Identification of neutrophils and eosinophils in upper airway mucosa with immunofluorescence multiplex image cytometry.

Characterization of inflammation in chronic rhinosinusitis with (CRSwNP) and without nasal polyps (CRSsNP) is an ongoing research process. To overcome limitations of current cytologic techniques, we investigated whether immunofluorescence multiplex image cytometry could quantify intact neutrophils, eosinophils, and other immune cells in solid upper airway mucosa. We used a four-channel immunofluorescence-microscopy technique for the simultaneous detection of the leukocyte marker CD45, the neutrophil marker myeloperoxidase, two eosinophil markers, i.e., major basic protein and eosinophil peroxidase, and DAPI (4',6-diamidin-2-phenylindole), in formalin-fixed paraffin-embedded upper airway tissue samples of patients with CRSwNP and CRSsNP, as well as of patients free of CRS with inferior turbinate hypertrophy (controls). Image acquisition and analysis were performed with TissueFAXS and StrataQuest (TissueGnostics, Vienna, Austria), respectively. Positive and negative immunostaining were differentiated with a specific fluorescence signal/background signal ratio. Isotype controls were used as negative controls. In six controls, nine patients with CRSsNP, and 11 patients with CRSwNP, the median area scanned and median cell count per patient were 14.2 mm2 and 34,356, respectively. In CRSwNP, the number of eosinophils was three times higher (23%) than that of neutrophils (7%). Three times more immune cells were encountered in CRSwNP (33%) compared to CRSsNP (11%). In controls, inflammation was balanced between the epithelial layer and lamina propria, in contrast to CRS (three times more pronounced inflammation in the lamina propria). The quantification of intact neutrophils, eosinophils, and other immune cells in solid tissue with undisrupted architecture seems feasible with immunofluorescence multiplex image cytometry.

Analysis of cells of epithelial, connective tissue and immune differentiation in HPV-positive-, HPV-negative oropharyngeal carcinoma and normal oropharyngeal tissue by immunofluorescence multiplex image cytometry: a preliminary report.

BACKGROUND: Epithelial, connective tissue and immune cells contribute in various ways to the pathophysiology of HPV positive (HPV+) and HPV negative (HPV-) oropharyngeal squamous cell carcinoma (OPSCC). We aimed to investigate the abundance of these cell lineages and their coexpression patterns in patients with HPV + and HPV- OPSCC. METHODS: We used a 4-channel immunofluorescence-microscopy technique for the simultaneous detection of three direct-conjugated antibodies (pancytokeratin, vimentin and CD45/CD18) and DAPI (4',6-Diamidin-2-phenylindole) in formalin fixed paraffin-embedded tissue samples (FFPE) of patients with HPV + and HPV- OPSCC, and of control patients. Image acquisition and analysis were performed with TissueFAXS and StrataQuest (TissueGnostics, Vienna, Austria), respectively, in tumor cell clusters/stroma in OPSCC specimens and epithelial layer/lamina propria in control specimens. Cell populations were created based on antibodies' coexpression patterns. Isotype and positive controls were examined for plausibility. RESULTS: The proportion of cells of epithelial differentiation in tumor cell clusters was higher in HPV + OPSCC (55%) than in HPV- OPSCC samples (44%). The proportion of connective tissue cells in tumor cell cluster was lower in HPV + OPSCC patients (18%) than in HPV- OPSCC patients (26%). The proportion of immune cells in tumor cell clusters was higher in HPV + OPSCC patients (25%) than in HPV- OPSCC patients (18%). The percentage of anaplastic, potentially de-differentiated cells, was 2% in control patients, and it was higher in HPV- OPSCC (21%) than in HPV + OPSCC samples (6%). CONCLUSIONS: This study provided the first quantitative data for the abundance of cells of epithelial, connective tissue and immune differentiation, in patients with OPSCC and control patients. The abundance of these different crucial cell populations was consistently originating from the same tissue sample. De-differentiation of tumor cells was higher in HPV- OPSCC than in HPV + OPSCC. In tumor cells clusters, the antitumoral host immune response was higher in HPV + OPSCC than in HPV- OPSCC, whereas the fibroblast response was higher in HPV- OPSCC than in HPV + OPSCC. This study contributed to the understanding of histopathologic differences between HPV + OPSCC and HPV- OPSCC patients.

