Predicting Lewis Acidity: Machine Learning the Fluoride Ion Affinity of p-Block-Atom-Based Molecules.

"How strong is this Lewis acid?" is a question researchers often approach by calculating its fluoride ion affinity (FIA) with quantum chemistry. Here, we present FIA49k, an extensive FIA dataset with 48,986 data points calculated at the RI-DSD-BLYP-D3(BJ)/def2-QZVPP//PBEh-3c level of theory, including 13 different p-block atoms as the fluoride accepting site. The FIA49k dataset was used to train FIA-GNN, two message-passing graph neural networks, which predict gas and solution phase FIA values of molecules excluded from training with a mean absolute error of 14 kJ mol-1 (r2=0.93) from the SMILES string of the Lewis acid as the only input. The level of accuracy is notable, given the wide energetic range of 750 kJ mol-1 spanned by FIA49k. The model's value was demonstrated with four case studies, including predictions for molecules extracted from the Cambridge Structural Database and by reproducing results from catalysis research available in the literature. Weaknesses of the model are evaluated and interpreted chemically. FIA-GNN and the FIA49k dataset can be reached via a free web app (www.grebgroup.de/fia-gnn).

Integration and viral oncogene expression of human papillomavirus type 16 in oropharyngeal squamous cell carcinoma and gastric cancer.

Persistent high-risk human papillomavirus (HR-HPV) infections cause cervical cancer and a fraction of head and neck cancer. To investigate whether HR-HPV infection might be also involved in the development of gastric cancer (GC), we developed a platform utilizing a rolling circle amplification (RCA)-based nested L1 polymerase chain reaction with Sanger sequencing to genotype the HPV DNA in cancer tissues of 361 GC and 89 oropharyngeal squamous cell carcinomas (OPSCC). HPV transcriptional activity was determined by E6/E7 mRNA expression and a 3' rapid amplification of cDNA ends was performed to identify HPV integration and expression of virus-host fusion transcripts. Ten of 361 GC, 2 of 89 OPSCC, and 1 of 22 normal adjacent tissues were HPV L1 DNA-positive. Five of the 10 HPV-positive GC were genotyped as HPV16 by sequencing and 1 of 2 GC with RCA/nested HPV16 E6/E7 DNA detection exhibited HPV16 E6/E7 mRNA. Two OPSCC displayed HPV16 L1 DNA and E6/E7 mRNA, of which 1 OPSCC tissue showed virus-host RNA fusion transcripts from an intron region of KIAA0825 gene. Together, our data reveal viral oncogene expression and/or integration in GC and OPSCC and a possible etiology role of HPV infections in gastric carcinogenesis.

Reactive oxygen species in cancer stem cells of head and neck squamous cancer.

One of the greatest challenges in systemic treatment of head and neck squamous cell carcinoma (HNSCC) is a small tumor cell population, namely, cancer stem-like cells (CSC). CSC can regenerate and maintain a heterogenic tumor by their self-renewal capacity. Their potential ability to be more resistant to and survival after chemo- and radiation therapy was also identified. Further studies have shown that reactive oxygen species (ROS) contribute to this CSC-associated resistance. In this review, we focus on the current knowledge of HNSCC-CSC, with regard to ROS as a possible and novel therapeutic approach in targeting CSC.

Heterogeneity of Head and Neck Squamous Cell Carcinoma Stem Cells.

Current systemic cancer treatment in head and neck squamous cell carcinoma (HNSCC) is moving toward more personalized approaches such as de-escalation protocols human-papilloma-virus dependent HNSCC or application of checkpoint inhibitors. However, these treatments have been challenged by cancer stem cells (CSC), a small population within the bulk tumor, which are leading to treatment failure, tumor recurrence, or metastases. This review will give an overview of the characteristics of HNSCC-CSC. Specifically, the mechanisms by which HNSCC-CSC induce tumor initiation, progression, recurrence, or metastasis will be discussed. Although evidence-based treatment options targeting HNSCC-CSC specifically are still being sought for, they warrant a promise for additional and sustainable treatment options where for HNSCC patients where others have failed.

pN status predicts outcomes in surgically treated pT1-pT2 patients of various disease stages with squamous cell carcinoma of the head and neck: a 17-year retrospective single center cohort study.

