CD4+ T cell-induced inflammatory cell death controls immune-evasive tumours.

Most clinically applied cancer immunotherapies rely on the ability of CD8+ cytolytic T cells to directly recognize and kill tumour cells1-3. These strategies are limited by the emergence of major histocompatibility complex (MHC)-deficient tumour cells and the formation of an immunosuppressive tumour microenvironment4-6. The ability of CD4+ effector cells to contribute to antitumour immunity independently of CD8+ T cells is increasingly recognized, but strategies to unleash their full potential remain to be identified7-10. Here, we describe a mechanism whereby a small number of CD4+ T cells is sufficient to eradicate MHC-deficient tumours that escape direct CD8+ T cell targeting. The CD4+ effector T cells preferentially cluster at tumour invasive margins where they interact with MHC-II+CD11c+ antigen-presenting cells. We show that T helper type 1 cell-directed CD4+ T cells and innate immune stimulation reprogramme the tumour-associated myeloid cell network towards interferon-activated antigen-presenting and iNOS-expressing tumouricidal effector phenotypes. Together, CD4+ T cells and tumouricidal myeloid cells orchestrate the induction of remote inflammatory cell death that indirectly eradicates interferon-unresponsive and MHC-deficient tumours. These results warrant the clinical exploitation of this ability of CD4+ T cells and innate immune stimulators in a strategy to complement the direct cytolytic activity of CD8+ T cells and natural killer cells and advance cancer immunotherapies.

Evaluation of tooth color change after a bleaching process with different lasers.

The aim of this in vitro study was to evaluate the efficiency of diode laser-activated bleaching systems for color change of teeth. 75 extracted teeth were studied in five different bleaching protocols. Group 1: diode laser 445 nm, 320 µm fiber, 0.5W, continuous wave mode, dose 53 J/cm2. Group 2: diode laser 970 nm, 320 µm fiber, 1W, continuous wave mode, dose 106.10 J/cm2. Group 3: diode laser 940 nm, bleaching handpiece, 7W, continuous wave mode, dose 105 J/cm2. Group 4: diode laser 940 nm, 300 µm fiber, 2W, continuous wave mode, dose 47.16 J/cm2. Group 5: bleaching process without laser activation. In groups 1, 2 and 5, teeth were bleached with Perfect Bleach Office + and in groups 3 and 4, LaserWhite20 bleaching gel was used. Tooth color was determined immediately after the bleaching process using a spectrophotometer. Color change data on the CIE L * a * b* system was analyzed statistically by the one-way ANOVA and Tukey's HSD test. All bleaching procedures resulted in a change of color. All laser groups (∆E * ab > 3) have statistically larger ∆E * ab values than the control group (∆E * ab = 0.73) (p < 0.05). The diode laser 445 nm has the largest ∆E * ab value (∆E * ab = 4.65) and results in a significantly higher color difference than all other groups. In terms of color score difference in VITA Shades, all laser-activated groups lead to a lightening effect while the control group leads to only a slight lightening effect. The diode laser 445 nm produced the greatest color difference. Laser-activated bleaching is more effective than conventional bleaching without light activation. The diode laser 445 nm performs best in this in vitro study.

Comprehensive analysis of body composition features in melanoma patients treated with tyrosine kinase inhibitors.

BACKGROUND: The introduction of tyrosine kinase inhibitors (TKI) has greatly improved the management of metastatic melanoma. Recent studies have uncovered a relationship between the body mass index (BMI) and outcome of patients with metastatic melanoma. However, conflicting results have challenged the relevance of this finding. In the current work, we aim to dissect body composition features of melanoma patients treated with TKI to evaluate their value as biomarkers. PATIENTS AND METHODS: We analyze body composition features via CT scans in a retrospective cohort of 57 patients with non-resectable stage III/IV melanoma receiving first-line treatment with TKI in our department, focusing on the impact of body composition on treatment efficacy and occurrence of adverse events. RESULTS: In uni- and multivariate analyses, we identify an association between the visceral adipose tissue gauge index (VATGI) and survival. We furthermore profile additional body composition features including sarcopenia, which was also associated with a shorter overall survival. Finally, we detected an enrichment of cases with fatigue in patients with low VATGI. CONCLUSIONS: Our study represents the first exploratory study evaluating the suitability of body composition measurements as biomarkers for melanoma patients treated with TKI. Our data suggest a putative use of VATGI as a biomarker predicting patient outcome.