EMT-related transcription factors and protein stabilization mechanisms involvement in cadherin switch of head and neck squamous cell carcinoma.

Epithelial to mesenchymal transition (EMT) describes a process where epithelial tumor cells acquire mesenchymal characteristics. EMT often correlates with invasion and an increased cell migration potential by losing cellular polarity and cell-cell junctions. It is mainly induced by tumor-microenvironment factors, such as TGF-beta 1 and IL-6, which activate the increased expression of the EMT-transcription factor (TF) Slug. We previously reported the Slug/Krüppel-like factor 4 (KLF4) switch in EMT in HNSCC, and found, that in human papilloma virus (HPV)-negative HNSCC Slug gene expression was significant higher represented, than in HPV-positive HNSCC. The purpose of this study was to investigate the impact of KLF4 and Slug on the regulation of the cadherin switch and on the EMT phenotype. Gene expression of KLF4 positive correlated with E-cadherin in 71 head and neck squamous cell carcinoma (HNSCC) patient tissue samples, which we also confirmed by the investigation of the Cancer Genome Atlas database (TCGA). HPV-transcripts contributed to stabilization of KLF4 at protein level, and simultaneously upregulated E-cadherin. Furthermore, ectopic KLF4 overexpression was associated with epithelial gene expression by induction of E-cadherin, ß-catenin and 70-kDa heat shock protein (HSP-70). The presence of HSP-70 ensures the membranous localization of E-cadherin, therefore, the ability of cells to form cadherin/catenin complexes and cellular linkages. In conclusion, KLF4 is a major regulator of the epithelial cadherin-adhesion in HNSCC.

CT analysis of the anterior nasal airway based on the direction of nasal airflow in patients with nasal obstruction and trauma controls.

PURPOSE: The anterior nose is the nasal segment with the highest resistance to airflow. In a hospital-based case-control study, we compared cross-sectional areas of the nasal cavities anterior to the piriform aperture determined by computed tomography (CT-CSA) in patients with nasal obstruction (cases) and unselected patients with trauma unrelated to the head and face (controls). METHODS: CT-CSA could be reproducibly identified at angles of 0o, 30°, 60°, and 90° to the nasal floor approximately perpendicular to the arcuate direction of nasal airflow using bony landmarks. CT-CSA were manually segmented and compared in cases and controls. In cases, we compared CT-CSA at 30° (CT-CSA30-narrow) with the minimum cross-sectional area determined by acoustic rhinometry (AR-MCA1-narrow), each on the narrower side. RESULTS: CT-CSA ranged from 7 to 250 mm2 with an average of 100 mm2 per nasal side. Side differences of the nasal airways indicating asymmetry of the nasal airways were greater in 40 cases than in 44 controls (p < 0.003). Moreover, bilateral CT-CSA were significantly smaller in cases than in controls (p < 0.001). CT-CSA30-narrow did not significantly correlate with AR-MCA1-narrow (r = 0.33; p = 0.07) and on average was 58% smaller than AR-MCA1-narrow. CONCLUSIONS: Cross-sectional areas of the anterior nose perpendicular to the direction of nasal airflow, which is considered relevant in terms of flow physics, can be reliably measured using CT. Anterior nasal cavities in patients with nasal obstruction were more asymmetric and, as a whole, narrower than in controls, the latter of which is not corrected by routine septoplasty.

[Oncolytic virotherapy in head and neck cancer]. / Onkolytische Virotherapie bei Kopf-Hals-Karzinomen.

OBJECTIVE: Oncolytic viruses (OV) infect and kill cancer cells and elicit an antitumoral immune response. With their potential to break through tumor immunoresistance, OV might be a future combination treatment option in patients with advanced head and neck cancer (HNC). Modes of action, biological modifications, handling and side effects of OV for treatment of HNC are reviewed. Results of preclinical and clinical trials are reported. METHODS: Publications and clinical trials dealing with OV and HNC were searched in PubMed and international platforms for clinical study records. Studies on preclinical and clinical trials regarding oncolytic Herpes Simplex Virus (HSV), Adenovirus, Vacciniavirus and Reovirus were selected. RESULTS: Enhanced infection and killing of tumor cells through capsid and genome modifications of OV were reported in recent preclinical studies. Most of the clinical studies were phase-I/II trials. In phase III studies, tumor regression and prolonged survival were observed after treatment with oncolytic HSV, Adenoviruses and Reoviruses. In most trials, OV were combined with chemoradiotherapy or immunotherapy. CONCLUSION: In the published studies, OV treatment of HNC patients was safe, often well tolerated and showed promising results with regard to response and survival, especially in combination with chemoradiotherapy or checkpoint inhibitors.

