High-confidence 3D template matching for cryo-electron tomography.

Visual proteomics attempts to build atlases of the molecular content of cells but the automated annotation of cryo electron tomograms remains challenging. Template matching (TM) and methods based on machine learning detect structural signatures of macromolecules. However, their applicability remains limited in terms of both the abundance and size of the molecular targets. Here we show that the performance of TM is greatly improved by using template-specific search parameter optimization and by including higher-resolution information. We establish a TM pipeline with systematically tuned parameters for the automated, objective and comprehensive identification of structures with confidence 10 to 100-fold above the noise level. We demonstrate high-fidelity and high-confidence localizations of nuclear pore complexes, vaults, ribosomes, proteasomes, fatty acid synthases, lipid membranes and microtubules, and individual subunits inside crowded eukaryotic cells. We provide software tools for the generic implementation of our method that is broadly applicable towards realizing visual proteomics.

Effect of the German tonsillitis guideline on indication for tonsil surgery in patients with recurrent acute tonsillitis: a population-based study.

Evidence-based indication for tonsil surgery in patients with recurrent acute tonsillitis (RAT) is an ongoing matter of debate. Since introduction of the German tonsillitis guideline in 2015, the indication criteria for tonsil surgery have become much stricter. It is unclear, if this has changed the indication policy. A retrospective population-based study was performed including all 1398 patients with RAT admitted for tonsil surgery in all Thuringian hospitals in 2011, 2015, and 2019. Changes over the years concerning patients' characteristics, number of tonsillitis episodes in the last 12 months treated with antibiotics (T12), and decision for tonsillectomy or tonsillotomy were analyzed using univariable and multivariable statistics. The surgical rates decreased from 28.56/100,000 population in 2011 to 23.57 in 2015, and to 11.60 in 2019. The relative amount of patients with ≥ 6 T12 increased from 14.1% in 2011 over 13.3% in 2015 to 35.9% in 2019. Most patients received a tonsillectomy (98% of all surgeries). Decision for tonsillotomy was seldom (1.2%). Multinomial logistic regression analysis with the year 2011 as reference showed that compared to the year 2015, the age of the patients undergoing surgery increased in 2015 (Odds ratio [OR] = 1.024; 95% confidence interval [CI] = 1.014-1.034; p < 0.001), and also in 2019 (OR 1.030: CI 1.017-1.043; p < 0.001). Compared to 2011, the number T12 was not higher in 2015, but in 2019 (OR 1.273; CI 1.185-1.367; p < 0.001). Stricter rules led to lower tonsil surgery rates but to a higher proportion of patients with ≥ 6 T12 before surgery. Tonsillectomy remained the dominating technique.

Treatment of recurrent acute tonsillitis-a systematic review and clinical practice recommendations.

Background: There is an ongoing debate on the indications for tonsil surgery in both children and adults with recurrent acute tonsillitis. The aim is to provide practical recommendations for diagnostics and treatment for recurrent acute tonsillitis including evidence-based decision making for tonsillectomy. Methods: A systematic literature search in PubMed, Embase, Web of Science, and ScienceDirect from 2014 until April 2023 resulted in 68 articles. These were the basis for the review and a comprehensive series of consensus statements on the most important diagnostics and indications for both non-surgical and surgical therapy. A consensus paper was circulated among the authors and members of the International Head and Neck Scientific Group until a final agreement was reached for all recommendations. Results: The differentiation between sore throat and tonsillitis patient episodes is mostly not feasible and hence is not relevant for diagnostic decision making. Diagnostics of a tonsillitis/sore throat episode should always include a classification with a scoring system (Centor, McIssac, FeverPAIN score) to estimate the probability of a bacterial tonsillitis, mainly due to group A streptococcus (GAS). In ambiguous cases, a point-of-care test GAS swab test is helpful. Consecutive counting of the tonsillitis/sore throat episodes is important. In addition, a specific quality of life score (Tonsillectomy Outcome Inventory 14 or Tonsil and Adenoid Health Status Instrument) should be used for each episode. Conservative treatment includes a combination of paracetamol and/or non-steroidal anti-inflammatory drugs. In case of high probability of bacterial tonsillitis, and only in such cases, especially in patients at risk, standard antibiotic treatment is initiated directly or by delayed prescription. Tonsillectomy is indicated and is highly effective if the patient has had ≥7 adequately treated episodes in the preceding year, ≥5 such episodes in each of the preceding 2 years, or ≥3 such episodes in each of the preceding 3 years. An essential part of surgery is standardized pain management because severe postoperative pain can be expected in most patients. Conclusion: It is necessary to follow a stringent treatment algorithm for an optimal and evidence-based treatment for patients with recurrent acute tonsillitis. This will help decrease worldwide treatment variability, antibiotic overuse, and avoid ineffective tonsillectomy.