Real-world outcomes using PD-1 antibodies and BRAF + MEK inhibitors for adjuvant melanoma treatment from 39 skin cancer centers in Germany, Austria and Switzerland.

BACKGROUND: Programmed death-1 (PD-1) antibodies and BRAF + MEK inhibitors are widely used for adjuvant therapy of fully resected high-risk melanoma. Little is known about treatment efficacy outside of phase III trials. This real-world study reports on clinical outcomes of modern adjuvant melanoma treatment in specialized skin cancer centers in Germany, Austria and Switzerland. METHODS: Multicenter, retrospective study investigating stage III-IV melanoma patients receiving adjuvant nivolumab (NIV), pembrolizumab (PEM) or dabrafenib + trametinib (D + T) between 1/2017 and 10/2021. The primary endpoint was 12-month recurrence-free survival (RFS). Further analyses included descriptive and correlative statistics, and a multivariate linear-regression machine learning model to assess the risk of early melanoma recurrence. RESULTS: In total, 1198 patients from 39 skin cancer centers from Germany, Austria and Switzerland were analysed. The vast majority received anti PD-1 therapies (n = 1003). Twelve-month RFS for anti PD-1 and BRAF + MEK inhibitor-treated patients were 78.1% and 86.5%, respectively (hazard ratio [HR] 1.998 [95% CI 1.335-2.991]; p = 0.001). There was no statistically significant difference in overall survival (OS) in anti PD-1 (95.8%) and BRAF + MEK inhibitor (96.9%) treated patients (p > 0.05) during the median follow-up of 17 months. Data indicates that anti PD-1 treated patients who develop immune-related adverse events (irAEs) have lower recurrence rates compared to patients with no irAEs (HR 0.578 [95% CI 0.443-0.754], p = 0.001). BRAF mutation status did not affect overall efficacy of anti PD-1 treatment (p > 0.05). In both, anti PD-1 and BRAF + MEK inhibitor treated cohorts, data did not show any difference in 12-month RFS and 12-month OS comparing patients receiving total lymph node dissection (TLND) versus sentinel lymph node biopsy only (p > 0.05). The recurrence prediction model reached high specificity but only low sensitivity with an AUC = 0.65. No new safety signals were detected. Overall, recorded numbers and severity of adverse events were lower than reported in pivotal phase III trials. CONCLUSIONS: Despite recent advances in adjuvant melanoma treatment, early recurrence remains a significant clinical challenge. This study shows that TLND does not reduce the risk of early melanoma recurrence and should only be considered in selected patients. Data further highlight that variables collected during clinical routine are unlikely to allow for a clinically relevant prediction of individual recurrence risk.

Wound healing after therapy of oral potentially malignant disorders with a 445-nm semiconductor laser: a randomized clinical trial.

OBJECTIVES: Oral potentially malignant disorders (OPMDs) are the most clinically relevant precursor lesions of the oral squamous cell carcinoma (OSCC). OSCC is one of the 15 most common cancers worldwide. OSCC is with its high rate of mortality an important cause of death worldwide. The diagnosis and therapy of clinically relevant precursor lesions of the OSCC is one of the main parts of prevention of this malignant disease. Targeted therapy is one of the main challenges concerning an oncologically safe tissue removal without overwhelming functional and aesthetic impairment. MATERIALS AND METHODS: In this randomized controlled trial, a newly introduced intraoral 445-nm semiconductor laser (2W; cw-mode; SIROLaser Blue, Dentsply Sirona, Bensheim, Germany) was used in the therapy of OPMDs. Duration and course of wound healing, pain, and scar tissue formation were compared to classical cold blade removal with primary suture by measuring remaining wound area, tissue colorimetry, and visual analogue scale. The study includes 40 patients randomized using a random spreadsheet sequence in two groups (n1 = 20; n2 = 20). RESULTS: This comparative analysis revealed a significantly reduced remaining wound area after 1, 2, and 4 weeks in the laser group compared to the cold blade group (p < 0.05). In the laser group, a significantly reduced postoperative pain after 1 week was measured (p < 0.05). CONCLUSION: Laser coagulation of OPMDs with the investigated 445-nm semiconductor laser is a safe, gentle, and predictable surgical procedure with beneficial wound healing and reduced postoperative discomfort. CLINICAL RELEVANCE: Compared to the more invasive and bloody cold blade removal with scalpel, the 445-nm semiconductor laser could be a new functional less traumatic tool in the therapy of OPMDs. The method should be further investigated with regard to the identification of further possible indications. TRAIL REGISTRATION: German Clinical Trials Register No: DRKS00032626.