OBJECTIVES: The optimal treatment for a substantial proportion of patients with pT1-pT2 squamous cell carcinomas of the head and neck (SCCHN) remains to be refined. The extent of surgery, role and potential benefit of adjuvant treatment are to be balanced against therapy-induced side effects. We compared the outcomes of surgery with or without adjuvant radiotherapy (RT) or chemotherapy (CRT) and investigated the prognostic value of established clinicopathological parameters. METHODS: Data were retrospectively collected for 227 patients who were treated by surgery alone (n = 31), RT (n = 87) and CRT (n = 109) in a single center. RESULTS: Patients with stage I/II disease who had received adjuvant RT showed a better disease-free survival (DFS) (P = 0.04) than those who had received adjuvant CRT treatment. Conversely, patients with stage III/IV disease who had received CRT showed a better overall survival (OS) (P = 0.003) and DFS (P = 0.03) than those who had received surgery alone or adjuvant RT without chemotherapy. Survival analysis demonstrated that patients with pN0 to pN1 had better OS (P = 0.02), disease-specific survival (DSS) (P = 0.003), DFS (P = 0.02) and metastases free survival (MFS) (P = 0.002) compared to patients with pN2 to pN3. Multivariate analysis showed that the pN status was an independent factor for OS (P = 0.03), DSS (P = 0.04), relapse-free survival (P = 0.03), DFS (P = 0.03). CONCLUSION: The pN status is the most important prognostic factor for pT1 to pT2 SCCHN. Adjuvant CRT was associated with significantly better survival outcomes in patients with pN1 and pN2-3 or more advanced stage, while adjuvant RT showed significantly better outcomes in patients with pN0.

Basal subtype is predictive for response to cetuximab treatment in patient-derived xenografts of squamous cell head and neck cancer.

Cetuximab is the single targeted therapy approved for the treatment of head and neck cancer (HNSCC). Predictive biomarkers have not been established and patient stratification based on molecular tumor profiles has not been possible. Since EGFR pathway activation is pronounced in basal subtype, we hypothesized this activation could be a predictive signature for an EGFR directed treatment. From our patient-derived xenograft platform of HNSCC, 28 models were subjected to Affymetrix gene expression studies on HG U133+ 2.0. Based on the expression of 821 genes, the subtype of each of the 28 models was determined by integrating gene expression profiles through centroid-clustering with previously published gene expression data by Keck et al. The models were treated in groups of 5-6 animals with docetaxel, cetuximab, everolimus, cis- or carboplatin and 5-fluorouracil. Response was evaluated by comparing tumor volume at treatment initiation and after 3 weeks of treatment (RTV). Tumors distributed over the 3 signature-defined subtypes: 5 mesenchymal/inflamed phenotype (MS), 15 basal type (BA), 8 classical type (CL). Cluster analysis revealed a strong correlation between response to cetuximab and the basal subtype. RTV MS 3.32 vs. BA 0.78 (MS vs. BA, unpaired t-test, p 0.0002). Cetuximab responders were distributed as following: 1/5 in MS, 5/8 in CL and 13/15 in the BA group. Activity of classical chemotherapies did not differ between the subtypes. In conclusion basal subtype was associated with response to EGFR directed therapy in head and neck squamous cell cancer patient-derived xenografts.

CO2 laser stapedotomy safety: influence of laser energy and time on bone-conduction hearing levels.

Total laser energy in CO2 stapedotomy depends on the laser settings and the amount of applications. It is unclear if the amount of total laser energy affects bone-conduction hearing thresholds and if possible effects are temporary or permanent. Alterations of bone-conduction hearing thresholds after single or multiple-shot CO2 laser stapedotomy were analyzed between 1 and 3 weeks and 1.5-6 months after primary (n = 501) or revision surgeries (n = 153) and correlated to time, laser energy, frequency, surgical technique, and pathology encountered in revision stapedotomy. In both time periods, most patients showed a lower bone-conduction threshold in the four-tone puretone average (PTA) at frequencies of 0.5, 1, 2, and 3 kHz that further improved over time. Between 1 and 3 weeks, the improvement was significant in subgroups with cumulative energies lower 1 J and successful one-shot technique or in revisions without laser application. The remaining subgroups with higher total energies showed significant improvements between 1.5 and 6 months. At 4 and 8 kHz, significant improvements were found during 1.5-6 months after primary and revision surgery independent of the used energy. Repeated CO2 laser applications showed no impairment in bone-conduction thresholds and can thus be considered as safe. In most patients, significant, yet unexplained, improvements in bone-conduction hearing thresholds were noticed in a time- and energy-related pattern.

