Efficient Gene Editing for Heart Disease via ELIP-Based CRISPR Delivery System.

Liposomes as carriers for CRISPR/Cas9 complexes represent an attractive approach for cardiovascular gene therapy. A critical barrier to this approach remains the efficient delivery of CRISPR-based genetic materials into cardiomyocytes. Echogenic liposomes (ELIP) containing a fluorescein isothiocyanate-labeled decoy oligodeoxynucleotide against nuclear factor kappa B (ELIP-NF-κB-FITC) were used both in vitro on mouse neonatal ventricular myocytes and in vivo on rat hearts to assess gene delivery efficacy with or without ultrasound. In vitro analysis was then repeated with ELIP containing Cas9-sg-IL1RL1 (interleukin 1 receptor-like 1) RNA to determine the efficiency of gene knockdown. ELIP-NF-κB-FITC without ultrasound showed limited gene delivery in vitro and in vivo, but ultrasound combined with ELIP notably improved penetration into heart cells and tissues. When ELIP was used to deliver Cas9-sg-IL1RL1 RNA, gene editing was successful and enhanced by ultrasound. This innovative approach shows promise for heart disease gene therapy using CRISPR technology.

Activation of interleukin 33-NFκB axis in granulosa cells during atresia and its role in disposal of atretic follicles†.

It has been previously shown that the cytokine interleukin 33 is required for two processes, i.e., autophagic digestion of granulosa cells and recruitment of macrophages into atretic follicles, for full disposal of atretic follicles. Now, this study shows that activation of interleukin 33-suppression of tumorigenicity 2-Nuclear Factor ĸB (NFκB) axis in granulosa in early atretic follicles may regulate those two events. Injection of human chorionic gonadotropin has been shown to induce a transient peak of interleukin 33 expression with synchronized atresia. In this model, interleukin 33-independent expression of suppression of tumorigenicity 2 in granulosa cells was detected in early atretic follicles before macrophage invasion. The activation of NFκB pathway in ovaries was further demonstrated in vivo in Tg mice with luciferase-reporter for NFκB activation; the activation was microscopically localized to granulosa cells in early atretic follicles. Importantly, antibody blockage of interleukin 33 or interleukin 33 Knock-out (KO) (Il33-/-) not only inhibited NFκB activity in ovaries, but it also altered expression of two key genes, i.e., reduction in proinflammatory interleukin6 (IL6) expression, and a surge of potential autophagy-inhibitory mammalian target of rapamycin (mTOR) expression in atretic follicles. By contrast, apoptosis and other genes, such as interleukin1ß (IL1ß) were not affected. In conclusion, in parallel to apoptosis, atresia signals also trigger activation of the interleukin 33-suppression of tumorigenicity 2-NFκB pathway in granulosa, which leads to (1) down-regulated expression of mTOR that is a negative regulator of autophagy and (2) up-regulated expression of proinflammatory IL6.

An evaluation of quantitative percussion diagnostics for determining the probability of a microgap defect in restored and unrestored teeth: A prospective clinical study.

STATEMENT OF PROBLEM: Current dental diagnostics are image based and cannot detect a structural microgap defect such as a crack in a tooth. Whether percussion diagnostics can effectively diagnose a microgap defect is unclear. PURPOSE: The purpose of the present study was to determine from a large multicenter prospective clinical study whether quantitative percussion diagnostics (QPD) could detect structural damage in teeth and whether a probability of its presence could be provided. MATERIAL AND METHODS: A nonrandomized prospective and multicenter clinical validation study with 224 participants was performed in 5 centers with 6 independent investigators. The study used QPD and the normal fit error to determine whether a microgap defect was present in a natural tooth. Teams 1 and 2 were blinded. Team 1 tested teeth scheduled for restoration with QPD, and Team 2 disassembled the teeth aided by a clinical microscope, transillumination, and a penetrant dye. Microgap defects were documented in written and video formats. Controls were participants without damaged teeth. The percussion response from each tooth was stored on a computer and analyzed. A total of 243 teeth were tested to provide approximately 95% power to test the performance goal of 70%, based on an assumed population overall agreement of 80%. RESULTS: Regardless of the collection method, tooth geometry, restoration material used, or restoration type, the data on detecting a microgap defect in a tooth were accurate. The data also reflected good sensitivity and specificity consistent with previously published clinical studies. The combined study data showed an overall agreement of 87.5% with a 95% confidence interval (84.2 to 90.3), beyond the 70% predetermined performance goal. The combined study data determined whether it was possible to predict the probability of a microgap defect. CONCLUSIONS: The results showed that the data on detecting microgap defects in a tooth site were consistently accurate and confirmed that QPD provided information to aid the clinician in treatment planning and early preventative treatment. QPD can also alert the clinician of probable diagnosed and undiagnosed structural problems via the use of a probability curve.

