The role of the m6A/m demethylase FTO in memory is both task and sex-dependent in mice.

Formation of long-term memories requires learning-induced changes in both transcription and translation. Epitranscriptomic modifications of RNA recently emerged as critical regulators of RNA dynamics, whereby adenosine methylation (m6A) regulates translation, mRNA stability, mRNA localization, and memory formation. Prior work demonstrated a pro-memory phenotype of m6A, as loss of m6A impairs and loss of the m6A/m demethylase FTO improves memory formation. Critically, these experiments focused exclusively on aversive memory tasks and were only performed in male mice. Here we show that the task type and sex of the animal alter effects of m6A on memory, whereby FTO-depletion impaired object location memory in male mice, in contrast to the previously reported beneficial effects of FTO depletion on aversive memory. Additionally, we show that female mice have no change in performance after FTO depletion, demonstrating that sex of the mouse is a critical variable for understanding how m6A contributes to memory formation. Our study provides the first evidence for FTO regulation of non-aversive spatial memory and sexspecific effects of m6A, suggesting that identification of differentially methylated targets in each sex and task will be critical for understanding how epitranscriptomic modifications regulate memory.

Therapeutic strategies: Surgery for human papillomavirus-associated oropharyngeal carcinoma.

Treatment of oropharyngeal cancer (OPC) has undergone considerable evolution since the discovery of human papillomavirus (HPV)-associated OPC. It is widely understood that HPV OPC affects a younger population and standard treatment offers improved oncologic outcomes compared with non-HPV OPC but can cause significant toxicities and long-term side effects. Surgery for treatment de-escalation is an active area of research. The purpose of this review is to explore surgery as it relates to the treatment of HPV OPC with a focus on the evolution of treatment, rationale for surgery, surgical techniques, outcomes, and the role of surgery in de-escalation of treatment.

Role of the histone variant H2A.Z.1 in memory, transcription, and alternative splicing is mediated by lysine modification.

Creating long-lasting memories requires learning-induced changes in gene expression, which are impacted by epigenetic modifications of DNA and associated histone proteins. Post-translational modifications (PTMs) of histones are key regulators of transcription, with different PTMs producing unique effects on gene activity and behavior. Although recent studies implicate histone variants as novel regulators of memory, effects of PTMs on the function of histone variants are rarely considered. We previously showed that the histone variant H2A.Z suppresses memory, but it is unclear if this role is impacted by H2A.Z acetylation, a PTM that is typically associated with positive effects on transcription and memory. To answer this question, we used a mutation approach to manipulate acetylation on H2A.Z without impacting acetylation of other histone types. Specifically, we used adeno-associated virus (AAV) constructs to overexpress mutated H2A.Z.1 isoforms that either mimic acetylation (acetyl-mimic) by replacing lysines 4, 7 and 11 with glutamine (KQ), or H2A.Z.1 with impaired acetylation (acetyl-defective) by replacing the same lysines with alanine (KA). Expressing the H2A.Z.1 acetyl-mimic (H2A.Z.1KQ) improved memory under weak learning conditions, whereas expressing the acetyl-defective H2A.Z.1KA generally impaired memory, indicating that the effect of H2A.Z.1 on memory depends on its acetylation status. RNA sequencing showed that H2A.Z.1KQ and H2A.Z.1KA uniquely impact the expression of different classes of genes in both females and males. Specifically, H2A.Z.1KA preferentially impacts genes involved in synaptic function, suggesting that acetyl-defective H2A.Z.1 impairs memory by altering synaptic regulation. Finally, we describe, for the first time, that H2A.Z is also involved in alternative splicing of neuronal genes, whereby H2A.Z depletion, as well as expression of H2A.Z.1 lysine mutants influence transcription and splicing of different gene targets, suggesting that H2A.Z.1 can impact behavior through effects on both splicing and gene expression. This is the first study to demonstrate that direct manipulation of H2A.Z post-translational modifications regulates memory, whereby acetylation adds another regulatory layer by which histone variants can fine tune higher brain functions through effects on gene expression and splicing.

New home for tiny symbionts: dinophytes determined as Zooxanthella are Peridiniales and distantly related to Symbiodinium.

