by lower RNA counts in variant carriers, strong SPINK5 localisation in outer ear skin, faint localisation to middle ear mucosa and eardrum and increased SPINK5 expression in human cholesteatoma. CONCLUSION: SPINK5 variants confer susceptibility to non-syndromic OM. These variants potentially contribute to middle ear pathology through breakdown of mucosal and epithelial barriers, immunodeficiency such as poor vaccination response, alteration of head and neck microbiota and facilitation of entry of opportunistic pathogens into the middle ear.

FUT2 Variants Confer Susceptibility to Familial Otitis Media.

Non-secretor status due to homozygosity for the common FUT2 variant c.461G>A (p.Trp154∗) is associated with either risk for autoimmune diseases or protection against viral diarrhea and HIV. We determined the role of FUT2 in otitis media susceptibility by obtaining DNA samples from 609 multi-ethnic families and simplex case subjects with otitis media. Exome and Sanger sequencing, linkage analysis, and Fisher exact and transmission disequilibrium tests (TDT) were performed. The common FUT2 c.604C>T (p.Arg202∗) variant co-segregates with otitis media in a Filipino pedigree (LOD = 4.0). Additionally, a rare variant, c.412C>T (p.Arg138Cys), is associated with recurrent/chronic otitis media in European-American children (p = 1.2 × 10-5) and US trios (TDT p = 0.01). The c.461G>A (p.Trp154∗) variant was also over-transmitted in US trios (TDT p = 0.01) and was associated with shifts in middle ear microbiota composition (PERMANOVA p < 10-7) and increased biodiversity. When all missense and nonsense variants identified in multi-ethnic US trios with CADD > 20 were combined, FUT2 variants were over-transmitted in trios (TDT p = 0.001). Fut2 is transiently upregulated in mouse middle ear after inoculation with non-typeable Haemophilus influenzae. Four FUT2 variants-namely p.Ala104Val, p.Arg138Cys, p.Trp154∗, and p.Arg202∗-reduced A antigen in mutant-transfected COS-7 cells, while the nonsense variants also reduced FUT2 protein levels. Common and rare FUT2 variants confer susceptibility to otitis media, likely by modifying the middle ear microbiome through regulation of A antigen levels in epithelial cells. Our families demonstrate marked intra-familial genetic heterogeneity, suggesting that multiple combinations of common and rare variants plus environmental factors influence the individual otitis media phenotype as a complex trait.

A2ML1 and otitis media: novel variants, differential expression, and relevant pathways.

A genetic basis for otitis media is established, however, the role of rare variants in disease etiology is largely unknown. Previously a duplication variant within A2ML1 was identified as a significant risk factor for otitis media in an indigenous Filipino population and in US children. In this report exome and Sanger sequencing was performed using DNA samples from the indigenous Filipino population, Filipino cochlear implantees, US probands, Finnish, and Pakistani families with otitis media. Sixteen novel, damaging A2ML1 variants identified in otitis media patients were rare or low-frequency in population-matched controls. In the indigenous population, both gingivitis and A2ML1 variants including the known duplication variant and the novel splice variant c.4061 + 1 G>C were independently associated with otitis media. Sequencing of salivary RNA samples from indigenous Filipinos demonstrated lower A2ML1 expression according to the carriage of A2ML1 variants. Sequencing of additional salivary RNA samples from US patients with otitis media revealed differentially expressed genes that are highly correlated with A2ML1 expression levels. In particular, RND3 is upregulated in both A2ML1 variant carriers and high-A2ML1 expressors. These findings support a role for A2ML1 in keratinocyte differentiation within the middle ear as part of otitis media pathology and the potential application of ROCK inhibition in otitis media.

Assessing ChatGPT's Competency in Addressing Interdisciplinary Inquiries on Chatbot Uses in Sports Rehabilitation: Simulation Study.