European otorhinolaryngology training programs: results of a European survey about training satisfaction, work environment and conditions in six countries.

ORL-students and residents have an ongoing debate about the "best" programme in Europe. Aim of this study was to comparatively assess differences among programmes in training, satisfaction, quality of life (QoL) of residents and recent otorhinolaryngologist (ORL) specialists in France, Germany, Spain, Italy, Austria, and Belgium. A self-administered anonymous questionnaire, structured in ten sections including general information, provided guidance, working environment, training structure, teaching of medical students, publication work, QoL, and satisfaction with training, were emailed to residents and recent ORL specialists. 476 returned questionnaires from 6 countries revealed that daily work hours were the highest in France and Belgium with 11 and 10.4 h on average, respectively. QoL, work conditions, and salary were best in Germany followed by Austria in terms of possibility of part-time contracts, better respect for post-duty day off, and compensation for overtime. Satisfaction with training including support and guidance of seniors was lowest in Italy, but, on the other hand, the publication work and support had a more important place than in other countries. In Belgium, there was some gap between the quality of teaching and feedback from seniors as well as apprenticeship. The highest satisfaction with training was in France and Spain followed by Austria. The study results provide guidance before choosing an ORL training programme in Europe. Country-specific strengths could be included into future harmonization efforts to improve all programmes, facilitate professional exchange and, finally, establish standards-of-care carried out by well-trained doctors also looking after a satisfying work-life balance.

[Severe Bleeding from the Middle Ear Cavity after Myringotomy: Review Based on a Case Report]. / Massive Blutung bei einer Parazentese: Übersicht anhand eines Fallberichts.

Report of a rare case of severe bleeding from the middle ear cavity after myringotomy. On the basis of the case report, the procedure for such bleeding is discussed in the context of the literature. A 6-year-old boy received a revision myringotomy in an ambulant setting. During the procedure a severe bleeding occurred. The external auditory canal was adequately packed. The patient was extubated and transferred to the clinic as an emergency. Computer tomography of the temporal bone showed the anatomical variant of a dehiscent high jugular bulb, which had been injured. Because no rebleeding occurred, the packing of the ear canal was removed and an explorative tympanoscopy was performed on the third postoperative day. When the tympanomeatal flap was lifted, the defect in the jugular bulb was found. The lesion was covered with Tutopatch® pads and fibrin glue and the auditory canal was packed again. After removal of the packing three weeks postoperatively a properly healed situs was found. No further measures were taken. The injury of a dehiscent jugular bulb in the course of ear surgeries leads to a massive hemorrhage. The case describes the diagnostic and therapeutic procedure for this relatively rare but severe complication.

[Immunotherapy Against Head and Neck Cancer Stem Cells]. / Immuntherapie gegen Tumorstammzellen bei Plattenepithelkarzinomen im Kopf-Hals-Bereich.

Immunotherapy against head and neck cancer stem cells Immunologic therapies like antibodies in solid tumors like squamous cell cancer of the head and neck are administered either alone or in combination with radiation and chemotherapy. Despite some respectable successes, the effect of this therapy reaches its limits due the ability of the tumor to escape the immune system. Cancer stem cells seem to play an important role in this process due to their intrinsic resistance to conventional therapy and the ability to regenerate tumor heterogeneity. This way they substantially contribute to the formation of recurrences and metastases. Therefore, future immunotherapies should target specifically this subpopulation, possibly in combination with other therapeutic modalities. In this review the immunologic features of cancer stem cells and their potential as target for immunotherapies is summarized.

Meta-analysis of survival in patients with HNSCC discriminates risk depending on combined HPV and p16 status.

