An Ensemble Learning Method for Detection of Head and Neck Squamous Cell Carcinoma Using Polarized Hyperspectral Microscopic Imaging.

Head and neck squamous cell carcinoma (HNSCC) has a high mortality rate. In this study, we developed a Stokes-vector-derived polarized hyperspectral imaging (PHSI) system for H&E-stained pathological slides with HNSCC and built a dataset to develop a deep learning classification method based on convolutional neural networks (CNN). We use our polarized hyperspectral microscope to collect the four Stokes parameter hypercubes (S0, S1, S2, and S3) from 56 patients and synthesize pseudo-RGB images using a transformation function that approximates the human eye's spectral response to visual stimuli. Each image is divided into patches. Data augmentation is applied using rotations and flipping. We create a four-branch model architecture where each branch is trained on one Stokes parameter individually, then we freeze the branches and fine-tune the top layers of our model to generate final predictions. Our results show high accuracy, sensitivity, and specificity, indicating that our model performed well on our dataset. Future works can improve upon these results by training on more varied data, classifying tumors based on their grade, and introducing more recent architectural techniques.

Polarized Hyperspectral Microscopic Imaging for Zebrafish.

Zebrafish is a well-established animal model for developmental and disease studies. Its optical transparency at early developmental stages is ideal for tissue visualization. Interaction of light with zebrafish tissues provides information on their structure and properties. In this study, we developed a microscopic imaging system for improving the visualization of unstained zebrafish tissues on tissue slides, with two different setups: polarized light imaging and polarized hyperspectral imaging. Based on the polarized light imaging setup, we collected the RGB images of Stokes vector parameters (S0, S1, S2, and S3), and calculated the Stokes vector derived parameters: the degree of polarization (DOP), the degree of linear polarization (DOLP)). We also calculated Stokes vector data based on the polarized hyperspectral imaging setup. The preliminary results demonstrate that Stokes vector data in two imaging setups (polarized light imaging and polarized hyperspectral imaging) are capable of improving the visualization of different types of zebrafish tissues (brain, muscle, skin cells, blood vessels, and yolk). Using the images collected from larval zebrafish samples by polarized light imaging, we found that DOP and DOLP could show clearer structural information of the brain and of skin cells, muscle and blood vessels in the tail. Furthermore, DOP and DOLP parameters derived from images collected by polarized hyperspectral imaging could show clearer structural information of skin cells developing around yolk as well as the surrounding blood vessel network. In addition, polarized hyperspectral imaging could provide complementary spectral information to the spatial information on Stokes vector data of zebrafish tissues. The polarized light imaging & polarized hyperspectral imaging systems provide a better insight into the microstructures of zebrafish tissues.

Polarized hyperspectral microscopic imaging system for enhancing the visualization of collagen fibers and head and neck squamous cell carcinoma.

Significance: Polarized hyperspectral microscopes with the capability of full Stokes vector imaging have potential for many biological and medical applications. Aim: The aim of this study is to investigate polarized hyperspectral imaging (PHSI) for improving the visualization of collagen fibers, which is an important biomarker related to tumor development, and improving the differentiation of normal and tumor cells on pathologic slides. Approach: We customized a polarized hyperspectral microscopic imaging system comprising an upright microscope with a motorized stage, two linear polarizers, two liquid crystal variable retarders (LCVRs), and a compact SnapScan hyperspectral camera. The polarizers and LCVRs worked in tandem with the hyperspectral camera to acquire polarized hyperspectral images, which were further used to calculate four Stokes vectors: S0, S1, S2, and S3. Synthetic RGB images of the Stokes vectors were generated for the visualization of cellular components in PHSI images. Regions of interest of collagen, normal cells, and tumor cells in the synthetic RGB images were selected, and spectral signatures of the selected components were extracted for comparison. Specifically, we qualitatively and quantitatively investigated the enhanced visualization and spectral characteristics of dense fibers and sparse fibers in normal stroma tissue, fibers accumulated within tumors, and fibers accumulated around tumors. Results: By employing our customized polarized hyperspectral microscope, we extract the spectral signatures of Stokes vector parameters of collagen as well as of tumor and normal cells. The measurement of Stokes vector parameters increased the image contrast of collagen fibers and cells in the slides. Conclusions: With the spatial and spectral information from the Stokes vector data cubes (S0, S1, S2, and S3), our PHSI microscope system enhances the visualization of tumor cells and tumor microenvironment components, thus being beneficial for pathology and oncology.