BACKGROUND: ChatGPT showcases exceptional conversational capabilities and extensive cross-disciplinary knowledge. In addition, it can perform multiple roles in a single chat session. This unique multirole-playing feature positions ChatGPT as a promising tool for exploring interdisciplinary subjects. OBJECTIVE: The aim of this study was to evaluate ChatGPT's competency in addressing interdisciplinary inquiries based on a case study exploring the opportunities and challenges of chatbot uses in sports rehabilitation. METHODS: We developed a model termed PanelGPT to assess ChatGPT's competency in addressing interdisciplinary topics through simulated panel discussions. Taking chatbot uses in sports rehabilitation as an example of an interdisciplinary topic, we prompted ChatGPT through PanelGPT to role-play a physiotherapist, psychologist, nutritionist, artificial intelligence expert, and athlete in a simulated panel discussion. During the simulation, we posed questions to the panel while ChatGPT acted as both the panelists for responses and the moderator for steering the discussion. We performed the simulation using ChatGPT-4 and evaluated the responses by referring to the literature and our human expertise. RESULTS: By tackling questions related to chatbot uses in sports rehabilitation with respect to patient education, physiotherapy, physiology, nutrition, and ethical considerations, responses from the ChatGPT-simulated panel discussion reasonably pointed to various benefits such as 24/7 support, personalized advice, automated tracking, and reminders. ChatGPT also correctly emphasized the importance of patient education, and identified challenges such as limited interaction modes, inaccuracies in emotion-related advice, assurance of data privacy and security, transparency in data handling, and fairness in model training. It also stressed that chatbots are to assist as a copilot, not to replace human health care professionals in the rehabilitation process. CONCLUSIONS: ChatGPT exhibits strong competency in addressing interdisciplinary inquiry by simulating multiple experts from complementary backgrounds, with significant implications in assisting medical education.

Interdisciplinary Inquiry via PanelGPT: Application to Explore Chatbot Application in Sports Rehabilitation.

Background: ChatGPT showcases exceptional conversational capabilities and extensive cross-disciplinary knowledge. In addition, it possesses the ability to perform multiple roles within a single chat session. This unique multi-role-playing feature positions ChatGPT as a promising tool to explore interdisciplinary subjects. Objective: The study intended to guide ChatGPT for interdisciplinary exploration through simulated panel discussions. As a proof-of-concept, we employed this method to evaluate the advantages and challenges of using chatbots in sports rehabilitation. Methods: We proposed a model termed PanelGPT to explore ChatGPTs' knowledge graph on interdisciplinary topics through simulated panel discussions. Applied to "chatbots in sports rehabilitation", ChatGPT role-played both the moderator and panelists, which included a physiotherapist, psychologist, nutritionist, AI expert, and an athlete. We act as the audience posed questions to the panel, with ChatGPT acting as both the panelists for responses and the moderator for hosting the discussion. We performed the simulation using the ChatGPT-4 model and evaluated the responses with existing literature and human expertise. Results: Each simulation mimicked a real-life panel discussion: The moderator introduced the panel and posed opening/closing questions, to which all panelists responded. The experts engaged with each other to address inquiries from the audience, primarily from their respective fields of expertise. By tackling questions related to education, physiotherapy, physiology, nutrition, and ethical consideration, the discussion highlighted benefits such as 24/7 support, personalized advice, automated tracking, and reminders. It also emphasized the importance of user education and identified challenges such as limited interaction modes, inaccuracies in emotion-related advice, assurance on data privacy and security, transparency in data handling, and fairness in model training. The panelists reached a consensus that chatbots are designed to assist, not replace, human healthcare professionals in the rehabilitation process. Conclusions: Compared to a typical conversation with ChatGPT, the multi-perspective approach of PanelGPT facilitates a comprehensive understanding of an interdisciplinary topic by integrating insights from experts with complementary knowledge. Beyond addressing the exemplified topic of chatbots in sports rehabilitation, the model can be adapted to tackle a wide array of interdisciplinary topics within educational, research, and healthcare settings.

Audiologic Measures in an Indigenous Community with A2ML1- and FUT2-Related Otitis Media.

Background: Many indigenous peoples are at elevated risk for otitis media, however there is limited information on hearing loss due to OM in these communities. An Indigenous Filipino community that has previously been described with an elevated prevalence of OM that is due to rare A2ML1 variants and a common FUT2 variant underwent additional phenological testing. In this study, we describe the audiologic profiles in A2ML1- and FUT2-related otitis media and the validity of otoscopy and genotyping for A2ML1 and FUT2 variants in screening for otitis media and hearing loss. Method: We analyzed A2ML1 and FUT2 genotypes together with demographic, otologic and audiologic data from tympanometry and hearing level assessments of 109 indigenous individuals. Results: We confirmed previous findings of a spectrum of nonsyndromic otitis media as associated with A2ML1 variants. A2ML1 and FUT2 variants were associated with high-frequency hearing loss at 4000 Hz. As expected, young age was associated with flat tympanograms, and eardrum perforations due to chronic otitis media were associated with severe-to-profound hearing loss across frequencies. Adding A2ML1 or FUT2 genotypes improved the validity of otoscopy as a screening test to rule out moderate-to-profound hearing loss. Conclusion: Continued multi-disciplinary management and audiologic follow-up using tympanometry and screening audiometry are needed to document and treat otitis media and prevent permanent hearing loss in the indigenous community.

