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ZusammenfassungSelektive PET- oder SPECT- Radiopharmaka sind inzwischen nicht nur für αvβ3, sondern auch weitere der 24 verschiedenen Integrine verfügbar, zum Beispiel α5β1, αvβ6, αvβ8 und α6. Da diese unter anderem auch von verschiedenen Karzinomen und im Zuge von Fibrose exprimiert werden, ist die Vorstellung, dass Integrine nur als Zielstrukturen für die Bildgebung von Angiogenese in Betracht kommen, endgültig überholt. Die derzeit besten Aussichten auf eine breite klinische Anwendung, sowohl diagnostisch als auch therapeutisch, haben derzeit αvβ6-Integrin-Radiopharmaka, da αvβ6 von vielen malignen Krebsarten (v. a. Pankreas-, Plattenepithel-, Basalzell-, Lungen- und Colonkarzinom) überexprimiert wird.

Background. Visual inspection and biopsy is the current standard of care for oral cancer diagnosis, but is subject to misinterpretation and consequently to misdiagnosis. Topically applied PARPi-FL is a molecularly specific, fluorescent contrast-based approach that may fulfil the unmet need for a simple, in vivo, non-invasive, cost-effective, point-of-care method for the early diagnosis of oral cancer. Here, we present results from a phase I safety and feasibility study on fluorescent, topically applied PARPi-FL. Twelve patients with a histologically proven squamous cell carcinoma of the oral cavity (OSCC) gargled a PARPi-FL solution for 60 seconds (15 mL, 100 nM, 250 nM, 500 nM, or 1000 nM), followed by gargling a clearing solution for 60 seconds. Fluorescence measurements of the lesion and surrounding oral mucosa were taken before PARPi-FL application, after PARPi-FL application and after clearing. Blood pressure, oxygen levels, clinical chemistry and CBC were obtained before and after tracer administration. Results. PARPi-FL was well-tolerated by all patients without any safety concerns. When analyzing the fluorescence signal, all malignant lesions showed a significant differential in contrast after administration of PARPi-FL, with the highest increase occurring at the highest dose level (1000 nM), where all patients had a tumor-to-margin fluorescence signal ratio of > 3. A clearing step was essential to increase signal specificity, as it clears unbound PARPi-FL trapped in normal anatomical structures. PARPi-FL tumor cell specificity was confirmed by ex vivo tabletop confocal microscopy. We have demonstrated that the fluorescence signal arose from the nuclei of tumor cells, endorsing our macroscopic findings.Conclusions. A PARPi-FL swish & spit solution is a rapid and non-invasive diagnostic tool that preferentially localizes fluorescent contrast to OSCC. This technique holds promise for the early detection of OSCC based on in vivo optical evaluation and targeted biopsy of suspicious lesions in the oral cavity. Clinicaltrials.gov - NCT03085147, registered on March 21st, 2017.

Purpose: Glioblastoma multiforme is a highly aggressive form of brain cancer whose location, tendency to infiltrate healthy surrounding tissue, and heterogeneity significantly limit survival, with scant progress having been made in recent decades. Experimental Design: 123I-MAPi (Iodine-123 Meitner-Auger PARP1 inhibitor) is a precise therapeutic tool composed of a PARP1 inhibitor radiolabeled with an Auger- and gamma-emitting iodine isotope. Here, the PARP inhibitor, which binds to the DNA repair enzyme PARP1, specifically targets cancer cells, sparing healthy tissue, and carries a radioactive payload within reach of the cancer cells' DNA. Results: The high relative biological efficacy of Auger electrons within their short range of action is leveraged to inflict DNA damage and cell death with high precision. The gamma ray emission of 123I-MAPi allows for the imaging of tumor progression and therapy response, and for patient dosimetry calculation. Here we demonstrated the efficacy and specificity of this small-molecule radiotheranostic in a complex preclinical model. In vitro and in vivo studies demonstrate high tumor uptake and a prolonged survival in mice treated with 123I-MAPi when compared with vehicle controls. Different methods of drug delivery were investigated to develop this technology for clinical applications, including convection enhanced delivery and intrathecal injection. Conclusions: Taken together, these results represent the first full characterization of an Auger-emitting PARP inhibitor which demonstrate a survival benefit in mouse models of GBM and confirm the high potential of 123I-MAPi for clinical translation.

For oral, oropharyngeal and oesophageal cancer, the early detection of tumours and of residual tumour after surgery are prognostic factors of recurrence rates and patient survival. Here, we report the validation, in animal models and a human, of the use of a previously described fluorescently labelled small-molecule inhibitor of the DNA-repair enzyme poly(ADP-ribose) polymerase 1 (PARP1) for the detection of cancers of the oral cavity, pharynx and oesophagus. We show that the fluorescent contrast agent can be used to quantify the expression levels of PARP1, and to detect oral, oropharyngeal and oesophageal tumours in mice, pigs and fresh human biospecimens when delivered topically or intravenously. The fluorescent PARP1 inhibitor can also detect oral carcinoma in a patient when applied as a mouthwash, and discriminate, between fresh biopsied samples of the oral tumour and the surgical resection margin with larger than 95% sensitivity and specificity. The PARP1 inhibitor could serve as the basis of a fast and sensitive assay for the early detection, and for the surgical-margin assessment, of epithelial cancers of the upper intestinal tract.

High levels of reactive oxygen species (ROS) in tumors have been shown to exert anti-tumor activity, leading to the concept of ROS induction as therapeutic strategy. The organometallic compound ferrocene (Fc) generates ROS through a reversible one-electron oxidation. Incorporation of Fc into a tumor-targeting, bioactive molecule can enhance its therapeutic activity and enable tumor specific delivery. Therefore, we conjugated Fc to five synthetic, Arg-Gly-Asp (RGD)-based integrin binding ligands to enable targeting of the cell adhesion and signaling receptor integrin subtypes αvβ3, α5β1, or αvβ6, which are overexpressed in various, distinct tumors. We designed and synthesized a library of integrin-ligand-ferrocene (ILF) derivatives and showed that ILF conjugates maintained the high integrin affinity and selectivity of their parent ligands. A thorough biological characterization allowed us to identify the two most promising ligands, an αvβ3 (L2b) and an αvβ6 (L3b) targeting ILF, which displayed selective integrin-dependent cell uptake and pronounced ferrocene-mediated anti-tumor effects in vitro, along with increased ROS production and DNA damage. Hence, ILFs are promising candidates for the selective, tumor-targeted delivery of ferrocene to maximize its anti-cancer efficacy and minimize systemic toxicity, thereby improving the therapeutic window of ferrocene compared to currently used non-selective anti-cancer drugs.

BackgroundWith increasing clinical use of NK-92 cells and their CAR-modified derivatives in cancer immunotherapy, there is a growing demand for efficient production processes of these “off-the-shelf” therapeutics. In order to ensure safety and prevent the occurrence of secondary tumors, (CAR-)NK-92 cell proliferation has to be inactivated before transfusion. This is commonly achieved by gamma irradiation. Recently, we showed proof of concept that low energy electron irradiation (LEEI) is a new method for NK-92 inactivation. LEEI has several advantages over gamma irradiation, including a faster reaction time, a more reproducible dose rate and much less requirements on radiation shielding. Here, LEEI was further evaluated as a promising alternative to gamma irradiation yielding cells with highly maintained cytotoxic effector function.MethodsEffectiveness and efficiency of LEEI and gamma irradiation were analyzed using NK-92 and CD123-directed CAR-NK-92 cells. LEE-irradiated cells were extensively characterized and compared to gamma-irradiated cells via flow cytometry, cytotoxicity assays, and comet assays, amongst others.ResultsOur results show that both irradiation methods caused a progressive decrease in cell viability and are, therefore, suitable for inhibition of cell proliferation. Notably, the NK-mediated specific lysis of tumor cells was maintained at stable levels for three days post-irradiation, with a trend towards higher activities after LEEI treatment as compared to gamma irradiation. Both gamma irradiation as well as LEEI led to substantial DNA damage and an accumulation of irradiated cells in the G2/M cell cycle phases. In addition, transcriptomic analysis of irradiated cells revealed approximately 12-fold more differentially expressed genes two hours after gamma irradiation, compared to LEEI. Analysis of surface molecules revealed an irradiation-induced decrease in surface expression of CD56, but no changes in the levels of the activating receptors NKp46, NKG2D, or NKp30.ConclusionsThe presented data show that LEEI inactivates (CAR-)NK-92 cells as efficiently as gamma irradiation, but with less impact on the overall gene expression. Due to logistic advantages, LEEI might provide a superior alternative for the manufacture of (CAR-)NK-92 cells for clinical application.

