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Oral squamous cell carcinoma (OSCC) is worldwide health problem associated with high morbidity and mortality. From both the patient and socioeconomic perspectives, prevention of progression of premalignant oral intraepithelial neoplasia (OIN) to OSCC is clearly the preferable outcome. Optimal OSCC chemopreventives possess a variety of attributes including high tolerability, bioavailability, efficacy and preservation of an intact surface epithelium. Terminal differentiation, which directs oral keratinocytes leave the proliferative pool to form protective cornified envelopes, preserves the protective epithelial barrier while concurrently eliminating growth-aberrant keratinocytes. This study employed human premalignant oral keratinocytes and an OSCC cell line to evaluate the differentiation-inducing capacity of the synthetic retinoid, fenretinide (4HPR). Full-thickness oral mucosal explants were evaluated for proof of concept differentiation studies. Results of this study characterize the ability of 4HPR to fulfill all requisite components for keratinocyte differentiation, i.e. nuclear import via binding to cellular RA binding protein-II (molecular modeling), binding to and subsequent activation of retinoic acid nuclear receptors (receptor activation assays), increased expression and translation of genes associated with keratinocyte differentiation [Reverse transcription polymerase chain reaction (RT-PCR), immunoblotting] upregulation of a transglutaminase enzyme essential for cornified envelope formation (transglutaminase 3, functional assay) and augmentation of terminal differentiation in human oral epithelial explants (image-analyses quantified corneocyte desquamation). These data build upon the chemoprevention repertoire of 4HPR that includes function as a small molecule kinase inhibitor and inhibition of essential mechanisms necessary for basement membrane invasion. An upcoming clinical trial, which will assess whether a 4HPR-releasing mucoadhesive patch induces histologic, clinical and molecular regression in OIN lesions, will provide essential clinical insights. [ABSTRACT FROM AUTHOR]

Basement membrane invasion defines malignant transformation of surface premalignancy. Treatment of oral squamous cell carcinoma (OSCC) cells with the synthetic vitamin A derivative, fenretinide (4HPR), induces numerous cancer-preventive effects including suppression of basement membrane invasion, elimination of anchorage-independent growth, disruption of actin cytoskeletal components and inhibition of the invasion-enabling focal adhesive kinase. The purpose of this study was to elucidate 4HPR's effects on additional invasion-relevant mechanisms including matrix metalloproteinase (MMP) activation and function, cell–extracellular matrix (ECM) attachments and interaction with a kinase that is essential for the epithelial–myoepithelial transformation i.e. c-Jun NH2-terminal kinase (JNK). Our data revealed that 4HPR binds with high affinity to the ATP-binding site of all three JNK isoforms with concurrent suppression of kinase function. Additional studies showed 4HPR treatment inhibited both OSCC cell–ECM adhesion and MMP activation and function. JNK downregulation and induced expression studies confirmed that the JNK3 isoform conveyed that largest impact on OSCC migration and invasion. Biodegradable polymeric implants formulated to preserve 4HPR's function and bioavailability were employed to assess 4HPR's chemopreventive impact on an OSCC tumor induction model. These studies revealed 4HPR local delivery significantly inhibited OSCC tumor size, mitotic indices and expression of the endothelial marker, erythroblast transformation-specific-related gene with concurrent increases in tumor apoptosis (cleaved caspase-3). Collectively, these data show that 4HPR suppresses invasion at multiple sites including 'outside-in' signaling, cell–ECM interactions and suppression of MMPs. These functions are also essential for physiologic function. Regulation is therefore essential and reinforces the pharmacologic advantage of local delivery chemopreventive formulations.. [ABSTRACT FROM AUTHOR]

Detecting when the underlying distribution changes for the observed time series is a fundamental problem arising in a broad spectrum of applications. In this paper, we study multiple change-point localization in the high-dimensional regression setting, which is particularly challenging as no direct observations of the parameter of interest is available. Specically, we assume we observe fxt; ytgn t=1 where fxtgn t=1 are p-dimensional covariates, fytgn t=1 are the univariate responses satisfying E(yt) = x> t for 1 tn and ft gn t=1 are the unobserved regression coefficients that change over time in a piecewise constant manner. We propose a novel projection-based algorithm, Variance Projected Wild Binary Segmentation (VPWBS), which transforms the original (difficult) problem of change-point detection in p-dimensional regression to a simpler problem of change-point detection in mean of a one-dimensional time series. VPWBS is shown to achieve sharp localization rate Op(1=n) up to a log factor, a signicant improvement from the best rate Op(1= p n) known in the existing literature for multiple change-point localization in high-dimensional regression. Extensive numerical experiments are conducted to demonstrate the robust and favorable performance of VPWBS over two state-of-the-art algorithms, especially when the size of change in the regression coefficients ft gn t=1 is small. [ABSTRACT FROM AUTHOR]