Automated Polarized Hyperspectral Imaging (PHSI) for ex-vivo and in-vivo Tissue Assessment.

Polarized light interactions with biological tissues can reveal information regarding tissue structure, while spectral characteristics are closely related to tissue composition. An integration of both modalities in a compact system could better assist tissue assessment. This study aims to develop a polarized hyperspectral imaging (PHSI) system that fulfills both linearly and circularly polarized hyperspectral imaging for in vivo and ex vivo applications. The system is comprised of a white LED, two linear polarizers, two liquid crystal variable retarders (LCVRs), and a hyperspectral snapshot camera. The system was calibrated to compute the full Stokes polarimetry. For tissue differentiation, fresh ex vivo mouse tissue specimens from kidney, liver, spleen, muscle, lung, and salivary gland of mice were imaged. The spectra of three features, named degree of polarization (DOP), degree of linear polarization (DOLP), and degree of circular polarization (DOCP), were generated. A k-nearest neighbor (k-NN) classifier was trained with multi-class spectra and 5-fold cross validation. It was found that DOP better differentiates tissue with an average accuracy of 0.87. Additionally, support vector machine (SVM) classifiers were trained to differentiate between each two of the organs, and it was determined that DOLP better identified kidney, liver, and spleen, whereas DOCP better identified muscle and lung tissues. Then, the setup was employed to image in vivo human fingers with and without a blood occlusion to qualitatively estimate oxygen saturation. Preliminary results demonstrate that both DOLP and DOCP reveal a distinction of oxygen saturation states. These results demonstrate the feasibility of the PHSI system for distinguishing between optical properties of tissues, which has the potential to reveal disease-related information for diverse medical applications.

Polarized Hyperspectral Microscopic Imaging for White Blood Cells on Wright-Stained Blood Smear Slides.

White blood cells, also called leukocytes, are hematopoietic cells of the immune system that are involved in protecting the body against both infectious diseases and foreign materials. The abnormal development and uncontrolled proliferation of these cells can lead to devastating cancers. Their timely recognition in the peripheral blood is critical to diagnosis and treatment. In this study, we developed a microscopic imaging system for improving the visualization of white blood cells on Wright's stained blood smear slides, with two different setups: polarized light imaging and polarized hyperspectral imaging. Based on the polarized light imaging setup, we collected the RGB images of Stokes vector parameters (S0, S1, S2, and S3) of five types of white blood cells (neutrophil, eosinophil, basophil, lymphocyte, and monocyte), and calculated the Stokes vector derived parameters: the degree of polarization (DOP), the degree of linear polarization (DOLP), and the degree of circular polarization (DOCP)). We also calculated Stokes vector data based on the polarized hyperspectral imaging setup. The preliminary results demonstrate that Stokes vector derived parameters (DOP, DOLP, and DOCP) could improve the visualization of granules in granulocytes (neutrophils, eosinophils, and basophils). Furthermore, Stokes vector derived parameters (DOP, DOLP, and DOCP) could improve the visualization of surface structures (protein patterns) of lymphocytes enabling subclassification of lymphocyte subpopulations. Finally, S2, S3, and DOCP could enhance the morphologic visualization of monocyte nucleus. We also demonstrated that the polarized hyperspectral imaging setup could provide complementary spectral information to the spatial information on different Stokes vector parameters of white blood cells. This work demonstrates that polarized light imaging & polarized hyperspectral imaging has the potential to become a strong imaging tool in the diagnosis of disorders arising from white blood cells.

Polarized hyperspectral microscopic imaging for collagen visualization on pathologic slides of head and neck squamous cell carcinoma.

