Breast cancers in monoallelic MUTYH germline mutation carriers have clinicopathological features overlapping with those in BRCA1 germline mutation carriers.

PURPOSE: Breast cancer patients referred to genetic counseling often undergo genetic testing with broad panels that include both breast cancer susceptibility genes as well as genes more specific for extramammary sites. As a result, patients are often incidentally found to have germline mutations in genes that are not necessarily related to breast cancer risk. One such gene is MUTYH. To understand the role MUTYH may play in breast cancer, the clinicopathological features of patients with monoallelic MUTYH germline mutation and breast cancer were examined. METHODS: The clinicopathological characteristics of the breast cancers from patients with monoallelic MUTYH mutation were compared to breast cancer patients with other germline mutations in known breast cancer susceptibility genes, including ATM, BRCA1/2, CHEK2, and PALB2. The breast cancer patients who received genetic counseling but tested negative for the aforementioned gene mutations were used as a control group. RESULTS: Histologic characteristics of the breast cancers arising in monoallelic MUTYH mutation carriers had significantly larger tumor size, higher tumor grade, and more high-risk biomarker profiles (i.e., Her2-positive and triple-negative) than breast cancer patients with susceptibility genes, except for BRCA1. MUTYH mutation carriers also showed a trend of more frequent intratumoral divergency in terms of tumor grade and biomarker profiles. CONCLUSION: Although germline monoallelic MUTYH mutation is not thought to confer a meaningfully increased risk of breast cancer development, it may contribute to pathological aggressiveness and diversity of breast cancers when they sporadically arise in MUTYH carriers.

Fabrication of waveguide directional couplers using 2-photon lithography.

Advances in 2-photon lithography have enabled in-lab production of sub-micron resolution and millimeter scale 3D optical components. The potential complex geometries are well suited to rapid prototyping and production of waveguide structures, interconnects, and waveguide directional couplers, furthering future development and miniaturization of waveguide-based imaging technologies. System alignment is inherent to the 2-photon process, obviating the need for manual assembly and allowing precise micron scale waveguide geometries not possible in traditional fused fiber coupler fabrication. Here we present the use of 2-photon lithography for direct printing of multi-mode waveguide couplers with air cladding and single mode waveguide couplers with uncured liquid photoresin cladding. Experimental results show reproducible coupling which can be modified by selected design parameters.

High-spatial density snapshot imaging spectrometer enabled by 2-photon fabricated custom fiber bundles.

We report on a proof-of-concept snapshot imaging spectrometer developed using an array of optical fibers fabricated with 2-photon polymerization (2PP). The dense input array maps to an output array with engineered void spaces for spectral information. Previously, the development and fabrication of custom fiber arrays for imaging spectrometers have been a complex, time-consuming, and costly process, requiring a semi-manual assembly of commercial components. This work applies an automatic development process based on 2PP additive manufacturing with the Nanoscribe GmbH Quantum X system. The technique allows printing of arbitrary optical quality structures with submicron resolution with less than 5 nm roughness, enabling small core fibers/integrated arrays. Specifically, we developed an array prototype of 40 × 80 with 6-micron pitch at the input and 80-micron pitch at the output. The air-clad fibers had a core diameter of 5 µm. Fabricated optical fiber arrays were incorporated into a prism-based imaging spectrometer system with 48 spectral channels to demonstrate multi-spectral imaging. Imaging of a USAF target and color printed letter C as well as spectral comparisons to a commercial spectrometer were used to validate the performance of the system. These results clearly demonstrate the functionality and potential applications of the 3D-printed fiber-based snapshot imaging spectrometer.

Ruggedized, field-ready snapshot light-guide-based imaging spectrometer for environmental and remote sensing applications.