Age-related changes of the innate immune system of the palatine tonsil in a healthy cohort.

Although tonsillectomy is performed frequently, the role of palatine tonsils in life long immune protection or tolerance is still debated and the consequences of their removal for the immune system are of general interest. We analysed the tonsillar myeloid compartment in healthy subjects across a wide range of age (64% male; age range: 3 - 85 years) and compared its composition to the peripheral blood. We could observe a strong accumulation of all granulocyte subsets in the aging tonsil, which was most pronounced for basophils and mast cells. On functional level, an increase of CD163 and CD206 expression among monocytes and an increase of neutrophils expressing the inhibitory FcγRIIb correlated with increasing age. While the age-related shift of the leukocyte composition towards monocytes in blood is not reflected in tonsils, the increasing immunoregulatory phenotype of tonsilar monocytes is potentially counteracting the phenomenon of inflammaging at higher age.

Association between preoperative serum C-reactive protein level and leukocyte count and postoperative pain after otolaryngological surgery.

PURPOSE: To determine whether the preoperative inflammatory serum C-reactive protein (CRP) and leukocyte count (LEUK) are associated with postoperative pain and complaints after otolaryngological surgery. METHODS: Retrospective evaluation of 680 patients (33% female, median age 50 years) receiving otolaryngological surgery between November 2008 and March 2017 in a tertiary university hospital. Postoperative pain on the first postoperative day was assessed using the validated questionnaire of the German-wide project Quality Improvement in Postoperative Pain Treatment (QUIPS) including a numeric rating scale for assessment of postoperative pain (NRS, 0-10). The influence of preoperative parameters including CRP and LEUK on patients' postoperative pain was estimated. RESULTS: Mean CRP value was 15.6 ± 34.6 mg/l and mean LEUK value 7.8 ± 3.2 Gpt/l. Patients with pharyngeal surgery had the highest CRP values (34.6 ± 52.9 mg/l), highest LEUK values (9.2 ± 4.2 Gpt/l) and the highest pain levels (3.1 ± 2.4 NRS) compared to all other surgical procedures (all p < 0.05). Higher postoperative pain was associated with LEUK values > 11.3 Gpt/l (r = 0.093, p = 0.016) and higher preoperative chronic pain (r = 0.127, p = 0.001). Multivariate analysis confirmed younger age, female gender, duration of surgery, preoperative chronic pain, type of surgery, and higher LEUK values > 11.3 as independent factors for postoperative pain. Perioperative antibiotics had no effect on the postoperative pain. CONCLUSION: Beyond known factors, preoperative LEUK as inflammation marker is an independent predictor for pain on the first postoperative day.

High-resolution MRI of the human palatine tonsil and its schematic anatomic 3D reconstruction.

The palatine tonsils form an important part of the human immune system. Together with the other lymphoid tonsils of Waldeyer's tonsillar ring, they act as the first line of defense against ingested or inhaled pathogens. Although histologically stained sections of the palatine tonsil are widely available, they represent the tissue only in two dimensions and do not provide reference to three-dimensional space. Such a representation of a tonsillar specimen based on imaging data as a 3D anatomical reconstruction is lacking both in scientific publications and especially in textbooks. As a first step in this direction, the objective of the present work was to image a resected tonsil specimen with high spatial resolution in a 9.4 T small-bore pre-clinical MRI and to combine these data with data from the completely sectioned and H&E stained same palatine tonsil. Based on the information from both image modalities, a 3D anatomical sketch was drawn by a scientific graphic artist. In perspective, such studies could help to overcome the difficulty of capturing the spatial extent and arrangement of anatomical structures from 2D images and to establish a link between three-dimensional anatomical preparations and two-dimensional sections or illustrations, as they have been found so far in common textbooks and anatomical atlases.