Data indicate a better prognosis for human papillomavirus (HPV)-associated head and neck squamous cell carcinoma (HNSCC). HPV and p16 detection are established markers for HPV-related HNSCC. Both are accepted as survival-independent predictors. Previous studies investigating the survival in HNSCC patients depending on HPV(+/-) and p16(+/-) status consistently found discordant results with p16(-)/HPV(+) and p16(+)/HPV(-). However, no meta-analysis regarding the survival according to combined HPV/p16 status has been performed yet. The objective of this study was to discriminate the impact of combined HPV(+/-) and p16(+/-) status on survival. Data sources were identification and review of publications assessing survival of the distinct subgroups with both p16 and HPV investigated in HNSCC until February, 2015. A meta-analysis was performed to classify survival and clinical outcomes. 18 out of 397 articles (4424 patients) were eligible for the meta-analysis. The percent proportion of the subgroups was 25 % for HPV(+)/p16(+), 61.2 % for HPV(-)/p16(-), 7.1 % for HPV(-)/p16(+) and 6.8 % for HPV(+)/P16(-). The meta-analysis showed a significantly improved 5-year overall survival (OS), 5-year disease-free survival and their corresponding hazard ratio for HPV(+)/p16(+) HNSCC in comparison to HPV(-)/p16(-), HPV(+)/p16(-) and HPV(-)/p16(+). The 5-year OS of the HPV(-)/p16(+) subgroup was intermediate while HPV(+)/p16(-) and HPV(-)/p16(-) HNSCC had the shortest survival. With current therapeutic strategies, survival of patients with HNSCC is better if associated with HPV(+)/p16(+) or HPV(-)/p16(+). Clinical trials are needed to confirm the distinct survival pattern and to investigate possible differences in survival for HPV(+)/p16(-) and HPV(-)/p16(+) HNSCC. To further differentiate p16(+) HNSCC, HPV testing may be advisable.

Feeling Normal? Long-Term Follow-up of Patients with a Cleft Lip-Palate after Rhinoplasty with the Derriford Appearance Scale (DAS-59).

The stigma of nasal deformity due to a congenital cleft lip-palate has an undeniable influence on the affected patient's life. It is therefore of interest to investigate if efforts to reduce esthetic and functional impairments by rhinoplasty (single or multiple) can result in an increased satisfaction with appearance and a self-perception similar to the noncleft population. Retrospective scoring before and after rhinoplasty using the validated Derriford Appearance Scale (DAS-59) and subsequent statistical evaluation and comparison to datasets available in the literature for further classification was used. Of the 61 patients who underwent at least one rhinoplasty, 26 responded to all questions. The mean age of responders was approximately 30 years of age and the male:female ratio was 1:1.2. The scale showed a significant overall improvement after surgery. The full scale and all subscale scores of the DAS-59 were significantly reduced after surgery demonstrating an improvement in the respective categories. Most importantly, if postoperative results were compared with a population concerned and unconcerned about appearance, no difference "facial self-consciousness" of appearance was apparent. Also postoperative subscores for "general self-consciousness" (GSC) and "social self-consciousness" of appearance (SSC) showed no difference from those obtained from the population concerned about appearance. The postoperative subscore for "sexual and bodily self-consciousness" of appearance (SBSC) indicated improvement beyond the level found in the concerned control population. Due to only a low improvement in the difference compared with the subscore representing a "negative self-concept," a statistically significant difference to the concerned population remained, possibly indicating that therapy beyond surgery is needed for improvement. After rhinoplasty, the investigated group of cleft lip-palate patients with nasal deformities showed an improvement in their self-conceived appearance as measured by the DAS-59. Their assessment of self-appearance was comparable to that of a group of noncleft persons with concern about their appearance. Taken together, rhinoplasties, primary and revision, add to the psychosocial well-being and an improved self-perception enhancing quality of life and enabling a more normal life. Further research is needed to clarify how the low reduction found in the "negative self-concept" may be addressed successfully.

A comprehensively characterized large panel of head and neck cancer patient-derived xenografts identifies the mTOR inhibitor everolimus as potential new treatment option.