Introduction Inhibitors of poly-(ADP)-ribose polymerase (PARP) are promising therapeutics for SCLC. We tested whether PARP inhibitor (PARPi) target engagement as measured by a fluorine 18–radiolabeled PARPi ([18F]PARPi) has the potential to predict drug efficacy in vivo. Methods Tumor growth inhibition during daily talazoparib treatment was evaluated in mice engrafted with SCLC patient-derived xenografts to evaluate talazoparib efficacy at multiple doses. Mice were intravenously injected with [18F]PARPi radiotracer at multiple timepoints after single doses of oral talazoparib to quantitatively assess the extent to which talazoparib could reduce tumor radiotracer uptake and positron-emission tomographic (PET)/computer tomographic activity. Tumors were harvested and tumor poly-(ADP) ribose level was measured by enzyme-linked immunosorbent assay. Results A dose range of talazoparib with differential therapeutic efficacy was established, with significant delay in time to reach 1000 mm3 for tumors treated with 0.3 mg/kg (p = 0.02) but not 0.1 mg/kg talazoparib. On PET/computed tomography with [18F]PARPi, reduction in [18F]PARPi uptake after talazoparib dosing was consistent with talazoparib clearance, with reduction in PET activity attenuating over 24 hours. Talazoparib target engagement, measured by maximum tumor PET uptake, increased in a dose-dependent manner (3.9% versus 2.1% injected dose/g for 0.1 and 0.3 mg/kg at 3 hours post-talazoparib, p = 0.003) and correlated with PARP enzymatic activity among individual tumors as measured by total tumor poly-(ADP) ribose (p = 0.04, R = 0.62 at 1 hour post-talazoparib). Conclusions PET imaging using [18F]PARPi has the potential to be a powerful tool in treatment monitoring by assessing PARPi target engagement in real-time.

In times of ongoing automatization of dairy cow husbandry, objective and reliable tools for mastitis diagnostic are highly in demand. The objective of this study was to investigate the diagnostic value of a handheld dynamometer and an infrared thermometer to diagnose and score clinical and subclinical mastitis and to compare those values with results from palpation of the udder tissue. Overall, 218 cows with clinical mastitis (i.e., 46 mild, 106 moderate, and 66 severe cases), 142 with subclinical mastitis, and 68 healthy cows were enrolled. Our data provide evidence that the dynamometer is an accurate diagnostic tool to differentiate between healthy udder quarters, and those with subclinical and clinical mastitis. Furthermore, the severity score of clinical mastitis can be estimated by dynamometer. The firmness threshold for the detection of clinical mastitis was 1.002 kg. Using a threshold of 1.175 kg in clinical mastitis quarters, it was possible to differentiate between negative and positive bacteriological results. A differentiation between healthy and clinical mastitis quarters with the infrared thermometer was possible, albeit udder surface temperatures were highly influenced by ambient temperature. Udder surface temperature increased by 0.15 to 0.18°C for each degree of ambient temperature. In conclusion, the utility of an infrared thermometer to estimate the udder health status of dairy cows is limited, whereas the handheld dynamometer appeared to be an accurate and objective method.

Oral cancer is one of the most common cancers globally. Survival rates for patients are directly correlated with stage of diagnosis; despite this knowledge, 60% of individuals are presenting with late-stage disease. Currently, the initial evaluation of a questionable lesion is performed by a conventional visual examination with white light. If a lesion is deemed suspicious, a biopsy is taken for diagnosis. However, not all lesions present suspicious under visual white light examination, and there is limited specificity in differentiating between benign and malignant transformations. Several vital dyes, light-based detection systems, and cytology evaluation methods have been formulated to aid in the visualization process, but their lack of specific biomarkers resulted in high false-positive rates and thus limits their reliability as screening and guidance tools. In this review, we will analyze the current methodologies and demonstrate the need for specific intraoral imaging agents to aid in screening and diagnosis to identify patients earlier. Several novel molecular imaging agents will be presented as, by result of their molecular targeting, they aim to have high specificity for tumor pathways and can support in identifying dysplastic/cancerous lesions and guiding visualization of biopsy sites. Imaging agents that are easy to use, inexpensive, noninvasive, and specific can be utilized to increase the number of patients who are screened and monitored in a variety of different environments, with the ultimate goal of increasing early detection.

The central paradigm of novel therapeutic approaches in cancer therapy is identifying and targeting molecular biomarkers. One such target is the nuclear DNA repair enzyme Poly-(ADP ribose) polymerase 1 (PARP1). Sensitivity to PARP inhibition in certain cancers such as gBRCAmut breast and ovarian cancers has led to its exploitation as a target. The overexpression of PARP1 in several types of cancer further evoked interest in its use as an imaging target. While PARP1-targeted inhibitors have fast developed and approved in this past decade, determination of PARP1 expression might help to predict the response to PARP inhibitor treatment. This has the potential of improving prognosis and moving towards tailored therapy options and/or dosages. This review summarizes the recent pre-clinical advancements in imaging and theranostic PARP1 targeted tracers. To assess PARP1 levels, several imaging probes with fluorescent or beta/gamma emitting radionuclides have been proposed and three have advanced to ongoing clinical evaluation. Apart from its diagnostic value in detection of primary tumors as well as metastases, this shall also help in delivering therapeutic radionuclides to PARP1 overexpressing tumors. Henceforth nuclear medicine has now advanced towards conjugating theranostic radionuclides to PARP1 inhibitors. This paves the way for a future of PARP1-targeted theranostics and personalized therapy.

With the ability to noninvasively image and monitor molecular processes within tumors, molecular imaging represents a fundamental tool for cancer scientists. In the current review, we describe emergent optical technologies for molecular imaging. We aim to provide the reader with an overview of the fundamental principles on which each imaging strategy is based, to introduce established and future applications, and to provide a rationale for selecting optical technologies for molecular imaging depending on disease location, biology, and anatomy. To accelerate clinical translation of imaging techniques, we also describe examples of practical applications in patients. Elevating these techniques into standard-of-care tools will transform patient stratification, disease monitoring, and response evaluation.

Timely and objective diagnosis and classification of mastitis is crucial to ensure adequate management and therapeutic decisions. Analyzing specific biomarkers in milk could be advantageous compared with subjective or semiquantitative criteria, such as palpation of the udder in clinical mastitis cases or evaluation of somatic cell count using cow side tests (e.g., California Mastitis Test) in subclinical mastitis quarters. The objective of this study was to investigate the diagnostic value of 3 biomarkers; cathelicidin, milk amyloid A, and haptoglobin for the diagnosis of subclinical and clinical mastitis. Furthermore, the suitability of these biomarkers to differentiate between mild, moderate, and severe clinical mastitis and the influence of different pathogens on biomarker levels was tested. A total of 67 healthy cows, 119 cows with subclinical mastitis, and 212 cows with clinical mastitis were enrolled in the study. Although cathelicidin, haptoglobin, and milk amyloid A were measured in all samples from healthy cows and those with subclinical mastitis, haptoglobin, and cathelicidin results were only available from 121 out of 212 cows with clinical mastitis. Milk amyloid A was measured in all samples. In cows with clinical mastitis, the mastitic quarter and a second healthy quarter serving as a healthy in-cow control quarter were sampled. It was possible to differentiate between healthy quarters, quarters with subclinical mastitis, and quarters with clinical mastitis using all 3 biomarkers. Concerning cathelicidin, thresholds were 0.000 [sensitivity (Se) = 0.83, specificity (Sp) = 0.97] and 0.053 (Se = 0.98, Sp = 0.99) for normalized optical density at 450 nm (NOD450) for differentiating between healthy quarters and quarters with subclinical or clinical mastitis, respectively. Thresholds of 1.28 µg/mL (Se = 0.65, Sp = 0.76) and 1.81 µg/mL (Se = 0.77, Sp = 0.83) for milk amyloid A and 3.65 µg/mL (Se = 0.92, Sp = 0.94) and 5.40 µg/mL mL (Se = 0.96, Sp = 0.99) for haptoglobin were calculated, respectively. Healthy in-cow control quarters from cows with CM showed elevated milk amyloid A and haptoglobin levels compared with healthy quarters from healthy cows. Only the level of milk amyloid A was higher in severe clinical mastitis cases compared with mild ones. In contrast to clinical mastitis, cathelicidin and haptoglobin in subclinical mastitis quarters were significantly influenced by different bacteriological results. The measurement of cathelicidin, milk amyloid A, and haptoglobin in milk proved to be a reliable method to detect quarters with subclinical or clinical mastitis.