We developed a polarized hyperspectral microscope to collect four types of Stokes vector data cubes (S0, S1, S2, and S3) of the pathologic slides with head and neck squamous cell carcinoma (HNSCC). Our system consists of an optical light microscope with a movable stage, two polarizers, two liquid crystal variable retarders (LCVRs), and a SnapScan hyperspectral camera. The polarizers and LCVRs work in tandem with the hyperspectral camera to acquire polarized hyperspectral images. Synthetic pseudo-RGB images are generated from the four Stokes vector data cubes based on a transformation function similar to the spectral response of human eye for the visualization of hyperspectral images. Collagen is the most abundant extracellular matrix (ECM) protein in the human body. A major focus of studying the ECM in tumor microenvironment is the role of collagen in both normal and abnormal function. Collagen tends to accumulate in and around tumors during cancer development and growth. In this study, we acquired images from normal regions containing normal cells and collagen fibers and from tumor regions containing cancerous squamous cells and collagen fibers on HNSCC pathologic slides. The preliminary results demonstrated that our customized polarized hyperspectral microscope is able to improve the visualization of collagen on HNSCC pathologic slides under different situations, including thick fibers of normal stroma, thin fibers of normal stroma, fibers of normal muscle cells, fibers accumulated in tumors, fibers accumulated around tumors. Our preliminary results also demonstrated that the customized polarized hyperspectral microscope is capable of extracting the spectral signatures of collagen based on Stokes vector parameters and can have various applications in pathology and oncology.

Automatic detection of head and neck squamous cell carcinoma on pathologic slides using polarized hyperspectral imaging and deep learning.

The study is to incorporate polarized hyperspectral imaging (PHSI) with deep learning for automatic detection of head and neck squamous cell carcinoma (SCC) on hematoxylin and eosin (H&E) stained tissue slides. A polarized hyperspectral imaging microscope had been developed in our group. In this paper, we firstly collected the Stokes vector data cubes (S0, S1, S2, and S3) of histologic slides from 17 patients with SCC by the PHSI microscope, under the wavelength range from 467 nm to 750 nm. Secondly, we generated the synthetic RGB images from the original Stokes vector data cubes. Thirdly, we cropped the synthetic RGB images into image patches at the image size of 96×96 pixels, and then set up a ResNet50-based convolutional neural network (CNN) to classify the image patches of the four Stokes vector parameters (S0, S1, S2, and S3) by application of transfer learning. To test the performances of the model, each time we trained the model based on the image patches (S0, S1, S2, and S3) of 16 patients out of 17 patients, and used the trained model to calculate the testing accuracy based on the image patches of the rest 1 patient (S0, S1, S2, and S3). We repeated the process for 6 times and obtained 24 testing accuracies (S0, S1, S2, and S3) from 6 different patients out of the 17 patients. The preliminary results showed that the average testing accuracy (84.2%) on S3 outperformed the average testing accuracy (83.5%) on S0. Furthermore, 4 of 6 testing accuracies of S3 (96.0%, 87.3%, 82.8%, and 86.7%) outperformed the testing accuracies of S0 (93.3%, 85.2%, 80.2%, and 79.0%). The study demonstrated the potential of using polarized hyperspectral imaging and deep learning for automatic detection of head and neck SCC on pathologic slides.

Automatic detection of head and neck squamous cell carcinoma on pathologic slides using polarized hyperspectral imaging and machine learning.

The aim of this study is to incorporate polarized hyperspectral imaging (PHSI) with machine learning for automatic detection of head and neck squamous cell carcinoma (SCC) on hematoxylin and eosin (H&E) stained tissue slides. A polarized hyperspectral imaging microscope had been developed in our group. In this paper, we imaged 20 H&E stained tissue slides from 10 patients with SCC of the larynx by the PHSI microscope. Several machine learning algorithms, including support vector machine (SVM), random forest, Gaussian naive Bayes, and logistic regression, were applied to the collected image data for the automatic detection of SCC on the H&E stained tissue slides. The performance of these methods was compared among the collected PHSI data, the pseudo-RGB images generated from the PHSI data, and the PHSI data after applying the principal component analysis (PCA) transformation. The results suggest that SVM is a superior classifier for the classification task based on the PHSI data cubes compared to the other three classifiers. The incorporate of four Stokes vector parameters improved the classification accuracy. Finally, the PCA transformed image data did not improve the accuracy as it might lose some important information from the original PHSI data. The preliminary results show that polarized hyperspectral imaging can have many potential applications in digital pathology.