[Prognostic factors for successful outcome in sialendoscopy for sialolithiasis]. / Prognostische Einflussfaktoren auf den Erfolg der Sialendoskopie bei Sialolithiasis.

BACKGROUND: The success of sialendoscopy depends on several factors. The 2008 introduced lithiasis-stenosis-dilatation (LSD) classification intended to describe more precise the stone-duct system. We investigated whether the LSD classification and additional pre- and intraoperative parameters could be used as prognostic factors for success. METHODS: We retrospectively assessed patients with primary sialendoscopy for sialolithiasis between September 2018 und March 2020. Among others, the outcome variables were the stone size, location and LSD classification. RESULTS: We included 37 patients. The success group included 12/37 (32 %) patients. The median stone size was 3.7 millimeters (mm) in the success group and 10.0 mm in the failure group (Mann-Whitney test; p < 0.0001). In the success group, 11/12 stones were distal in contrast to the failure group (13/25 stones proximal; Pearson's chi-square test; p = 0.010). We noted 10 L1S0D0 stones in the success group in contrast to the failure group (15 L3aS0D0 stones; Pearson chi-square test; p = 0.001). For distal stones smaller than 5 mm, the success rate was 100 % and for proximal stones larger than 4 mm, it was 0 %. For stones free in the duct lumen (L1S0D0), the success rate was 60-100 %. CONCLUSION: The distal stone location and the smaller stone size in a normal duct should be beneficial prognostic factors for success. Future studies should focus on the LSD classification, stone volume, stone duct orientation and stone distance from the papilla.

Cancer stem cells and their unique role in metastatic spread.

Cancer stem cells (CSC) possess abilities generally associated with embryonic or adult stem cells, especially self-renewal and differentiation, but also dormancy and cellular plasticity that allow adaption to new environmental circumstances. These abilities are ideal prerequisites for the successful establishment of metastasis. This review highlights the role of CSCs in every step of the metastatic cascade from cancer cell invasion into blood vessels, survival in the blood stream, attachment and extravasation as well as colonization of the host organ and subsequent establishment of distant macrometastasis.

Characterization of epithelial cells, connective tissue cells and immune cells in human upper airway mucosa by immunofluorescence multichannel image cytometry: a pilot study.

Epithelial, connective tissue and immune cells contribute in various ways to the pathophysiology of chronic rhinosinusitis (CRS). However, data of their distribution in upper airway mucosa are sparse. We aimed to provide quantitative, purely informative data on the distribution of these cell lineages and their coexpression patterns, which might help identifying, e.g., cells in the epithelium undergoing through epithelial-mesenchymal transition (EMT). For this purpose, we used immunofluorescence multichannel image cytometry (IMIC). We examined fixed paraffin-embedded tissue samples (FFPE) of six patients with chronic rhinosinusitis (CRS) and of three patients without CRS (controls). The direct-conjugated antibodies pancytokeratin, vimentin and CD45/CD18 were used for coexpression analysis in epithelial layer and lamina propria. Image acquisition and analysis were performed with TissueFAXS and StrataQuest, respectively. To distinguish positive from negative expression, a ratio between cell-specific immunostaining intensity and background was developed. Isotype controls were used as negative controls. Per patient, a 4.5-mm2 tissue area was scanned and a median of 14,875 cells was recognized. The most common cell types were cytokeratin-single-positive (26%), vimentin-single-positive (13%) and CD45/CD18-single-positive with CD45/CD18-vimentin-double-positive cells (29%). In the patients with CRS, CD45/CD18-single-positive cells were 3-6 times higher compared to the control patients. In the epithelial layer, cytokeratin-vimentin-double-positive EMT cells were observed 3-5 times higher in the patients with CRS than in the control patients. This study provided quantitative data for the distribution of crucial cell types in CRS. Future studies may focus on the distribution and coexpression patterns of different immune cells in CRS or even cancer tissue.

Does low-level laser therapy affect the survival of patients with head and neck cancer?