30-day readmission rate in pediatric otorhinolaryngology inpatients: a retrospective population-based cohort study.

OBJECTIVES: Analysis of frequency and reasons for planned and unplanned 30-day readmission in hospitalized pediatric otorhinolaryngology patients using German Diagnosis Related Group (G-DRG) system data. METHODS: A retrospective population-based cohort study in Thuringia, Germany, was performed for the year 2015 with 2440 cases under 18 years (55.6% male) out of a total number of 15.271 inpatient cases. The majority of pediatric patients were from 2 to 5 years old (54.5%). The most frequent diagnoses were hyperplasia of adenoids or/and tonsils (26.6%). 36 cases (1.5%) experienced readmission within 30-days. RESULTS: 30-day readmission was planned in 9 cases (25% of all readmission) and was unplanned in 27 cases (75%). The median interval between index and readmission treatment was 8 days. Postoperative bleeding after adenoidectomy, tonsillotomy/tonsillectomy or tracheostomy (33.4%) and infectious complications after surgery like acute otitis media, abscess formation or fever (36.2%) were the most frequent reasons for 30-day readmission. Compared to adults treated in 2015 in Thuringia, the readmission rate was higher in adult patients (8.9%) than in this pediatric cohort. In contrast to children, readmissions in adults were mainly planned (65.1%) with a different spectrum of underlying diseases and reasons for readmission. CONCLUSION: The 30-day readmission rate seemed to be lower for pediatric otolaryngology patients compared to adult patients. Unplanned readmissions dominated in pediatric patients, whereas planned readmissions dominated in adults.

Tonsillectomy versus tonsillotomy for recurrent acute tonsillitis in children and adults (TOTO): study protocol for a randomized non-inferiority trial.

BACKGROUND: Tonsillectomy is one of the most frequently performed surgeries in children and young adults worldwide. For decades, tonsillectomy was the surgical treatment of choice for recurrent acute tonsillitis. Tonsillotomy was used in some countries as an alternative to tonsillectomy only for the treatment of obstructive sleep apnea in young children. In recent years, an increase of tonsillotomy also to treat recurrent acute tonsillitis can be observed. Therefore, the German Institute for Quality and Efficiency in Health Care (IQWiG) was commissioned by the Federal Joint Committee (G-BA) to investigate whether tonsillotomy offers advantages compared to tonsillectomy. The meta-analysis of the IQWiG including studies until 2016 revealed that the long-term benefits and harms of tonsillotomy compared to tonsillectomy are unclear. Consequently, the G-BA performed a European call for a clinical trial. A consortium of the German Professional Association of ENT-surgeons (BVHNO), the German Society of Oto-Rhino-Laryngology, Head and Neck Surgery (DGHNO-KHC), and the Jena University Hospital were finally selected to perform the TOTO study. METHODS: TOTO is a multicenter, 1:1 two-arm, randomized non-blinded non-inferiority trial. Four hundred fifty-four patients ≥ 3 years of age will be randomly allocated to undergo either tonsillotomy or tonsillectomy as surgical treatment of recurrent acute tonsillitis. All participants will be followed up for a total of 24 months. The primary outcome is the number of sore throat days experienced over the 24-month follow-up. DISCUSSION: TOTO is designed to evaluate the effectiveness and efficiency of tonsillectomy versus tonsillectomy for the management of patients with recurrent acute tonsillitis. Tonsil disease and surgery have a major impact on preschool and school children as well as on economically active young adults, with individual and societal costs through loss of school visits, earnings, and productivity. If tonsillotomy is at least as effective as tonsillectomy but with reduced morbidity, this would reduce costs to the healthcare system and society. TRIAL REGISTRATION: German Clinical Trials Register DRKS00020823 . Registered on 04 September 2020.