Endosymbiotic dinophytes are diverse and are found in a large variety of aquatic partners. They are colloquially coined zooxanthellae, and knowledge about those dinophytes with a coral as partner (i.e., Symbiodinium) is extensive. However, Zooxanthella nutricula has been specifically described based on material isolated from radiolarians, and its phylogenetic position within the dinophyte tree is unclear at present. We isolated genomic DNA and sequenced the ribosomal RNA genes from an endosymbiotic dinophyte found in a radiolarian. In phylogenetic analyses, the endosymbiont was distantly related to Symbiodinium and the Suessiales, but clustered together with members of the Peridiniales. Specifically, it was the sister lineage of a small group, whose members host a diatom as endosymbiont (i.e., the dinotoms). Endosymbiosis is thus of multiple origin within the dinophytes, and more research is necessary to work out ecological and morphological character traits that are congruent to the DNA trees.

Urinary and dialysate losses of vitamin D-binding protein in children on chronic peritoneal dialysis.

BACKGROUND: Vitamin D deficiency is widely prevalent in chronic kidney disease [CKD] patients. The aim of our study was to determine whether losses of vitamin D binding protein [VDBP] in urine and dialysate contribute to circulating 25-hydroxyvitamin D [25OHD] levels in chronic peritoneal dialysis [PD] patients. METHODS: Dialysate, serum, and urine VDBP levels were measured in 16 children on PD and compared with serum and urine VDBP in ten CKD4-5 patients. Serum VDBP levels were correlated with total circulating 25OHD and peritoneal VDBP losses. RESULTS: The mean age of the study population was 9.4 ± 3.8 years and the median time on dialysis 7.5 (1-18) months. In CKD4-5 patients, urinary VDBP losses were >300-fold higher than seen in age-matched healthy children and correlated with urinary albumin loss (p = 0.0008). There was a significant correlation between serum VDBP and total dialysate and urine losses of VDBP (p = 0.03, r = -0.53). Dialysate VDBP losses correlate with dialysate albumin loss (p = 0.01). VDBP losses in the long daytime dwell were higher than in the overnight drain (p = 0.04). Serum VDBP levels were lower in children with a longer dialysis vintage (p = 0.0004, r = -0.77). In PD patients, the mean total loss of VDBP in dialysate and urine was 1.91 ± 1.6 µmol/day, equivalent to ~7% of the total circulating level of VDBP in healthy controls. There was no correlation between 25(OH)D and VDBP. CONCLUSIONS: Peritoneal VDBP losses mirror both dialysate and urinary albumin losses, and are associated with a longer dialysis vintage but do not contribute to vitamin D deficiency in children on PD.

Use of telemedicine consultations in head and neck cancer: patient perceptions, acceptability and accessibility.

BACKGROUNDS: Telemedicine has been effective in the management of various medical conditions, however, there is limited knowledge of its use in head and neck oncosurveillance. This is of importance currently while trying to mitigate the risk of transmission during the COVID-19 pandemic. This study aims to evaluate acceptability, satisfaction and perceptions of telemedicine technology among outpatients for head and neck oncologic surveillance. METHODS: A cross-sectional study was conducted for head and neck surgical oncology patients who attended telemedicine consultations between March and October 2020 at the Peter MacCallum Cancer Centre. Data on demographic, socioeconomic and acceptability variables was collected, utilizing Likert scale questions. The primary outcome measures were patient satisfaction and perceptions, while the secondary outcome was access to technology. RESULTS: One hundred and fifteen patients were invited to participate, and 100 were included in the final analysis; 95% of patient's had a positive experience with telemedicine appointments and were willing to have future telemedicine appointments. Regional and rural patients were more accepting of telemedicine consultations, citing savings in travel time and the minimal disruption to normal day-to-day activities. All participants had access to telecommunication devices, with 63% having three or more devices. Issues identified include a lack of physical examination by clinician for 65% of participants and the inability to self-examine for 88% of participants. CONCLUSION: The study demonstrates patients' acceptance of telemedicine appointments as a component of outpatient surveillance for head and neck surgical oncology. This has benefits during the COVID-19 pandemic in addition to increasing accessibility for rural patients.

Use of Intraoperative Frozen Section to Assess Final Tumor Margin Status in Patients Undergoing Surgery for Oral Cavity Squamous Cell Carcinoma.