Patient-derived xenograft (PDX) models have shown to reflect original patient tumors better than any other preclinical model. We embarked in a study establishing a large panel of head and neck squamous cell carcinomas PDX for biomarker analysis and evaluation of established and novel compounds. Out of 115 transplanted specimens 52 models were established of which 29 were characterized for response to docetaxel, cetuximab, methotrexate, carboplatin, 5-fluorouracil and everolimus. Further, tumors were subjected to sequencing analysis and gene expression profiling of selected mTOR pathway members. Most frequent response was observed for docetaxel and cetuximab. Responses to carboplatin, 5-fluorouracil and methotrexate were moderate. Everolimus revealed activity in the majority of PDX. Mutational profiling and gene expression analysis did not reveal a predictive biomarker for everolimus even though by trend RPS6KB1 mRNA expression was associated with response. In conclusion we demonstrate a comprehensively characterized panel of head and neck cancer PDX models, which represent a valuable and renewable tissue resource for evaluation of novel compounds and associated biomarkers.

Acquisition of diagnostic and surgical skills in otorhinolaryngology: a comparison of France and Germany.

With the coalescing of the European countries, among many others, official initiatives have been launched to propose minimum requirements for Otolaryngology, Head and Neck surgery (ORL) specialty training by creating an European log book. This study was initiated to assess and compare the acquisition of basic medical key diagnostic and surgical skills by residents and recent ORL specialists in France (FRA) and Germany (GER) and to determine whether gender-specific differences exist. For this, an anonymous questionnaire with questions to basic medical, diagnostic and surgical procedures specific to ORL was developed. 120 FRA and 125 GER questionnaires were returned from participants with a median training experience of 4 years. The female to male ratio was 1.3:1 and 78% of respondents were residents and 22% recent specialists. Concerning diagnostic procedures, there was no significant overall difference. Germans performed better in basic medical skills, while study participants from FRA had performed surgical procedures significantly more often in a more independent manner than German respondents in the areas of otology, rhinology and head and neck. Only in septoplasty, as part of rhinology, the Germans had a light advantage compared to the French. No difference was found for trauma surgery. No gender-specific difference became apparent. Taken together, in FRA, ORL training is far more surgically orientated than in GER. It remains unclear at what time, the Germans may catch up with their skills. Initiatives should be taken in GER to secure an adequate acquisition of surgical skill and experience to maintain a high level of ORL-specific competence.

Training satisfaction and work environment in Otorhinolaryngology, Head and Neck surgery: a comparison between France and Germany.

With the coalescing of Europe, increased mobility of professionals emerges. Initiatives to harmonize medical education were launched. In Otolaryngology, Head and Neck surgery (ORL) an European board examination was created to ensure standards. Quality of training, satisfaction and quality of life of residents and recent ORL specialists were compared to assess different aspects of work and hierarchical relationships in France (FRA) and Germany (GER) by means of an anonymous questionnaire. 120 FRA and 125 GER questionnaires were included. 78 % of respondents were residents. 86 % would choose the same training again. In both countries, a majority felt well considered with responsibilities adapted to their level of training and with supportive supervisors. Germans reported average daily work hours of 9.6 versus 11 in FRA with compensated overtime (76 %) and a possibility of part-time work (62 %), both nearly inexistent in FRA. In GER, the day-off after duty was more often respected. French attributed their seniors better pedagogic skills, taking time for explanations and providing better teaching. Offering a good training was a more important objective in French training centers (77 vs. 51 %). In both countries, surgical training relied on coaching. Research activities were comparable. The overall satisfaction with ORL training was high. Differences concerned structure of training, guidance by senior doctors and the working conditions. The study results provide guidance before choosing a program and may help to improve current training by identifying positive aspects that, if combined could lead to a convergence of programs. However, present high standards of education must be maintained.

Function but not phenotype of melanoma peptide-specific CD8(+) T cells correlate with survival in a multiepitope peptide vaccine trial (ECOG 1696).

