Correlations of diffusion tensor imaging and clinical measures with spinal cord cross-sectional area measurements in pediatric spinal cord injury patients.

PURPOSE: The purpose of this work was to employ a semi-automatic method for measuring spinal cord cross-sectional area (SCCSA) and investigate the correlations between diffusion tensor imaging (DTI) metrics and SCCSA for the cervical and thoracic spinal cord for typically developing pediatric subjects and pediatric subject with spinal cord injury. METHODS: Ten typically developing (TD) pediatric subjects and ten pediatric subjects with spinal cord injury (SCI) were imaged using a Siemens Verio 3 T MR scanner to acquire DTI and high-resolution anatomic scans covering the cervical and thoracic spinal cord (C1-T12). SCCSA was measured using a semi-automated edge detection algorithm for the entire spinal cord. DTI metrics were obtained from whole cord axial ROIs at each vertebral level. SCCSA measures were compared to DTI metrics by vertebral level throughout the entire cord, and above and below the injury site. Correlation analysis was performed to compare SCCSA, DTI and clinical measures as determined by the International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI) examination. RESULTS: In subjects with SCI, FA and SCCSA had a positive correlation (r = 0.81, P < 0.01), while RD and SCCSA had a negative correlation (r = -0.68, P = 0.02) for the full spinal cord. FA and SCCSA were correlated above (r = 0.56, P < 0.01) and below (r = 0.54, P < 0.01) the injury site. TD subjects showed negative correlations between AD and SCCSA (r = -0.73, P = 0.01) and RD and SCCSA (r = -0.79, P < 0.01). CONCLUSION: The ability to quickly and effectively measure SCCSA in subjects with SCI has the potential to allow for a better understanding of the progression of atrophy following a SCI. Correlations between cord cross section and DTI metrics by vertebral level suggest that imaging inferior and superior to lesion may yield useful information for diagnosis and prognosis.

An fMRI Study of the Effects of Vibroacoustic Stimulation on Functional Connectivity in Patients with Insomnia.

BACKGROUND: It is well known that vibratory and auditory stimuli from vehicles such as cars and trains can help induce sleep. More recent literature suggests that specific types of vibratory and acoustic stimulation might help promote sleep, but this has not been tested with neuroimaging. Thus, the purpose of this study was to observe the effects of vibroacoustic stimulation (providing both vibratory and auditory stimuli) on functional connectivity changes in the brain using resting state functional magnetic resonance imaging (rs-fMRI), and compare these changes to improvements in sleep in patients with insomnia. METHODS: For this study, 30 patients with insomnia were randomly assigned to receive one month of a vibroacoustic stimulation or be placed in a waitlist control. Patients were evaluated pre- and postprogram with qualitative sleep questionnaires and measurement of sleep duration with an actigraphy watch. In addition, patients underwent rs-fMRI to assess functional connectivity. RESULTS: The results demonstrated that those patients receiving the vibroacoustic stimulation had significant improvements in measured sleep minutes as well as in scores on the Insomnia Severity Index questionnaire. In addition, significant changes were noted in functional connectivity in association with the vermis, cerebellar hemispheres, thalamus, sensorimotor area, nucleus accumbens, and prefrontal cortex. CONCLUSIONS: The results of this study show that vibroacoustic stimulation alters the brain's functional connectivity as well as improves sleep in patients with insomnia.

Fundamentals of Preoperative Task Functional Brain Mapping.

Blood oxygenation level-dependent (BOLD) imaging is gaining traction in the clinical realm as a measure for quantifying changes in regional blood flow in response to external stimuli. Through the evoked signal changes that are a consequence of hemoglobin's intrinsic paramagnetic properties, this technique allows for the statistical mapping of brain regions associated with a given task, which has broad applications in preneurosurgical planning for tumor resection. From an acquisition perspective, collection of BOLD data most commonly requires the use of echo planar imaging readout schemes. These sequences are currently widely available on most clinical scanners and at various field strengths. However, while the BOLD acquisition protocol is relatively straightforward, additional hardware and rigorous image processing are needed to correlate the time-dependent signal changes associated with a specific and well defined task. This manuscript will provide the necessary information to detail the physiologic underpinning of acquiring BOLD sensitized images and the important technical aspects of processing the data for use in a surgical environment.

