Impacts of vascular comorbidities on free flap perfusion in microvascular head and neck reconstruction.

PURPOSE: Arterial hypertension (AHTN), type 2 diabetes mellitus (DM), and atherosclerotic vascular disease (ASVD) are common vascular comorbidities in patients undergoing reconstruction of the head and neck region with a microvascular free flap. These conditions may affect flap perfusion (microvascular blood flow and tissue oxygenation), which is a prerequisite for flap survival and thus reconstruction success. This study aimed to investigate the impacts of AHTN, DM, and ASVD on flap perfusion. METHODS: Data from 308 patients who underwent successful reconstruction of the head and neck region with radial free forearm flaps, anterolateral thigh flaps, or fibula free flaps between 2011 and 2020 were retrospectively analyzed. Flap perfusion was measured intraoperatively and postoperatively with the O2C tissue oxygen analysis system. Flap blood flow, hemoglobin concentration, and hemoglobin oxygen saturation were compared between patients with and without AHTN, DM, and ASVD. RESULTS: Intraoperative hemoglobin oxygen saturation and postoperative blood flow were lower in patients with ASVD than in patients without ASVD (63.3% vs. 69.5%, p = 0.046; 67.5 arbitrary units [AU] vs. 85.0 AU, p = 0.036; respectively). These differences did not persist in the multivariable analysis (all p > 0.05). No difference was found in intraoperative or postoperative blood flow or hemoglobin oxygen saturation between patients with and without AHTN or DM (all p > 0.05). CONCLUSION: Perfusion of microvascular free flaps used for head and neck reconstruction is not impaired in patients with AHTN, DM, or ASVD. Unrestricted flap perfusion may contribute to the observed successful use of microvascular free flaps in patients with these comorbidities.

Flap perfusion monitoring with an attached surface probe in microvascular reconstruction of the oral cavity.

OBJECTIVES: Postoperative flap monitoring is essential in oral microvascular reconstruction for timely detection of vascular compromise. This study investigated the use of attached surface probes for the oxygen-2-see (O2C) analysis system (LEA Medizintechnik, Germany) for intraoral flap perfusion monitoring. MATERIALS AND METHODS: The study included 30 patients who underwent oral reconstruction with a microvascular radial-free forearm flap (RFFF) or anterolateral thigh flap (ALTF) between 2020 and 2022. Flap perfusion was measured with attached (3-mm measurement depth) and unattached surface probes (2- and 8-mm measurement depths) for the O2C analysis system at 0, 12, 24, 36, and 48 h postoperatively. Flap perfusion monitoring with attached surface probes was evaluated for cut-off values for flap blood flow, hemoglobin concentration, and hemoglobin oxygen saturation indicative of vascular compromise and for accuracy and concordance with unattached surface probes. RESULTS: Three RFFFs were successfully revised, and one ALTF was unsuccessfully revised. The cut-off values indicative of vascular compromise for flap perfusion monitoring with attached surface probes were for RFFF and ALTF: blood flow < 60 arbitrary units (AU) and < 40AU, hemoglobin concentration > 100AU and > 80AU (both > 10% increase), and hemoglobin oxygen saturation < 40% and < 30%. Flap perfusion monitoring with attached surface probes yielded a 97.1% accuracy and a Cohen's kappa of 0.653 (p < 0.001). CONCLUSIONS: Flap perfusion monitoring with attached surface probes for the O2C analysis system detected vascular compromise accurately and concordantly with unattached surface probes. CLINICAL RELEVANCE: Attached surface probes for the O2C analysis system are a feasible option for intraoral flap perfusion monitoring.

Evaluation of perfusion parameters of gingival inflammation using laser Doppler flowmetry and tissue spectrophotometry- a prospective comparative clinical study.