ECOG 1696 was a Phase II multi-center trial testing vaccination with melanoma peptides, gp100, MART-1 and tyrosinase delivered alone, with GM-CSF, IFN-α2b or both cytokines to HLA-A2(+) patients with metastatic melanoma. Here, the frequency of circulating CD8(+) tetramer(+) (tet(+) ) T cells and maturation stages of responding T cells were serially monitored and compared with baseline values in a subset of patients (n = 37) from this trial. Multiparameter flow cytometry was used to measure the frequency of CD8(+) T cells specific for gp100, MART-1, tyrosinase and influenza (FLU) peptides. Expression of CD45RA/CCR7 on CD8(+) tet(+) T cells and CD25, CD27, CD28 on all circulating T cells was determined. Vaccine-induced changes in the CD8(+) tet(+) T cell frequency and phenotype were compared with results of IFN-γ ELISPOT assays and with clinical responses. The frequency of CD8(+) tet(+) T cells in the circulation was increased for the melanoma peptides (p < 0.03-0.0001) but not for FLU (p < 0.9). Only gp100- and MART-1-specific T cells differentiated to CD45RA(+) CCR7(-) effector/memory T cells. In contrast to the IFN-γ ELISPOT frequency, previously correlated with overall survival (Kirkwood et al., Clin Cancer Res 2009;15:1443-51), neither the frequency nor differentiation stage of CD8(+) tet(+) T cells correlated with clinical responses. Delivery of GM-CSF and/or IFN-α2b had no effects on the frequency or differentiation of CD8(+) tet(+) , CD8+ or CD4+ T cells. Phenotypic analyses of CD8(+) tet(+) T cells did not correlate with clinical responses to the vaccine, indicating that functional assessments of peptide-specific T cells are preferable for monitoring of anti-tumor vaccines.

Human skeletal muscle-derived stem cells retain stem cell properties after expansion in myosphere culture.

Human skeletal muscle contains an accessible adult stem-cell compartment in which differentiated myofibers are maintained and replaced by a self-renewing stem cell pool. Previously, studies using mouse models have established a critical role for resident stem cells in skeletal muscle, but little is known about this paradigm in human muscle. Here, we report the reproducible isolation of a population of cells from human skeletal muscle that is able to proliferate for extended periods of time as floating clusters of rounded cells, termed "myospheres" or myosphere-derived progenitor cells (MDPCs). The phenotypic characteristics and functional properties of these cells were determined using reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry and immunocytochemistry. Our results showed that these cells are clonogenic, express skeletal progenitor cell markers Pax7, ALDH1, Myod, and Desmin and the stem cell markers Nanog, Sox2, and Oct3/4 significantly elevated over controls. They could be maintained proliferatively active in vitro for more than 20 weeks and passaged at least 18 times, despite an average donor-age of 63 years. Individual clones (4.2%) derived from single cells were successfully expanded showing clonogenic potential and sustained proliferation of a subpopulation in the myospheres. Myosphere-derived cells were capable of spontaneous differentiation into myotubes in differentiation media and into other mesodermal cell lineages in induction media. We demonstrate here that direct culture and expansion of stem cells from human skeletal muscle is straightforward and reproducible with the appropriate technique. These cells may provide a viable resource of adult stem cells for future therapies of disease affecting skeletal muscle or mesenchymal lineage derived cell types.

Strategies for Advanced Oncolytic Virotherapy: Current Technology Innovations and Clinical Approaches.

Oncolytic virotherapy is a type of nanomedicine with a dual antitumor mechanism. Viruses are engineered to selectively infect and lyse cancer cells directly, leading to the release of soluble antigens which induce systemic antitumor immunity. Representative drug Talimogene laherparepvec has showed promising therapeutic effects in advanced melanoma, especially when combined with immune checkpoint inhibitors with moderate adverse effects. Diverse viruses like herpes simplex virus, adenovirus, vaccina virus, and so on could be engineered as vectors to express different transgenic payloads, vastly expanding the therapeutic potential of oncolytic virotherapy. A number of related clinical trials are under way which are mainly focusing on solid tumors. Studies about further optimizing the genome of oncolytic viruses or improving the delivering system are in the hotspot, indicating the future development of oncolytic virotherapy in the clinic. This review introduces the latest progress in clinical trials and pre-clinical studies as well as technology innovations directed at oncolytic viruses. The challenges and perspectives of oncolytic virotherapy towards clinical application are also discussed.