We investigate sequential change point estimation and detection in univariate nonparametric settings, where a stream of independent observations from sub-Gaussian distributions with a common variance factor and piecewise-constant but otherwise unknown means are collected. We develop a simple CUSUM-based methodology that provably control the probability of false alarms or the average run length while minimizing, in a minimax sense, the detection delay. We allow for all the model parameters to vary in order to capture a broad range of levels of statistical hardness for the problem at hand. We further show how our methodology is applicable to the case in which multiple change points are to be estimated sequentially. [ABSTRACT FROM AUTHOR]

To realize the potential of large molecular weight substances to treat neurological disorders, novel approaches are required to surmount the blood—brain barrier (BBB). We investigated whether fusion of a receptor-binding peptide from apolipoprotein E (apoE) with a potentially therapeutic protein can bind to LDL receptors on the BBB and be transcytosed into the CNS. A lysosomal enzyme, α-L-iduronidase (IDUA), was used for biological and therapeutic evaluation in a mouse model of mucopolysaccharidosis (MPS) type I, one of the most common lysosomal storage disorders with CNS deficits. We identified two fusion candidates, IDUAe1 and IDUAe2, by in vitro screening, that exhibited desirable receptor-mediated binding, endocytosis, and transendothelial transport as well as appropriate lysosomal enzyme trafficking and biological function. Robust peripheral IDUAe1 or IDUAe2 generated by transient hepatic expression led to elevated enzyme levels in capillary-depleted, enzyme-deficient brain tissues and protein delivery into nonendothelium perivascular cells, neurons, and astrocytes within 2 d of treatment. Moreover, 5 mo after long-term delivery of moderate levels of IDUAe1 derived from maturing red blood cells, 2% to 3% of normal brain IDUA activities were obtained in MPS I mice, and IDUAe1 protein was detected in neurons and astrocytes throughout the brain. The therapeutic potential was demonstrated by normalization of brain glycosaminoglycan and β-hexosaminidase in MPS I mice 5 mo after moderate yet sustained delivery of IDUAe1. These findings provide a noninvasive and BBB-targeted procedure for the delivery of large-molecule therapeutic agents to treat neurological lysosomal storage disorders and potentially other diseases that involve the brain. [ABSTRACT FROM AUTHOR]

We study the multivariate nonparametric change point detection problem, where the data are a sequence of independent $p$ -dimensional random vectors whose distributions are piecewise-constant with Lipschitz densities changing at unknown times, called change points. We quantify the size of the distributional change at any change point with the supremum norm of the difference between the corresponding densities. We are concerned with the localization task of estimating the positions of the change points. In our analysis, we allow for the model parameters to vary with the total number of time points, including the minimal spacing between consecutive change points and the magnitude of the smallest distributional change. We provide information-theoretic lower bounds on both the localization rate and the minimal signal-to-noise ratio required to guarantee consistent localization. We formulate a novel algorithm based on kernel density estimation that nearly achieves the minimax lower bound, save possibly for logarithm factors. We have provided extensive numerical evidence to support our theoretical findings. [ABSTRACT FROM AUTHOR]