Importance: Methods of assessing final margin status in patients undergoing surgery for oral cavity squamous cell carcinoma, such as intraoperative frozen section histopathology (IFSH) taken from the tumor bed, may have limitations in accuracy. Objective: To evaluate the accuracy and implications of using IFSH samples to assess tumor bed margins in patients undergoing surgery for oral cavity squamous cell carcinoma (SCC). Design, Setting, and Participants: This cross-sectional study included 1257 patients who underwent surgery for oral cavity SCC between January 1, 2000, and December 31, 2015, at an academic cancer center. A total of 4821 IFSH samples were examined from 1104 patients (87.8%) who had at least 1 IFSH sample. Institutional practice is to harvest margins for IFSH from the tumor bed. Statistical analysis was performed from August 1, 2021, to April 4, 2022. Main Outcomes and Measures: Sensitivity and specificity were calculated for IFSH samples of margins compared with the permanent pathology samples of the same tissue and for IFSH compared with the final tumor specimen histopathology (FTSH). Results were classified using a binary method, with histopathologic reports interpreted as either negative (including negative or atypia or dysplasia) or positive (including carcinoma in situ, suspicious, or positive). Results: A total of 1257 patients met the inclusion criteria, including 709 men (56.4%), with a median age of 62 years (IQR, 52-73 years); 1104 patients (627 men [56.8%]; median age, 62 years [IQR, 52-72 years]) had IFHS samples. For IFSH relative to permanent sections of the IFSH tissue, sensitivity and specificity of IFSH were high (sensitivity, 76.5% [95% CI, 67.5%-85.5%]; specificity, 99.9% [95% CI, 99.8%-100%]), with discordant results in 24 of 4821 total specimens (0.5%). Final specimen margins were positive in 11.7% of patients (147 of 1257). Compared with FTSH, the sensitivity of IFSH for defining margin status was 10.8% (95% CI, 5.8%-15.8%), and the specificity was 99.1% (95% CI, 98.8%-99.4%). The rate of discordance was 4.0% (171 of 4284 specimens) between IFSH and FTSH. Conclusions and Relevance: The findings of this cross-sectional study suggest that IFSH is accurate compared with permanent pathologic characteristics of the same tissue, but less reliable at assessing final margin status on the tumor specimen. Despite a high specificity, the sensitivity of IFSH compared with FTSH is low, which may be associated with the inherent inability of tumor bed IFSH margin analysis to accurately account for the 3-dimensional association of tumor margins with the periphery of the specimen and the overall low rate of positive final tumor margins. Although tumor bed IFSH is widely used in the management of oral cavity cancer, this study suggests that there are limitations of this modality in assessing the final surgical margin status.

Histone macroH2A1 is a stronger regulator of hippocampal transcription and memory than macroH2A2 in mice.

Histone variants H2A.Z and H3.3 are epigenetic regulators of memory, but roles of other variants are not well characterized. macroH2A (mH2A) is a structurally unique histone that contains a globular macrodomain connected to the histone region by an unstructured linker. Here we assessed if mH2A regulates memory and if this role varies for the two mH2A-encoding genes, H2afy (mH2A1) and H2afy2 (mH2A2). We show that fear memory is impaired in mH2A1, but not in mH2A2-deficient mice, whereas both groups were impaired in a non-aversive spatial memory task. However, impairment was larger for mH2A1- deficient mice, indicating a preferential role for mH2A1 over mH2A2 in memory. Accordingly, mH2A1 depletion in the mouse hippocampus resulted in more extensive transcriptional de-repression compared to mH2A2 depletion. mH2A1-depleted mice failed to induce a normal transcriptional response to fear conditioning, suggesting that mH2A1 depletion impairs memory by altering transcription. Using chromatin immunoprecipitation (ChIP) sequencing, we found that both mH2A proteins are enriched on transcriptionally repressed genes, but only mH2A1 occupancy was dynamically modified during learning, displaying reduced occupancy on upregulated genes after training. These data identify mH2A as a regulator of memory and suggest that mH2A1 supports memory by repressing spurious transcription and promoting learning-induced transcriptional activation.

The Effect of Different Round Window Sealants on Cochlear Mechanics Over Time.

BACKGROUND: This project investigated the effects of round window membrane (RWM) sealants after surgical incision, with a focus on audiological thresholds, ossicular mechanics, and the impact upon cochlear function and pathology. METHODS: Twenty-eight guinea pigs were randomly allocated to one of three sealant groups (muscle, n = 7; fascia, n = 7, Tisseel, n = 8) or an unsealed control group (n = 6). Preoperative hearing was measured using auditory brainstem responses (ABRs). The ossicular chain and RWM were exposed surgically, and Laser Doppler Vibrometry (LDV) measurements were obtained from the long process of the incus. The RWM was incised then sealed (or left unsealed) according to group. ABR testing and LDV measurements were repeated 4 and 12 weeks after surgery. At 12 weeks all cochleae were harvested. RESULTS: ABR thresholds deteriorated over time in all groups. Overall, group was not statistically significant (p = 0.064). There was no significant effect by group on LDV measurements (p = 0.798). Histopathological analyses of the RWM showed that the fascia group had more extensive fibrosis than other groups (Independent-Samples Median Test, p = 0.001). However, there were minimal differences in the outer hair cell counts between the different intervention groups. CONCLUSIONS: All the interventions appeared to be safe while none affected the cochlear mechanics or hearing thresholds in a statistically significant manner.