Prevention of work-related musculoskeletal disorders among dental professionals: A scoping review.

BACKGROUND: Musculoskeletal disorders (MSDs) are highly prevalent among dental professionals. Studies examining the prevention of dental work-related MSDs have been completed globally. OBJECTIVE: To identify and evaluate the available evidence regarding MSD prevention in dental practice, and to identify knowledge gaps. METHODS: An electronic search was conducted across multiple scientific databases. Identified articles were assessed according to inclusion/exclusion criteria. Systematic reviews, interventions and published expert opinion were included. RESULTS: Five systematic reviews, 27 intervention studies and 21 expert opinion articles were included. There was overall consensus that use of ergonomic equipment is beneficial in MSD prevention. Knowledge gaps identified included: a lack of systematic reviews on the role of exercise and therapy in MSD prevention; a lack of high-quality intervention studies; a lack of research in general outside of ergonomic equipment use. Expert opinion articles have been published despite a lack of dental-specific research. CONCLUSIONS: Whilst more robust research is required, current evidence supports the use of optical enhancement, saddle chairs, and wide-diameter silicone-handled instruments in MSD prevention. Knowledge gaps were identified relating to the use of exercise, therapy, and education in the prevention of dental work-related MSDs. Further research is required, specifically high-quality intervention studies.

Characterization, isolation, and in vitro culture of leptomeningeal fibroblasts.

Meninges, or the membranous coverings of the brain and spinal cord, play host to dozens of morbid pathologies. In this study we provide a method to isolate the leptomeningeal cell layer, identify leptomeninges in histologic slides, and maintain leptomeningeal fibroblasts in in vitro culture. Using an array of transcriptomic, histological, and cytometric analyses, we identified ICAM1 and SLC38A2 as two novel markers of leptomeningeal cells in vivo and in vitro. Our results confirm the fibroblastoid nature of leptomeningeal cells and their ability to form a sheet-like layer that covers the brain and spine parenchyma. These findings will enable researchers in central nervous system barriers to describe leptomeningeal cell functions in health and disease.

Requirement of brain interleukin33 for aquaporin4 expression in astrocytes and glymphatic drainage of abnormal tau.

Defective aquaporin4 (AQP4)-mediated glymphatic drainage has been linked to tauopathy and amyloid plaque in Alzheimer's disease. We now show that brain interleukin33 (IL33) is required for regulation of AQP4 expression in astrocytes, especially those at neuron-facing membrane domain (n-AQP4). First, IL33-deficient (Il33-/-) mice showed a loss of n-AQP4 after middle age, which coincided with a rapid accumulation of abnormal tau in neurons and a reduction in drainage of abnormal tau to peripheral tissues. Second, injection of recombinant IL33 induced robust expression of AQP4 at perivascular endfoot (p-AQP4) of astrocytes, but not n-AQP4, in Il33-/- brains. Although the increased p-AQP4 greatly accelerated drainage of intracerebroventricularly injected peptides, it did not substantially accelerate drainage of abnormal tau. These results suggest that p-AQP4 drives overall convective flow toward perivenous space, i.e., glymphatics, whereas n-AQP4 may generate an aqueous flow away from neurons to remove neuronal wastes, e.g., abnormal tau. We have previously shown the role of brain IL33 in DNA repair and autophagy in neurons with oxidative stress. Now, we show that IL33 deficiency also impairs glymphatic drainage. Defects in those mechanisms together may lead to chronic neurodegeneration and tauopathy at old age in IL33-deficient mice.