The maturity of fruits and vegetables such as tomatoes significantly impacts indicators of their quality, such as taste, nutritional value, and shelf life, making maturity determination vital in agricultural production and the food processing industry. Tomatoes mature from the inside out, leading to an uneven ripening process inside and outside, and these situations make it very challenging to judge their maturity with the help of a single modality. In this paper, we propose a deep learning-assisted multimodal data fusion technique combining color imaging, spectroscopy, and haptic sensing for the maturity assessment of tomatoes. The method uses feature fusion to integrate feature information from images, near-infrared spectra, and haptic modalities into a unified feature set and then classifies the maturity of tomatoes through deep learning. Each modality independently extracts features, capturing the tomatoes' exterior color from color images, internal and surface spectral features linked to chemical compositions in the visible and near-infrared spectra (350 nm to 1100 nm), and physical firmness using haptic sensing. By combining preprocessed and extracted features from multiple modalities, data fusion creates a comprehensive representation of information from all three modalities using an eigenvector in an eigenspace suitable for tomato maturity assessment. Then, a fully connected neural network is constructed to process these fused data. This neural network model achieves 99.4% accuracy in tomato maturity classification, surpassing single-modal methods (color imaging: 94.2%; spectroscopy: 87.8%; haptics: 87.2%). For internal and external maturity unevenness, the classification accuracy reaches 94.4%, demonstrating effective results. A comparative analysis of performance between multimodal fusion and single-modal methods validates the stability and applicability of the multimodal fusion technique. These findings demonstrate the key benefits of multimodal fusion in terms of improving the accuracy of tomato ripening classification and provide a strong theoretical and practical basis for applying multimodal fusion technology to classify the quality and maturity of other fruits and vegetables. Utilizing deep learning (a fully connected neural network) for processing multimodal data provides a new and efficient non-destructive approach for the massive classification of agricultural and food products. [ABSTRACT FROM AUTHOR]

Accurate data acquisition from flexible sensors placed on deformable 3D freeform surfaces is of critical importance for many applications, such as wearable electronics, human‐machine interfaces, and soft robotics. However, the mechanical coupling between the sensor and the deformable subject surface to bending and stretching deformations can significantly reduce the accuracy of the acquired data. This study combines a polyimide (PI) micropore isolation layer (PIL) and serpentine electrodes with a flexible piezoresistive sensor to mitigate the issue of mechanical coupling. As a mechanical buffer to distribute the external pressure and reduce the strain concentration, the PIL can avoid the bending interference for bending curvature up to 256 m−1, while maintaining a high sensitivity of S > 21.5 kPa−1. The serpentine electrode design further allows the sensor to reliably acquire data in the presence of stretching up to 45% without cross‐talk. The versatility of the developed sensor is demonstrated in several human‐machine interaction scenarios, including gesture recognition and motion detection. The design strategies on materials and structures from this study can also be applied for the development of other flexible sensors with high sensitivity and low deformation interference to avoid the mechanical coupling between the sensor and the deformable surface. [ABSTRACT FROM AUTHOR]

Background: An emerging metabolic theory of pulmonary hypertension (PH) suggests that cellular and mitochondrial metabolic dysfunction underlies the pathology of this disease. We and others have previously demonstrated the existence of hyperproliferative, apoptosis-resistant, proinflammatory adventitial fibroblasts from human and bovine hypertensive pulmonary arterial walls (PH-Fibs) that exhibit constitutive reprogramming of glycolytic and mitochondrial metabolism, accompanied by an increased ratio of glucose catabolism through glycolysis versus the tricarboxylic acid cycle. However, the mechanisms responsible for these metabolic alterations in PH-Fibs remain unknown. We hypothesized that in PH-Fibs microRNA-124 (miR-124) regulates PTBP1 (polypyrimidine tract binding protein 1) expression to control alternative splicing of pyruvate kinase muscle (PKM) isoforms 1 and 2, resulting in an increased PKM2/PKM1 ratio, which promotes glycolysis and proliferation even in aerobic environments.Methods: Pulmonary adventitial fibroblasts were isolated from calves and humans with severe PH (PH-Fibs) and from normal subjects. PTBP1 gene knockdown was achieved via PTBP1-siRNA; restoration of miR-124 was performed with miR-124 mimic. TEPP-46 and shikonin were used to manipulate PKM2 glycolytic function. Histone deacetylase inhibitors were used to treat cells. Metabolic products were determined by mass spectrometry-based metabolomics analyses, and mitochondrial function was analyzed by confocal microscopy and spectrofluorometry.Results: We detected an increased PKM2/PKM1 ratio in PH-Fibs compared with normal subjects. PKM2 inhibition reversed the glycolytic status of PH-Fibs, decreased their cell proliferation, and attenuated macrophage interleukin-1β expression. Furthermore, normalizing the PKM2/PKM1 ratio in PH-Fibs by miR-124 overexpression or PTBP1 knockdown reversed the glycolytic phenotype (decreased the production of glycolytic intermediates and byproducts, ie, lactate), rescued mitochondrial reprogramming, and decreased cell proliferation. Pharmacological manipulation of PKM2 activity with TEPP-46 and shikonin or treatment with histone deacetylase inhibitors produced similar results.Conclusions: In PH, miR-124, through the alternative splicing factor PTBP1, regulates the PKM2/PKM1 ratio, the overall metabolic, proliferative, and inflammatory state of cells. This PH phenotype can be rescued with interventions at various levels of the metabolic cascade. These findings suggest a more integrated view of vascular cell metabolism, which may open unique therapeutic prospects in targeting the dynamic glycolytic and mitochondrial interactions and between mesenchymal inflammatory cells in PH. [ABSTRACT FROM AUTHOR]