The immune function within the tumor microenvironment has become a prominent therapeutic target, with tumor-associated macrophages (TAMs) playing a critical role in immune suppression. We propose an 89Zr-labeled high-density lipoprotein (89Zr-HDL) nanotracer as a means of monitoring response to immunotherapy. Methods: Female MMTV-PyMT mice were treated with pexidartinib, a colony-stimulating factor 1 receptor (CSF1R) inhibitor, to reduce TAM density. The accumulation of 89Zr-HDL within the tumor was assessed using PET/CT imaging and autoradiography, whereas TAM burden was determined using immunofluorescence. Results: A significant reduction in 89Zr-HDL accumulation was observed in PET/CT images, with 2.9% ± 0.3% and 3.7% ± 0.2% injected dose/g for the pexidartinib- and vehicle-treated mice, respectively. This reduction was corroborated ex vivo and correlated with decreased TAM density. Conclusion: These results support the potential use of 89Zr-HDL nanoparticles as a PET tracer to quickly monitor the response to CSF1R inhibitors and other therapeutic strategies targeting TAMs. We thank the Small Animal Imaging Core, the Radiochemistry and Molecular Imaging Probes Core, and the Molecular Cytology Core at Memorial Sloan Kettering Cancer Center. This work was supported by National Institutes of Health grants R01 CA204441, P30 CA008748 and R01 CA220234. The authors thank the Tow Foundation and Memorial Sloan Kettering Cancer Center's Center for Molecular Imaging & Nanotechnology (CMINT), the Imaging and Radiation Sciences Program and the MSK Molecularly Targeted Intraoperative Imaging Fund. Sí

OBJECTIVES: The evaluation of disease extent and post-therapy surveillance of head and neck cancer using 2-deoxy-2-[(18)F]fluoro-D-glucose ([(18)F]FDG) PET is often complicated by physiological uptake in normal tissues of the head and neck region, especially after surgery or radiotherapy. However, irrespective of low positive predictive values, [(18)F]FDG PET remains the standard of care to stage the disease and monitor recurrences. Here, we report the preclinical use of a targeted poly (ADP-ribose) polymerase1 (PARP1) binding PET tracer, fluorine-18 labeled poly (ADP-ribose) polymerase1 inhibitor ([(18)F]PARPi), as a potential alternative with greater specificity. METHODS: Using an orthotopic xenograft mouse model injected with either FaDu or Cal 27 (human squamous cell carcinoma cell lines) we performed PET/CT scans with the 2 tracers and compared the results. Gamma counts and autoradiography were also assessed and correlated with histology. RESULTS: The average retained activity of [(18)F]PARPi across cell lines in tumor-bearing tongues was 0.9 ± 0.3 %ID/g, 4.1 times higher than in control (0.2 ± 0.04 %ID/g). Autoradiography and histology confirmed that the activity arose almost exclusively from the tumor areas, with a signal/normal tissue around a ratio of 42.9 ± 21.4. In vivo, [(18)F]PARPi-PET allowed delineation of tumor from healthy tissue (p < 0.005), whereas [(18)F]FDG failed to do so (p = 0.209). CONCLUSIONS AND IMPLICATIONS FOR PATIENT CARE: We demonstrate that [(18)F]PARPi is more specific to tongue tumor tissue than [(18)F]FDG. [(18)F]PARPi PET allows for the straightforward delineation of oral cancer in mouse models, suggesting that clinical translation could result in improved imaging of head and neck cancer when compared to [(18)F]FDG.

Background We report preclinical and first-in-human-brain-cancer data using a targeted poly (ADP-ribose) polymerase 1 (PARP1) binding PET tracer, [18F]PARPi, as a diagnostic tool to differentiate between brain cancers and treatment-related changes. Methods We applied a glioma model in p53-deficient nestin/tv-a mice, which were injected with [18F]PARPi and then sacrificed 1 h post-injection for brain examination. We also prospectively enrolled patients with brain cancers to undergo dynamic [18F]PARPi acquisition on a dedicated positron emission tomography/magnetic resonance (PET/MR) scanner. Lesion diagnosis was established by pathology when available or by Response Assessment in Neuro-Oncology (RANO) or RANO-BM response criteria. Resected tissue also underwent PARPi-FL staining and PARP1 immunohistochemistry. Results In a preclinical mouse model, we illustrated that [18F]PARPi crossed the blood–brain barrier and specifically bound to PARP1 overexpressed in cancer cell nuclei. In humans, we demonstrated high [18F]PARPi uptake on PET/MR in active brain cancers and low uptake in treatment-related changes independent of blood–brain barrier disruption. Immunohistochemistry results confirmed higher PARP1 expression in cancerous than in noncancerous tissue. Specificity was also corroborated by blocking fluorescent tracer uptake with an excess unlabeled PARP inhibitor in patient cancer biospecimen. Conclusions Although larger studies are necessary to confirm and further explore this tracer, we describe the promising performance of [18F]PARPi as a diagnostic tool to evaluate patients with brain cancers and possible treatment-related changes.

Background Radiation injury can be indistinguishable from recurrent tumor on standard imaging. Current protocols for this differential diagnosis require one or more follow-up imaging studies, long dynamic acquisitions, or complex image post-processing; despite much research, the inability to confidently distinguish between these two entities continues to pose a significant dilemma for the treating clinician. Using mouse models of both glioblastoma and radiation necrosis, we tested the potential of poly(ADP-ribose) polymerase (PARP)-targeted PET imaging with [18F]PARPi to better discriminate radiation injury from tumor. Results In mice with experimental radiation necrosis, lesion uptake on [18F]PARPi-PET was similar to contralateral uptake (1.02 ± 0.26 lesion/contralateral %IA/ccmax ratio), while [18F]FET-PET clearly delineated the contrast-enhancing region on MR (2.12 ± 0.16 lesion/contralateral %IA/ccmax ratio). In mice with focal intracranial U251 xenografts, tumor visualization on PARPi-PET was superior to FET-PET, and lesion-to-contralateral activity ratios (max/max, p = 0.034) were higher on PARPi-PET than on FET-PET. Conclusions A murine model of radiation necrosis does not demonstrate [18F]PARPi avidity, and [18F]PARPi-PET is better than [18F]FET-PET in distinguishing radiation injury from brain tumor. [18F]PARPi-PET can be used for discrimination between recurrent tumor and radiation injury within a single, static imaging session, which may be of value to resolve a common dilemma in neuro-oncology.

The DNA repair enzyme poly(ADP-ribose) polymerase 1 (PARP-1) is overexpressed in glioblastoma, with overall low expression in healthy brain tissue. Paired with the availability of specific small molecule inhibitors, PARP-1 is a near-ideal target to develop novel radiotherapeutics to induce DNA damage and apoptosis in cancer cells, while sparing healthy brain tissue. Methods: We synthesized an 131I-labeled PARP-1 therapeutic and investigated its pharmacology in vitro and in vivo. A subcutaneous tumor model was used to quantify retention times and therapeutic efficacy. A potential clinical scenario, intratumoral convection-enhanced delivery, was mimicked using an orthotopic glioblastoma model combined with an implanted osmotic pump system to study local administration of 131I-PARPi (PARPi is PARP inhibitor). Results: 131I-PARPi is a 1(2H)-phthalazinone, similar in structure to the Food and Drug Administration–approved PARP inhibitor AZD-2281. In vitro studies have shown that 131I-PARPi and AZD-2281 share similar pharmacologic profiles. 131I-PARPi delivered 134.1 cGy/MBq intratumoral injected activity. Doses to nontarget tissues, including liver and kidney, were significantly lower. Radiation damage and cell death in treated tumors were shown by p53 activation in U87-MG cells transfected with a p53-bioluminescent reporter. Treated mice showed significantly longer survival than mice receiving vehicle (29 vs. 22 d, P < 0.005) in a subcutaneous model. Convection-enhanced delivery demonstrated efficient retention of 131I-PARPi in orthotopic brain tumors, while quickly clearing from healthy brain tissue. Conclusion: Our results demonstrate 131I-PARPi’s high potential as a therapeutic and highlight PARP’s relevance as a target for radionuclide therapy. Radiation plays an integral role in brain tumor therapy, and radiolabeled PARP therapeutics could ultimately lead to improvements in the standard of care.

The objective of this study was to evaluate the dental color exposed to acute cigarette smoke treatment and quantify the amount of nicotine in samples exposed to cigarette smoke, after dental prophylaxis and after in-office bleaching. Sixty-nine healthy human molars were subjected to cigarette smoke in a cigarette machine. The teeth were divided into three groups: positive control, prophylaxis, and bleaching. Forty cycles of smoke exposition with duration of 15 min each were performed using 10 cigarettes (positive control). Dental prophylaxis was performed with a rotating brush and prophylaxis paste; in-office bleaching was performed with 35% hydrogen peroxide, in two sessions of three 15-min applications, with a 1-week interval between sessions. The color was evaluated at the baseline, after exposure to cigarette smoke, after dental prophylaxis, and after in-office bleaching. Teeth from each group were powdered and analyzed by gas chromatography–mass spectrometry in order to measure the amount of nicotine present in each group. Data from quantification of nicotine and color change were analyzed by one-way ANOVA and Tukey’s test (α = 0.05). Data for subjective and objective color evaluation, a perceptible dental darkening occurred in teeth after exposure to cigarette smoke. Dental prophylaxis was able to recover the original color of teeth however, only after bleaching teeth became whiter than at the baseline (p

Aim The purpose of this study was to evaluate the dental effect and sensitivity of at-home dental bleaching with 10% hydrogen peroxide (H2O2). Materials and methods Twenty volunteers with A2 or darker central incisors were selected for this study. Was used 10% H2O2 for thirty minutes twice a day, for two weeks. Shade evaluation was assessed visually by the value-oriented shade guide Vita Classical shade guide, Vita Bleachedguide 3D-MASTER and by the Easyshade spectrophotometer at baseline, during bleaching (first and second weeks), and post-bleaching (one month). The perceptions of sensitivity were recorded by the patients through the numerical rating scale (0 to 4) and 0 to 10 visual analog scales daily. Data from the shade guide units was subjected to a one way repeated measures (RM) analysis of variance (ANOVA) test (a = 5%). The overall AE, absolute risk and intensity of tooth sensitivity were calculated as well as the 95% confidence interval. Results The absolute risk of tooth sensitivity was 65% and the intensity was mild. Data from ASGU and AE after 1 month of bleaching for H2O2 10% showed significant whitening, 4 units for Vita Classical, 5 units for Vita Bleachedguide and 9.7 units for spectrophotometer. Conclusion At-home bleaching using 10% hydrogen peroxide is effective in 14 days of bleaching. The most common adverse events were mild tooth sensitivity, and no subjects discontinued use early because of a treatment-related adverse event. Clinical significance At-home dental bleaching with 10% hydrogen peroxide gel reduces the time of use of the tray, maintaining the effectiveness with low intensity of dental sensitivity. Brazilian clinical trials registry (REBEC) RBR-45xmzj.