The histone chaperone Anp32e regulates memory formation, transcription, and dendritic morphology by regulating steady-state H2A.Z binding in neurons.

Rapid removal of histone H2A.Z from neuronal chromatin is a key step in learning-induced gene expression and memory formation, but mechanisms underlying learning-induced H2A.Z removal are unclear. Anp32e was recently identified as an H2A.Z-specific histone chaperone that removes H2A.Z from nucleosomes in dividing cells, but its role in non-dividing neurons is unclear. Moreover, prior studies investigated Anp32e function under steady-state rather than stimulus-induced conditions. Here, we show that Anp32e regulates H2A.Z binding in neurons under steady-state conditions, with lesser impact on stimulus-induced H2A.Z removal. Functionally, Anp32e depletion leads to H2A.Z-dependent impairment in transcription and dendritic arborization in cultured hippocampal neurons, as well as impaired recall of contextual fear memory and transcriptional regulation. Together, these data indicate that Anp32e regulates behavioral and morphological outcomes by preventing H2A.Z accumulation in chromatin rather than by regulating activity-mediated H2A.Z dynamics.

Comparison of consumer information on the internet to the current evidence base for minimally invasive parathyroidectomy.

BACKGROUND: The Internet is increasingly used as a source of health information by patients. Under these circumstances, the opportunity exists for Internet sites ostensibly providing patient information to act to promote surgical referrals based on exaggerated claims. This study aims to assess quantitatively and qualitatively the Internet-based consumer health information for minimally invasive parathyroidectomy (MIP) techniques. METHODS: This is a prospective analysis of Internet web sites. Descriptive information about specific published claims on each of the web sites was documented and compared to the published evidence base. Web sites were then rated using a validated composite score (CS) tool and an MIP score tool developed specifically for the study. RESULTS: The search yielded 308 web sites, which, after assessment by the inclusion criteria left 44 unique web sites suitable for analysis. "Exaggerated," "misleading," or "false" claims were present in 27.3% of the web sites analyzed. The false claims category had a high negative item-total correlation with the overall score, and accuracy was found to have a statistically significant (p < 0.05) negative correlation with quality. However, analysis performed for country of origin and the organization responsible for the web site found no significant difference. CONCLUSIONS: Web sites offering information in relation to MIP have a surprisingly high rate of claims that are not in accord with the evidence. Such claims may be posted to attract surgical referrals. It is difficult for consumers to differentiate quality consumer health web sites from poor ones as there are no hard and fast rules to differentiate them.

Prophylactic dissection of level V in primary mucosal SCC in the clinically N positive neck: A systematic review.

OBJECTIVE: To review the evidence for level V dissection in the management of previously untreated mucosal squamous cell carcinoma (SCC) of the head and neck presenting with nodal metastasis when level V is clinically uninvolved. DATA SOURCE: The Preferred Reporting Items for Systematic Review and Meta-Analyses (PRISMA) were used to conduct a systematic review of the current literature, including all English language articles published after 1990. A literature search was performed on November 29, 2015, of Medline, EMBASE, Cumulative Index to Nursing and Allied Health Literature (CINAHL), and the Cochrane Library. REVIEW METHODS: The search yielded a total of 270 papers. Strict inclusion and exclusion criteria were applied, leaving 20 eligible papers. Overall prevalence was calculated using random effect meta-analysis. RESULTS: The overall prevalence of level V occult disease in the node (N)-positive neck, irrespective of subsite, was 2.56% (95% confidence interval 1.29-3.84) (2,368 patients and 2,533 necks). The prevalence of occult level V metastasis was up to 7.7% for oral cavity and 8.3% for oropharyngeal tumors. Five studies reported regional recurrence rates over variable time periods. There is exceedingly limited data on outcomes, such as spinal accessory nerve function, quality of life, and perioperative complications. CONCLUSION: Mucosal head and neck SCC presenting with nodal metastasis but with level V clinically uninvolved has a low prevalence of occult level V disease. Routine dissection of level V does not appear to be warranted; however, a definitive conclusion is unable to be drawn due to limited data on morbidity and oncological outcomes. Laryngoscope, 127:2074-2080, 2017.

The Sophono bone-conduction system: Surgical, audiologic, and quality-of-life outcomes.