Wireless passive sensors based on inductor-capacitor (LC) circuits have been an area of intense research in wearable electronics because of their battery-free and wirelessly connected operation. However, significant challenges remain in achieving high sensitivity and long-range performance for such device. In this study, we present a bilayer coils LC wireless passive iontronic pressure sensor (BWIPS) that overcomes these limitations. The bilayer coils strategy enhances the coupling effect and extends the operating distance up to 20 mm. Meanwhile, the capacitive iontronic pressure sensor (IPS) achieves high sensitivity (0.54 kPa−1) and a broad sensing range (400 kPa). By integrating bilayer coils and IPS, the BWIPS exhibits a remarkable sensitivity of −37.4 kHz kPa−1. Furthermore, the flexibility of the sensor allows it to maintain its resonant frequency regardless of various bending radius, making it suitable for use in prosthetic devices. Our BWIPS technology holds great potential for advancing the development of wireless passive sensors in the future, particularly in applications that require high sensitivity, broad sensing range, and non-contact detection at a distance. [ABSTRACT FROM AUTHOR]

Local, long-acting release fenretinide (4HPR) millicylindrical implants were prepared and evaluated for their release kinetics in vivo and their ability to suppress oral cancer tumor explant growth. Poly(lactic- co -glycolic acid)(PLGA) implants were prepared as a function of drug loading and the presence of various excipients (pore-formers, solubilizers, crystallization inhibitors) to enhance release of the insoluble 4HPR. Release kinetics and bioerosion of PLGA were monitored both in vitro in a PBS/Tween 80 buffer and in vivo by recovery of the drug remaining at the injection site. 4HPR was released from PLGA implants much slower in vivo than in the drug solubilizing media in vitro , with a 3-week lag phase and continuous release of >2 months, but showed some release enhancement by addition of solubilizers. Water-soluble PVA/sucrose implants for release of 4HPR served to determine if drug dissolution provided suitable controlled release without the PLGA, and this formulation showed continuous drug release over 6 weeks in vivo. Placement of PLGA-4HPR implants adjacent to oral cancer tumor murine xenografts showed inhibition of tumor growth relative to sham implants, indicating the potential for the local 4HPR delivery approach to be useful for oral cancer chemoprevention. [ABSTRACT FROM AUTHOR]

Introduction: Oral squamous cell carcinoma (OSCC), is associated with high morbidity and mortality. Preemptive interventions have been postulated to provide superior therapeutic options, but their implementation has been restricted by the availability of broadly applicable local delivery systems. Methods: We address this challenge by engineering a delivery vehicle, Janus nanoparticles (JNP), that combine the dual mucoadhesive properties of a first cationic chitosan compartment with a second hydrophobic poly(lactide-co-glycolide) release compartment. JNP are designed to avoid rapid mucus clearance while ensuring stable loading and controlled release of the IL-6 receptor antagonist, tocilizumab (TCZ). Results: The JNP featured defined and monodispersed sizes with an average diameter of 327 nm and a PDI of 0.245, high circularities above 0.90 and supported controlled release of TCZ and effective internalization by oral keratinocytes. TCZ released from JNP retained its biological activity and effectively reduced both, soluble and membrane-bound IL-6Rα (71% and 50%). In full-thickness oral mucosal explants, 76% of the JNP breached the stratum corneum and in 41% were observed in the basal cell layer indicating excellent mucopenetrating properties. When tested in an aggressive OSCC xenograft model, TCZ-loaded JNP showed high levels of xenograft inhibition and outperformed all control groups with respect to inhibition of tumor cell proliferation, reduction in tumor size and reduced expression of the proto-oncogene ERG. Conclusion: By combining critically required, yet orthogonal properties within the same nanoparticle design, the JNP in this study, demonstrate promise as precision delivery platforms for intraoral field-coverage chemoprevention, a vastly under-researched area of high clinical importance. [ABSTRACT FROM AUTHOR]