Low-level laser therapy (LLLT) is used in patients with head and neck cancer (HNC) for treatment-related mucositis. There is conflicting evidence as to whether LLLT leads to the proliferation of tumor cells and whether it interferes with the tumoricidal effect of radiotherapy or chemoradiotherapy, if the tumor lies within the LLLT field. Using fuzzy matching, 126 HNC patients who had received LLLT including the tumor region and 126 matching HNC patients without LLLT (controls) treated at the Department of Otorhinolaryngology, Head & Neck Surgery, Medical University of Innsbruck, were identified. The overall survival was compared using the Kaplan-Meier analysis. Fuzzy matching yielded 2 patient samples well comparable in terms of risk of death. The survival did not significantly differ between patients with and without LLLT (p = 0.18). An increased risk of death in HNC patients who received LLLT covering the tumor region was not observed in our study.

Real-life assessment of chronic rhinosinusitis patients using mobile technology: The mySinusitisCoach project by EUFOREA.

BACKGROUND: Chronic rhinosinusitis (CRS) is a chronic inflammatory disease associated with a substantial personal and socioeconomic burden. Monitoring of patient-reported outcomes by mobile technology offers the possibility to better understand real-life burden of CRS. METHODS: This study reports on the cross-sectional evaluation of data of 626 users of mySinusitisCoach (mSC), a mobile application for CRS patients. Patient characteristics of mSC users were analysed as well as the level of disease control based on VAS global rhinosinusitis symptom score and adapted EPOS criteria. RESULTS: The mSC cohort represents a heterogeneous group of CRS patients with a diverse pattern of major symptoms. Approximately half of patients reported nasal polyps. 47.3% of all CRS patients were uncontrolled based on evaluation of VAS global rhinosinusitis symptom score compared to 40.9% based on adapted EPOS criteria. The impact of CRS on sleep quality and daily life activities was significantly higher in uncontrolled versus well-controlled patients. Half of patients had a history of FESS (functional endoscopic sinus surgery) and reported lower symptom severity compared to patients without a history of FESS, except for patients with a history of more than 3 procedures. Patients with a history of FESS reported higher VAS levels for impaired smell. CONCLUSION: Real-life data confirm the high disease burden in uncontrolled CRS patients, clearly impacting quality of life. Sinus surgery improves patient-reported outcomes, but not in patients with a history of more than 3 procedures. Mobile technology opens a new era of real-life monitoring, supporting the evolution of care towards precision medicine.

Nasal Floor Asymmetry Is Associated With Nasal Obstruction.

PURPOSE: The role of morphologic characteristics of the nasal cavity in nasal obstruction is not yet sufficiently understood. The aim of this study was to determine which morphometric parameters of the nasal cavity severely impair nasal breathing and when. PATIENTS AND METHODS: In a hospital-based, computed tomography-morphometric cross-sectional study, we evaluated computed tomography coronal scans of patients with known nasal obstruction scheduled to undergo functional nasal surgical procedures (cases) and trauma patients without facial involvement or known nasal obstruction (controls). The primary predictor variable was case versus control. In both groups, we measured and compared the piriform aperture width; nasal floor canting; piriform aperture vertical height, height-width ratio, and total cross-sectional area; height difference between the right and left nasal floors; and nasal septal thickness; as well as age and gender differences. Metric data means, standard deviations, and 95% confidence intervals were calculated and analyzed. RESULTS: The sample was composed of 60 patients evenly divided between cases and controls. Of these, 30 were men. The average age of the cases and controls was 27.4 ± 7.8 years and 38.5 ± 18.6 years, respectively (P < .001). The differences in piriform aperture width were not statistically significant between cases and controls (23.3 ± 1.9 mm and 23.8 ± 1.7 mm, respectively; P > .2). In contrast, we noted statistically significant differences between cases and controls in nasal floor canting (5.4° ± 4.6° and 1.8° ± 1.5°, respectively; P < .001) and height difference between the right and left nasal floors (1.8 ± 1.2 mm and 1.0 ± 0.7 mm, respectively; P = .002). CONCLUSIONS: Nasal floor canting of 3° or greater and a height difference between the right and left nasal floors of 1.5 mm or greater may contribute to the etiology of clinically relevant nasal obstruction. A piriform aperture width of 22 mm or less may be considered narrow. Future studies can determine when and how exactly to surgically address a clinically relevant narrow piriform aperture and nasal floor canting.