Hepatobiliary and pancreatic cancers along with other gastrointestinal malignancies remain the leading cause of cancer-related deaths worldwide. Strategies developed in the recent years on immunotherapy and cancer vaccines in the setting of primary liver cancer as well as in pancreatic cancer are the scope of this review. Significance of orthotopic and autochthonous animal models which mimic and/or closely reflect human malignancies allowing for a prompt and trustworthy analysis of new therapeutics is underlined. Combinational approaches that on one hand, specifically target a defined cancer-driving pathway, and on the other hand, restore the functions of immune cells, which effector functions are often suppressed by a tumor milieu, are shown to have the strongest perspectives and future directions. Among combinational immunotherapeutic approaches a personalized- and individual cancer case-based therapy is of special importance.

SUMMARY Objective: This clinical study evaluated the color longevity after 30 months of at-home bleaching with 10% carbamide peroxide (CP) in smokers and nonsmokers. Methods: Sixty patients, 30 smokers and 30 nonsmokers, were subjected to bleaching with 10% CP (Whiteness Perfect–FGM) for three hours daily for three weeks. The color was measured at baseline and at one month and 30 months after the completion of dental bleaching using the shade guide Vita classical organized by value (ΔSGU) and the shade guide Vita Bleachedguide 3D-MASTER. At the 30-month recall, the color was assessed before and after dental prophylaxis. Data from color evaluation were analyzed by two-way repeated-measures analysis of variance and Tukey test for the contrast of means (α=0.05). Results: Twenty-one smokers and 22 nonsmokers attended the 30 month recall. For both shade guides, only the main factor of assessment time was statistically significant (p Conclusion: Thirty months after at-home bleaching with 10% CP gel, dental darkening was detected in both groups, which cannot be solely attributed to stains caused by extrinsic staining from daily food, drinks, and smoke (in smokers).

Background The implantation of rotary blood pumps as ventricular assist devices (VADs) has become a viable therapy for quite a number of patients with end-stage heart failure. However, these rotary blood pumps cause adverse events that are related to blood trauma. It is currently believed that turbulence in the pump flow plays a significant role. But turbulence has not been measured to date because there is no optical access to the flow space in rotary blood pumps because of their opaque casings. Methods This difficulty is overcome with a scaled-up model of the HeartMate II (HM II) rotary blood pump with a transparent acrylic housing. A 2-component laser Doppler velocimetry (LDV) system was used for the measurement of time resolved velocity profiles and velocity spectra upstream and downstream of the rotor blades. Observing similarity laws, the speed and pump head were adjusted to correspond closely to the design point of the original pump – 10,600 rpm speed and 80 mmHg pressure head. A model fluid consisting of a water-glycerol mixture was used. Results The measured velocity spectra were scalable by the Kolmogorov length and the Kolmogorov length was estimated to be between 14 and 24 μm at original scale, thus being about 1.5 to 3 times the size of a red blood cell. Conclusions It can be concluded that turbulence is indeed present in the investigated blood pump and that it can be described by Kolmogorov's theory of turbulence. The size of the smallest vortices compares well to the turbulence length scales as found in prosthetic heart valves, for example.

Introduction Bleached enamel surfaces may undergo changes and retain more dye, which is a reason to recommend the reduction/suspension of foods with dyes during dental bleaching. Aim Evaluate the effects of the action of natural and artificial dyes on the bleached enamel of extracted human teeth. Materials and methods Fifty human premolars were used, which were distributed in 5 groups (n = 10) according to the following staining solutions: GW (distilled water); GB (beet); Gca (caramel); GC (carmine); and GR (red 40). After the removal of the root and pulp section, the teeth were embedded in acrylic resin blocks, stored in artificial saliva, and kept at 37°C. At-home bleaching was performed using 16% carbamide peroxide (CP) for 3 hours daily for 3 weeks. After each daily session of bleaching, the specimens were exposed to the dye solution twice a day for 5 min; one of these exposures was performed immediately after bleaching. The color was recorded using a spectrophotometer according to the CIE Lab system (Δ E) for the following periods: baseline, during bleaching (after 1st, 2nd, and 3rd week) and post-bleaching (after 1 week and 1 month). The color was evaluated by two-way analysis of variance and Tukey's test (α=0.05). Results There was effective dental bleaching for all groups: GW (18.5 ± 6.1), GB (19.9 ± 4.4), Gca (18.9 ± 6.1), GC (20.2 ± 4.6), and GR (19.3 ± 4.2), p 0.05). Conclusion The exposure to beet, carmine, caramel, and red 40 dyes did not interfere with the effectiveness of dental bleaching using 16% CP. Clinical significance Dyes consumption during bleaching did not affect the effectiveness of dental bleaching.

Major determining factors for survival of patients with oral, oropharyngeal, and esophageal cancer are early detection, the quality of surgical margins, and the contemporaneous detection of residual tumor. Intuitively, the exposed location at the epithelial surface qualifies these tumor types for utilization of visual aids to assist in discriminating tumor from healthy surrounding tissue. Here, we explored the DNA repair enzyme PARP1 as imaging biomarker and conducted optical imaging in animal models, human tissues and as part of a first-in-human clinical trial. Our data suggests that PARP1 is a quantitative biomarker for oral, oropharyngeal, and esophageal cancer and can be visualized with PARPi-FL, a fluorescently labeled small molecule contrast agent for topical or intravenous delivery. We show feasibility of PARPi-FL-assisted tumor detection in esophageal cancer, oropharyngeal and oral cancer. We developed a contemporaneous PARPi-FL topical staining protocol for human biospecimens. Using fresh oral cancer tissues within 25 min of biopsy, tumor and margin samples were correctly identified with >95% sensitivity and specificity without terminal processing. PARPi-FL imaging can be integrated into clinical workflows, potentially providing instantaneous assessment of the presence or absence of microscopic disease at the surgical margin. Additionally, we showed first-in-human PARPi-FL imaging in oral cancer. In aggregate, our preclinical and clinical studies have the unifying goal of verifying the clinical value of PARPi-FL-based optical imaging for early detection and intraoperative margin assignment.

Breast cancer is the most common type of malignant growth in women. Early detection of breast cancer, as well as the identification of possible metastatic spread poses a significant challenge because of the structural and genetic heterogeneity that occurs during the progression of the disease. Currently, mammographies, biopsies and MRI scans are the standard of care techniques used for breast cancer diagnosis, all of which have their individual shortfalls, especially when it comes to discriminating tumors and benign growths. With this in mind, we have developed a non-invasive optoacoustic imaging strategy that targets the acidic environment of breast cancer. A pH low insertion peptide (pHLIP) was conjugated to the dark quencher QC1, yielding a non-fluorescent sonophore with high extinction coefficient in the near infrared that increases signal as a function of increasing amounts of membrane insertion. In an orthotopic murine breast cancer model, pHLIP-targeted optoacoustic imaging allowed us to differentiate between healthy and breast cancer tissues with high signal/noise ratios. In vivo, the sonophore QC1-pHLIP could detect malignancies at higher contrast than its fluorescent analog ICG-pHLIP, which was developed for fluorescence-guided surgical applications. PHLIP-type optoacoustic imaging agents in clinical settings are attractive due to their ability to target breast cancer and a wide variety of other malignant growths for diagnostic purposes. Intuitively, these agents could also be used for visualization during surgery.

Glioblastoma multiforme is a highly aggressive form of brain cancer whose location, tendency to infiltrate healthy surrounding tissue, and heterogeneity significantly limit survival, with scant progress having been made in recent decades.123I-MAPi (Iodine-123 Meitner-Auger PARP1 inhibitor) is a precise therapeutic tool composed of a PARP1 inhibitor radiolabeled with an Auger- and gamma-emitting iodine isotope. Here, the PARP inhibitor, which binds to the DNA repair enzyme PARP1, specifically targets cancer cells, sparing healthy tissue, and carries a radioactive payload within reach of the cancer cells’ DNA. The high relative biological efficacy of Auger electrons within their short range of action is leveraged to inflict DNA damage and cell death with high precision. The gamma ray emission of123I-MAPi allows for the imaging of tumor progression and therapy response, and for patient dosimetry calculation. Here we demonstrated the efficacy and specificity of this small molecule radiotheranostic in a complex preclinical model.In vitroandin vivostudies demonstrate high tumor uptake and a prolonged survival in mice treated with123I-MAPi when compared to vehicle controls. Different methods of drug delivery were investigated to develop this technology for clinical applications, including convection enhanced delivery (CED) and intrathecal injection. Taken together, these results represent the first full characterization of an Auger-emitting PARP inhibitor, demonstrate a survival benefit in mouse models of GBM, and confirm the high potential of123I-MAPi for clinical translation.One Sentence SummaryA novel PARP1-targeted Auger radiotherapeutic shows translational potential as a theranostic tool for imaging and killing cancer cells, resulting in tumor delineation and prolonged survival in a glioblastoma model.