Tumor cell intrinsic Toll-like receptor 4 signaling promotes melanoma progression and metastatic dissemination.

Most melanoma-associated deaths result from the early development of metastasis. Toll-like receptor 4 (TLR4) expression on nontumor cells is well known to contribute to tumor development and metastatic progression. The role of TLR4 expression on tumor cells however is less well understood. Here we describe TLR4 as a driver of tumor progression and metastatic spread of melanoma cells by employing a transplantable mouse melanoma model. HCmel12 melanoma cells lacking functional TLR4 showed increased sensitivity to tumor necrosis factor α induced cell killing in vitro compared to cells with intact TLR4. Interestingly, TLR4 knockout melanoma cells also showed impaired migratory capacity in vitro and a significantly reduced ability to metastasize to the lungs after subcutaneous transplantation in vivo. Finally, we demonstrate that activation of TLR4 also promotes migration in a subset of human melanoma cell lines. Our work describes TLR4 as an important mediator of melanoma migration and metastasis and provides a rationale for therapeutic inhibition of TLR4 in melanoma.

Generalized Nodules and Sclerosis of the Skin: A Quiz.

is missing (Quiz).

The aryl hydrocarbon receptor promotes inflammation-induced dedifferentiation and systemic metastatic spread of melanoma cells.

The aryl hydrocarbon receptor (AHR) is a ligand binding-transcription factor of the basic helix-loop-helix family regulating multiple cellular functions such as differentiation, cell cycle, apoptosis, and inflammatory reactions. In neoplastic diseases, the AHR has been described to modulate proliferation and differentiation in dichotomous ways, either inhibiting or augmenting the growth of tumors. The precise role of AHR in melanoma is mostly unknown. Here, we report a functional effect of AHR activation on inflammation-induced melanoma cell dedifferentiation and the development of lung metastases in a mouse model. Via in silico analyses of "The Cancer Genome Atlas" human melanoma cohort, we detected a correlation between AHR expression levels and a dedifferentiated melanoma cell phenotype with an invasive gene signature, which we were able to functionally recapitulate in a panel of human melanoma cell lines. Both human and mouse melanoma cell lines upregulated AHR expression after inflammatory stimulation with tumor necrosis factor-α (TNF-α). Activation of AHR in human and mouse melanoma cell lines with the endogenous ligand formylindolo(3,2-b)carbazole (FICZ) promoted inflammation-induced dedifferentiation in vitro. Importantly, mouse melanoma cells with CRISPR/Cas9-mediated disruption of the AHR gene showed impaired in vivo tumor growth after transplantation in the skin as well as decreased numbers of spontaneous lung metastases. Taken together, our results demonstrate a functional role for AHR expression in melanoma development and metastatic progression. This provides a scientific basis for future experiments that further dissect the underlying molecular mechanisms and assess the potential for AHR inhibition as part of multimodal melanoma treatment strategies.

Acceptability and understanding of the Ages & Stages Questionnaires®, Third Edition, as part of the Healthy Child Programme 2-year health and development review in England: Parent and professional perspectives.

BACKGROUND: The Healthy Child Programme is the universal public health system in England to assess and monitor child health from 0 to 19. Following a review of measures for closer monitoring at age 2 years, the Department of Health for England implemented the Ages & Stages Questionnaires®, Third Edition (ASQ-3™; Hereon, ASQ-3). AIM: The aim of this study was to evaluate the acceptability and understanding of the ASQ-3 in England by health professionals and parents. METHOD: A mixed-methods approach was used. This paper reports on the qualitative data drawn from interviews with 40 parents and 12 focus groups with 85 health professionals. The data were analysed using applied thematic analysis. FINDINGS: Overall, parents and health professionals found the ASQ-3 acceptable and understandable and could use it as a measure at age 2 years. The ability to work in partnership was valued. Some limitations included potential to cause anxiety, concerns around the safety of some of the items, and use of Americanized language. Health professional's training in the use the ASQ-3 was inconsistent. CONCLUSION: The ASQ-3 is an acceptable and understandable measure to use as part of the 2-year assessment with some adaptations to the English context and some standardized training for health professionals.