Hyperspectral Microscopic Imaging for the Detection of Head and Neck Squamous Cell Carcinoma on Histologic Slides.

The purpose of this study is to investigate hyperspectral microscopic imaging and deep learning methods for automatic detection of head and neck squamous cell carcinoma (SCC) on histologic slides. Hyperspectral imaging (HSI) cubes were acquired from pathologic slides of 18 patients with SCC of the larynx, hypopharynx, and buccal mucosa. An Inception-based two-dimensional convolutional neural network (CNN) was trained and validated for the HSI data. The automatic deep learning method was tested with independent data of human patients. This study demonstrated the feasibility of using hyperspectral microscopic imaging and deep learning classification to aid pathologists in detecting SCC on histologic slides.

Development of a polarized hyperspectral microscope for cardiac fiber orientation imaging.

Myocardial fiber orientation is closely related to the functions of the heart. The development of imaging tools for depicting myocardial fiber orientation is important. We developed a polarized hyperspectral imaging microscope (PHSIM) for cardiac fiber orientation imaging, which is capable of polarimetric imaging and hyperspectral imaging. Polarimetric imaging is realized by the integration of two polarizers. Hyperspectral imaging is realized by snapscan Preliminary imaging experiments were implemented on an unstained paraffin embedded tissue slides of a chicken heart. We also set up a Monte Carlo simulation program based on the cylinder optical model to simulate the cardiac fiber structure of the sample and the optical setup of the PHSIM system, in which we can calculate the system output light intensity related to cardiac fiber orientation. According to the imaging and simulation results, there exists a variation of intensity of acquired images with the polar angles from the maximum to the minimum under different wavelengths, which should relate to the orientation of cardiac fibers. In addition, there is a shift of the polar angle where the maximum intensity appears when a rotation of the sample happened both in the simulation and imaging experiments. Further work is required for imaging more types of myocardial tissues at different parts and the design of a complete quantitative model to describe the relations among polar angles, wavelengths, and cardiac fiber orientations.

Development of a new polarized hyperspectral imaging microscope.

In this study, we proposed and designed a transmission mode polarized hyperspectral imaging microscope (PHSIM). The hyperspectral imaging (HSI) component is based on the snapscan with a hyperspectral camera. The HSI wavelength range is from 467-700 nm. Polarized light imaging is realized by the integration of two polarizers and two liquid crystal variable retarders (LCVR), which is capable of full Stokes polarimetric imaging. The new imaging device was tested for the detection of squamous cell carcinoma (SCC) in H&E stained oral tissue slides of 8 patients. One normal area and one cancerous area on each slide are selected to make the comparison. The preliminary results indicated that the spectral curves of the Stokes vector parameters (S0, S1, S2, S3) of the normal area on the H&E stained oral tissue slides are different from those of SCC in certain wavelength range. Further work is required to apply the new polarized hyperspectral imaging microscope to a large number of patient samples and to test the PHSIM system in different cancer types.

Hyperspectral Microscopic Imaging for Automatic Detection of Head and Neck Squamous Cell Carcinoma Using Histologic Image and Machine Learning.