Purpose: Clinically available intraoperative imaging tools to assist surgeons in identifying occult lesions are limited and partially responsible for the high rate of disease recurrence in patients with neuroendocrine tumors (NET). Using the established clinical efficacy of radiolabeled somatostatin analogs as a model, we demonstrate the ability of a fluorescent somatostatin analog to selectively target tumors that overexpress somatostatin receptor subtype-2 (SSTR2) and demonstrate utility for fluorescence-guided surgery (FGS). Experimental Design: A multimodality chelator (MMC) was used as a “radioactive linker” to synthesize the fluorescently labeled somatostatin analog, 67/68Ga-MMC(IR800)-TOC. In vivo studies were performed to determine the pharmacokinetic profile, optimal imaging time point, and specificity for SSTR2-expressing tissues. Meso- and microscopic imaging of resected tissues and frozen sections were also performed to further assess specific binding, and binding to human NETs was examined using surgical biospecimens from patients with pancreatic NETs. Results: Direct labeling with 67Ga/68Ga provided quantitative biodistribution analysis that was in agreement with fluorescence data. Receptor-mediated uptake was observed in vivo and ex vivo at the macro-, meso-, and microscopic scales. Surgical biospecimens from patients with pancreatic NETs also displayed receptor-specific agent binding, allowing clear delineation of tumor boundaries that matched pathology findings. Conclusions: The radioactive utility of the MMC allowed us to validate the binding properties of a novel FGS agent that could have a broad impact on cancer outcomes by equipping surgeons with real-time intraoperative imaging capabilities.

Resumo: Objetivo: Investigar os principais fatores de risco para o desenvolvimento da retinopatia da prematuridade (ROP) nos prematuros da Unidade de Terapia Intensiva (UTI) Neonatal da Santa Casa de Misericórdia de Ponta Grossa. Métodos: Estudo retrospectivo, tipo caso controle, através da revisão de prontuários entre Janeiro/2010 à Dezembro/2014. Foram incluídas crianças com peso ao nascer ?1.500 gramas, idade gestacional ?37 semanas e, excluídos portadores de malformação congênita, não submetidos à fundoscopia e os com prontuários incompletos. Os fatores de risco foram comparados com análises uni e multivarida. Para as variáveis quantitativas foi empregado o Teste t, para as qualitativas o Teste Exato de Fisher e, após análise univariada utilizou-se regressão logística. Resultados: Foram selecionados 172 neonatos, divididos em grupos: controle com 154 (89,5%) e caso com 18 (10,5%). Os fatores estudados foram: idade gestacional, peso ao nascer, APGAR 1º e 5º minuto, uso de surfactante, tempo de oxigenoterapia, fração inspirada de oxigênio (FiO2) utilizada, icterícia, necessidade de fototerapia, desenvolvimento de broncodisplasia pulmonar, enterocolite necrotizante, hemorragia intracraniana e cirurgias. Perante a análise univariada a idade gestacional, peso ao nascer, APGAR 1º e 5º minuto, tempo de oxigenoterapia, FiO2 máxima, necessidade de surfactante, tempo de início da icterícia e nível de bilirrubina indireta apresentaram significância estatística (p

The aim of this study was to identify predictor factors associated with the whitening outcome and risk and intensity of bleaching-induced tooth sensitivity from pooled data of 11 clinical trials of dental bleaching performed by the same research group.The individual patient data of several published and ongoing studies about dental bleaching was collected and retrospectively analyzed. At the patient-level, independent variables (bleaching techniques [at-home and in-office protocols], sex, age and baseline tooth color in shade guide unit [SGU]) as well as dependent variables (color change in shade guide units (ΔSGU), color change in the CIEL*a*b* system (ΔE), risk and intensity of TS in a visual analog scale) were collected. Multivariable linear regression and multivariable logistic regression models were carried out using backward elimination whenever the p-values were higher than 0.05.A significant relationship between baseline color and age on color change estimates was detected (p0.001). Every increase of one SGU in the baseline color resulted in an increase of approximate 0.66 in the final ΔSGU and 2.48 for the ΔE. For every increase of one year in the participant's age we observed a decrease of the whitening degree of 0.07 for the final ΔSGU and 0.69 for the ΔE. The bleaching technique was shown to be a significant predictor of ΔSGU (p0.001) but not of ΔE. In regard to TS, baseline color and bleaching technique are significant predictors (p0.001). The risk of TS for at-home bleaching was 51% (95% CI 41.4-60.6) and for the in-office 62.9% (95% CI 56.9-67.3).Younger patients with darker teeth reach a higher degree of whitening. Patient with darker teeth and submitted to at-home bleaching presents lower risk and intensity of TS.The baseline color of the teeth and the patient's age is directly related to the effectiveness of dental bleaching and TS.

Despite efforts in prevention, cervical cancer still presents with a high worldwide incidence and remains a great problem in public health, especially in low-income countries. Screening programs, such as colposcopy with Papanicolaou testing, have greatly improved mortality rates. However, the agents currently used to delineate those lesions (topical application of acetic acid or Lugol iodine) lack specificity and sometimes can lead to unnecessary biopsies or even cervical excisions. A tool to enable in vivo histology to quickly and quantitatively distinguish between tumor, dysplastic tissue, and healthy tissue would be of great clinical interest. Methods: Here, we describe the use of PARPi-FL, a fluorescent inhibitor of poly[adenosine diphosphate-ribose]polymerase 1 (PARP1), which is a nuclear enzyme that is overexpressed in cancer when compared with the normal surrounding tissues. We exploit its use as an optical imaging agent to specifically target PARP1 expression, which was demonstrated to be higher in cervical cancer than the normal surrounding tissue. Results: After topical application of PARPi-FL on freshly excised cone biopsy samples, the nuclei of tumor cells emitted a specific fluorescent signal that could be visualized using a handheld fluorescence confocal microscope. Conclusion: This approach has the potential to improve in vivo identification of tumor cells during colposcopy examination, allowing a rapid, noninvasive, and accurate histopathologic assessment.

The preclinical potential of many diagnostic and therapeutic small molecules is limited by their rapid washout kinetics and consequently modest pharmacological performances. In several cases, these could be improved by loading the small molecules into nanoparticulates, improving blood half-life, in vivo uptake and overall pharmacodynamics. In this study, we report a nanoemulsion (NE) encapsulated form of PARPi-FL. As a proof of concept, we used PARPi-FL, which is a fluorescently labeled sensor for olaparib, a FDA-approved small molecule inhibitor of the nuclear enzyme poly(ADP-ribose)polymerase 1 (PARP1). Encapsulated PARPi-FL showed increased blood half-life, and delineated subcutaneous xenografts of small cell lung cancer (SCLC), a fast-progressing disease where efficient treatment options remain an unmet clinical need. Our study demonstrates an effective method for expanding the circulation time of a fluorescent PARP inhibitor, highlighting the pharmacokinetic benefits of nanoemulsions as nanocarriers and confirming the value of PARPi-FL as an imaging agent targeting PARP1 in small cell lung cancer.

Poly(ADP-ribose)polymerase (PARP) inhibitors have emerged as potent antitumor drugs. Here, we describe the intrinsic fluorescence properties of the clinically approved PARP inhibitor rucaparib and its potential to directly measure drug distribution and target engagement—a critical factor for understanding drug action and improving efficacy. Methods: We characterized the photophysical properties of rucaparib and determined its quantum yield and lifetime. Using confocal microscopy and flow cytometry, we imaged the intracellular distribution of rucaparib and measured uptake and release kinetics. Results: Rucaparib has an excitation/emission maximum of 355/480 nm and a quantum yield of 0.3. In vitro time-lapse imaging showed accumulation in cell nuclei within seconds of administration. Nuclear rucaparib uptake increased with higher PARP1 expression, and we determined an intracellular half-life of 6.4 h. Conclusion: The label-free, intrinsic fluorescence of rucaparib can be exploited to interrogate drug distribution and target binding, critical factors toward improving treatment efficacy and outcome.