A PIK3CA transgenic mouse model with chemical carcinogen exposure mimics human oral tongue tumorigenesis.

Oral cancer causes significant global mortality and has a five-year survival rate of around 64%. Poor prognosis results from late-stage diagnosis, highlighting an important need to develop better approaches to detect oral premalignant lesions (OPLs) and identify which OPLs are at highest risk of progression to oral squamous cell carcinoma (OSCC). An appropriate animal model that reflects the genetic, histologic, immunologic, molecular and gross visual features of human OSCC would aid in the development and evaluation of early detection and risk assessment strategies. Here, we present an experimental PIK3CA + 4NQO transgenic mouse model of oral carcinogenesis that combines the PIK3CA oncogene mutation with oral exposure to the chemical carcinogen 4NQO, an alternate experimental transgenic mouse model with PIK3CA as well as E6 and E7 mutations, and an existing wild-type mouse model based on oral exposure to 4NQO alone. We compare changes in dorsal and ventral tongue gross visual appearance, histologic features and molecular biomarker expression over a time course of carcinogenesis. Both transgenic models exhibit cytological and architectural features of dysplasia that mimic human disease and exhibit slightly increased staining for Ki-67, a cell proliferation marker. The PIK3CA + 4NQO model additionally exhibits consistent lymphocytic infiltration, presents with prominent dorsal and ventral tongue tumours, and develops cancer quickly relative to the other models. Thus, the PIK3CA + 4NQO model recapitulates the multistep genetic model of human oral carcinogenesis and host immune response in carcinogen-induced tongue cancer, making it a useful resource for future OSCC studies.

Ten-year retrospective study of the effectiveness of quantitative percussion diagnostics as an indicator of the level of structural pathology in teeth.

STATEMENT OF PROBLEM: Conventional dental diagnostic aids are only partially effective in diagnosing structural defects such as cracks in teeth. A more predictable diagnostic for structural instability in the mouth is needed. PURPOSE: The purpose of this clinical study with an increased population size was to evaluate the effectiveness of diagnosing structural instability by using the quantitative percussion diagnostics (QPD) system and to evaluate the influence of independent variables on the relationship between normal fit error (NFE) and observed structural instability found during the clinical disassembly of teeth. MATERIAL AND METHODS: Twenty-two participants with 264 sites needing restoration were enrolled in an institutional review board-approved 10-year retrospective clinical study. Each site had been tested with the QPD system before being disassembled microscopically with video documentation, and the clinical disassembly results were recorded on a defect-assessment sheet. The NFE data were separately recorded from the preexisting records. The classification of structural pathology based on the disassembly observations for each of the 264 sites was conducted by the clinical researcher (C.G.S.) who was blinded to the NFE values. RESULTS: The 264 sites from 22 patients were classified as 8 in the none group, 87 in the moderate group, and 169 in the severe group based on the disassembly findings. The NFE data for the sites were analyzed by using the predefined NFE cutoffs that were independently generated from the previous cumulative logistic regression and decision tree model. For the cumulative logistic regression, 235 out of 264 sites were correctly classified with an agreement of 0.89 (adjusted 95% CI: 0.83-0.95). The number of correctly classified sites for the decision tree model was 234, and the agreement was also 0.89 (adjusted 95% CI: 0.83-0.94). For both cumulative logistic regression and decision tree models, the overall misclassification rate was less than 20% for any restoration material or restoration type. Therefore, the overall performance of NFE classification was consistently good, regardless of restoration material or type. In addition, the sensitivity of the severe category was above 90% for any restoration material or type for the decision tree model. CONCLUSIONS: The QPD system was found to be a reliable diagnostic aid for classifying structural damage in the categories of none, moderate, or severe based on clinical disassembly findings under the clinical microscope and NFE values. Furthermore, it was determined that restoration type and restoration design were not significant factors in correlating structural pathology with NFE.

Inter-molecular epitope spreading does not lead to extension of autoimmunity beyond target tissue in autoimmune glomerulonephritis.