The purpose of this study is to develop hyperspectral imaging (HSI) for automatic detection of head and neck cancer cells on histologic slides. A compact hyperspectral microscopic system is developed in this study. Histologic slides from 15 patients with squamous cell carcinoma (SCC) of the larynx and hypopharynx are imaged with the system. The proposed nuclei segmentation method based on principle component analysis (PCA) can extract most nuclei in the hyperspectral image without extracting other sub-cellular components. Both spectra-based support vector machine (SVM) and patch-based convolutional neural network (CNN) are used for nuclei classification. CNNs were trained with both hyperspectral images and pseudo RGB images of extracted nuclei, in order to evaluate the usefulness of extra information provided by hyperspectral imaging. The average accuracy of spectra-based SVM classification is 68%. The average AUC and average accuracy of the HSI patch-based CNN classification is 0.94 and 82.4%, respectively. The hyperspectral microscopic imaging and classification methods provide an automatic tool to aid pathologists in detecting SCC on histologic slides.

Freeform fabrication of tissue-simulating phantom for potential use of surgical planning in conjoined twins separation surgery.

Preoperative assessment of tissue anatomy and accurate surgical planning is crucial in conjoined twin separation surgery. We developed a new method that combines three-dimensional (3D) printing, assembling, and casting to produce anatomic models of high fidelity for surgical planning. The related anatomic features of the conjoined twins were captured by computed tomography (CT), classified as five organ groups, and reconstructed as five computer models. Among these organ groups, the skeleton was produced by fused deposition modeling (FDM) using acrylonitrile-butadiene-styrene. For the other four organ groups, shell molds were prepared by FDM and cast with silica gel to simulate soft tissues, with contrast enhancement pigments added to simulate different CT and visual contrasts. The produced models were assembled, positioned firmly within a 3D printed shell mold simulating the skin boundary, and cast with transparent silica gel. The produced phantom was subject to further CT scan in comparison with that of the patient data for fidelity evaluation. Further data analysis showed that the produced model reassembled the geometric features of the original CT data with an overall mean deviation of less than 2 mm, indicating the clinical potential to use this method for surgical planning in conjoined twin separation surgery.

Factors influencing the risk of recurrence in patients with esophageal carcinoma treated with surgery: A single institution analysis consisting of 1002 cases.

The aim of this study was to identify the risk of recurrence in patients with thoracic esophageal squamous cell carcinoma (TESCC) treated with curative surgery. The retrospective analysis included 1002 consecutive patients with TESCC who had been treated with curative surgery in Zhejiang Cancer Hospital, China, between 2003 and 2008. Univariate and multivariate analyses using the Kaplan-Meier method and Cox proportional hazards model, respectively, were performed to identify the independent risk factors for locoregional recurrence and all the recurrence events. The 2-and 5-year recurrence rates were 39.0 and 59.2%, respectively. More than 85% of recurrences occurred within 36 months. The variables associated with a higher rate of recurrence in the univariate analysis were gender (male), length of tumor (≥5 cm), depth of invasion (deeper), lymph node metastasis (greater), histological grade (higher) and vessel involvement (positive). By multivariate analyses, gender (HR, 1.7; 95% CI, 1.2-2.5; P=0.002), depth of invasion (HR, 1.4; 95% CI, 1.2-1.6; P<0.001) and lymph node involvement (HR, 1.4; 95% CI, 1.3-1.5; P<0.001) were independent predictive factors of recurrence. Post-operative radiotherapy or chemotherapy did not significantly prolong failure-free survival (FFS), particularly in patients with early-stage disease. Information regarding the depth of primary tumor invasion and the number of lymph nodes involved may help in evaluating the recurrence risk in patients with TESCC treated with curative surgery. Further studies are required to clarify the correlation between recurrence and the different multidisciplinary treatment approaches.

Longevity pathways: HSF1 and FoxO pathways, a new therapeutic target to prevent age-related diseases.