A field-ready, fiber-based high spatial sampling snapshot imaging spectrometer was developed for applications such as environmental monitoring and smart farming. The system achieves video rate frame transfer and exposure times down to a few hundred microseconds in typical daylight conditions with ∼63,000 spatial points and 32 spectral channels across the 470nm to 700nm wavelength range. We designed portable, ruggedized opto-mechanics to allow for imaging from an airborne platform. To ensure successful data collection prior to flight, imaging speed and signal-to-noise ratio was characterized for imaging a variety of land covers from the air. The system was validated by performing a series of observations including: Liriope Muscari plants under a range of water-stress conditions in a controlled laboratory experiment and field observations of sorghum plants in a variety of soil conditions. Finally, we collected data from a series of engineering flights and present reassembled images and spectral sampling of rural and urban landscapes.

HER2 Expression in Endometrial Cancers Diagnosed as Clear Cell Carcinoma.

There is increasing evidence that many endometrial cancers (EC) diagnosed as clear cell carcinoma (CCC) have substantial overlap with both serous carcinoma (SC) and endometrioid carcinoma (EmC), not only in terms of morphology and immunophenotype but also by molecular characterization. Now with use of HER2-based therapy in SC, a CCC diagnosis in serous-like tumors has the potential to exclude patients from receiving beneficial therapy. To assess HER2 in CCC in relation to other characteristics, a tissue microarray of archived CCC, EmC, and SC was stained for HER2 alongside a battery of immunostains used in EC. Cases with equivocal HER2 IHC were also assessed by in situ hybridization. HER2 status was assessed in 229 cases (23 CCC, 74 SC, 132 EmC). HER2 was positive in 48% of cases diagnosed as CCC, 19% of SC, and 0% of EmC. Rigorous morphologic and immunophenotypic review by 5 gynecologic pathologists revealed diagnostic disagreement in 8/11 HER2+ cases diagnosed as CCC, with SC as the other major diagnostic consideration. All HER2+ (n=25) cases were MMR-intact and most HER2+ EC had aberrant p53 staining (22/25, 88%); the 3 cases with a wild type pattern for p53 (12%) were all negative for ER. Based on these findings, patients with a diagnosis of CCC should be included in future clinical trials of HER2-targeted therapy. Moreover, given the diagnostic difficulty surrounding CCC, immunohistochemistry-based algorithms that include aberrant p53 and/or the absence of ER expression may provide a more objective means of establishing eligibility criteria than is currently possible using traditional histologic classification.

Radiometric and design model for the tunable light-guide image processing snapshot spectrometer (TuLIPSS).

The tunable light-guide image processing snapshot spectrometer (TuLIPSS) is a novel remote sensing instrument that can capture a spectral image cube in a single snapshot. The optical modelling application for the absolute signal intensity on a single pixel of the sensor in TuLIPSS has been developed through a numerical simulation of the integral performance of each optical element in the TuLIPSS system. The absolute spectral intensity of TuLIPSS can be determined either from the absolute irradiance of the observed surface or from the tabulated spectral reflectance of various land covers and by the application of a global irradiance approach. The model is validated through direct comparison of the simulated results with observations. Based on tabulated spectral reflectance, the deviation between the simulated results and the measured observations is less than 5% of the spectral light flux across most of the detection bandwidth for a Lambertian-like surface such as concrete. Additionally, the deviation between the simulated results and the measured observations using global irradiance information is less than 10% of the spectral light flux across most of the detection bandwidth for all surfaces tested. This optical modelling application of TuLIPSS can be used to assist the optimal design of the instrument and explore potential applications. The influence of the optical components on the light throughput is discussed with the optimal design being a compromise among the light throughput, spectral resolution, and cube size required by the specific application under consideration. The TuLIPSS modelling predicts that, for the current optimal low-cost configuration, the signal to noise ratio can exceed 10 at 10 ms exposure time, even for land covers with weak reflectance such as asphalt and water. Overall, this paper describes the process by which the optimal design is achieved for particular applications and directly connects the parameters of the optical components to the TuLIPSS performance.

High nuclear TPX2 expression correlates with TP53 mutation and poor clinical behavior in a large breast cancer cohort, but is not an independent predictor of chromosomal instability.