We prospectively evaluated the surgical, audiologic, and quality-of-life outcomes in 5 patients-2 men and 3 women, aged 22 to 64 years (mean: 41.8)-who were implanted with the Sophono Alpha 2 MPO Processor. The indications for implantation of this bone-conduction device included recurrent ear canal infections with hearing aids (n = 3), single-sided deafness (n = 1), and patient preference in view of difficulty using a conventional hearing aid (n = 1). In addition to the patient with single-sided deafness, 3 patients had a bilateral mixed hearing loss and 1 had a bilateral conductive hearing loss. Outcomes measures included surgical complications, functional gain (FG), speech discrimination in quiet and noise, and patient satisfaction as determined by the Glasgow Benefit Inventory (GBI) and the Entific Medical Systems bone-anchored hearing aid questionnaire (BAQ). The only postsurgical complication noted was a minor skin reaction and pain in 1 patient that resolved with conservative management. In the 3 patients with the mixed hearing loss, the mean FG was 13.3, 20.0, 11.7, and 11.7 dB at 0.5, 1, 2, and 4 kHz, respectively; in the patient with the bilateral conductive hearing loss, the FG was 10, 25, 10, and 15 dB at the same frequencies. Speech discrimination scores with the Sophono device were comparable to those seen with conventional hearing aids. After implantation, all 5 patients experienced a positive quality-of-life outcome according to the GBI, although 1 of them had only a marginal improvement. On follow-up, all patients reported that they remained satisfied with their implant and that they used their device all day long. We conclude that the Sophono bone-conduction system is a safe and effective option that should be considered for patients with a mixed or conductive hearing loss who are unable to use a conventional hearing aid, as well as for those with single-sided deafness.

Concurrent primary and metastatic cutaneous head and neck squamous cell carcinoma: Analysis of prognostic factors.

BACKGROUND: There is a small subgroup of patients with head and neck cutaneous squamous cell carcinoma (cSCC) in which nodal metastases present concurrently with the primary lesion. There is evidence that these tumors may represent a more aggressive subset of cSCC. The aim of this study was to determine whether alternative clinicopathologic prognostic factors should be applied to this patient cohort. METHODS: A retrospective analysis of data from prospective databases of 2 large head and neck cancer units in Sydney, Australia, was performed. Ninety-five patients with concurrent primary and nodal metastatic head and neck cSCC were suitable for inclusion in the study. RESULTS: Univariable analysis was performed for overall survival (OS) and disease-specific survival (DSS). OS was adversely affected by immunosuppression (p = .011) and nodal extracapsular spread (ECS) (p = .006). Similarly, immunosuppression (p = .005) and ECS (p = .005) indicated a worse outcome for DSS. ECS and immunosuppression remained significant in the multivariable analysis. CONCLUSIONS: This study found that adverse prognostic factors were similar to the current evidence for nonconcurrent metastatic cSCC. In particular, the primary lesion had no significant influence on survival.

Changes in active ankle dorsiflexion range of motion after acute inversion ankle sprain.

CONTEXT: Posterior calf stretching is believed to improve active ankle dorsiflexion range of motion (AADFROM) after acute ankle-inversion sprain. OBJECTIVE: To describe AADFROM at baseline (postinjury) and at 2-wk time periods for 6 wk after acute inversion sprain. DESIGN: Randomized trial. SETTING: Sports clinic. PARTICIPANTS: 11 men and 11 women (age range 11-54 y) with acute inversion sprain. INTERVENTION: Standardized home exercise program for acute inversion sprain. MAIN OUTCOME MEASURE: AADFROM with the knee extended. RESULTS: Time main effect on AADFROM was significant (F3,57 = 108, P < .001). At baseline, mean active sagittal-plane motion of the ankle was 6 degrees of plantar flexion, whereas at 2, 4, and 6 wk AADFROM was 7 degrees, 11 degrees, and 11 degrees, respectively. CONCLUSIONS: AADFROM increased significantly from baseline to week 2 and from week 2 to week 4. Normal AADFROM was restored within 4 wk after acute inversion sprain.

Full-thickness burn formation after the use of electrical stimulation for rehabilitation of unicompartmental knee arthroplasty.

Electrical stimulation and interferential current are commonly used modalities in the physical rehabilitation of patients who have undergone joint arthroplasty surgery. Sparse data are available in the literature regarding potential complications from using these modalities. We report on a patient who underwent a unicompartmental knee arthroplasty with a subsequent full-thickness skin burn over the metal implant after electrical stimulation and interferential current modalities in physical therapy.