BACKGROUND: The aim of this study was to determine the values of different perfusion parameters- such as oxygen saturation, the relative amount of hemoglobin, and blood flow- in healthy subjects compared to patients with gingivitis as a non-invasive measurement method. METHODS: A total of 114 subjects were enrolled in this study and separated into subjects with gingivitis (50) and without gingivitis (64) based on clinical examination. Gingival perfusion was measured at 22 points in the maxilla and mandible using laser Doppler flowmetry and tissue spectrophotometry (LDF-TS) with the "oxygen to see" device. All patients underwent measurement of gingival perfusion, followed by the clinical evaluation (measurement of probing depths, evaluation of bleeding on probing, plaque level, and biotype). Perfusion parameters were compared between the groups, associations between the non-invasive and clinical measurements were analyzed, and theoretical optimal cut-off values for predicting gingivitis were calculated with receiver operating characteristics. RESULTS: The mean oxygen saturation, mean relative amount of hemoglobin, and mean blood flow all significantly differed between the groups with and without gingivitis (p = 0.005, p < 0.001, and p < 0.001, respectively). The cut-off value for predicting gingivitis was > 40 AU (p < 0.001; sensitivity 0.90, specificity 0.67). CONCLUSIONS: As a non-invasive method, LDF-TS can help determine gingival hyperemia. Flow values above 40 AU indicate a higher risk of hyperemia, which can be associated with inflammation. The LDF-TS method can be used for the objective evaluation of perfusion parameters during routine examinations and can signal the progression of hyperperfusion before any change in clinical parameters is observed. TRIAL REGISTRATION: All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. The study was approved by the institutional Clinical Research Ethics Committee (Ethik-Kommission der Medizinischen Fakultät der RWTH Aachen, Decision Number 286/20) and retrospectively registered by the German Clinical Trials Register (File Number DRKS00024048, registered on the 15th of October 2021).

Evaluation of the long-term results of vascular anastomosis using polyurethane adhesive and shape-memory stent in the rat carotid artery model.

INTRODUCTION: In free flaps, 5%-10% of complications are related to failure of sutured vascular anastomoses. Adhesive-based microvascular anastomoses are potential alternatives but are associated with failure rates of 70% in research studies. VIVO is a new adhesive with slow biodegradation within 6 months that has shown a 100% patency rate in research studies over 2 h observation time but long-term patency has not been evaluated. The authors hypothesize that VIVO will enable a reliable microvascular procedure comparable to sutured anastomoses over a 28-day period. MATERIALS AND METHODS: The right common carotid artery of 60 male Sprague Dawley rats, ~450 g, were used for microvascular end-to-end anastomosis. VIVO was applied with reduced sutures with a temporary catheter in one group and in the other with a custom-shaped memory stent. Anastomoses with eight interrupted sutures served as control. All groups were n = 20. Anastomosis time and bleeding were recorded for each procedure. Doppler flowmetry was performed 20 min, 1, 10, and 28 days postoperatively. Postmortem toluidine staining was used for semi-quantitative analysis of stenosis, thrombosis, necrosis, and aneurysm formation by histologic evaluation. RESULTS: No occlusion was detected 20 min and 1 day postoperative, and after 28 days of observation in all anastomoses. The anastomosis time of the VIVO with catheter group was about 32% significantly faster than the VIVO with stent group. In the VIVO group with stent, the bleeding time was ~80% shorter than in the control group with 2.1 ± 0.3 and VIVO with catheter 2.0 ± 0.5 (p ≤ .001 each). Minor and nonsignificant stent-associated thrombus formation and stent-typical intraluminal stenosis were detected exclusively in the VIVO with stent group. CONCLUSION: Within the limitations of a rat study, the use of VIVO in anastomosis showed promising results. VIVO with catheter was found to be advantageous.

Effect of drain application on postoperative complaints after surgical removal of impacted wisdom teeth-a randomized observer-blinded split-mouth clinical trial.