BACKGROUND: Targeting Protein for Xenopus Kinesin Like Protein 2 (TPX2) is a microtubule associated protein that functions in mitotic spindle assembly. TPX2 also localizes to the nucleus where it functions in DNA damage repair during S-phase. We and others have previously shown that TPX2 RNA levels are strongly associated with chromosomal instability (CIN) in breast and other cancers, and TPX2 RNA levels have been demonstrated to correlate with aggressive behavior and poor clinical outcome across a range of solid malignancies, including breast cancer. METHODS: We perform TPX2 IHC on a cohort of 253 primary breast cancers and adopt a clinically amenable scoring system to separate tumors into low, intermediate, or high TPX2 expression. We then correlate TPX2 expression against diverse pathologic parameters and important measures of clinical outcome, including disease-specific and overall survival. We link TPX2 expression to TP53 mutation and evaluate whether TPX2 is an independent predictor of chromosomal instability (CIN). RESULTS: We find that TPX2 nuclear expression strongly correlates with high grade morphology, elevated clinical stage, negative ER and PR status, and both disease-specific and overall survival. We also show that increased TPX2 nuclear expression correlates with elevated ploidy, supernumerary centrosomes, and TP53 mutation. TPX2 nuclear expression correlates with CIN via univariate analyses but is not independently predictive when compared to ploidy, Ki67, TP53 mutational status, centrosome number, and patient age. CONCLUSIONS: Our findings demonstrate a strong correlation between TPX2 nuclear expression and aggressive tumor behavior, and show that TPX2 overexpression frequently occurs in the setting of TP53 mutation and elevated ploidy. However, TPX2 expression is not an independent predictor of CIN where it fails to outperform existing clinical and pathologic metrics.

Expression of the Fusarium graminearum terpenome and involvement of the endoplasmic reticulum-derived toxisome.

The sesquiterpenoid deoxynivalenol (DON) is an important trichothecene mycotoxin produced by the cereal pathogen Fusarium graminearum. DON is synthesized in specialized subcellular structures called toxisomes. The first step in DON synthesis is catalyzed by the sesquiterpene synthase (STS), Tri5 (trichodiene synthase), resulting in the cyclization of farnesyl diphosphate (FPP) to produce the sesquiterpene trichodiene. Tri5 is one of eight putative STSs in the F. graminearum genome. To better understand the F. graminearum terpenome, the volatile and soluble fractions of fungal cultures were sampled. Stringent regulation of sesquiterpene accumulation was observed. When grown in trichothecene induction medium, the fungus produces trichothecenes as well as several volatile non-trichothecene related sesquiterpenes, whereas no volatile terpenes were detected when grown in non-inducing medium. Surprisingly, a Δtri5 deletion strain grown in inducing conditions not only ceased accumulation of trichothecenes, but also failed to produce the non-trichothecene related sesquiterpenes. To test whether Tri5 from F. graminearum may be a promiscuous STS directly producing all observed sesquiterpenes, Tri5 was cloned and expressed in E. coli and shown to produce primarily trichodiene in addition to minor, related cyclization products. Therefore, while Tri5 expression in F. graminearum is necessary for non-trichothecene sesquiterpene biosynthesis, direct catalysis by Tri5 does not explain the sesquiterpene deficient phenotype observed in the Δtri5 strain. To test whether Tri5 protein, separate from its enzymatic activity, may be required for non-trichothecene synthesis, the Tri5 locus was replaced with an enzymatically inactive, but structurally unaffected tri5N225D S229T allele. This allele restores non-trichothecene synthesis but not trichothecene synthesis. The tri5N225D S229T allele also restores toxisome structure which is lacking in the Δtri5 deletion strain. Our results indicate that the Tri5 protein, but not its enzymatic activity, is also required for the synthesis of non-trichothecene related sesquiterpenes and the formation of toxisomes. Toxisomes thus not only may be important for DON synthesis, but also for the synthesis of other sesquiterpene mycotoxins such as culmorin by F. graminearum.