Inter-molecular epitope spreading during autoimmune pathogenesis leads to generation of new pathogenic epitopes on other autoantigens beyond the original one. It raises an important question as whether autoimmunity extends beyond the target tissues if new epitopes are on the molecules shared with other tissues. This study is aimed addressing this question in a rat anti-glomerular basement membrane (GBM) glomerulonephritis model induced by a T cell epitope of glomerulus-specific collagen4α3. We have demonstrated inter-molecular B cell epitope spreading. Four novel epitopes were first identified by screening a phage display random peptide library against autoantibodies isolated from the GBM of immunized rats. All four epitopes were derived from GBM proteins with three from laminins and one from collagen4α4. Three out of four synthetic peptides were nephritogenic. Importantly, two peptides from lamininα1 and lamininß1, respectively, induced severe inflammation in glomeruli but not in the interstitial tissues, despite the presence of more abundant laminins in the tubular basement membranes. Our study suggests that surrounding tissues may display a lower or altered susceptibility to autoimmune inflammation. Thus, preventing extension of autoimmune inflammation beyond the original target tissue.

Glymphatic Efficiency Is a Critical Factor for Using Abnormal Tau in Peripheral Tissues as Biomarker for Alzheimer's Disease.

Alzheimer's Disease or other dementias are characterized by the accumulation of abnormal tau and amyloid ß peptides in brains. Therefore, abnormal tau and amyloid peptides in peripheral tissues or blood have been explored as diagnostic biomarkers. On the other hand, recent studies have revealed glymphatics a special drainage system for brain's wastes. We aimed to investigate whether effectiveness of glymphatic system affects the quantity of abnormal tau in the peripheral tissues. We have previously shown that aged IL33 KO (Il33 -/-) mice develop Alzheimer's like disease. Despite a large quantity of abnormal tau in brains, Il33 -/- mice showed a much lower amount of abnormal tau drained to the peripheral tissues kidneys than in wild type mice. Our further study showed that it was caused by defective glymphatic drainage since Il33 KO impaired glymphatics. Thus, it is necessary to identify biomarkers, which can evaluate efficiency of glymphatic drainage. Simultaneous measurement of these biomarkers and abnormal tau in peripheral tissues or blood may be critical for accurate diagnosis of Alzheimer's disease.

Quantitative percussion diagnostics as an indicator of the level of the structural pathology of teeth: Retrospective follow-up investigation of high-risk sites that remained pathological after restorative treatment.

STATEMENT OF PROBLEM: Structural damage may remain even after a tooth is restored. Conventional diagnostic aids do not quantify the severity of structural damage or allow the monitoring of structural changes after restoration. PURPOSE: The purpose of this retrospective clinical study was to provide an in-depth analysis of 9 high-risk sites after restoration. The analysis followed structural defects found upon disassembly, restorative materials used, therapeutic procedures provided, current longevity, and long-term quantitative percussion diagnostics (QPD) to monitor results. The hypothesis was that QPD can be used to quantify positive and negative changes in structural stability. MATERIAL AND METHODS: Sixty sites requiring restoration were part of an institutional review board-approved clinical study. Each participant was examined comprehensively, including QPD testing, at each follow-up. Long-term changes in normal fit error (NFE) values after restoration were evaluated according to a pathology rating system established in an earlier publication. Nine highly compromised sites were chosen for further analysis and monitored for an additional 6 years. RESULTS: Of the 9 high-risk sites (NFE>0.04), 7 sites improved and 2 sites deteriorated. Potential causes for each trend were documented. CONCLUSIONS: The data support the hypothesis that QPD can be used to monitor changes in structural stability after restoration. Knowledge of changes in advance of any symptoms allows further preventive or therapeutic intervention before serious structural damage can occur. Follow-up QPD indications of site improvement can also assure the clinician of the desired structural outcome.

In vivo study of the effectiveness of quantitative percussion diagnostics as an indicator of the level of structural pathology of teeth after restoration.