Lack of Methylation Changes in GJB2 and RB1 Non-coding Regions of Cochlear Implant Patients with Sensorineural Hearing Loss.

Objective: Recent advances in epigenetic studies continue to reveal novel mechanisms of gene regulation and control, however little is known on the role of epigenetics in sensorineural hearing loss (SNHL) in humans. We aimed to investigate the methylation patterns of two regions, one in RB1 and another in GJB2 in Filipino patients with SNHL compared to hearing control individuals. Methods: We investigated an RB1 promoter region that was previously identified as differentially methylated in children with SNHL and lead exposure. Additionally, we investigated a sequence in an enhancer-like region within GJB2 that contains four CpGs in close proximity. Bisulfite conversion was performed on salivary DNA samples from 15 children with SNHL and 45 unrelated ethnically-matched individuals. We then performed methylation-specific real-time PCR analysis (qMSP) using TaqMan® probes to determine percentage methylation of the two regions. Results: Using qMSP, both our cases and controls had zero methylation at the targeted GJB2 and RB1 regions. Conclusion: Our study showed no changes in methylation at the selected CpG regions in RB1 and GJB2 in the two comparison groups with or without SNHL. This may be due to a lack of environmental exposures to these target regions. Other epigenetic marks may be present around these regions as well as those of other HL-associated genes.

Identification of Novel Candidate Genes and Variants for Hearing Loss and Temporal Bone Anomalies.

Background: Hearing loss remains an important global health problem that is potentially addressed through early identification of a genetic etiology, which helps to predict outcomes of hearing rehabilitation such as cochlear implantation and also to mitigate the long-term effects of comorbidities. The identification of variants for hearing loss and detailed descriptions of clinical phenotypes in patients from various populations are needed to improve the utility of clinical genetic screening for hearing loss. Methods: Clinical and exome data from 15 children with hearing loss were reviewed. Standard tools for annotating variants were used and rare, putatively deleterious variants were selected from the exome data. Results: In 15 children, 21 rare damaging variants in 17 genes were identified, including: 14 known hearing loss or neurodevelopmental genes, 11 of which had novel variants; and three candidate genes IST1, CBLN3 and GDPD5, two of which were identified in children with both hearing loss and enlarged vestibular aqueducts. Patients with variants within IST1 and MYO18B had poorer outcomes after cochlear implantation. Conclusion: Our findings highlight the importance of identifying novel variants and genes in ethnic groups that are understudied for hearing loss.

Impact of low-dose heparin on duration of peripherally inserted central catheter at the Neonatal Intensive Care Unit: A meta-analysis

OBJECTIVES@#To determine efficacy of continuous heparin infusion vs placebo on maintenance of peripherally inserted central catheter line among neonates admitted at the NICU.@*METHODS@#This is a meta-analysis of randomized controlled trials reported in accordance with PRISMA checklist. Cochrane Risk-of-bias tool was used in assessment of reporting biases. Pooled risk ratios were estimated using random- or fixed-effects model.@*RESULTS@#Of 4519 studies identified, 4 studies were included, and all have low risk of bias. Meta-analysis showed that continuous heparin infusion on PICCs had significantly higher duration of catheter patency compared to the placebo group (MD=2.22, 95%CI=1.03-3.14, pvalue<0.00001). Heparin group also had decreased risk of occlusion (RR=0.47, 95%CI=0.94, pvalues=0.03) compared to control. The risk for other adverse events such as thrombosis, infection, IVH progression, and mortality was comparable between the two groups. @*CONCLUSION@#Continuous heparin infusion in PICC fluids can prolong duration of catheter patency by 2.2 days and reduce risk of catheter-related occlusion by 50%, without having significant effect on incidence of other adverse events.@*RECOMMENDATIONS@#Continuous heparin infusion on PICC fluids should be part of maintenance and care policy at the NICU, but precautions should be followed to prevent adverse outcomes. Systematic review of intermittent heparin flushing can be a window of opportunity.

Genetic and Environmental Determinants of Otitis Media in an Indigenous Filipino Population.