OBJECTIVES: The aim of this randomized observer-blinded split-mouth-study is to objectively assess the influence of a rubber drain on postoperative swelling using 3D face scans as measurement method and additionally evaluate pain, trismus and complications after the osteotomy of lower third molars. MATERIAL AND METHODS: Seventy-two patients with symmetrically impacted lower wisdom teeth were recruited. Before the operation, patients rated pain using the visual analogue scale, the interincisal distance was measured, and 3D face scans were taken with an optical scanner. Each patient underwent two procedures which were at least 30 days apart. On one side, a rubber drain was inserted randomly before closure, the contralateral control side was closed without drainage. On the third and tenth postoperative day, face scans to quantify the swelling, pain evaluation and trismus measurements were performed. Due to loss of follow-up, 32 patients were excluded which resulted in 40 out of 72 patients remaining in the study. RESULTS: There was no statistical difference in using a drain on swelling and trismus on the third and tenth day (p > 0.05). Pain was slightly worse on the third day on the treatment side, but the difference was not significant (p > 0.05). We observed no differences in the number of wound infections. CONCLUSIONS: The insertion of a rubber drain does not have any influence on swelling, pain or trismus and has no impact on the number of wound infections. CLINICAL RELEVANCE: The use of a rubber drain cannot be recommended as no reduction of postoperative discomfort was detected.

Postoperative Morbidity and Complications in Elderly Patients after Harvesting of Iliac Crest Bone Grafts.

Background and objectives: In oral and maxillofacial operations, the iliac crest is a commonly used donor site from which to harvest bone for augmentation prior to dental implantation or for reconstruction of jaw defects caused by trauma or pathological lesions. In an aging society, the proportion of elderly patients undergoing iliac crest bone grafting for oral augmentation is growing. Although postoperative morbidity is usually moderate to low, the age and health of the patient should be considered as risk factors for complications and delayed mobilization after the operation. The aim of this retrospective study was to evaluate the postoperative morbidity and complications in elderly patients after the harvesting of iliac crest bone grafts for oral surgery. Material and Methods: Data were collected from a total of 486 patients (aged 7-85) who had a surgical procedure that included the harvesting of iliac crest bone grafts for intraoral transplantation. All patients were operated on between 2005 and 2021 in the Department for Oral and Maxillofacial Surgery of the University Hospital in Aachen, Germany. As parameters for postoperative morbidity and complications, gait disturbances, hypesthesia of cutaneous nerves, incision hernias, iliac crest fractures, delayed wound healing, and unfavorable scar formation at the donor site were all evaluated. Results: The study was performed with 485 patients due to the exclusion of one patient as the only one from whom grafts were taken from both sides. When younger and older patients were compared, neither gait disturbances (p = 0.420), nor hernias (p = 0.239), nor fractures (p = 0.239), nor hypesthesia (p = 0.297), nor wound healing delay (p = 0.294), nor scar problems (p = 0.586) were significantly different. However, the volume of the graft was significantly correlated with the duration of the hospital stay (ρ = 0.30; p < 0.01) but not with gait disturbances (ρ = 0.60; p = 0.597). Additionally, when controlling for age (p = 0.841), sex (p = 0.031), ASA class (p = 0.699), preexisting orthopedic handicaps (p = 0.9828), and the volume of the bone graft (p = 0.770), only male sex was associated with the likelihood of suffering gait disturbances (p = 0.031). Conclusions: In conclusion, harvesting bone grafts from the anterior iliac crest for intraoral augmentation is a safe procedure for both young and elderly patients. Although there is some postoperative morbidity, such as gait disturbances, hypesthesia, scar formation, or delayed wound healing at the donor site, rates for these minor complications are low and mostly of short duration. Major complications, such as fractures or incision hernias, are very rare. However, in our study, the volume of the bone graft was associated with a longer stay in hospital, and this should be considered in the planning of iliac crest bone graft procedures.

Providing Professionally Meaningful Recognition to Enhance Frontline Engagement.

To achieve transformation in care delivery, frontline nursing staff must be committed to their organization's mission, engaged in their work, and capable of delivering high-quality care. However, data from Advisory Board Survey Solutions show that, when compared with other frontline staff, nurses are the least engaged and most disengaged. In this article, the authors describe strategies for addressing a top opportunity for improving nurse engagement-ensuring nurses feel meaningfully recognized for their professional impact.

Translating Market Forces Into Frontline Terms.

To transform healthcare delivery, frontline nursing staff must be committed to their work and to driving their organization's mission forward. However, Advisory Board Survey Solutions data show that nurses are the least engaged as compared with other frontline staff. In this article, the authors identify a top opportunity for improving nurse engagement: ensuring that nurses understand how executives' actions reflect their organization's missions and values.