Stable finite element schemes are developed for the solution of the equations modeling the flow of viscoelastic fluids. In contrast with classical statements of these equations, which introduce the stress as a primary variable, these schemes explicitly involve the deformation tensor and elastic energy. Energy estimates and existence of solutions to the discrete problem are established for schemes of arbitrary order without any restrictions on the time step, mesh size, or Weissenberg number. Convergence to smooth solutions is established for the classical Oldroyd–B fluid. Numerical experiments for two classical benchmark problems verify the robustness of this approach. [ABSTRACT FROM AUTHOR]

In situ stem cell gene transfer, mediated by intra-bone marrow (iBM) injection of a lentiviral vector (LV), presents a novel approach that could potentially take full advantage of any stem cells in bone cavity and overcome many difficulties encountered in ex vivo gene transfer. Our recent studies have demonstrated significant gene transfer into hematopoietic stem cells (HSC) and mesenchymal stem/ progenitor cells (MSC) after intrafemoral injection of LV in adult mice without any pre-conditioning. In this study, we sought to determine if HSC and MSC gene transfer can be enhanced by pretreatment with 5-fluorouracil (5-FU) in combination with in vivo selection. An advanced 3rd-generation LV was constructed expressing a variant of O6-methylguanine DNA methyltransferase (MGMTP140K) and GFP under regulation of elongation factor 1- alpha promoter. VSVG-pseudotyped vector (7 × 107 TU/ml) was injected into femurs of adult C57/Bl6 mice that were pretreated with a sublethal dose of 5-FU either 2- (n=6) or 5- (n=6) days before injection, in comparison with non-conditioning mice (n=7) or buffer-injected controls (n=6). Two days post injection, BM of femurs from injected side of some mice (2-3 per group) were harvested and transplanted into lethally irradiated mice. Only one (out of 4) BMT recipient derived from 2-D treatment group demonstrated significant level of GFP+ PBL (9.7%) two months post transplantation, although successful engraftment was indicated in recipients from all LV-injected groups (n=13). To monitor gene transfer in progenitor cells, CFC assay was also performed using BM of femurs from the short-term mice. Similar levels of GFPcontaining colonies were observed in both 2-D pretreated (mean of 8.1%) and un-conditioned (mean of 8.2%) mice, while less GFP colonies (mean of 3.6%) were detected in 5-D pretreated mice. To evaluate stem cell gene transfer and monitor potential hematologic toxicity, we performed complete blood count and FACS analysis periodically in primary injected mice. All blood parameters were normal except that mild monocytosis and leukocytosis were detected in most of LV-injected (8 of 12) and all buffer-injected (3 of 3) mice up to two month post injection, suggesting acute inflammation related to iBM injection procedure. Four-weeks post injection, various levels of GFP-expressing cells were detected in lymphoid and myeloid subpopulations of all 2D-pretreated (mean of 1.9% and 2.5% respectively), all 5D-pretreated (mean of 1% and 0.5%) and two (out of 4) un-conditioned mice. However, 9-weeks after injection, the highest GFP labeling was observed in one of the un-conditioned mice (7% and 11.7%). Further studies are underway to charaterize gene transfer in HSC and MSC, and enhance transgene frequency by MGMTP140K-mediated in vivo selection. Our preliminary data suggested that timing of preconditioning is critical for improving LV-mediated in vivo gene transfer. This study will provide valuable information to optimize in situ BM stem cell gene transfer which might lead toward a novel approach for treatment of human diseases.Molecular Therapy (2006) 13, S40–S40; doi: 10.1016/j.ymthe.2006.08.116 [ABSTRACT FROM AUTHOR]