STATEMENT OF PROBLEM: Conventional diagnostic aids based upon imagery and patient symptoms do not indicate whether restorative treatments have eliminated structural pathology. PURPOSE: The purpose of this clinical study was to evaluate quantitative percussion diagnostics (QPD), a mechanics-based methodology that tests the structural integrity of teeth noninvasively. The study hypothesis was that QPD would provide knowledge of the structural instability of teeth after restorative work. MATERIAL AND METHODS: Eight participants with 60 sites needing restoration were enrolled in an IRB-approved clinical study. Each participant was examined comprehensively, including QPD testing. Each site was disassembled and microscopically video documented, and the results were recorded on a defect assessment sheet. A predictive model was developed for the pathology rating based on normalized fit error (NFE) values using data from the before treatment phase of the study published previously. Each restored site was then tested using QPD. The mean change in NFE values after restoration was evaluated by the pathology rating before treatment. The model was then used to predictively classify the rating after restoration based on the NFE values after treatment. The diagnostic potential of the rating was explored as a marker for risk of pathology after restoration. RESULTS: After restoration, 51 of the 60 sites fell below an NFE of 0.04, representing a greatly stabilized tooth site sample group. Several sites remained in the high-risk category and some increased in pathologic micromovement. Two models were used to determine severity with indicative cutoff points to group sites with similar values. CONCLUSIONS: The data support the hypothesis that QPD can indicate a revised level of structural instability of teeth after restoration.

A Phase I/II Study of Nab-Paclitaxel, Cisplatin, and Cetuximab With Concurrent Radiation Therapy for Locally Advanced Squamous Cell Cancer of the Head and Neck.

Nab-paclitaxel might impact efficacy of radiation for head and neck (H&N) cancer. Nab-paclitaxel, cisplatin, cetuximab, and radiation were evaluated in patients with locally advanced head and neck cancer in this phase I/II trial. Median follow-up was 24 months for 34 patients. The maximum tolerated dose of nab-paclitaxel was 20 mg/m2 with 20 mg/m2 cisplatin and 250 mg/m2 cetuximab. The 2-year progression-free survival (PFS) was 60% (95% confidence interval (CI) 0.42, 0.78), local control 71% (95% CI 0.55, 0.87), and overall survival 68% (95% CI 0.50, 0.86). This is the first study evaluating these agents with radiation in humans, with similar 2-year PFS as historic control.

CD8αα+MHC Class II+ Cell with the Capacity To Terminate Autoimmune Inflammation Is a Novel Antigen-Presenting NK-like Cell in Rats.

Discovery of immune tolerance mechanisms, which inhibit pre-existing autoimmune inflammation, may provide us with new strategies for treating autoimmune diseases. We have identified a CD8αα+MHC class II+ cell with professional APC capacity during our investigation on spontaneous recovery from autoimmune glomerulonephritis in a rat model. This cell actively invades inflamed target tissue and further terminates an ongoing autoimmune inflammation by selective killing of effector autoreactive T cells. In this study, we show that this cell used a cytotoxic machinery of Ly49s+ NK cells in killing of target T cells. Thus, this CD8αα+MHC class II+ cell was a dually functional Ag-presenting NK-like (AP-NK) cell. Following its coupling with target T cells through Ag presentation, killing stimulatory receptor Ly49s6 and coreceptor CD8αα on this cell used rat nonclassic MHC class I C/E16 on the target T cells as a ligand to initiate killing. Thus, activated effector T cells with elevated expression of rat nonclassic MHC class I C/E16 were highly susceptible to the killing by the CD8αα+ AP-NK cell. Granule cytolytic perforin/granzyme C from this cell subsequently mediated cytotoxicity. Thus, inhibition of granzyme C effectively attenuated the killing. As it can recognize and eliminate effector autoreactive T cells in the inflamed target tissue, the CD8αα+ AP-NK cell not only represents a new type of immune cell involved in immune tolerance, but it also is a potential candidate for developing a cell-based therapy for pre-existing autoimmune diseases.

Conjugate-SELEX: A High-throughput Screening of Thioaptamer-liposomal Nanoparticle Conjugates for Targeted Intracellular Delivery of Anticancer Drugs.