Modern medicine is directed towards the prevention, detection and cure of individual diseases. Yet, current medical models inadequately describe aging-associated diseases. We now know that failure in longevity pathways including oxidative stress, multisystem dysregulation, inflammation, sarcopenia, protein deposition and atherosclerosis are associated with age-related diseases. Such longevity pathways are potential targets for therapeutic intervention. Interventions in specific pathways have been shown to ameliorate and postpone the aging phenotype by activation of multiple genes. The strategy that we propose in this paper is to apply interventions simultaneously on complementary longevity pathways to achieve a synergistic result. For instance, aging is known to attenuate the HSF1 pathway leading to production of very toxic beta-amyloid fibrils. Consequently, the FoxO pathway is activated, resulting in the formation of less toxic high molecular weight aggregates as a defense mechanism. Thus the simultaneous upregulation of the HSF1 and FoxO pathways could potentially decrease protein deposition and proteotoxicity, thereby retarding or possibly preventing the onset of neurodegenerative diseases. Modulating these two pathways may also delay the onset of other age-related pathologies including cognitive decline, cancer, diabetes and cardiovascular disease due to its multi-gene effect. In this paper, we will discuss the role of several agents on the simultaneous modulation of these two central longevity pathways. The aging of western societies makes prevention of age-related diseases a pressing priority.

Acteylcholinesterase inhibitor rivastigmine enhances cellular defenses in neuronal and macrophage-like cell lines.

Neuroprotection mediated by the cellular heat shock response offers 1 clinical strategy to prevent, stabilize, and possibly reverse neurodegenerative processes. Although damaged proteins are thought to be the primary stimulus for the heat shock response, several studies indicate that pharmaceutical agents can either directly induce the heat shock transcription factor (Hsf1) or enhance its activation during different forms of cellular stress. Because Hsf1 is now known to combat the proteotoxicity of aging and has a central role in modulating amyloid aggregation, pharmacologic interventions to strengthen Hsf1 action may have important implications for preventing neurodegeneration linked to altered and damaged proteins such as observed in Alzheimer's disease. Given reports that some agents for the treatment of Alzheimer's disease have neuroprotective properties, this project investigated whether rivastigmine, which is an acetyl and butaryl cholinesterase inhibitor, mediates the neuroprotection of the neuronal-like cell line SH-SY5Y. The cells were exposed to various concentrations of rivastigmine to determine whether the drug protected cells from toxic injury and induced the 1st phase of the cellular heat shock response. In all, 100-micromol/L rivastigmine decreases cell death by 40% compared with untreated cells. This concentration enhances Hsf1 activation by strengthening both its multimerization and its phosphorylation, which leads to increased messenger RNA (mRNA) for hsp70. Therefore, one of the putative neuroprotective mechanisms of rivastigmine seems to be mediated through the heat shock response. These results also are observed in cultured macrophage-like cells, which suggests a future clinical tool for monitoring pharmacologically improved stress responses in peripheral blood mononuclear cells during treatment of Alzheimer disease.

Electromagnetic field therapy delays cellular senescence and death by enhancement of the heat shock response.

Hormesis may result when mild repetitive stress increases cellular defense against diverse injuries. This process may also extend in vitro cellular proliferative life span as well as delay and reverse some of the age-dependent changes in both replicative and non-replicative cells. This study evaluated the potential hormetic effect of non-thermal repetitive electromagnetic field shock (REMFS) and its impact on cellular aging and mortality in primary human T lymphocytes and fibroblast cell lines. Unlike previous reports employing electromagnetic radiation, this study used a long wave length, low energy, and non-thermal REMFS (50MHz/0.5W) for various therapeutic regimens. The primary outcomes examined were age-dependent morphological changes in cells over time, cellular death prevention, and stimulation of the heat shock response. REMFS achieved several biological effects that modified the aging process. REMFS extended the total number of population doublings of mouse fibroblasts and contributed to youthful morphology of cells near their replicative lifespan. REMFS also enhanced cellular defenses of human T cells as reflected in lower cell mortality when compared to non-treated T cells. To determine the mechanism of REMFS-induced effects, analysis of the cellular heat shock response revealed Hsp90 release from the heat shock transcription factor (HSF1). Furthermore, REMFS increased HSF1 phosphorylation, enhanced HSF1-DNA binding, and improved Hsp70 expression relative to non-REMFS-treated cells. These results show that non-thermal REMFS activates an anti-aging hormetic effect as well as reduces cell mortality during lethal stress. Because the REMFS configuration employed in this study can potentially be applied to whole body therapy, prospects for translating these data into clinical interventions for Alzheimer's disease and other degenerative conditions with aging are discussed.