Sesquiterpene Synthase-3-Hydroxy-3-Methylglutaryl Coenzyme A Synthase Fusion Protein Responsible for Hirsutene Biosynthesis in Stereum hirsutum.

The wood-rotting mushroom Stereum hirsutum is a known producer of a large number of namesake hirsutenoids, many with important bioactivities. Hirsutenoids form a structurally diverse and distinct class of sesquiterpenoids. No genes involved in hirsutenoid biosynthesis have yet been identified or their enzymes characterized. Here, we describe the cloning and functional characterization of a hirsutene synthase as an unexpected fusion protein of a sesquiterpene synthase (STS) with a C-terminal 3-hydroxy-3-methylglutaryl-coenzyme A (3-hydroxy-3-methylglutaryl-CoA) synthase (HMGS) domain. Both the full-length fusion protein and truncated STS domain are highly product-specific 1,11-cyclizing STS enzymes with kinetic properties typical of STSs. Complementation studies in Saccharomyces cerevisiae confirmed that the HMGS domain is also functional in vivo Phylogenetic analysis shows that the hirsutene synthase domain does not form a clade with other previously characterized sesquiterpene synthases from Basidiomycota. Comparative gene structure analysis of this hirsutene synthase with characterized fungal enzymes reveals a significantly higher intron density, suggesting that this enzyme may be acquired by horizontal gene transfer. In contrast, the HMGS domain is clearly related to other fungal homologs. This STS-HMGS fusion protein is part of a biosynthetic gene cluster that includes P450s and oxidases that are expressed and could be cloned from cDNA. Finally, this unusual fusion of a terpene synthase to an HMGS domain, which is not generally recognized as a key regulatory enzyme of the mevalonate isoprenoid precursor pathway, led to the identification of additional HMGS duplications in many fungal genomes, including the localization of HMGSs in other predicted sesquiterpenoid biosynthetic gene clusters.IMPORTANCE Hirsutenoids represent a structurally diverse class of bioactive sesquiterpenoids isolated from fungi. Identification of their biosynthetic pathways will provide access to this chemodiversity for the discovery and synthesis of molecules with new bioactivities. The identification and successful cloning of the previously elusive hirsutene synthase from the S. hirsutum provide important insights and strategies for biosynthetic gene discovery in Basidiomycota. The finding of a terpene synthase-HMGS fusion, the discovery of other sesquiterpenoid biosynthetic gene clusters with dedicated HMGS genes, and HMGS gene duplications in fungal genomes give new importance to the role of HMGS as a key regulatory enzyme in isoprenoid and sterol biosynthesis that should be exploited for metabolic engineering.

Programming characteristics of cochlear implants in children: effects of aetiology and age at implantation.

OBJECTIVE: We investigated effects of aetiology and age at implantation on changes in threshold (T) levels, comfortable (C) levels and dynamic range (DR) for cochlear implants (CIs) in children over the first five years of life. DESIGN: Information was collected at 6 months post-activation of CIs, and at 3 and 5 years of age. STUDY SAMPLE: One hundred and sixty-one children participating in the Longitudinal Outcomes of Children with Hearing Impairment (LOCHI) study. RESULTS: Children with neural and structural cochlear lesions had higher T-levels and C-levels as compared to those without these conditions. Parameter settings varied from manufacturer's defaults more often in the former than in the latter group. Investigation of the effect of age at implantation for children without neural and structural cochlear lesions showed that those implanted at ≤12 months of age had higher T-levels and narrower DR at 6 months post-activation, as compared to the later-implanted group. For both early- and later-implanted groups, the C-levels at 6 months post-activation were lower than those at age 3 and 5 years. There were no significant differences in T-levels, C-levels, or DR between age 3 and 5 years. CONCLUSIONS: Aetiology and age at implantation had significant effects on T-levels and C-levels.

Factors influencing speech perception in noise for 5-year-old children using hearing aids or cochlear implants.