Treatment-related features improve machine learning prediction of prognosis in soft tissue sarcoma patients.

BACKGROUND AND PURPOSE: Current prognostic models for soft tissue sarcoma (STS) patients are solely based on staging information. Treatment-related data have not been included to date. Including such information, however, could help to improve these models. MATERIALS AND METHODS: A single-center retrospective cohort of 136 STS patients treated with radiotherapy (RT) was analyzed for patients' characteristics, staging information, and treatment-related data. Therapeutic imaging studies and pathology reports of neoadjuvantly treated patients were analyzed for signs of response. Random forest machine learning-based models were used to predict patients' death and disease progression at 2 years. Pre-treatment and treatment models were compared. RESULTS: The prognostic models achieved high performances. Using treatment features improved the overall performance for all three classification types: prediction of death, and of local and systemic progression (area under the receiver operatoring characteristic curve (AUC) of 0.87, 0.88, and 0.84, respectively). Overall, RT-related features, such as the planning target volume and total dose, had preeminent importance for prognostic performance. Therapy response features were selected for prediction of disease progression. CONCLUSIONS: A machine learning-based prognostic model combining known prognostic factors with treatment- and response-related information showed high accuracy for individualized risk assessment. This model could be used for adjustments of follow-up procedures.

Efficiency of soft tissue incision with a novel 445-nm semiconductor laser.

Using a 445-nm semiconductor laser for tissue incision, an effective cut is expected due to the special absorption properties of blue laser light in soft tissues. The aim of the present study was the histological evaluation of tissue samples after incision with a 445-nm diode laser. Forty soft tissue specimens were obtained from pork oral mucosa and mounted on a motorized linear translation stage. The handpiece of a high-frequency surgery device, a 970-nm semiconductor laser, and a 445-nm semiconductor laser were connected to the slide, allowing a constant linear movement (2 mm/s) and the same distance of the working tip to the soft tissue's surface. Four incisions were made each: (I) 970-nm laser with conditioned fiber tip, contact mode at 3-W cw; (II-III): 445-nm laser with non-conditioned fiber tip, contact mode at 2-W cw, and non-contact mode (1 mm) at 2 W; and (IV): high-frequency surgery device with straight working tip, 90° angulation, contact mode at 50 W. Histological analysis was performed after H&E staining of the embedded specimens at 35-fold magnification. The comparison of the incision depths showed a significant difference depending on the laser wavelength and the selected laser parameters. The highest incision depth was achieved with the 445-nm laser contact mode (median depth 0.61 mm, min 0.26, max 1.17, interquartile range 0.58) (p < 0.05) with the lowest amount of soft tissue denaturation (p < 0.05). The lowest incision depth was measured for the high-frequency surgical device (median depth 0.36 mm, min 0.12, max 1.12, interquartile range 0.23) (p < 0.05). Using a 445-nm semiconductor laser, a higher cutting efficiency can be expected when compared with a 970-nm diode laser and high-frequency surgery. Even the 445-nm laser application in non-contact mode shows clinically acceptable incision depths without signs of extensive soft tissue denaturation.

Mid-infrared beam splitter for ultrashort pulses.

A design is presented for a beam splitter suitable for ultrashort pulses in the mid-infrared and terahertz spectral range consisting of a structured metal layer on a diamond substrate. Both the theory and experiment show that this beam splitter does not distort the temporal pulse shape.

The Ol1mpiad: concordance of behavioural faculties of stage 1 and stage 3 Drosophila larvae.