Diffusion Tensor Imaging Assessment of Regional White Matter Changes in the Cervical and Thoracic Spinal Cord in Pediatric Subjects.

There are no studies to date,describing changes in the diffusion tensor imaging (DTI) metrics of the white matter (WM) regions of the entire cervical and thoracic spinal cord (SC) remote from the lesion in pediatric spinal cord injury (SCI) subjects. The purpose of this study was to determine whether DTI at sites cephalad and caudal to a lesion provides measures of cord abnormalities in children with chronic SCI. A retrospective study included 10 typically developing subjects (TD) and 10 subjects with chronic SCI who underwent SC imaging in 2014-2017. Axial diffusion tensor images using an inner field of view DTI sequence were acquired to cover the entire cervical and thoracic SC. Regions of interest were drawn on the SC WM: right and left lateral (motor), ventral (motor), and dorsal (sensory) tracts. To detect differences in DTI metrics between TD and SCI of the cord, a one way analysis of variance with pooled t test was performed. A stepwise regression analysis was performed to assess the correlation between DTI metrics and clinical scores. In motor and sensory tracts, fractional anisotropy (FA) and axial diffusivity (AD) were significantly decreased in the proximal segments of the caudal cord. In motor tracts cephalad to the lesion, FA was significantly decreased whereas AD was significantly increased in the proximal segment; however, AD was decreased in the distal and middle segments. International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI) total score was significantly correlated with FA and AD of the motor and sensory tracts cephalad to the lesion. This study demonstrates that FA and AD have the potential to be sensitive biomarkers of the full extent of cord injury and might be useful in detecting remote injuries to the SC and in guiding new treatments.

Age related diffusion and tractography changes in typically developing pediatric cervical and thoracic spinal cord.

Background and objective: Diffusion tensor imaging (DTI) and diffusion tensor tractography (DTT) are two techniques that can measure white matter integrity of the spinal cord. Recently, DTI indices have been shown to change with age. The purpose of this study is (a) to evaluate the maturational states of the entire pediatric spinal cord using DTI and DTT indices including fractional anisotropy (FA), mean diffusivity (MD), mean length of white matter fiber tracts and tract density and (b) to analyze the DTI and DTT parameters along the entire spinal cord as a function of spinal cord levels and age. Method: A total of 23 typically developing (TD) pediatric subjects ranging in age from 6 to 16 years old (11.94 ±â€¯3.26 (mean ±â€¯standard deviation), 13 females and 10 males) were recruited, and scanned using 3.0 T MR scanner. Reduced FOV diffusion tensor images were acquired axially in the same anatomical location prescribed for the T2-weighted images to cover the entire spinal cord (C1-mid L1 levels). To mitigate motion induced artifacts, diffusion directional images were aligned with the reference image (b0) using a rigid body registration algorithm performed by in-house software developed in Matlab (MathWorks, Natick, Massachusetts). Diffusion tensor maps (FA and MD) and streamline deterministic tractography were then generated from the motion corrected DTI dataset. DTI and DTT parameters were calculated by using ROIs drawn to encapsulate the whole cord along the entire spinal cord by an independent board certified neuroradiologist. These indices then were compared between two age groups (age group A = 6-11 years (n = 11) and age group B = 12-16 years (n = 12)) based on similar standards and age definitions used for reporting spinal cord injury in the pediatric population. Standard least squared linear regression based on a restricted maximum likelihood (REML) method was used to evaluate the relationship between age and DTI and DTT parameters. Results: An increase in FA (group A = 0.42 ±â€¯0.097, group B = 0.49 ±â€¯0.116), white matter tract density (group A = 368.01 ±â€¯236.88, group B = 440.13 ±â€¯245.24) and mean length of fiber tracts (group A = 48.16 ±â€¯20.48 mm, group B = 60.28 ±â€¯23.87 mm) and a decrease in MD (group A = 1.06 ±â€¯0.23 × 10-3 mm2/s, group B = 0.82 ±â€¯0.24 × 10-3 mm2/s) were observed with age along the entire spinal cord. Statistically significant increases have been shown in FA (p = 0.004, R2 = 0.57), tract density (p = 0.0004, R2 = 0.58), mean length of fiber tracts (p < 0.001, R2 = 0.5) and a significant decrease has been shown in MD (p = 0.002, R2 = 0.59) between group A and group B. Also, it has been shown DTI and DTT parameters vary along the spinal cord as a function of intervertebral disk and mid-vertebral body level. Conclusion: This study provides an initial understanding of age related changes of DTI values as well as DTT metrics of the spinal cord. The results show significant differences in DTI and DTT parameters which may result from decreasing water content, myelination of fiber tracts, and the thickening diameter of fiber tracts during the maturation process. Consequently, when quantitative DTI and DTT of the spinal cord is undertaken in the pediatric population an age and level matched normative dataset should be used to accurately interpret the quantitative results.