Enhanced differentiation of functional human T cells in NSGW41 mice with tissue-specific expression of human interleukin-7.

Humanized mouse models have become increasingly valuable tools to study human hematopoiesis and infectious diseases. However, human T-cell differentiation remains inefficient. We generated mice expressing human interleukin-7 (IL-7), a critical growth and survival factor for T cells, under the control of murine IL-7 regulatory elements. After transfer of human cord blood-derived hematopoietic stem and progenitor cells, transgenic mice on the NSGW41 background, termed NSGW41hIL7, showed elevated and prolonged human cellularity in the thymus while maintaining physiological ratios of thymocyte subsets. As a consequence, numbers of functional human T cells in the periphery were increased without evidence for pathological lymphoproliferation or aberrant expansion of effector or memory-like T cells. We conclude that the novel NSGW41hIL7 strain represents an optimized mouse model for humanization to better understand human T-cell differentiation in vivo and to generate a human immune system with a better approximation of human lymphocyte ratios.

A prospective cohort register-based study of chronic postsurgical pain and long-term use of pain medication after otorhinolaryngological surgery.

Data on chronic postsurgical pain (CPSP) after otorhinolaryngological surgery are sparse. Adult in-patients treated in 2017 were included into the prospective PAIN OUT registry. Patients' pain on the first postoperative day (D1), after six months (M6) and 12 months (M12) were evaluated. Determining factor for CPSP was an average pain intensity ≥ 3 (numeric rating scale 0-10) at M6. Risk factors associated with CPSP were evaluated by univariate and multivariate analyses. 10% of 191 included patients (60% male, median age: 52 years; maximal pain at D1: 3.5 ± 2.7), had CPSP. Average pain at M6 was 0.1 ± 0.5 for patients without CPSP and 4.2 ± 1.2 with CPSP. Average pain with CPSP still was 3.7 ± 1.1 at M12. Higher ASA status (Odds ratio [OR] = 4.052; 95% confidence interval [CI] = 1.453-11.189; p = 0.007), and higher minimal pain at D1 (OR = 1.721; CI = 1.189-2.492; p = 0.004) were independent predictors of CPSP at M6. Minimal pain at D1 (OR = 1.443; CI = 1.008-2.064; p = 0.045) and maximal pain at M6 (OR = 1.665; CI = 1.340-2.069; p < 0.001) were independent predictors for CPSP at M12. CPSP is an important issue after otorhinolaryngological surgery. Better instrument for perioperative assessment should be defined to identify patients at risk for CPSP.

Role of Body Dysmorphic Disorder in Patients With Postparalytic Facial Synkinesis.

OBJECTIVES/HYPOTHESIS: To evaluate the role of body dysmorphic disorder (BDD) in patients with postparalytic facial nerve syndrome with synkinesis (PFS). STUDY DESIGN: A single-center retrospective cohort study. METHODS: A total of 221 adults (74% women; median age: 44 years; median duration since onset of facial palsy: 1.6 years) were included. To diagnose BDD, the BDD Munich Module was used. Associations with House-Brackmann grading, Stennert index grading, Facial Clinimetric Evaluation (FaCE) survey, Facial Disability Index (FDI), general quality of life (SF-36), Beck Depression Inventory (BDI), and the Liebowitz Social Anxiety Scale (LSAS) was analyzed. RESULTS: A total of 59 patients (27%) were classified as patients with BDD. Significant associations were found between the diagnosis of BDD and female gender and lower FDI, FaCE, and SF-36 scores and higher BDI and LSAS scores. Multivariate analysis revealed BDI, FaCE total score, and FaCE social function subscore as independent factors associated with BDD. CONCLUSION: BDD was a relevant diagnosis in patients with PFS. A higher BDD level was associated with general and facial-specific quality of life and more psychosocial disabilities. Optimal treatment of PFS has to include these nonmotor dysfunctions. LEVEL OF EVIDENCE: 3 Laryngoscope, 131:E2518-E2524, 2021.