OBJECTIVE: We investigated the factors influencing speech perception in babble for 5-year-old children with hearing loss who were using hearing aids (HAs) or cochlear implants (CIs). DESIGN: Speech reception thresholds (SRTs) for 50% correct identification were measured in two conditions - speech collocated with babble, and speech with spatially separated babble. The difference in SRTs between the two conditions give a measure of binaural unmasking, commonly known as spatial release from masking (SRM). Multiple linear regression analyses were conducted to examine the influence of a range of demographic factors on outcomes. STUDY SAMPLE: Participants were 252 children enrolled in the Longitudinal Outcomes of Children with Hearing Impairment (LOCHI) study. RESULTS: Children using HAs or CIs required a better signal-to-noise ratio to achieve the same level of performance as their normal-hearing peers but demonstrated SRM of a similar magnitude. For children using HAs, speech perception was significantly influenced by cognitive and language abilities. For children using CIs, age at CI activation and language ability were significant predictors of speech perception outcomes. CONCLUSIONS: Speech perception in children with hearing loss can be enhanced by improving their language abilities. Early age at cochlear implantation was also associated with better outcomes.

Hearing aid fitting and developmental outcomes of children fit according to either the NAL or DSL prescription: fit-to-target, audibility, speech and language abilities.

OBJECTIVE: This study examined the influence of prescription on hearing aid (HA) fitting characteristics and 5-year developmental outcomes of children. DESIGN: A randomised controlled trial implemented as part of a population-based study on Longitudinal Outcomes of Children with Hearing Impairment (LOCHI). STUDY SAMPLE: Two-hundred and thirty-two children that were fit according to either the National Acoustic Laboratories (NAL) or Desired Sensation Level (DSL) prescription. RESULTS: Deviation from targets and root-mean-square error in HA fitting revealed no significant difference between fitting prescriptions. Aided audibility quantified by using the Speech Intelligibility Index (SII) model showed that DSL provided higher audibility than NAL at low and medium input levels but not at high input level. After allowing for hearing loss desensitisation, differences in audibility between prescription groups were significant only at low input level. The randomised trial of prescription that was implemented for 163 children revealed no significant between-group differences in speech production, perception, and language; but parent-rated functional performance was higher for the DSL than for the NAL group. CONCLUSIONS: Proximity to prescriptive targets was similar between fitting prescriptions. The randomised trial revealed differences in aided audibility at low input level between prescription groups, but no significant differences in speech and language abilities.

Traversing the fungal terpenome.

Fungi (Ascomycota and Basidiomycota) are prolific producers of structurally diverse terpenoid compounds. Classes of terpenoids identified in fungi include the sesqui-, di- and triterpenoids. Biosynthetic pathways and enzymes to terpenoids from each of these classes have been described. These typically involve the scaffold generating terpene synthases and cyclases, and scaffold tailoring enzymes such as e.g. cytochrome P450 monoxygenases, NAD(P)+ and flavin dependent oxidoreductases, and various group transferases that generate the final bioactive structures. The biosynthesis of several sesquiterpenoid mycotoxins and bioactive diterpenoids has been well-studied in Ascomycota (e.g. filamentous fungi). Little is known about the terpenoid biosynthetic pathways in Basidiomycota (e.g. mushroom forming fungi), although they produce a huge diversity of terpenoid natural products. Specifically, many trans-humulyl cation derived sesquiterpenoid natural products with potent bioactivities have been isolated. Biosynthetic gene clusters responsible for the production of trans-humulyl cation derived protoilludanes, and other sesquiterpenoids, can be rapidly identified by genome sequencing and bioinformatic methods. Genome mining combined with heterologous biosynthetic pathway refactoring has the potential to facilitate discovery and production of pharmaceutically relevant fungal terpenoids.

Mushroom hunting by using bioinformatics: application of a predictive framework facilitates the selective identification of sesquiterpene synthases in basidiomycota.