[Sexual Behavior and High-Risk HPV Infection in the Head and Neck]. / High-Risk-HPV-Infektionen im Kopf-Hals-Bereich ­ Welche Bedeutung hat das Sexualverhalten?

In the second half of the 20th century, the sexual revolution went hand in hand with changes in common sexual practices. At the turn of the millennium, an increase in the detection rate of HPV-positive oropharyngeal carcinomas (OPC) in the USA was observed for the first time. An increase in the OPC was also observed in Europe. It was shown that this increase was due to HPV-positive OPC. However, the detection rate of the HPV-positive OPC has national and regional differences. Within Europe, the highest detection rate is in Northern Europe (56,5 %), followed by Central Europe (37,6 %). An association between patients with OPC and ≥ 6 sexual partners (OR = 1,25) and ≥ 4 oral sex partners (OR = 2,25) has been described. An HPV infection is usually asymptomatic and eliminated by the immune system within a few months. The persistence of the virus is oncogene. Smoking favors virus persistence, which is why the combination of smoking and oral sex with ≥ 5 partners is a particular risk factor. It was also examined whether partners from patients with cervical cancer have an increased oropharyngeal HPV infection rate. There was no definite declaration on this, but further investigations are required. Oral condoms or dental dams are suitable for the prevention of an oral HPV infection. The studies regarding the effectiveness of HPV vaccination for the prevention of an OPC are poor. Authors described a significantly lower detection rate of oral HPV infection in vaccinated people than in non-vaccinated people (0,11 % vs. 1,61 %; p = 0,008).

[Biologics for Chronic Rhinosinusitis]. / Biologika bei chronischer Rhinosinusitis.

Chronic rhinosinusitis (CRS) is an inflammation of the nasal and paranasal mucosa, lasting for more than 12 weeks. By now approximately 15 % of the European and American population are affected, which indicates that CRS is a serious health problem. Beside other subgroups the most important classification is CRS with nasal polyps (CRSwNP) and without nasal polyps (CRSsNP). Standard-of-care therapies include nasal saline solution as well as topic or systemic corticosteroids. If this does not lead to a sufficient recovery, surgical therapy is a good option. A new therapy option is represented by biologics, particularly monoclonal antibodies (mAB). They are well-established for treatment of asthma bronchiale. Due to the fact that asthma is often associated with nasal polyps, and mAB may could also lead to improvements in CRS, studies were conducted. In the meantime there is a number of mAB which have emerged as an alternative treatment for patients with CRSwNP.

[Fistula prophylaxis with muscle-sparing, myofascial pectoralis major lobe in salvage laryngectomy]. / Fistelprophylaxe mit muskelsparendem, myofaszialem Pectoralis-major-Lappen bei Salvage-Laryngektomie.

BACKGROUND: In recurrent hypopharyngeal/laryngeal squamous cell carcinoma after radiation salvage-laryngectomy (salvage-LE) represents the best curative treatment option. Pre-irradiation promotes development of pharyngocutaneous fistulas (pc-fistula). Transfer of unirradiated tissue to the surgical site reduces fistula rate. Frequently, a myocutaneous Pectoralis-major-flap (PMF) is applied. We describe a muscle sparing, myofascial PMF (ms-PMF) and its functional & aesthetic results. METHODS: For the ms-PMF, the pectoralis major is exposed via two 8 cm long subclavicular & submammary horizontal incisions. The pectoral branch of the thoracoacromial artery, which represents the blood supply for the ms-PMF, is identified between the clavicular and sternocostal aspect of the muscle. A craniocaudal stripe of the pectoralis major is recovered and supraclaviculary transposed towards the pharynx. The clavicular and sternocostal part of the muscle remain untouched.The clear-margin resection rate (CMRR), mean overall survival (OS), complications, median length of hospital stay, & functional & aesthetic outcome were assessed. RESULTS: In 25 patients, salvage-LE with ms-PMF was performed. The CMRR was 100 %, median OS 1.3 (follow-up 2.8) years, complications rate 16 % (4 complete lobe necroses, 3 pc-fistulas) and median inpatient stay 20 (11-78) days. Good functional outcome were observed: limitations of oral food intake occurred in 2-, limitations in voice rehabilitation in 3 patients. Wound healing disorders without lobe necrosis were not observed. The aesthetic results were appealing. CONCLUSION: The ms-PMF is a less invasive & safe procedure for fistula prophylaxis in salvage-LE with favorable functional & aesthetic results.