Broadening Access to Nontraditional Development Opportunities to Drive Frontline Engagement.

The nursing workforce is at the center of many changes associated with care delivery transformation. To achieve this transformation, frontline staff must be engaged in their work, committed to their organization's mission, and capable of delivering high-quality care. In this article, the authors describe strategies for addressing 1 of the greatest opportunities for improving nurse engagement identified using these data: ensuring nurses feel professional development and promotion opportunities offered at their organization help them to improve.

Regulating the Flow of Change to Reduce Fontline Nurse Stress and Burnout.

The nursing workforce is at the center of many changes associated with care delivery transformation. To achieve this transformation, frontline nursing staff must be engaged in their work, committed to their organization's mission, and capable of delivering high-quality care. To identify top opportunities for driving nursing engagement, researchers from The Advisory Board Company analyzed engagement survey responses from more than 343 000 employees at 575 healthcare organizations. In this article, the authors describe 3 strategies for addressing 1 of the greatest opportunities for improving nurse engagement: ensuring nurses feel their organization helps them reduce stress and burnout.

Does the Anastomosis Recipient Vessel Have an Influence on Free Flap Perfusion in Microvascular Head and Neck Reconstruction-A Retrospective Analysis of 338 Cases with Comparison of Flap Perfusion between Different Arterial and Venous Recipient Vessels in Radial Free Forearm Flaps, Anterolateral Thigh Flaps, and Fibula Free Flaps.

Background: Flap perfusion is a prerequisite for microvascular free flap survival and a parameter routinely used for flap monitoring. The aim of this study was to investigate the influence of the anastomosis recipient vessel on flap perfusion. Methods: Flap perfusion was retrospectively analyzed in 338 patients who underwent head and neck reconstruction with microvascular free flaps between 2011 and 2020. The Oxygen-to-see tissue oxygen analysis system measurements for intraoperative and postoperative flap blood flow, hemoglobin concentration, and hemoglobin oxygen saturation at 8 and 2 mm tissue depths were compared between arterial anastomosis recipient vessels (external carotid artery [ECA], facial artery [FAA], lingual artery [LIA], and superior thyroid artery [STA]) and venous anastomosis recipient vessels (internal jugular vein [IJV], combination of IJV and IJV branches, IJV branches, and external jugular vein). Results: The postoperative hemoglobin concentration at 2 mm tissue depth differed significantly between arterial anastomosis recipient vessels (ECA, 41.0 arbitrary units [AU]; FAA, 59.0 AU; LIA, 51.5 AU; STA, 59.0 AU; p = 0.029). This difference did not persist in the multivariable testing (p = 0.342). No other differences in flap blood flow, hemoglobin concentration, or hemoglobin oxygen saturation were observed between the arterial and venous anastomosis recipient vessels (p > 0.05 for all). Conclusions: The arterial and venous recipient vessels used for anastomosis did not influence microvascular free flap perfusion. This underlines the capability of the studied recipient vessels to adequately perfuse free flaps, may explain the observed indifferent flap survival rates between commonly used anastomosis recipient vessels, and implies that the recipient vessel is not a confounding variable for flap monitoring with the Oxygen-to-see tissue oxygen analysis system. Further prospective studies are needed to confirm the findings.

Optical Three-Dimensional Imaging for Objective Evaluation of the Donor Site after Anterolateral Thigh Flap Surgery.