Engineered repeated electromagnetic field shock therapy for cellular senescence and age-related diseases.

A new consensus of gerontologists proposes that delay of biological senescence is the most potent public health measure for preventing chronic disease in late life. At the most fundamental level, cellular aging is characterized by a decline in repair and homeostatic systems. Thus, interventions that protect or rejuvenate these cellular systems hold significant promise for preventing or delaying the onset of age-related diseases. The most likely candidates for delaying senescence are the longevity-linked transcription factors DAF16 and HSF1. If one were to engineer negligible senescence, a key target would be the heat shock protein axis regulated by HSF1. This pathway is the preferred pathway to prevent protein damage or aggregation, whereas DAF-16/FOXO is a backup pathway activated during stress. Reduced HSF1 activity appears to accelerate tissue aging and shortens life span. Conversely, over-expression of HSF1 increases life span and decreases amyloid toxicity in animal models. This paper describes enhancement of the HSR/HSF1 pathway engineered by repeated electromagnetic field shock (REMFS). In a recent study, we demonstrated that REMFS therapy upregulates the HSR/HSF1 pathway, delays cellular senescence in young cells, and transiently reverses it in senescent cells, thus altering cellular mortality. The technology of applying certain beneficial EMF energy to the human body to stimulate the natural stress response and activate the repair and maintenance systems is a new strategy for engineered negligible senescence. We discuss the exciting clinical implications of REMFS therapy in human aging and disease.

Pathomolecular effects of homocysteine on the aging process: a new theory of aging.

Homocysteine has been associated with the most common age-related diseases but never associated with the acceleration of the aging process. This theoretical paper will try to demonstrate the pro-aging effects of homocysteine at the molecular, cellular, and organ level. High homocysteine levels in homocystinuria are associated with premature disease of the cardiovascular, skeletal, neurological, and other systems. These observations are similar to those noted in the aging process and should be considered as a progeroid syndrome. There is enough scientific evidence to support that homocysteine accelerates the aging process at the cellular and at the organism level. Most importantly, decreasing homocysteine levels by dietary or pharmacological interventions could prolong maximum life span in humans and/or delay the onset of the most common age-related diseases.

Clonal senescence alters endothelial ICAM-1 function.

Little is known how age alters the dynamics and function of cell adhesion molecules, especially under inflammatory and stressful conditions. One membrane constituent, intercellular adhesion molecule-1 (ICAM-1) is a transmembrane glycoprotein of the immunoglobulin (Ig) superfamily that regulates key outside-->in and inside-->out signals associated with cell-to-cell interactions. If conditions such as age and inflammation change usual ICAM-1 action then important downstream effects ultimately perturb endothelial cell function. In this report, ICAM-1 accumulates in late passage endothelial cells when compared to early passage endothelial cells, yet ICAM-1 protein expression is attenuated when senescent cells are challenged by TNF-alpha (10ng/ml). Importantly, age alters ICAM-1 dynamic properties from directed to random receptor motion within the membrane. Single particle tracking reveals that the average ICAM-1 mobility is 44% less in late than early passage cells after its motion is stimulated by the Protein Kinase C (PKC) activator, phorbol myristate acetate (PMA). The mechanism for altered ICAM-1 mobility partly can be explained by a reduced rate of alpha-actinin linking with ICAM-1 in late passage Human Pulmonary Artery Endothelial Cells (HPAECs). Furthermore, tyrosine phosphorylation of alpha-actinin, a requirment for ICAM-1 clustering, is markedly reduced in senescent cells. These findings support a hypothesis that senescence results in changes of ICAM-1 activation and clustering, thus resulting in an age-dependent transmembrane signaling disorder. Therefore, further understanding of age-dependent disturbances of ICAM-1 regulation during inflammation can provide important clues as to appropriate targets for therapeutic interventions and prevention of vascular disorders in elderly at the level of the endothelial surface membrane.