Patients with advanced head and neck squamous cell carcinoma receiving chemotherapy have a poor prognosis partly due to normal tissue toxicity; therefore, development of a tumor-targeted drug delivery platform to minimize collateral toxicity is a goal of cancer nanomedicine. Aptamers can achieve this purpose. While conventional Systematic Evolution of Ligands by Exponential Enrichment (SELEX) screens aptamer-only libraries and conjugates them to delivery vehicles after selection, we hypothesized that specific delivery requires screening libraries with aptamer-nanoparticle conjugates. We designed a procedure called, "Conjugate-SELEX", where liposomal nanoparticles (LNP) conjugated with aptamers is screened to identify aptamers that carried attached LNPs to the human head and neck squamous cell carcinoma cell cytosol. Aptamer-LNPs were simultaneously selected for a low affinity to human hepatocytes, minimizing hepatoxicity and LNP clearance. Post-SELEX Next Generation sequencing demonstrated convergence to a family of sequences with one base difference. Affinity pulldown and proteomics analysis identified the uptake-mediating surface receptor as the neuroblast differentiation-associated protein AHNAK (Desmoyokin), a ubiquitous intracellular protein expressed in certain epithelial cell types. Uptake studies with the lead aptamer-conjugates showed enhanced uptake and increased cytotoxicity induced by doxorubicin in cells treated with aptamer-conjugated LNPs over LNP controls. Conjugate-SELEX identifies aptamers capable of targeted cytosolic delivery of attached LNPs payload, while minimizing off-target delivery. The technique lends itself to identification of uptake-mediating surface receptors.

In vivo study of the effectiveness of quantitative percussion diagnostics as an indicator of the level of the structural pathology of teeth.

STATEMENT OF PROBLEM: Conventional dental diagnostic aids based upon imagery and patient symptoms are at best only partially effective for the detection of fine structural defects such as cracks in teeth. PURPOSE: The purpose of this clinical study was to determine whether quantitative percussion diagnostics (QPD) provided knowledge of the structural instability of teeth before restorative work begins. QPD is a mechanics-based methodology that tests the structural integrity of teeth noninvasively. MATERIAL AND METHODS: Eight human participants with 60 sites needing restoration were enrolled in an institutional review board-approved clinical study. Comprehensive examinations were performed in each human participant, including QPD testing. Each site was disassembled and microscopically video documented, and the results were recorded on a defect assessment sheet. Each restored site was then tested using QPD. The normal fit error (NFE), which corresponds to the localized defect severity, was correlated with any pretreatment structural pathology. RESULTS: QPD agreed with clinical disassembly in 55 of 60 comparisons (92% agreement). Moreover, the method achieved 98% specificity and 100% sensitivity for detecting structural pathologies found later upon clinical disassembly. Overall, the NFE was found to be highly predictive of advanced structural pathology. CONCLUSIONS: The data from the present in vivo study support the hypothesis that QPD can provide the clinician with advance knowledge of the structural instability of teeth before restorative work begins.

Differentiating Glomerular Inflammation from Fibrosis in a Bone Marrow Chimera for Rat Anti-Glomerular Basement Membrane Glomerulonephritis.

BACKGROUND: Many types of glomerulonephritis (GN) undergo tandem connected phases: inflammation and fibrosis. Fibrosis in human GNs leads to irreversible end-stage disease. This study investigated how these 2 phases were controlled. METHODS: Using a rat anti-glomerular basement membrane GN model, we established bone marrow (BM) chimeras between GN-resistant Lewis (LEW) and GN-susceptible Wistar Kyoto (WKY) rats. Glomerular inflammation and fibrosis were compared between chimeras. RESULTS: LEW's BM to WKY chimeras with or without co-transfer of host WKY's T cells were GN-resistant. On the other hand, WKY's BM to LEW (LEW(WKY)) chimeras developed glomerular inflammation and albuminuria upon immunization. Quantitative analysis showed that the number and composition of inflammatory cells in glomeruli of immunized LEW(WKY) chimeras were similar to those in immunized WKY rats at their inflammatory peak. Thus, glomerular inflammation was controlled by BM-derived non-T cell populations. However, unlike WKY rats, LEW(WKY) rats did not develop fibrosis until the end of experiments (84 days) in spite of persistent inflammation and albuminuria. CONCLUSION: Inflammation alone was not sufficient to trigger fibrosis, suggesting a critical role of glomerular cells in the fibrotic process. As LEW(WKY) chimera allows us to separate glomerular inflammation from fibrosis, this model provides a useful tool to study how fibrosis is initiated following inflammation.