Background: The high volume of the fasciomyocutaneous anterolateral thigh flaps (ALT) is suitable for the reconstruction of pronounced soft tissue defects. At the same time, harvesting ALT results in a drastic change in thigh shape. Here, we present an optical three-dimensional imaging method for thigh comparison, which can be an objective and reproducible method for evaluating donor sites after ALT harvesting. Methods: In total, 128 thighs were scanned with an optical three-dimensional scanner, Vectra XT ®. Sixty-eight non-operated right and left thighs were compared and served as a control. Sixty thighs were scanned in the ALT group. The average surface area deviations, thigh volume, thigh circumference, and flap ratio to thigh circumference were calculated. The results were correlated with Δthigh circumference and Δvolume of the unoperated thighs of the control group. Results: No significant difference between the thigh volumes of the right and left thighs was found in the control group. Removal of an ALT flap showed a significant (p < 0.007) volume reduction compared to unoperated thighs (2.7 ± 0.8 L and 3.3 ± 0.9 L, respectively). Flap area correlated strongly with the Δthigh circumference (r = 0.66, p < 0.001) and Δvolume (r = 0.68, p < 0.001). Strong correlations were observed between flap ratio and thigh circumference with Δhigh circumference (r = 0.57, p < 0.001) and Δvolume (r = 0.46, p < 0.05). Conclusions: Optical three-dimensional imaging provides an objective and reproducible tool for detecting changes in thigh morphology volume differences after ALT harvesting.

Dental Implant Placement in the Maxilla Following Ridge Augmentation with Free Iliac Bone Graft and Oral Rehabilitation with Fixed Prosthesis: a Three-Year Follow-Up Study.

Objectives: This prospective follow-up study aimed to evaluate the stability values of tapered titanium implants inserted into maxilla following ridge augmentation with free iliac bone graft and crestal bone changes up to three years of follow-up. Material and Methods: A total of seven patients with 34 tapered titanium implants in the maxilla with fixed protheses were enrolled in this prospective follow-up study. Patients with previously augmented maxillae using free iliac bone grafts were included. Implant stability was measured (Osstell™) for up to three months of healing. Peri-implant bone resorption was measured using radiographic images taken immediately after implant surgery and after three years. Using a clinical and radiological examination survival and success rates were evaluated. Results: After implant insertion, the stability was 60.93, whereas the stability increased significantly (P = 0.0192) to 64.97 at implant exposure (after 3 months). The mean bone loss around the implants was 1.13 mm after three years. Clinical parameters revealed a mean sulcus depth of 2.76 (1.18) mm and a bleeding on probing score of 0.29 (0.58). The survival rate was 100%, and the success rate was 67.65% at the end of the study. Conclusions: Tapered implants can be used in free iliac bone grafts for fixed dentures. Implant stability values were high after insertion. In terms of a success rate of 67.65%, the patient's jaw reconstruction indicated a reduced implant success when comparing the data with healthy patients without any augmentation procedures.

Impact of age on microvascular free flap perfusion in head and neck reconstruction.

OBJECTIVE: Aging is associated with structural and functional cardiovascular changes that may affect microvascular free flap perfusion, which is a prerequisite for flap viability and flap success. This study is aimed to investigate the impact of age on flap perfusion. STUDY DESIGN: Intraoperative and postoperative flap perfusion of 348 patients who underwent successful reconstruction with a radial free forearm flap (RFFF), anterolateral thigh flap (ALTF), or fibular free flap (FFF) was retrospectively analyzed and compared between older (>70 years) and younger patients (≤70 years) using the Mann-Whitney test and linear regression models. RESULTS: Intraoperative flap blood flow and hemoglobin concentration (55.5 arbitrary units [AU] vs 69.3 AU, P = .004; 42.5 AU vs 47.3 AU, P = .016, respectively) were reduced in ALTFs in older patients compared with younger patients. These 2 differences did not persist in multivariable testing (P = .097 and P = .323, respectively). No other differences were observed between the older and younger patients in terms of intraoperative and postoperative flap blood flow, hemoglobin concentration, and hemoglobin oxygen saturation in RFFFs, ALTFs, and FFFs (all P > .05). CONCLUSIONS: Age has no impact on microvascular free flap perfusion in RFFFs, ALTFs, and FFFs. This could contribute to the observed equal success of free flaps in older patients.

Evaluation of the Hemostatic Effect of an Innovative Tissue Adhesive during Extraction Therapy under Rivaroxaban in a Rodent Model.