The Basidiomycota fungi represent a diverse source of natural products, particularly the sesquiterpenoids. Recently, genome sequencing, genome mining, and the subsequent discovery of a suite of sesquiterpene synthases in Omphalotus olearius was described. A predictive framework was developed to facilitate the discovery of sesquiterpene synthases in Basidiomycota. Phylogenetic analyses indicated a conservation of both sequence and initial cyclization mechanisms used. Here, the first robust application of this predictive framework is reported. It was used to selectively identify sesquiterpene synthases that follow 1,6-, 1,10-, and 1,11-cyclization mechanisms in the crust fungus Stereum hirsutum. The successful identification and characterization of a 1,6- and a 1,10-cyclizing sesquiterpene synthase, as well as three 1,11-cyclizing Δ(6) -protoilludene synthases, is described. This study verifies the accuracy and utility of the predictive framework as a roadmap for the discovery of specific sesquiterpene synthases from Basidiomycota, and thus represents an important step forward in natural product discovery.

Outcomes of early- and late-identified children at 3 years of age: findings from a prospective population-based study.

OBJECTIVE: To address the question of whether, on a population level, early detection and amplification improve outcomes of children with hearing impairment. DESIGN: All families of children who were born between 2002 and 2007, and who presented for hearing services below 3 years of age at Australian Hearing pediatric centers in New South Wales, Victoria, and Southern Queensland were invited to participate in a prospective study on outcomes. Children's speech, language, functional, and social outcomes were assessed at 3 years of age, using a battery of age-appropriate tests. Demographic information relating to the child, family, and educational intervention was solicited through the use of custom-designed questionnaires. Audiological data were collected from the national database of Australian Hearing and records held at educational intervention agencies for children. Regression analysis was used to investigate the effects of each of 15 predictor variables, including age of amplification, on outcomes. RESULTS: Four hundred and fifty-one children enrolled in the study, 56% of whom received their first hearing aid fitting before 6 months of age. On the basis of clinical records, 44 children (10%) were diagnosed with auditory neuropathy spectrum disorder. There were 107 children (24%) reported to have additional disabilities. At 3 years of age, 317 children (70%) were hearing aid users and 134 children (30%) used cochlear implants. On the basis of parent reports, about 71% used an aural/oral mode of communication, and about 79% used English as the spoken language at home. Children's performance scores on standardized tests administered at 3 years of age were used in a factor analysis to derive a global development factor score. On average, the global score of hearing-impaired children was more than 1 SD below the mean of normal-hearing children at the same age. Regression analysis revealed that five factors, including female gender, absence of additional disabilities, less severe hearing loss, higher maternal education, and (for children with cochlear implants) earlier age of switch-on were associated with better outcomes at the 5% significance level. Whereas the effect of age of hearing aid fitting on child outcomes was weak, a younger age at cochlear implant switch-on was significantly associated with better outcomes for children with cochlear implants at 3 years of age. CONCLUSIONS: Fifty-six percent of the 451 children were fitted with hearing aids before 6 months of age. At 3 years of age, 134 children used cochlear implants and the remaining children used hearing aids. On average, outcomes were well below population norms. Significant predictors of child outcomes include: presence/absence of additional disabilities, severity of hearing loss, gender, maternal education, together with age of switch-on for children with cochlear implants.

A comparison of NAL and DSL prescriptive methods for paediatric hearing-aid fitting: predicted speech intelligibility and loudness.

OBJECTIVE: To examine the impact of prescription on predicted speech intelligibility and loudness for children. DESIGN: A between-group comparison of speech intelligibility index (SII) and loudness, based on hearing aids fitted according to NAL-NL1, DSL v4.1, or DSL m[i/o] prescriptions. A within-group comparison of gains prescribed by DSL m[i/o] and NAL-NL2 for children in terms of SII and loudness. STUDY SAMPLE: Participants were 200 children, who were randomly assigned to first hearing-aid fitting with either NAL-NL1, DSL v4.1, or DSL m[i/o]. Audiometric data and hearing-aid data at 3 years of age were used. RESULTS: On average, SII calculated on the basis of hearing-aid gains were higher for DSL than for NAL-NL1 at low input level, equivalent at medium input level, and higher for NAL-NL1 than DSL at high input level. Greater loudness was associated with DSL than with NAL-NL1, across a range of input levels. Comparing NAL-NL2 and DSL m[i/o] target gains revealed higher SII for the latter at low input level. SII was higher for NAL-NL2 than for DSL m[i/o] at medium- and high-input levels despite greater loudness for gains prescribed by DSL m[i/o] than by NAL-NL2. CONCLUSION: The choice of prescription has minimal effects on speech intelligibility predictions but marked effects on loudness predictions.