Background Current imaging techniques may not detect all prostate cancer (PCa) lesions. Objective To evaluate positron emission tomography (PET)/computed tomography (CT) using the radiolabeled GRPR antagonist probe BAY86-7548 (68Ga-RM2) for localization of newly diagnosed PCa in comparison with multiparametric magnetic resonance imaging (mpMRI), histopathology, and immunohistochemistry (IHC). Design, setting, and participants This was a prospective study of 16 men with biopsy-proven PCa (2 low, 8 intermediate, and 6 high risk). 68Ga-RM2 PET/CT was performed within 4 wk after mpMRI and within 2 wk before radical prostatectomy and extended bilateral pelvic lymph node dissection. Outcome measurements and statistical analysis The presence of cancer was evaluated by blinded specialists using a 5-point Likert scale, with lesions scoring 4 or 5 considered positive, on 68Ga-RM2 PET/CT, mpMRI, and 68Ga-RM2 PET/CT-mpMRI fused images for each of 12 anatomic areas of the prostate. Whole-mount, step-section pathology served as the reference standard. Expression of GRPR and prostate-specific membrane antigen (PSMA) was analyzed via IHC of tumor paraffin sections. Results and limitations Of 192 areas analyzed, 128 contained cancer. The sensitivity, specificity, and accuracy of 68Ga-RM2 PET/CT imaging and mpMRI did not differ significantly; fusing the images maximized the sensitivity and accuracy (85.2% and 83.9%, respectively) and averaged the specificity (81.3%). The area under the receiver operating characteristic curve was 0.76 for PET visual analysis, 0.72 for PET quantitative analysis, 0.76 for mpMRI, and 0.85 for combined PET/CT and mpMRI analysis. 68Ga-RM2 uptake did not correlate with Gleason score. IHC analysis revealed weaker staining for GRPR than for PSMA, and the expression of these markers was not correlated (r = 0.3882). The major limitation is the small sample size. Conclusions 68Ga-RM2 PET/CT is promising for detection and localization of primary PCa, and complements mpMRI. GRPR expression appears to be independent from PSMA expression, suggesting that GRPR- and PSMA-targeted PET imaging may be complementary. Patient summary This pilot prospective study shows that a positron emission tomography probe that binds to a marker of prostate cancer, GRPR, improves the ability of magnetic resonance imaging to detect prostate cancer.

The PPAR-α agonist, oleoylethanolamide (OEA) has neuroprotective properties in stroke models. However, its rapid degradation represents a limitation for an effective therapeutic approach. In this study, we evaluated the effects of a stable OEA-modeled compound, octadecylpropyl sulfamide (SUL) on the cognitive, behavioral, cellular and molecular alterations associated with hypoxia-ischemia (HI) in mice. Mice subjected to HI were treated with the PPAR-α antagonist GW6471 (GW) (1 mg/kg) followed 15 min later by SUL (3 and 10 mg/kg). Behavioral, motor, and cognitive tests were carried out 24 h and 7 days after the HI. The levels of microglia, reactive astrocytes and neuronal nuclei were studied using immunofluorescence, and the expression of genes related to the N-acyl-ethanolamides/endocannabinoid signaling systems was determined by qRT-PCR at the end of the experimental sequence. HI induced brain damage in the ipsilateral hippocampus and cortex, which lead to severe memory impairments, and motor coordination deficits. Significant neuronal loss, increased microglia and reactive astrocytes, and compensatory changes in genes associated with the inflammation/immune and endocannabinoid systems were observed in these brain structures of lesioned mice. SUL reversed the memory and motor deficits, decreased the overexpression of microglia and astrocytes, and reduced neurodegeneration induced by HI. Cnr1 and Cnr2 gene expression was modulated by SUL in both sham and HI mice, while Pparα and Faah expression was regulated in HI mice. GW completely blocked the beneficial actions of SUL. These findings suggest that treatment with SUL reduces brain damage and the associated motor and memory deficits induced by HI probably by normalizing the changes in neuroinflammation/immune system mediators. This work was supported by Fundació La Marató de TV3 (#2011/111830) to RM and FF, AGAUR (#-ICREA Acadèmia-2015), AGAUR (#SGR2009-00731), and MINECO (#SAF2014-59648-P) to RM. Instituto de Salud Carlos III-RETICS/RTA (#RD16/0017) to RM and FF, and (PI16/01689) to FF. JS holds a “Miguel Servet II” research contract from ISCIII, FIMABIS and ERDF-EU (CPII17/00024).

SUMMARYBleaching-induced tooth sensitivity (TS) is highly prevalent. Objective: This study aimed to determine if the combination of opioids and nonopioids analgesics (Tylex) may provide a better analgesic effect. Method: A triple-blind, parallel, randomized two-center clinical trial was conducted with 105 healthy patients who received either a placebo or a combination of acetaminophen/codeine. The first dose of Tylex 30 mg (acetaminophen 500 mg/codeine 30 mg) or placebo was administered one hour before the in-office bleaching (35% hydrogen peroxide), and extra doses were administered every six hours for 48 hours. The TS was recorded using a visual analog scale of 0 to 10 and a numeric rating scale of 0 to 4 in different periods: during bleaching, one hour up to 24 hours, and 24 hours up to 48 hours postbleaching. The color was measured before and one month after dental bleaching with a visual shade guide (Vita Classical), Vita Bleachedguide 3D-MASTER, and the spectrophotometer Vita Easyshade. The absolute risk of TS was evaluated using the Fisher exact test. Data of TS intensity with numeric rating scale of the two groups were compared with the Mann-Whitney U-test and the Friedman test, while data from the visual analog scale were evaluated by two-way repeated measures analysis of variance and the Tukey test for pairwise comparison. The color changes between groups were compared using the Student t-test (α=0.05). Results: No significant differences between the groups were observed in the risk and intensity of TS. The overall absolute risk of TS was approximately 96%. No significant differences between groups were observed in terms of color change (p>0.05) for any scale. Conclusion: The use of an acetaminophen/codeine combination prior to in-office bleaching does not reduce the risk and intensity of bleaching-induced TS.

Inducing graft acceptance without chronic immunosuppression remains an elusive goal in organ transplantation. Using an experimental transplantation mouse model, we demonstrate that local macrophage activation through dectin-1 and toll-like receptor 4 (TLR4) drives trained immunity-associated cytokine production during allograft rejection. We conducted nanoimmunotherapeutic studies and found that a short-term mTOR-specific high-density lipoprotein (HDL) nanobiologic treatment (mTORi-HDL) averted macrophage aerobic glycolysis and the epigenetic modifications underlying inflammatory cytokine production. The resulting regulatory macrophages prevented alloreactive CD8+ T cell-mediated immunity and promoted tolerogenic CD4+ regulatory T (Treg) cell expansion. To enhance therapeutic efficacy, we complemented the mTORi-HDL treatment with a CD40-TRAF6-specific nanobiologic (TRAF6i-HDL) that inhibits co-stimulation. This synergistic nanoimmunotherapy resulted in indefinite allograft survival. Together, we show that HDL-based nanoimmunotherapy can be employed to control macrophage function in vivo. Our strategy, focused on preventing inflammatory innate immune responses, provides a framework for developing targeted therapies that promote immunological tolerance. Funding for this research was provided by: National Institutes of Health (R01 EB009638, R01 HL144072, P01 HL131478, EB009638, R01 HL118440, R01 HL125703, R01 AI139623) Netherlands Organisation for Scientific Research (ZonMW Veni 016156059, ZonMW Vidi 91713324, ZonMW Vici 91818622) Comunidad de Madrid (B2017/BMD-3731) American Heart Association (16SDG27250090) NIH Program of Excellence in Nanotechnology (HHSN368201000045C, K25 EB016673, P30 CA008748) Harold S. Geneen Charitable Trust Spanish Ministry of Science (SAF2016-80031-R) ERC Consolidator (310372) Sí

Significance Diffuse large B-cell lymphoma (DLBCL) is the most common adult lymphoma, accounting for 37% of all non-Hodgkin lymphoma cases in the United States. Despite an approximate 50% cure rate, refractory or relapsed cases have a poor prognosis and require timely medical interventions. Therefore, accurate diagnostic methods play a pivotal role in managing DLBCL. 18 F- fluorodeoxyglucose positron emission tomography ([ 18 F]FDG-PET) imaging, the current standard imaging modality for diagnosing DLBCL, often fails to differentiate inflamed from malignant lymph nodes in patients with DLBCL. To address this urgent medical need, we have developed a targeted PET imaging method that accurately distinguishes malignancy from inflammation in the lymph nodes. Our targeted PET imaging approach could play an essential role in the clinical development of therapies that induce significant inflammation in DLBCL.

More sensitive and specific methods for early detection are imperative to improve survival rates in oral cancer. However, oral cancer detection is still largely based on visual examination and histopathology of biopsy material, offering no molecular selectivity or spatial resolution. Intuitively, the addition of optical contrast could improve oral cancer detection and delineation, but so far no molecularly targeted approach has been translated. Our fluorescently labeled small-molecule inhibitor PARPi-FL binds to the DNA repair enzyme poly(ADP-ribose)polymerase 1 (PARP1) and is a potential diagnostic aid for oral cancer delineation. Based on our preclinical work, a clinical phase I/II trial opened in March 2017 to evaluate PARPi-FL as a contrast agent for oral cancer imaging. In this commentary, we discuss why we chose PARP1 as a biomarker for tumor detection and which particular characteristics make PARPi-FL an excellent candidate to image PARP1 in optically guided applications. We also comment on the potential benefits of our molecularly targeted PARPi-FL-guided imaging approach in comparison to existing oral cancer screening adjuncts and mention the adaptability of PARPi-FL imaging to other environments and tumor types.