An increase in rivaroxaban therapies is associated with increased numbers of postoperative bleeding despite the use of hemostatic sponges, which are currently the gold standard treatment. VIVO has shown promising hemostatic results, favorable tissue properties, and ease of application, although it has not yet been used in the oral cavity. The aim of this study was to evaluate the hemostatic properties of VIVO in the extraction sockets of 31 rodents and compare this to gelatin sponge (GSP) therapy. At rivaroxaban concentrations of 264.10 ± 250.10 ng/mL, 62 extraction sockets were generated, of which 31 were treated with VIVO and 31 with GSP. The duration time, early and late bleeding events, and wound healing score were determined. Histologic examinations of the tissues were performed after 5 days. VIVO presented a longer procedure, 1.26 ± 0.06 min, but a significantly shorter bleeding time, 0.14 ± 0.03 min. There was no difference between the two groups in terms of the severity and timing of bleeding. More minor early bleeding events were observed for GSP. VIVO showed a significantly better healing score, with favorable histological results. In an animal study, VIVO showed promising hemostatic properties after tooth extraction under ongoing anticoagulative therapy.

Attached compared with unattached surface probes for monitoring flap perfusion in microvascular head and neck reconstruction: a feasibility study.

Unattached surface probes are commonly used with the O2C analysis system (LEA Medizintechnik, Germany) to monitor microvascular free flap perfusion. This study compared attached and unattached surface probes for extraoral free flaps. The study included 34 patients who underwent extraoral microvascular head and neck reconstruction between 2020 and 2022. Flap perfusion was monitored postoperatively using the O2C analysis system at 0, 12, 24, 36, and 48 h, with an attached surface probe at 3 mm tissue depth and an unattached surface probe at 2 mm and 8 mm tissue depths. Clinical complications, technical errors, and perfusion measurement values were compared. No clinical complications (attachment suture infections) or technical errors (probe detachment) occurred. Flap blood flow values of the probes were partially different (3 mm vs. 2 and 8 mm: p < 0.001; p = 0.308) and moderately correlated (3 mm with 2 and 8 mm: r = 0.670, p < 0.001; r = 0.638, p < 0.001). Hemoglobin concentration and oxygen saturation values were generally different (3 mm vs. 2 and 8 mm: all p < 0.001) and variably correlated (3 mm with 2 and 8 mm: r = 0.756, r = 0.645; r = 0.633, r = 0.307; all p < 0.001). Both probes are comparable in terms of technical feasibility and patient safety, with flap perfusion values dependent on tissue measurement depth.

Impact of secondary tumor-free resection margins in soft tissue on local, regional, and distant recurrence in R0-resected oral squamous cell carcinoma.

BACKGROUND: The impact of additional soft tissue resection on recurrence of oral squamous cell carcinoma (OSCC) remains controversial. The study aim was to compare recurrence between patients with secondary tumor-free resection margins after intraoperative additional resection (STF-RM) and patients with primary tumor-free resection margins without additional resection (PTF-RM). METHODS: Forty-five patients with STF-RM were matched with patients with PTF-RM according to Union for International Cancer Control stage, tumor location, and treatment modality and compared for local, regional, and distant recurrence. RESULTS: Patients with STF-RM showed lower local and distant control rates compared to patients with PTF-RM (66.2% vs. 82.8%; p = 0.045 and 86.3% vs. 100.0%; p = 0.021). STF-RM was the only predictor of local recurrence accounting for tumor (T) status, nodal (N) status, tumor grade, margin distance, and extracapsular extension (hazard ratio 4.21 [95% confidence interval 1.26-14.04]; p = 0.019). CONCLUSIONS: STF-RM have an adverse impact on local and distant recurrence of OSCC.

Cross-sectional analysis of spatial distribution and total amount of harvestable calvarian split bone.