Therapy resistance mediated by cancer stem cells.

Cancer stem cells (CSC) possess abilities generally associated with embryonic or adult stem cells, especially self-renewal and differentiation. The CSC model assumes that this subpopulation of cells sustains malignant growth, which suggests a hierarchical organization of tumors in which CSCs are on top and responsible for the generation of intratumoral heterogeneity. Effective tumor therapy requires the eradication of CSC as they can support regrowth of the tumor resulting in recurrence. However, eradication of CSC is difficult because they frequently are therapy resistant. Therapy resistance is mediated by the acquisition of dormancy, increased DNA repair and drug efflux capacity, decreased apoptosis as well as the interaction between CSC and their supporting microenvironment, the CSC niche. This review highlights the role of CSC in chemo- and radiotherapy resistance as well as possible ways to overcome CSC mediated therapy resistance.

Therapy resistance mediated by exosomes.

Therapy resistance can arise within tumor cells because of genetic or phenotypic changes (intrinsic resistance), or it can be the result of an interaction with the tumor microenvironment (extrinsic resistance). Exosomes are membranous vesicles 40 to 100 nm in diameter constitutively released by almost all cell types, and mediate cell-to-cell communication by transferring mRNAs, miRNAs, DNAs and proteins causing extrinsic therapy resistance. They transfer therapy resistance by anti-apoptotic signalling, increased DNA-repair or delivering ABC transporters to drug sensitive cells. As functional mediators of tumor-stroma interaction and of epithelial to mesenchymal transition, exosomes also promote environment-mediated therapy resistance.Exosomes may be used in anticancer therapy exploiting their delivery function. They may effectively transfer anticancer drugs or RNAs in the context of gene therapy reducing immune stimulatory effects of these drugs and hydrophilic qualities facilitating crossing of cell membranes.

A long-term follow-up study on otoacoustic emissions testing in paediatric patients with severe malaria in Gabon.

BACKGROUND: In a previous study, severe and cerebral malaria have been connected with acute cochlear malfunction in children, demonstrated by a decrease of transitory evoked otoacoustic emissions (TEOAEs) reproducibility. This study aims to determine whether cochlear malfunction persists for 4 years after recovery from severe malaria in a subset of the previous study's collective. Follow-up TEOAEs were performed on site (CERMEL, Hôpital Albert Schweitzer, Lambaréné, Gabon) or at the participants' homes; 33 out of 90 participants included in the initial investigation by Schmutzhard et al. could be retrieved and were re-examined, 31/33 could be included. Of the 57 missing participants, 51 could not be contacted, 1 had moved away, 4 refused to cooperate, and 1 had died. METHODS: As in the initial investigation, participants of this prospective follow-up study were subjected to TEOAE examination on both ears separately. A wave correlation rate of > 60% on both ears was considered a "pass"; if one ear failed to pass, the examination was considered a "fail". The results were compared to the primary control group. Additionally, a questionnaire has been applied focusing on subsequent malaria infections between the primary inclusion and follow-up and subjective impairment of hearing and/or understanding. RESULTS: The cohort's mean age was 9 years, 14 children were female, 18 male. 31 had been originally admitted with severe, one with cerebral malaria. 83.8% of participants (n = 26) presented with a TEOAE correlation rate of > 60% on both ears (the cut-off for good cochlear function); in the control group, 92.2% (n = 83) had passed TEOAE examination on both ears. Recurrent severe malaria was associated with a worse TEOAE correlation rate. Age at infection and gender had no influence on the outcome. CONCLUSIONS: Cochlear malfunction seems to be persistent after 4 years in more than 16% of children hospitalized for malaria. In a healthy control group, this proportion was 7.8%. Yet, the severity of the initial TEOAE-decrease did not predict a worse outcome.

The Pectoralis Major Island Flap: Short Scar Modified Muscle-Sparing Harvesting Technique Improves Aesthetic Outcome in Reconstructive Head and Neck Surgery.