Mapping brain function to brain structure is a fundamental task for neuroscience. For such an endeavour, the Drosophila larva is simple enough to be tractable, yet complex enough to be interesting. It features about 10,000 neurons and is capable of various taxes, kineses and Pavlovian conditioning. All its neurons are currently being mapped into a light-microscopical atlas, and Gal4 strains are being generated to experimentally access neurons one at a time. In addition, an electron microscopic reconstruction of its nervous system seems within reach. Notably, this electron microscope-based connectome is being drafted for a stage 1 larva - because stage 1 larvae are much smaller than stage 3 larvae. However, most behaviour analyses have been performed for stage 3 larvae because their larger size makes them easier to handle and observe. It is therefore warranted to either redo the electron microscopic reconstruction for a stage 3 larva or to survey the behavioural faculties of stage 1 larvae. We provide the latter. In a community-based approach we called the Ol1mpiad, we probed stage 1 Drosophila larvae for free locomotion, feeding, responsiveness to substrate vibration, gentle and nociceptive touch, burrowing, olfactory preference and thermotaxis, light avoidance, gustatory choice of various tastants plus odour-taste associative learning, as well as light/dark-electric shock associative learning. Quantitatively, stage 1 larvae show lower scores in most tasks, arguably because of their smaller size and lower speed. Qualitatively, however, stage 1 larvae perform strikingly similar to stage 3 larvae in almost all cases. These results bolster confidence in mapping brain structure and behaviour across developmental stages.

Histological validation of ICDAS II and radiological assessment of occlusal carious lesions in permanent teeth.

The International Caries Detection and Assessment System (ICDAS) was introduced for a detailed evaluation of dental caries. The aim of the present study was to compare the ICDAS scores and radiologically evaluated caries depths to the histologically evaluated carious lesions in permanent teeth. 84 freshly extracted human teeth were included. Visual examination and scoring of the occlusal aspect were performed according to the ICDAS II criteria after completing a respective e-learning programme to support training in the use of ICDAS. Bucco-lingual digital X-ray images of the teeth were taken. Specimens were then fixed in formalin and embedded in a photocuring one-component methacrylate-based resin. Longitudinal sections were cut and stained with rhodamine B, fuchsin and acetic light green dye to assess the caries extension by light microscopic analysis. Assessing ICDAS II scores and histological findings, a rank correlation coefficient of r = 0.890 could be found. ICDAS II/radiology and histology/radiology showed correlation coefficients of r = 0.658 and 0.661, respectively. Evaluating receiver operating characteristic (ROC) curves, no exact predictability could be found for caries lesions in enamel for both ICDAS II and radiological evaluation. Focussing on deep dentin lesions, values of 0.940 (ICDAS II) and 0.845 (radiology) showed high predictability with respect to the histologically observed caries extension. The present study indicates an acceptable validity of the ICDAS II criteria when applied to permanent teeth. Especially, dentin lesions can be reliably detected. Thus, ICDAS assessment provides the possibility of reducing X-ray exposure for caries detection.

The enhancing effect of a laser photochemotherapy with cisplatin or zolendronic acid in primary human osteoblasts and osteosarcoma cells in vitro.

BACKGROUND: Photodynamic therapies (PDT) have become increasingly popular in the adjuvant treatment of different tumour entities. Chemotherapeutic agents, such as cisplatin may be used in combination with low-level laser therapy (LLLT) as laser photochemotherapy. The aim of this study was to investigate the effect of LLLT on cell bioviability of normal and malignant bone cells under chemotherapeutic conditions with either cisplatin or zolendronic acid in vitro. METHODS: Primary human osteoblasts (HOB) and an osteosarcoma cell line (Saos-2) were treated with different concentrations of zolendronic acid or cisplatin and irradiated twice with a diode laser (wavelength 670 nm, 120 s, energy outputs of 100mW/cm2 , continuous wave mode). Cell viability was tested by XTT-assay and via histomorphological analysis. RESULTS: LLLT alone increased bioviability for both cell lines. LLLT lowered HOB viability at the three highest concentrations of cisplatin and zolendronic acid. For Saos-2, LLLT reduced cell viability at every concentration of cisplatin. In cases of incubation with zolendronic acid, similar to osteoblasts, LLLT lowered cell viability at the highest concentration only. CONCLUSIONS: Based on the conditions of this study, laser photochemotherapy may be able to raise the cytotoxicity of cisplatin and zolendronic acid in benign and malignant bone cells. This could be of interest in the development of new therapeutic treatment modalities against neoplastic bone diseases like osteosarcoma.