Insufficient chemotherapy response and rapid disease progression remain concerns for small-cell lung cancer (SCLC). Oncologists rely on serial CT scanning to guide treatment decisions, but this cannot assess in vivo target engagement of therapeutic agents. Biomarker assessments in biopsy material do not assess contemporaneous target expression, intratumoral drug exposure, or drug-target engagement. Here, we report the use of PARP1/2-targeted imaging to measure target engagement of PARP inhibitors in vivo. Using a panel of clinical PARP inhibitors, we show that PARP imaging can quantify target engagement of chemically diverse small molecule inhibitors in vitro and in vivo. We measure PARP1/2 inhibition over time to calculate effective doses for individual drugs. Using patient-derived xenografts, we demonstrate that different therapeutics achieve similar integrated inhibition efficiencies under different dosing regimens. This imaging approach to non-invasive, quantitative assessment of dynamic intratumoral target inhibition may improve patient care through real-time monitoring of drug delivery., Treatment of small-cell lung cancer remains a challenge due to multiple mechanisms of resistance to current therapies; measuring patient response is crucial in adapting and choosing adequate treatment. Here the authors develop a strategy to visualise in vivo dynamics of a class of widely used PARP inhibitors.

Background Chronic cardiac rejection is intimately associated with cardiac allograft vasculopathy and fibrosis, both causing severe complications that cannot be reversed. Thus, there is an urgent need for early diagnosis and for development of therapeutic agents. Chronic rejection is accompanied by the dramatic upregulation of EDA + fibronectin (EDA + Fn), which is virtually undetectable in the normal healthy adult. Methods In this study, we evaluated the potential of the monoclonal antibody F8, specific to that molecule, to selectively accumulate in chronically rejected allografts. Results A syngeneic immunocompetent heterotopic rat heart transplantation model was used to induce chronic rejection within 70 days. The F8 antibody or an antibody of irrelevant specificity, labeled with the dye DY-682, was administered and near-infrared fluorescence (NIRF) imaging was performed. Targeting performance was assessed by macroscopic organ imaging and fluorescence microscopy. A selective accumulation of the F8 antibody (but not of the negative control antibody) was observed by NIRF imaging in cardiac allografts. The antibody localized to diseased blood vessels as well as to fibrotic regions, where the cognate antigen is abundantly expressed. Conclusions This is the first example of antibody-mediated imaging of chronic cardiac rejection. The findings pave the way to immuno-positron emission tomography (immuno-PET) imaging of this clinical condition in patients using the human F8 antibody labeled with a suitable radionuclide (e.g., iodine-124). Furthermore, it would be conceivable to use the F8 antibody as a delivery vehicle to assess experimentally whether a bioactive payload (e.g., drug or cytokine) may be able to reduce disease progression.

A systematic review of clinical studies to evaluate the frequency of micronuclei in the oral mucosa of smokers and non-smokers in adult patients was performed. A comprehensive search was carried out on MEDLINE via PubMeb, Scopus, Web of Science, LILACS, BBO, and Cochrane Library and SIGLE without restrictions. Dissertations and thesis were searched using the ProQuest Dissertations and Periodicos Capes Thesis Databases. We included only cross-sectional clinical trials that compared the frequency of micronuclei in the oral mucosa of smokers and non-smokers in adult patients. After the removal of duplicates, 1338 articles were identified. After title and abstract screening, 35 studies remained. Eighteen studies were further excluded, whereas 17 studies remained for qualitative analysis and 16 for the meta-analysis of the primary and secondary outcomes. A significant difference in the frequency of micronuclei in smokers when compared to non-smokers was observed in the present study. Despite the high variation in the methodology of the assessed studies, this study showed a higher frequency of micronuclei in exfoliated cells of smokers compared to non-smokers. The use of tobacco is associated with cytotoxic and genotoxic effects because a higher frequency of micronuclei in exfoliated cells of smokers was observed.

SUMMARY Objective: The aim of this study was to evaluate the genotoxicity of in-office bleaching with 35% hydrogen peroxide in epithelial cells from the gingival and lip tissues. Methods and Materials: Thirty volunteers with central incisors shade A1 or darker were selected for this study. The gingival tissue of the teeth to be bleached was isolated with a light-polymerized resin dam, and the 35% hydrogen peroxide gel was administered during three 15-minute applications over the course of the 45-minute application period. Two bleaching sessions with a one-week interval in between were performed. Exfoliated oral mucosa gingival epithelial cells and upper lip lining were collected at baseline and one month after the in-office dental bleaching. The scraped cells were placed on clean glass slides and smears were prepared. After staining with Giemsa solution, two blinded examiners performed cell and micronuclei counts under a 100× optical microscope. Tooth sensitivity was evaluated using the Visual Analogue Scale (VAS). Shade evaluation was recorded before and one month after the bleaching treatment with the value-oriented shade guide Vita Bleachedguide 3D-MASTER and the spectrophotometer Vita Easyshade. Data from the shade guide units and the micronuclei (MN) frequency were subjected to a Mann-Whitney test (α=0.05). The overall difference between before and one month after the bleaching treatment (ΔE and ΔSGU), absolute risk, and intensity of tooth sensitivity (TS) were calculated, as was the 95% confidence interval (CI). Results: The frequency of MN was not increased after bleaching with 35% hydrogen peroxide in both study groups (p>0.05). The absolute risk of TS of the participants was 93% (95% CI, 79%-98%), with a mean VAS intensity of 5.7 ± 2.9 (95% CI, 4.6-6.8). Meaningful whitening was observed after bleaching. The change in shade guide units in the Bleachedguide 3D-MASTER was 2.3 ± 1.4. In terms of ΔE, the change in color was 7.7 ± 3.5. Conclusions: The in-office bleaching did not induce DNA damage to the gingival and lip tissue during the bleaching period. Although effective whitening was observed, most of the participants experienced TS.

SUMMARYObjective: A systematic review and meta-analysis were performed to evaluate the risk and intensity of tooth sensitivity during in-office and at-home bleaching in adult patients. The efficacy of dental bleaching was also evaluated.Methods: A comprehensive search was performed in the MEDLINE via PubMed, Scopus, Web of Science, Latin American and Caribbean Health Sciences Literature database, Brazilian Library in Dentistry, Cochrane Library, and System for Information on Grey Literature in Europe without restrictions. The annual conference of the International Association for Dental Research abstracts (1990-2014) and unpublished and ongoing trials registry were also searched. Dissertations and theses were searched using the ProQuest Dissertations and Periódicos Capes Theses databases. Only randomized clinical trials that compared the prevalence or intensity of tooth sensitivity during in-office and at-home bleaching in adult patients were included and studies that evaluated the efficacy of these dental bleaching techniques, in terms of shade guide units (ΔSGU) and in terms of color difference measured with a spectrophotometer (ΔE*).Results: After the removal of duplicates, 1139 articles were identified. After title and abstract screening, 29 studies remained. Fifteen studies were further excluded, whereas 12 studies remained for qualitative analyses and 8 for the meta-analysis of the primary and secondary outcomes. No significant difference in the risk/intensity of tooth sensitivity or in bleaching efficacy was observed in the present study.Conclusion: In an overall comparison of at-home and in-office bleaching, no differences were detected, either regarding risk/intensity of tooth sensitivity or the effectiveness of the bleaching treatment. This comparison, however, does not take into consideration variations in the protocols (daily usage time, number of bleaching sessions, and product concentration) of the bleaching techniques in the studies included.

SUMMARY Objectives: The aim of this study was to evaluate the efficacy, color stability, risk, and intensity of tooth sensitivity (TS) of combined bleaching techniques performed with 20% or 35% hydrogen peroxide for an in-office protocol. Methods: Thirty patients were randomly divided into two groups and submitted to a single 45-minute in-office bleaching session with 35% hydrogen peroxide or 20% hydrogen peroxide. At-home bleaching was performed with 10% carbamide peroxide for two hours daily over the course of two weeks. The color was evaluated with the value-oriented shade guide Vita Classical at different periods up to 12 months after bleaching. Patients recorded the intensity of TS using a five-point verbal scale. Color change data were submitted to a two-way repeated-measures analysis of variance and Tukey test (α=0.05). The absolute risk and intensity of TS were compared with the Fisher exact test and Mann-Whitney test, respectively (α=0.05). Results: On average, an effective and similar whitening of three units in shade guide was observed for both groups, which remained stable for 12 months. When both protocols were compared, the one with hydrogen peroxide 35% showed a higher risk (p=0.02) and intensity of TS (p=0.04). In regard to the TS intensity, no significant difference was observed up to 48 hours after in-office bleaching (p=0.09) and during the at-home bleaching phase of the study (p=0.71). Conclusion: The combined bleaching technique using at-home bleaching associated with in-office bleaching was effective and stable over the course of 12 months, regardless of the concentration of the hydrogen peroxide used for in-office bleaching. However, the protocol with 20% hydrogen peroxide produced lower risk and intensity of TS.

Targeting and inhibiting DNA repair pathways is a powerful strategy of controlling malignant growth. One such strategy includes the inhibition of PARP1, a central element in the intracellular DNA damage response. To determine and visualize the expression and intercellular distribution of PARP1 in vivo, and to monitor the pharmacokinetics of PARP1 targeted therapeutics, fluorescent small probes were developed. To date, however, it is unclear how these probes behave in a more realistic clinical setting, where DNA damage has been induced through one or more prior lines of therapy. Here, we use one such imaging agent, PARPi-FL, in tissues both with and without prior DNA damage, and investigate its value as a probe for PARP1 imaging. We show that PARP1 expression in oral cancer is high, and that the uptake of PARPi-FL is selective, irrespective of whether cells were exposed to irradiation or not. We also show that PARPi-FL uptake increases in response to DNA damage, and that this increase is reflected in higher enzyme expression. Our findings provide a framework for measuring exposure of cells to external beam radiation, and could help to elucidate the effects of such treatments non-invasively in mouse models of cancer.