A randomized controlled comparison of NAL and DSL prescriptions for young children: hearing-aid characteristics and performance outcomes at three years of age.

OBJECTIVE: To determine the influence of choice of prescription and other child-, family- and intervention-related factors on speech, language, and functional performance of hearing-impaired children by three years of age. DESIGN AND STUDY SAMPLE: A randomized controlled design was implemented as part of a population-based, longitudinal study on outcomes of children with hearing impairment (LOCHI) in Australia. Two hundred and eighteen children were randomly assigned to either the NAL or the DSL prescription for first fitting of hearing aids. Their performance outcomes were evaluated. RESULTS: Prescriptive targets were closely matched in children's hearing aids. There were no significant differences in children's language, speech production, or functional performance between prescriptions. Parents' ratings of children's device usage and loudness discomfort were not significantly different between prescription groups. Functional performance within the first year of fitting together with degree of hearing loss, presence of additional disabilities, and maternal education explained 44% of variation in language ability of children by three years of age. CONCLUSIONS: There was no significant association between choice of hearing-aid prescription and variance in children's outcomes at three years of age. In contrast, additional disability, maternal educational level, and early functional performance were significant predictive factors of children's outcomes.

A randomized controlled trial of nonlinear frequency compression versus conventional processing in hearing aids: speech and language of children at three years of age.

OBJECTIVE: To determine the effect of nonlinear frequency compression (NLFC) on children's development of speech and language at three years of age. DESIGN: A randomized controlled trial was conducted as part of the population-based longitudinal study on outcomes of children with hearing impairment (LOCHI). Participants were randomly assigned to fitting with NLFC (Phonak Naida V SP or UP) or with conventional processing in hearing aids, prescribed by using either the NAL or the DSL formula. Standardized tests of speech production, receptive and expressive language were administered, and parent ratings were collected. All assessments were double-blinded. STUDY SAMPLE: Participants were 44 of the 450 children in the LOCHI cohort. RESULTS: Compared to children using conventional processing, receptive and expressive language was higher but receptive vocabulary and consonant articulation scores were lower for children who use NLFC. There was increased substitution of affricates by fricatives for children using NLFC, compared to children using conventional amplification. After allowing for the effect of multiple demographic variables, the difference in global language scores between groups was not significant (effect: 0.8 [95% confidence interval: - 6.7, 8.3]). CONCLUSIONS: There is insufficient evidence to indicate a difference in language ability between children using NLFC and those using conventional amplification.

Audiological characteristics of children with congenital unilateral hearing loss: insights into Age of reliable behavioural audiogram acquisition and change of hearing loss.