Stiffening of large arteries is increasingly used as an independent predictor of risk and therapeutic outcome for small artery dysfunction in many diseases including pulmonary hypertension. The molecular mechanisms mediating downstream vascular cell responses to large artery stiffening remain unclear. We hypothesize that high pulsatility flow, induced by large artery stiffening, causes inflammatory responses in downstream pulmonary artery endothelial cells (PAECs) through toll-like receptor (TLR) pathways. To recapitulate the stiffening effect of large pulmonary arteries that occurs in pulmonary hypertension, ultrathin silicone tubes of variable mechanical stiffness were formulated and were placed in a flow circulatory system. These tubes modulated the simulated cardiac output into pulsatile flows with different pulsatility indices, 0.5 (normal) or 1.5 (high). PAECs placed downstream of the tubes were evaluated for their expression of proinflammatory molecules (ICAM-1, VCAM-1, E-selectin and MCP-1), TLR receptors and intracellular NF-κB following flow exposure. Results showed that compared to flow with normal pulsatility, high pulsatility flow induced proinflammatory responses in PAECs, enhanced TLR2 expression but not TLR4, and caused NF-κB activation. Pharmacologic (OxPAPC) and siRNA inhibition of TLR2 attenuated high pulsatility flow-induced pro-inflammatory responses and NF-κB activation in PAECs. We also observed that PAECs isolated from small pulmonary arteries of hypertensive animals exhibiting proximal vascular stiffening demonstrated a durable ex-vivo proinflammatory phenotype (increased TLR2, TLR4 and MCP-1 expression). Intralobar PAECs isolated from vessels of IPAH patients also showed increased TLR2. In conclusion, this study demonstrates for the first time that TLR2/NF-κB signaling mediates endothelial inflammation under high pulsatility flow caused by upstream stiffening, but the role of TLR4 in flow pulsatility-mediated endothelial mechanotransduction remains unclear. [ABSTRACT FROM AUTHOR]

The K-Cl cotransporter (KCC) regulates red blood cell (RBC) volume, especially in reticulocytes. Western blot analysis of RBC membranes revealed KCC1, KCC3, and KCC4 proteins in mouse and human cells, with higher levels in reticulocytes. KCC content was higher in sickle versus normal RBC, but the correlation with reticulocyte count was poor, with inter-individual variability in KCC isoform ratios. Messenger RNA for each isoform was measured by real time RT-quantitative PCR. In human reticulocytes, KCC3a mRNA levels were consistently the highest, 1-7-fold higher than KCC4, the second most abundant species. Message levels for KCC1 and KCC3b were low. The ratios of KCC RNA levels varied among individuals but were similar in sickle and normal RBC. During in vivo maturation of human erythroblasts, KCC3a RNA was expressed consistently, whereas KCC1 and KCC3b levels declined, and KCC4 message first increased and then decreased. In mouse erythroblasts, a similar pattern for KCC3 and KCC1 expression during in vivo differentiation was observed, with low KCC4 RNA throughout despite the presence of KCC4 protein in mature RBC. During differentiation of mouse erythroleukemia cells, protein levels of KCCs paralleled increasing mRNA levels. Functional properties of KCCs expressed in HEK293 cells were similar to each other and to those in human RBC. However, the anion dependence of KCC in RBC resembled most closely that of KCC3. The results suggest that KCC3 is the dominant isoform in erythrocytes, with variable expression of KCC1 and KCC4 among individuals that could result in modulation of KCC activity. [ABSTRACT FROM AUTHOR]

Background: Epigenetic programming, dynamically regulated by histone acetylation, is a key mechanism regulating cell proliferation and survival. Little is known about the contribution of histone deacetylase (HDAC) activity to the development of pulmonary arterial hypertension, a condition characterized by profound structural remodeling of pulmonary arteries and arterioles.Methods and Results: HDAC1 and HDAC5 protein levels were elevated in lungs from human idiopathic pulmonary arterial hypertension and in lungs and right ventricles from rats exposed to hypoxia. Immunohistochemistry localized increased expression to remodeled vessels in the lung. Both valproic acid, a class I HDAC inhibitor, and suberoylanilide hydroxamic acid (vorinostat), an inhibitor of class I, II, and IV HDACs, mitigated the development of and reduced established hypoxia-induced pulmonary hypertension in the rat. Both valproic acid and suberoylanilide hydroxamic acid inhibited the imprinted highly proliferative phenotype of fibroblasts and R-cells from pulmonary hypertensive bovine vessels and platelet-derived growth factor-stimulated growth of human vascular smooth muscle cells in culture. Exposure to valproic acid and suberoylanilide hydroxamic acid was associated with increased levels of p21 and FOXO3 and reduced expression of survivin. The significantly higher levels of expression of cKIT, monocyte chemoattractant protein-1, interleukin-6, stromal-derived factor-1, platelet-derived growth factor-b, and S100A4 in R-cells were downregulated by valproic acid and suberoylanilide hydroxamic acid treatment.Conclusions: Increased HDAC activity contributes to the vascular pathology of pulmonary hypertension. The effectiveness of HDAC inhibitors, valproic acid, and suberoylanilide hydroxamic acid, in models of pulmonary arterial hypertension supports a therapeutic strategy based on HDAC inhibition in pulmonary arterial hypertension. [ABSTRACT FROM AUTHOR]