The current study presents [(18)F]PARPi as imaging agent for PARP1 expression.[(18)F]PARPi was generated by conjugating a 2H-phthalazin-1-one scaffold to 4-[(18)F]fluorobenzoic acid. Biochemical assays, optical in vivo competition, biodistribution analysis, positron emission tomography (PET)/X-ray computed tomography, and PET/magnetic resonance imaging studies were performed in subcutaneous and orthotopic mouse models of glioblastoma.[(18)F]PARPi shows suitable pharmacokinetic properties for brain tumor imaging (IC50 = 2.8 ± 1.1 nM; logPCHI = 2.15 ± 0.41; plasma-free fraction = 63.9 ± 12.6 %) and accumulates selectively in orthotopic brain tumor tissue. Tracer accumulation in subcutaneous brain tumors was 1.82 ± 0.21 %ID/g, whereas in healthy brain, the uptake was only 0.04 ± 0.01 %ID/g.[(18)F]PARPi is a selective PARP1 imaging agent that can be used to visualize glioblastoma in xenograft and orthotopic mouse models with high precision and good signal/noise ratios. It offers new opportunities to non-invasively image tumor growth and monitor interventions.

Background Although the understanding of the genetic and molecular basis of cancer has advanced significantly over the past several decades, imaging and treatment options for glioblastoma patients have been more limited (N Engl J Med 359:492-507, 2008). This is in part due to difficulties in diagnosing this disease early, combined with its diffuse, infiltrative growth. This study was aimed at the development of a novel diagnostic tool for glioblastoma through the synthesis of a small molecule based on radioiodinated poly(ADP-ribose)polymerase 1 (PARP1) targeted tracers. This PARP1 is a biomarker that is overexpressed in glioblastoma tissue, but has only low expression levels in the healthy brain (Neoplasia 16:432-40, 2014). Methods A library of PARP1 inhibitors (iodo-PARPis) was synthesized. Based on their pharmacokinetic properties and nuclear PARP1 binding, the most successful inhibitor was radiolabeled with 131I and 124I. Biodistribution as well as imaging experiments were performed in orthotopic and subcutaneous mouse models of glioblastoma. Results One member of our iodo-poly(ADP-ribose)polymerase 1 (PARP1) inhibitor library, I2-PARPi, shows promising biophysical properties for in vivo application. All synthesized tracers have IC50 values in the nanomolar range (9 ± 2–107 ± 4 nM) and were able to inhibit the uptake of a fluorescent PARP1 inhibitor analog (PARPi-FL). I2-PARPi was able to reduce the uptake of PARPi-FL by 78 ± 4 % in vivo. In mouse models of glioblastoma, we show that the radioiodinated inhibitor analog has high uptake in tumor tissue (U251 MG xenograft, tumor, 0.43 ± 0.06 %ID/g; brain, 0.01 ± 0.00 %ID/g; muscle, 0.03 ± 0.01 %ID/g; liver, 2.35 ± 0.57 %ID/g; thyroid, 0.24 ± 0.06 %ID/g). PET and SPECT imaging performed in orthotopic glioblastoma models with [124I]- and [131I]-I2-PARPi showed selective accumulation in the tumor tissue. These results were also verified using autoradiography of tumor sections, which displayed focal selective uptake of the tracer in the tumor regions as confirmed by histology. The uptake could be blocked through pre-injection of excess unlabeled PARP1 inhibitor (Olaparib). Conclusions We have successfully synthesized and radioiodinated the PARP1 selective tracer I2-PARPi. The novel tracer shows selective binding to tumor tissue, both in vitro and in models of glioblastoma, and has the potential to serve as a selective PET imaging agent for brain tumors. Electronic supplementary material The online version of this article (doi:10.1186/s13550-015-0123-1) contains supplementary material, which is available to authorized users.

[Purpose] Tumor cells can be effectively inactivated by heating mediated by magnetic nanoparticles. However, optimized nanomaterials to supply thermal stress inside the tumor remain to be identified. The present study investigates the therapeutic effects of magnetic hyperthermia induced by superparamagnetic iron oxide nanoparticles on breast (MDA-MB-231) and pancreatic cancer (BxPC-3) xenografts in mice in vivo., [Methods] Superparamagnetic iron oxide nanoparticles, synthesized either via an aqueous (MF66; average core size 12 nm) or an organic route (OD15; average core size 15 nm) are analyzed in terms of their specific absorption rate (SAR), cell uptake and their effectivity in in vivo hyperthermia treatment., [Results] Exceptionally high SAR values ranging from 658 ± 53 W*gFe −1 for OD15 up to 900 ± 22 W*gFe −1 for MF66 were determined in an alternating magnetic field (AMF, H = 15.4 kA*m−1 (19 mT), f = 435 kHz). Conversion of SAR values into system-independent intrinsic loss power (ILP, 6.4 ± 0.5 nH*m2*kg−1 (OD15) and 8.7 ± 0.2 nH*m2*kg−1 (MF66)) confirmed the markedly high heating potential compared to recently published data. Magnetic hyperthermia after intratumoral nanoparticle injection results in dramatically reduced tumor volume in both cancer models, although the applied temperature dosages measured as CEM43T90 (cumulative equivalent minutes at 43°C) are only between 1 and 24 min. Histological analysis of magnetic hyperthermia treated tumor tissue exhibit alterations in cell viability (apoptosis and necrosis) and show a decreased cell proliferation., The described work was carried out within the project “Multifunctional Nanoparticles for the Selective Detection and Treatment of Cancer” (Multifun), which is funded by the European Seventh Framework Program (FP7/2007–2013) under grant agreement no. 262943. F.J.T acknowledges financial support from Ramon y Cajal subprogram (RYC-2011-09617).

Hepatocellular carcinoma (HCC) represents the second leading cause of cancer-related deaths and is reported to be resistant to chemotherapy caused by tumor-initiating cells. These tumor-initiating cells express stem cell markers. An accumulation of tumor-initiating cells can be found in 2% to 50% of all HCC and is correlated with a poor prognosis. Mechanisms that mediate chemoresistance include drug export, increased metabolism, and quiescence. Importantly, the mechanisms that regulate quiescence in tumor-initiating cells have not been analyzed in detail so far. In this research we have developed a single cell tracking method to follow up the fate of tumor-initiating cells during chemotherapy. Thereby, we were able to demonstrate that mCXCL1 exerts cellular state-specific effects regulating the resistance to chemotherapeutics. mCXCL1 is the mouse homolog of the human IL8, a chemokine that correlates with poor prognosis in HCC patients. We found that mCXCL1 blocks differentiation of premalignant cells and activates quiescence in tumor-initiating cells. This process depends on the activation of the mTORC1 kinase. Blocking of the mTORC1 kinase induces differentiation of tumor-initiating cells and allows their subsequent depletion using the chemotherapeutic drug doxorubicin. Our work deciphers the mCXCL1–mTORC1 pathway as crucial in liver cancer stem cell maintenance and highlights it as a novel target in combination with conventional chemotherapy. Cancer Res; 76(18); 5550–61. ©2016 AACR.

The role of CB2 cannabinoid receptors (CB2R) in global brain lesions induced by hypoxia-ischemia (HI) insult is still unresolved. The aim of this study was to evaluate the involvement of CB2R in the behavioural and biochemical underpinnings related to brain damage induced by HI in adult mice, and the mechanisms involved. CB2R knockout (KO) mice and wild-type littermates (WT) underwent permanent ligation of the left common carotid artery and hypoxia. Behavioural measurements in the rotarod, beam walking, object recognition, open field, and Irwin tests were carried out 24h, 72h and 7 days. In KO mice, more extensive brain injury was observed. Behavioural deficits in the Irwin test were observed in both genotypes; while WT mice showed progressive recovery by day 7, KO mice did not. Only KO mice showed alterations in motor learning, coordination and balance, and did not recover over time. A higher expression of microglia and astrocytes was observed in several brain areas of lesioned WT and KO mice. The possible alteration of the inflammatory-related factors HIF-1α and TIM-3 was evaluated in these animals. In both genotypes, HIF-1α and TIM-3 expression was observed in lesioned areas associated with activated microglia. However, the expression levels of these proteins were exacerbated in KO mice in several lesioned and non-lesioned brain structures. Our results indicate that CB2R may have a crucial neuroprotective role following HI insult through the modulation of the inflammatory-related HIF-1α/TIM-3 signalling pathway in microglia. This work was supported by the European Commission, FP7 (#HEALTH-F2-2013-602891), the Spanish Ministerio de Economía y Competitividad-MINECO (#SAF2014-59648-P), the Spanish Instituto de Salud Carlos III, RETICS-RTA (#RD12/0028/0023) and the Generalitat de Catalunya, AGAUR (#2014-SGR-1547). C.L.P. and R.N. are recipients of a predoctoral fellowship from the Spanish Ministerio de Educación, Cultura y Deporte (MECD) and the Spanish MINECO, respectively. I.M.L.M. is a recipient of a predoctoral fellowship from the Mexican Consejo Nacional de Ciencia y Tecnología (CONACyT). FEDER partial funding is also acknowledged