IL-33 is required for disposal of unnecessary cells during ovarian atresia through regulation of autophagy and macrophage migration.

Physiological processes such as ovarian follicle atresia generate large amounts of unnecessary cells or tissue detritus, which needs to be disposed of rapidly. IL-33 is a member of the IL-1 cytokine gene family. Constitutive expression of IL-33 in a wide range of tissues has hinted at its role beyond immune defense. We have previously reported a close correlation between IL-33 expression patterns and ovarian atresia. In this study, we demonstrated that IL-33 is required for disposal of degenerative tissue during ovarian atresia using Il33(-/-) mice. Deletion of the Il33 gene impaired normal disposal of atretic follicles, resulting in massive accumulations of tissue wastes abundant with aging-related catabolic wastes such as lipofuscin. Accumulation of tissue wastes in Il33(-/-) mice, in turn, accelerated ovarian aging and functional decline. Thus, their reproductive life span was shortened to two thirds of that for Il33(+/-) littermates. IL-33 orchestrated disposal mechanism through regulation of autophagy in degenerating tissues and macrophage migration into the tissues. Our study provides direct evidence supporting an expanded role of IL-33 in tissue integrity and aging through regulating disposal of unnecessary tissues or cells.

Cancer stem cells are enriched in Fanconi anemia head and neck squamous cell carcinomas.

Fanconi anemia (FA) patients have an increased risk of head and neck squamous cell carcinoma (HNSCC) at a higher rate with no apparent risk factors. HNSCC of FA patients is an aggressive tumor characterized by multifocal origin, early metastases and frequent recurrences. Given that cancer stem cells (CSC) drive tumorigenesis, tumor recurrence and metastasis, in this study, we characterized the CSC population in FA and sporadic HNSCC. The Aldefluor assay was used to characterize and isolate CSC with high aldehyde dehydrogenase (ALDH) activity (ALDHpos) in cell lines derived from FA and sporadic HNSCC. Isolated ALDHpos and ALDHneg cells were examined for the expression of stemness genes using reverse transcription-polymerase chain reaction (RT-PCR) array. Tumor cell-derived FA and sporadic HNSCC were examined for their ability to form tumorspheres in vitro. Stem-like cell population in FA and sporadic HNSCC in human and mouse xenograft tumors were evaluated using ALDH isoform 1 (ALDH1) immunohistochemistry. FA­HNSCC cell lines harbor a greater proportion of ALDHpos cells (15-31%) compared to sporadic HNSCC (10%). Expression of Nanog, Oct-3/4 and Stella, molecular markers of undifferentiated embryonic stem (ES) cells were detected in the ALDHpos FA­HNSCC cells and not in the ALDHneg cells. FA­HNSCC cell lines revealed enhanced in vitro tumorsphere formation compared to sporadic HNSCC cells. A higher percentage of ALDH1pos tumor cells are noted in the human and mouse xenograft tumors of FA­HNSCC compared to sporadic HNSCC tumors. FA­HNSCC are highly enriched for CSC and may serve as a model to develop CSC-targeted therapies for HNSCC.

Nonliteral understanding of number words.

One of the most puzzling and important facts about communication is that people do not always mean what they say; speakers often use imprecise, exaggerated, or otherwise literally false descriptions to communicate experiences and attitudes. Here, we focus on the nonliteral interpretation of number words, in particular hyperbole (interpreting unlikely numbers as exaggerated and conveying affect) and pragmatic halo (interpreting round numbers imprecisely). We provide a computational model of number interpretation as social inference regarding the communicative goal, meaning, and affective subtext of an utterance. We show that our model predicts humans' interpretation of number words with high accuracy. Our model is the first to our knowledge to incorporate principles of communication and empirically measured background knowledge to quantitatively predict hyperbolic and pragmatic halo effects in number interpretation. This modeling framework provides a unified approach to nonliteral language understanding more generally.