OBJECTIVE: To identify genetic and environmental risk factors for otitis media in an indigenous Filipino population. STUDY DESIGN: Cross-sectional study. SETTING: Indigenous Filipino community. SUBJECTS AND METHODS: Clinical history and information on breastfeeding, tobacco smoke exposure, and swimming were obtained from community members. Heads of households were interviewed for family history and personal beliefs on ear health. Height and weight were measured. Otoscopic findings were described for the presence and character of perforation or discharge. An A2ML1 duplication variant that confers otitis media susceptibility was Sanger sequenced in all DNA samples. Co-occurrence of middle ear bacteria detected by 16S rRNA gene sequencing was determined according to A2ML1 genotype and social cluster. RESULTS: The indigenous Filipino population has a ~50% prevalence of otitis media. Young age was associated with otitis media (4 age strata; P = .004); however, age was nonsignificant as a bistratal or continuous variable. There was no association between otitis media and sex, body mass index, breastfeeding, tobacco exposure, or deep swimming. In multivariate analyses, A2ML1 genotype is the strongest predictor of otitis media, with an odds ratio of 3.7 (95% confidence interval: 1.3-10.8; P = .005). When otitis media diagnoses were plotted across ages, otitis media was observed within the first year of life, and chronic otitis media persisted up to adulthood, particularly in A2ML1-variant carriers. CONCLUSION: Among indigenous Filipinos, A2ML1 genotype is the primary risk factor for otitis media and main determinant of disease progression, although age, the middle ear microbiome, and social clusters might modulate the effect of the A2ML1 genotype.

Middle ear microbiome differences in indigenous Filipinos with chronic otitis media due to a duplication in the A2ML1 gene.

BACKGROUND: Previously rare A2ML1 variants were identified to confer otitis media susceptibility in an indigenous Filipino community and in otitis-prone US children. The goal of this study is to describe differences in the middle ear microbiome between carriers and non-carriers of an A2ML1 duplication variant that increases risk for chronic otitis media among indigenous Filipinos with poor health care access. METHODS: Ear swabs were obtained from 16 indigenous Filipino individuals with chronic otitis media, of whom 11 carry the A2ML1 duplication variant. Ear swabs were submitted for 16S rRNA gene sequencing. RESULTS: Genotype-based differences in microbial richness, structure, and composition were identified, but were not statistically significant. Taxonomic analysis revealed that the relative abundance of the phyla Fusobacteria and Bacteroidetes, and genus Fusobacterium were nominally increased in carriers compared to non-carriers, but were non-significant after correction for multiple testing. We also detected rare bacteria including Oligella that was reported only once in the middle ear. CONCLUSIONS: These findings suggest that A2ML1-related otitis media susceptibility may be mediated by changes in the middle ear microbiome. Knowledge of middle ear microbial profiles according to genetic background can be potentially useful for therapeutic and prophylactic interventions for otitis media and can guide public health interventions towards decreasing otitis media prevalence within the indigenous Filipino community.

Rare A2ML1 variants confer susceptibility to otitis media.

A duplication variant within the middle ear-specific gene A2ML1 cosegregates with otitis media in an indigenous Filipino pedigree (LOD score = 7.5 at reduced penetrance) and lies within a founder haplotype that is also shared by 3 otitis-prone European-American and Hispanic-American children but is absent in non-otitis-prone children and >62,000 next-generation sequences. We identified seven additional A2ML1 variants in six otitis-prone children. Collectively, our studies support a role for A2ML1 in the pathophysiology of otitis media.

Lack of methylation changes in GJB2 and RB1 non-coding regions of cochlear implant patients with sensorineural hearing loss

Objective@#Recent advances in epigenetic studies continue to reveal novel mechanisms of gene regulation and control, however little is known on the role of epigenetics in sensorineural hearing loss (SNHL) in humans. We aimed to investigate the methylation patterns of two regions, one in RB1 and another in GJB2 in Filipino patients with SNHL compared to hearing control individuals. @*Methods@#We investigated an RB1 promoter region that was previously identified as differentially methylated in children with SNHL and lead exposure. Additionally, we investigated a sequence in an enhancer-like region within GJB2 that contains four CpGs in close proximity. Bisulfite conversion was performed on salivary DNA samples from 15 children with SNHL and 45 unrelated ethnically-matched individuals. We then performed methylation-specific real-time PCR analysis (qMSP) using TaqMan® probes to determine percentage methylation of the two regions. @*Results@#Using qMSP, both our cases and controls had zero methylation at the targeted GJB2 and RB1 regions. @*Conclusion@#Our study showed no changes in methylation at the selected CpG regions in RB1 and GJB2 in the two comparison groups with or without SNHL. This may be due to a lack of environmental exposures to these target regions. Other epigenetic marks may be present around these regions as well as those of other HL-associated genes.