Background--Epigenetic programming, dynamically regulated by histone acetylation, is a key mechanism regulating cell proliferation and survival. Little is known about the contribution of histone deacetylase (HDAC) activity to the development of pulmonary arterial hypertension, a condition characterized by profound structural remodeling of pulmonary arteries and arterioles. Methods and Results--HDACl and HDAC5 protein levels were elevated in lungs from human idiopathic pulmonary arterial hypertension and in lungs and right ventricles from rats exposed to hypoxia. Immunohistochemistry localized increased expression to remodeled vessels in the lung. Both valproic acid, a class I HDAC inhibitor, and suberoylanilide hydroxamic acid (vorinostat), an inhibitor of class I, II, and IV HDACs, mitigated the development of and reduced established hypoxia-induced pulmonary hypertension in the rat. Both valproic acid and suberoylanilide hydroxamic acid inhibited the imprinted highly proliferative phenotype of fibroblasts and R-cells from pulmonary hypertensive bovine vessels and platelet-derived growth factor-stimulated growth of human vascular smooth muscle cells in culture. Exposure to valproic acid and suberoylanilide hydroxamic acid was associated with increased levels of p21 and FOX03 and reduced expression of survivin. The significantly higher levels of expression of cKIT, monocyte chemoattractant protein-1, interleukin-6, stromal-derived factor-1, platelet-derived growth factor-b, and S100A4 in R-cells were downregulated by valproic acid and suberoylanilide hydroxamic acid treatment. Conclusions--Increased HDAC activity contributes to the vascular pathology of pulmonary hypertension. The effectiveness of HDAC inhibitors, valproic acid, and suberoylanilide hydroxamic acid, in models of pulmonary arterial hypertension supports a therapeutic strategy based on HDAC inhibition in pulmonary arterial hypertension. [ABSTRACT FROM AUTHOR]

Summary: Detection of DNA methylation has produced promising results as biomarkers for head and neck squamous cell carcinoma (HNSCC). However, current panels are limited by an insufficient number of sensitive and specific tumor markers. MicroRNAs (miR) play an important role in tumorigenesis, and may represent a novel panel of molecules for the development of cancer biomarkers. We investigated methylation of three miRNA promoter sites of miR-9 (miR-9-1, miR-9-2, miR-9-3) in 107 human head and neck tissue samples and controls. We found methylations of miR-9-1 and miR-9-3 were higher in oral and oropharyngeal carcinomas than that in laryngeal carcinoma, achieving a combined sensitivity of 63% and 56%, respectively, for these two tumor types, compared to 21% for the laryngeal carcinoma. Quantitative PCR of miR-9 showed reduced expression associated with methylation of miR-9 in tumor tissues. To investigate the functional consequences of miR-9 methylation, we found that miR-9 methylation is correlated with miR-9 expression level in human HNSCC cell lines. Demethylation treatment using 5-aza-deoxycytidine restored its expression in a miR-9 methylated human HNSCC cell line UM-SCC22A. Furthermore, cell proliferation and viability was significantly inhibited, while PTEN expression was elevated after transfection of miR-9 into the UM-SCC22A cell line. In summary, our results suggest that methylations of miR-9-1 and miR-9-3 are sensitive and specific biomarkers for HNSCC, particularly for oral and oropharyngeal squamous cell carcinomas. In addition, miR-9 may function as a tumor suppressor in HNSCC through inhibition of cell proliferation and elevation of tumor suppressor PTEN. [ABSTRACT FROM AUTHOR]