Effect of an Intensified Combined Electromyography and Visual Feedback Training on Facial Grading in Patients With Post-paralytic Facial Synkinesis.

Background: There is no current standard for facial synkinesis rehabilitation programs. The benefit and stability of effect of an intensified 10-day facial training combining electromyography and visual biofeedback training was evaluated. Methods: Fifty-four patients (77.8% female; median age: 49.5 years) with post-paralytic facial synkinesis (median time to onset of paralysis: 31.1 months) were included in retrospective longitudinal study between January 2013 and June 2016. Facial function was assesses at baseline (T0), first days of training (T1), last day of training (T2), and follow-up visit (T3) at a median time of 6 months later using the House-Brackmann (HB) facial nerve grading system, Stennert index (SI), Facial Nerve Grading System 2.0 (FNGS 2.0), and Sunnybrook Facial Grading System (SFGS). Pairwise comparisons between the time points with post-hoc Bonferroni correction were performed. Results: No significant changes of the gradings and subscores were seen between T0 and T1 (all p > 0.01). The 10-day combined and intensified feedback training between T1 and T2 improved facial symmetry and decreased synkinetic activity. Facial grading with the FNGS 2.0 or the SFGS were most suited to depict the training effect. FNGS 2.0, regional score, FNGS 2.0, synkinesis score, and FNGS 2.0 total score improved significantly (all p ≤ 0.0001). Both, the FNGS 2.0 and the SFGS showed the strongest improvement in the nasolabial fold/zygomatic and the oral region. Neither the age of the patient (r = 0.168; p = 0.224), the gender (r = 0.126; p = 0.363) nor the length of the interval between onset of the palsy and training start (r = 0.011; p = 0.886) correlated with the changes of the SFGS between T1 and T2. The results remained stable between T2 and T3 without any further significant change. Conclusion: Intensified daily combined electromyography and visual biofeedback training over 10 days was effective in patients with facial synkinesis and benefits were stable 6 months after therapy.

Non-idiopathic peripheral facial palsy: prognostic factors for outcome.

OBJECTIVES: There is a lack of data on patients' and diagnostic factors for prognostication of complete recovery in patients with non-idiopathic peripheral facial palsy (FP). METHODS: Cohort register-based study of 264 patients with non-idiopathic peripheral FP and uniform diagnostics and standardized treatment in a university hospital from 2007 to 2017 (47% female, median age: 57 years). Clinical data, facial grading, electrodiagnostics, motor function tests, non-motor function tests, and onset of prednisolone therapy were assessed for their impact on the probability of complete recovery using univariable and multivariable statistics. RESULTS: The most frequent reason for a non-idiopathic peripheral FP was a reactivation of Varicella Zoster Virus (VZV; 36.4%). Traumatic origin had a higher proportion of complete FP (52.9%). Furthermore, in traumatic FP, the mean interval between onset and start of prednisolone therapy was longer than in other cases (5.6 ± 6.2 days). Patients with reactivation of VZV, Lyme disease or otogenic FP had a significant higher recovery rate (p = 0.002, p < 0.0001, p = 0.018, respectively), whereas patients with post-surgery FP and other reasons had a significant lower recovery rate (p < 0.0001). After multivariate analyses voluntary activity in first EMG, Lyme disease and post-surgery cause were identified as independent diagnostic and prognostic factors on the probability of complete recovery (all p < 0.05). CONCLUSION: Infectious causes for non-idiopathic FP like VZV reactivation and Lyme disease had best probability for complete recovery. Post-surgery FP had a worse prognosis. LEVEL OF EVIDENCE: 2.

Ratings of Emotional Expressions in Static and Dynamic Facial Recordings Before and After Upper Face Botulinum Injections-An Observational Prospective Study.