BACKGROUND: Pectoralis major flap reconstruction is often associated with large unappealing scars in head-neck surgery. We recently established an alternative harvesting technique that improves aesthetic outcome. OBJECTIVES: The objective of this study was to demonstrate a modified surgical technique that harvests the pectoralis major muscle as an island flap and focuses on minimizing incision lines and applying a muscle-sparing approach. METHODS: A retrospective analysis covering the period 2008-2018 was conducted. Patients who underwent pectoralis major island flap reconstruction for fistula prophylaxis after salvage laryngectomy at the Medical University of Innsbruck were included. Flap harvesting was performed subcutaneously using two small incision lines, thereby sparing the clavicular and upper sternocostal aspects of the pectoralis muscle. RESULTS: Twenty-three patients with squamous cell carcinoma of the larynx (n = 19) or pharynx (n = 4) underwent salvage laryngectomy and consecutive reconstruction using a pectoralis major island flap and our muscle-sparing technique. Mean overall operation time was 147 ± 48.6 min. The majority (70%) of patients had an uneventful course of recovery and showed satisfying aesthetic results and low donor site morbidity as compared to traditional harvesting techniques. Six (26%) patients had major complications that required surgical revision. Three (13%) cases were complicated with fistulas. CONCLUSION: The pectoralis major island flap is shown to be reliable in otolaryngeal surgery. This surgical technique minimizes scarring and preserves thoracic wall architecture by taking a less invasive approach, thereby improving aesthetic outcome and reducing overall patient morbidity.

Age-Dependent Calcium-Binding Protein Expression in the Spiral Ganglion and Hearing Performance of C57BL/6J and 129/SvJ Mice.

BACKGROUND/AIMS: Calcium-binding proteins in neurons buffer intracellular free Ca2+ ions, which interact with proteins controlling enzymatic and ion channel activity. The heterogeneous distribution of calretinin, calbindin, and parvalbumin influences calcium homeostasis, and calcium-related neuronal processes play an important role in neuronal aging and degeneration. This study evaluated age-related changes in calretinin, calbindin, and parvalbumin immune reactivity in spiral ganglion cells. METHODS: A total of 16 C57BL/6J and 16 129/SvJ mice at different ages (2, 4, 7, and 12 months) were included in the study. Hearing thresholds were assessed using auditory brainstem response before inner ears were excised for further evaluation. Semiquantitative immunohistochemistry for the aforementioned calcium-binding proteins was performed at the cellular level. RESULTS: The hearing thresholds of C57BL/6J and 129/SvJ mice increased significantly by 7 months of age. The average immune reactivity of calbin-din as well as the relative number of positive cells increased significantly with aging, but no significant alterations in calretinin or parvalbumin were observed. CONCLUSIONS: Upregulation of calbindin could serve as a protection to compensate for functional deficits that occur with aging. Expression of both calretinin and parvalbumin seem to be stabilizing factors in murine inner ears up to the age of 12 months in C57BL/6J and 129/SvJ mice.

Brain-Derived Neurotrophin and TrkB in Head and Neck Squamous Cell Carcinoma.

We hypothesized that in head and neck squamous cell carcinoma (HNSCC), the neurotrophin brain-derived neurotrophic factor (BDNF) and its high affinity receptor TrkB regulate tumor cell survival, invasion, and therapy resistance. We used in situ hybridization for BDNF and immunohistochemistry (IHC) for TrkB in 131 HNSCC samples. Brain-derived neurotrophic factor was highly expressed in normal mucosa in HNSCC tissue and in cell lines, whereas only 42.74% of HNSCC tissue was TrkB⁺. One fourth of HNSCC cases was human papilloma virus (HPV)- positive, but the TrkB IHC frequency was not different in HPV-positive (HPV⁺) and negative cases. The UPCI-SCC090 cells expressed constitutive levels of TrkB. Transforming-growth-factor-ß1 (1 ng/mL TGF-ß1) induced TrkB in a subpopulation of SCC-25 cells. A single 10-µg/mL mitomycin C treatment in UPCI-SCC090 cells induced apoptosis and BDNF did not rescue them. The SCC-25 cells were resistant to the MMC treatment, and their growth decreased after TGF-ß1 treatment, but was restored by BDNF if it followed TGF-ß1. Taken together, BDNF might be ineffective in HPV⁺ HNSCC patients. In HPV- HNSCC patients, tumor cells did not die after chemotherapeutic challenge and BDNF with TGF-ß1 could improve tumor cell survival and contribute to worse patient prognosis.