Objectives: The aims of this study were to report the audiological characteristics of children with congenital unilateral hearing loss (UHL), examine the age at which the first reliable behavioural audiograms can be obtained, and investigate hearing changes from diagnosis at birth to the first reliable behavioural audiogram. Method: This study included a sample of 91 children who were diagnosed with UHL via newborn hearing screening and had reliable behavioural audiograms before 7 years of age. Information about diagnosis, audiological characteristics and etiology were extracted from clinical reports. Regression analysis was used to explore the potential reasons influencing the age at which first reliable behavioural audiograms were obtained. Correlation and ANOVA analyses were conducted to examine changes in hearing at octave frequencies between 0.5 and 4 kHz. The proportions of hearing loss change, as well as the clinical characteristics of children with and without progressive hearing loss, were described according to two adopted definitions: Definition 1: criterion (1): a decrease in 10 dB or greater at two or more adjacent frequencies between 0.5 and 4 kHz, or criterion (2): a decrease in 15 dB or greater at one octave frequency in the same frequency range. Definition 2: a change of ≥20 dB in the average of pure-tone thresholds at 0.5, 1, and 2 kHz. Results: The study revealed that 48 children (52.7% of the sample of 91 children) had their first reliable behavioural audiogram by 3 years of age. The mean age at the first reliable behavioural audiogram was 3.0 years (SD 1.4; IQR: 1.8, 4.1). We found a significant association between children's behaviour and the presence or absence of ongoing middle ear issues in relation to the delay in obtaining a reliable behavioural audiogram. When comparing the hearing thresholds at diagnosis with the first reliable behavioural audiogram across different frequencies, it was observed that the majority of children experienced deterioration rather than improvement in the initial impaired ear at each frequency. Notably, there were more instances of hearing changes (either deterioration or improvement), in the 500 Hz and 1,000 Hz frequency ranges compared to the 2,000 Hz and 4,000 Hz ranges. Seventy-eight percent (n = 71) of children had hearing deterioration between the diagnosis and the first behavioural audiogram at one or more frequencies between 0.5 and 4 kHz, with a high proportion of them (52 out of the 71, 73.2%) developing severe to profound hearing loss. When using the averaged three frequency thresholds (i.e., definition 2), only 26.4% of children (n = 24) in the sample were identified as having hearing deterioration. Applying definition 2 therefore underestimates the proportion of children that experienced hearing changes. The study also reported diverse characteristics of children with or without hearing deterioration. Conclusion: The finding that 78% of children diagnosed with UHL at birth had a decrease in hearing loss between the hearing levels at first diagnosis and their first behavioural audiogram highlights the importance of monitoring hearing threshold levels after diagnosis, so that appropriate intervention can be implemented in a timely manner. For clinical management, deterioration of 15 dB at one or more frequencies that does not recover warrants action.

Stress Keratin 17 Is a Predictive Biomarker Inversely Associated with Response to Immune Check-Point Blockade in Head and Neck Squamous Cell Carcinomas and Beyond.

Low response rates in immune check-point blockade (ICB)-treated head and neck squamous cell carcinoma (HNSCC) drive a critical need for robust, clinically validated predictive biomarkers. Our group previously showed that stress keratin 17 (CK17) suppresses macrophage-mediated CXCL9/CXCL10 chemokine signaling involved in attracting activated CD8+ T cells into tumors, correlating with decreased response rate to pembrolizumab-based therapy in a pilot cohort of ICB-treated HNSCC (n = 26). Here, we performed an expanded analysis of the predictive value of CK17 in ICB-treated HNSCC according to the REMARK criteria and investigated the gene expression profiles associated with high CK17 expression. Pretreatment samples from pembrolizumab-treated HNSCC patients were stained via immunohistochemistry using a CK17 monoclonal antibody (n = 48) and subjected to spatial transcriptomic profiling (n = 8). Our findings were validated in an independent retrospective cohort (n = 22). CK17 RNA expression in pembrolizumab-treated patients with various cancer types was investigated for predictive significance. Of the 48 patients (60% male, median age of 61.5 years), 21 (44%) were CK17 high, and 27 (56%) were CK17 low. A total of 17 patients (35%, 77% CK17 low) had disease control, while 31 patients (65%, 45% CK17 low) had progressive disease. High CK17 expression was associated with a lack of disease control (p = 0.037), shorter time to treatment failure (p = 0.025), and progression-free survival (PFS, p = 0.004), but not overall survival (OS, p = 0.06). A high CK17 expression was associated with lack of disease control in an independent validation cohort (p = 0.011). PD-L1 expression did not correlate with CK17 expression or clinical outcome. CK17 RNA expression was predictive of PFS and OS in 552 pembrolizumab-treated cancer patients. Our findings indicate that high CK17 expression may predict resistance to ICB in HNSCC patients and beyond.