Otoscopic and audiologic findings in an ati community in Boracay

Background: Certain indigenous populations have been noted by the World Health Organization (WHO) to havethe highest prevalence ratesforchronicsuppurativeotitis media (CSOM), including the Australian Aborigines (28-43%), Greenlanders (2-10%) and Alaskan Eskimos (2-10%). Objectives: To determine the prevalence of common ear problems, particularly CSOM, among the indigenous Ati or Aeta community in Bolabog, Boracay, and to determine their hearing sensitivity using screening audiometry. Methods: Study Design - Descriptive cross-sectional study. Setting - A small Ati community in Bolabog, Boracay. Population - A total of 63 adults and children underwent medical interview and otoscopy. Additionally 24 had their hearing screened by audiometry. Results: About a quarter of the population participated in the study, including 41 children (40 percent of all children) and 22 adults (18 percent of all adults). Forty-six percent of children and 23 percent of adults who were examined had previous history of ear discharge, while 22 percent of children and 45 percent of adults who were examined had history of hearing loss. Seventeen percent of children had history of hearing loss in the family. CSOM was found in 18 (43.90 percent) children and 8 (36.36 percent) adults. Impacted cerumen was found in 17.1 percent of children. Eleven female children underwent screening audiometry. Of these, eight had normal hearing and three had abnormal findings. Thirteen adults were also tested, five of whom were male and had normal hearing bilaterally. Four of eight female adults had abnormal hearing, of which three were unilateral. Conclusions: The Ati population in Bolabog, Boracay belongs to a group with the highest prevalence rates for CSOM (27.0 percent). A bigger sample for screening audiometry is required for proper estimation of hearing loss prevalence. Both environmental and genetic factors may have increased the prevalence of CSOM in the Ati population of Boracay. (Author)

Accuracy of Siemens hearcheck™ navigator as a screening tool for hearing loss

Objective@#To calculate the accuracy, sensitivity, specificity and positive predictive values of the Siemens HearCheck™ Navigator in detecting hearing loss and to compare values of these parameters when the examination is done in a soundproof booth and in a quiet room.@*Methods@#Design: Analytical, cross-sectional study Setting: Tertiary Public University Hospital Patients: Patients seen at the Ear Unit of a tertiary public university hospital from June 2009 to August 2010 were tested using the Siemens HearCheck™ Navigator and pure tone audiometry, inside a soundproof audiometry booth and in a quiet room with an ambient noise of 50dB, with a different investigator for each examination. Each ear was treated as a separate subject. Results obtained from the HearCheck™ Navigator were designated as observed values and were classified as “no hearing loss” for green light, and “with hearing loss” for yellow or red lights. Results were compared with pure tone air conduction averages designated as gold standard values. Normal hearing acuity (0-25 dB) was classified as no hearing loss. Pure tone air conduction averages of 26dB and above were classified as “with hearing loss” and were further stratified as mild hearing loss (26-40dB) and moderate or worse hearing loss (>41 dB). Observed and gold standard values were compared and tabulated in a 2x2 table for all levels of hearing loss, mild hearing loss, and moderate or worse hearing loss. Accuracy, sensitivity, specificity, positive and negative predictive values of the Siemens HearCheck™ Navigator inside a soundproof audiometry booth and in a quiet room were determined using pure tone audiometry as the gold standard.@*Results@#100 patients (200 ears) were tested, with a median age of 43 years old (range 15-75), and an almost equal number of male and female participants (52 males, 48 females). Accuracy rate of the Siemens HearCheck™ Navigator inside the soundproof audiometry booth and in a quiet room were 82.5% and 84% respectively for all levels of hearing loss. Sensitivity, specificity, positive and negative predictive values were similar whether the examination was done inside the soundproof audiometry booth or in a quiet room. These values were notably higher in patients with moderate or worse hearing loss compared to patients with mild hearing loss.@*Conclusion@#The Siemens HearCheck™ Navigator shows potential as an accurate, portable, easy-to-use tool to screen for hearing loss, especially for cases of moderate or worse hearing loss, without the need for soundproof audiometry booths or special training. It is recommended that further studies be done to differentiate degrees of hearing loss, and to evaluate its usefulness in other target populations, including school children and the elderly.