BACKGROUND: Botulinum toxin A (BTX), a neurotoxin widely used for facial aesthetics, causes dose-dependent muscle paralysis. It was hypothesized that treatment of mimic muscles with BTX might have a positive impact on emotional expression in static images (photographs), but a negative impact in dynamic recordings (videos). OBJECTIVES: The aim of this study was to compare of emotional expression recorded in photographs and videos before and after treatment with BTX. METHODS: Twenty healthy women (mean age, 45 years) received a dose of 19 mouse units of XEOMIN (Merz, Frankfurt am Main, Germany) into the procerus, occipitofrontalis, and orbicularis oculi muscles. Photographs and videos of the participants' faces with neutral and happy expressions were recorded before treatment and 2 weeks later. Recordings were rated by naive raters blind to the conditions and in balanced order. RESULTS: Videos were generally rated as more pleasant, arousing, attractive, and genuine than photographs (all Ps > 0.001). This was especially the case for videos with neutral expression (P = 0.003). Independent of presentation mode and facial expression, women were rated as more attractive after BTX treatment (P = 0.03). CONCLUSIONS: In contrast to the hypothesis, the reduced mobility had no detectable negative impact on dynamic emotional expression, but videos received more positive ratings, particularly for neutral expressions. It is thus recommended to assess emotional expression with dynamic recordings to evaluate the effects of treatment with BTX. BTX seems to improve perceived attractiveness, although the cause of this effect remains unclear.

Cytokine production in patients with recurrent acute tonsillitis: analysis of tonsil samples and blood.

The aim of this study was to examine T cell function in tonsils of patients with recurrent acute tonsillitis (RAT) or peritonsillar abscess (PTA) by analyzing the cytokine production following T cell receptor (TCR) and co-receptor stimulation with a combination of anti-CD3 and anti-CD28 antibodies. The release of IFN-γ, TNF-α, IL-2, IL-4, IL-6, IL-10 and IL-17A from isolated, stimulated T cells of 27 palatine tonsils (10 RAT, 7 PTA, 10 tonsils without inflammation) was measured via a bead-based flow cytometric analysis. The results were compared with the cytokine release of isolated peripheral T cells in a subset of the same patients (6 PTA, 4 patients without signs of inflammation in the blood). TCR stimulation increased the concentration of released cytokines in tonsil and blood as well as in different forms of inflammation and tissue with no inflammation. Stimulation increased the pro-inflammatory cytokines TNF-α, IFN-γ, and IL-2 more than the anti-inflammatory cytokines IL-4 and IL-10 in tonsil and blood samples in RAT, PTA, and samples without inflammation. Blood of patients with PTA showed a higher pro-inflammatory cytokine level compared to the samples of patients without inflammation. T cells in tonsils are fully responsive and competent for antigen-induced cytokine production in RAT and PTA. One should be aware that tonsillectomy, if indicated, might remove a functioning immune organ. Tonsillotomy might be an alternative even in adults to maintain immunological function.

[Current Consideration of Tonsillectomy and Tonsillotomy]. / Aktuelle Betrachtung der Tonsillektomie und Tonsillotomie.

What is right? Tonsillotomy or tonsillectomy? Moreover, when and what has to be preferred? Is the discussion never ending? No, and this is good. Tonsil surgery is and will remain a frequent surgical procedure for the otolaryngologist. Especially, because many children are operated, and because - also rare - significant complications can occur, a quality-controlled benefit-risk-analysis is important. Part of this is systematic analysis of the evidence of the benefits and risks of tonsillotomy and tonsillectomy, as it was performed recently by the Institute for Quality and Efficiency in Health Care (Institut für Qualität und Wirtschaftlichkeit im Gesundheitswesen, IQWiG) by order of the Federal Joint Committee (Gemeinsamer Bundesausschuss, G-BA). One result was the implementation of the tonsillotomy as a regular service paid by the insurance companies. Nevertheless, the discussion is not yet concluded. Actually, a randomized controlled trial is in preparation sponsored by the G-BA to proof the non-inferiority of tonsillotomy against tonsillectomy for the treatment of patients with recurrent acute tonsillitis. The present article on ENT specialist knowledge should put the reader, especially ENT residents, in a position to advice patients and their relatives on the pros and cons of tonsil surgery.