BACKGROUND: Bone resorption of the jaw leads to challenging implant placement. Frequently, augmentation of the jaw is necessary. Is calvarian split bone an alternative to other extraoral donor sites and what volume of bone is harvestable? The aim was to evaluate the spatial distribution and the total amount of harvestable calvarian split bone. MATERIAL AND METHODS: Computerized tomographies of 600 patients were divided into four groups (male and female: ≤45 years and >45 years). The skull was segmented and cut into the harvestable compartments (Os frontale, Ossa parietalia). The volume and thickness of the harvestable bone were calculated. RESULTS: The overall harvestable bone was 110.644 ± 25.429 cm³. The bone from the Os frontale was significantly less than harvestable bone from the Os parietale (p < 0.001). More bone could be harvested from the right Os parietale. In younger males, significantly more bone could be harvested than in females (females ≤45 years: p = 0.001; females >45 years: p = 0.003). A weak negative correlation existed between the participants' age and the harvestable bone volume of the left Os parietale (r = -0.087; p = 0.033). The thickness of the harvestable bone from the Ossa parietalia is greater in females than in males. CONCLUSION: A great amount of calvarian bone can be harvested to augment the jaw. Surgeons must acknowledge that more bone is harvestable from males than females while the female bone is thicker. Calculating the volume leads to accurate results of the available bone.

The deubiquitylase USP9X controls ribosomal stalling.

When a ribosome stalls during translation, it runs the risk of collision with a trailing ribosome. Such an encounter leads to the formation of a stable di-ribosome complex, which needs to be resolved by a dedicated machinery. The initial stalling and the subsequent resolution of di-ribosomal complexes requires activity of Makorin and ZNF598 ubiquitin E3 ligases, respectively, through ubiquitylation of the eS10 and uS10 subunits of the ribosome. We have developed a specific small-molecule inhibitor of the deubiquitylase USP9X. Proteomics analysis, following inhibitor treatment of HCT116 cells, confirms previous reports linking USP9X with centrosome-associated protein stability but also reveals a loss of Makorin 2 and ZNF598. We show that USP9X interacts with both these ubiquitin E3 ligases, regulating their abundance through the control of protein stability. In the absence of USP9X or following chemical inhibition of its catalytic activity, levels of Makorins and ZNF598 are diminished, and the ribosomal quality control pathway is impaired.

Immunomonitoring of Stage IV Relapsed Neuroblastoma Patients Undergoing Haploidentical Hematopoietic Stem Cell Transplantation and Subsequent GD2 (ch14.18/CHO) Antibody Treatment.

Haploidentical stem cell transplantation (haplo SCT) in Stage IV neuroblastoma relapsed patients has been proven efficacious, while immunotherapy utilizing the anti-GD2 antibody dinutuximab beta has become a standard treatment for neuroblastoma. The combinatorial therapy of haplo SCT and dinutuximab may potentiate the efficacy of the immunotherapy. To gain further understanding of the synergistic effects, functional immunomonitoring was assessed during the clinical trial CH14.18 1021 Antibody and IL2 After haplo SCT in Children with Relapsed Neuroblastoma (NCT02258815). Rapid immune reconstitution of the lymphoid compartment was confirmed, with clinically relevant dinutuximab serum levels found in all patients over the course of treatment. Only one patient developed human anti-chimeric antibodies (HACAs). In-patient monitoring revealed highly functional NK cell posttransplant capable of antibody-dependent cellular cytotoxicity (ADCC). Degranulation of NK cell subsets revealed a significant response increased by dinutuximab. This was irrespective of the KIR receptor-ligand constellation within the NK subsets, defined by the major KIR receptors CD158a, CD158b, and CD158e. Moreover, complement-dependent cytotoxicity (CDC) was shown to be an extremely potent effector-cell independent mechanism of tumor cell lysis, with a clear positive correlation to GD2 expression on the cancer cells as well as to the dinutuximab concentrations. The ex vivo testing of patient-derived effector cells and the sera collected during dinutuximab therapy demonstrated both high functionality of the newly established lymphoid immune compartment and provided confidence that the antibody dosing regimen was sufficient over the duration of the dinutuximab therapy (up to nine cycles in a 9-month period). During the course of the dinutuximab therapy, proinflammatory cytokines and markers (sIL2R, TNFa, IL6, and C reactive protein) were significantly elevated indicating a strong anti-GD2 immune response. No impact of FcGR polymorphism on event-free and overall survival was found. Collectively, this study has shown that in-patient functional immunomonitoring is feasible and valuable in contributing to the understanding of anti-cancer combinatorial treatments such as haplo SCT and antibody immunotherapy.