Characterization of spinal cord diffusion tensor imaging metrics in clinically asymptomatic pediatric subjects with incidental congenital lesions.

STUDY DESIGN: Retrospective study. OBJECTIVES: To perform quantitative DTI measurements of the entire cervical and thoracic spinal cord (SC) in typically developing (TD) pediatric subjects with incidental findings of syringomyelia or hydromyelia on conventional MRI and in a TD population without any abnormalities. SETTING: USA. METHODS: 26 TD recruited as part of large SC DTI study, four of these had incidental findings. Axial DTI images were acquired on 3T MR scanner to cover the cervical and thoracic SC. We performed group analysis of DTI values in the cord above and below the MR-defined lesion. For single-subject analysis, the cord above and below the lesion was compared to average values of TD population. A standard least squares regression model was used to compare DTI parameters fractional anisotropy (FA), mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) between TD population and subjects with hydromyelia and syringomyelia. A p value of 0.05 was used for statistical significance. RESULTS: In group analysis, MD and AD were significantly different in cord above the lesion in subjects with hydromyelia and syringomyelia (n = 4) compared to TD population (n = 22). For single-subject analysis, DTI parameters were significantly different in cord above the syringomyelia and below the syringomyelia; MD, AD, and RD were significantly different. A subject with hydromyelia showed significant difference in FA below the lesion. CONCLUSIONS: This study demonstrates that DTI has the potential to be used as an imaging biomarker to evaluate SC above and below the congenital lesion in syringohydromyelia subjects.

Application of Color Transformation Techniques in Pediatric Spinal Cord MR Images: Typically Developing and Spinal Cord Injury Population.

The purpose of this study was to evaluate an improved and reliable visualization method for pediatric spinal cord MR images in healthy subjects and patients with spinal cord injury (SCI). A total of 15 pediatric volunteers (10 healthy subjects and 5 subjects with cervical SCI) with a mean age of 11.41 years (range 8-16 years) were recruited and scanned using a 3.0T Siemens Verio MR scanner. T2-weighted axial images were acquired covering entire cervical spinal cord level C1 to C7. These gray-scale images were then converted to color images by using five different techniques including hue-saturation-value (HSV), rainbow, red-green-blue (RGB), and two enhanced RGB techniques using automated contrast stretching and intensity inhomogeneity correction. Performance of these techniques was scored visually by two neuroradiologists within three selected cervical spinal cord intervertebral disk levels (C2-C3, C4-C5, and C6-C7) and quantified using signal to noise ratio (SNR) and contrast to noise ratio (CNR). Qualitative and quantitative evaluation of the color images shows consistent improvement across all the healthy and SCI subjects over conventional gray-scale T2-weighted gradient echo (GRE) images. Inter-observer reliability test showed moderate to strong intra-class correlation (ICC) coefficients in the proposed techniques (ICC > 0.73). The results suggest that the color images could be used for quantification and enhanced visualization of the spinal cord structures in addition to the conventional gray-scale images. This would immensely help towards improved delineation of the gray/white and CSF structures and further aid towards accurate manual or automatic drawings of region of interests (ROIs).

Changes in cerebellar functional connectivity and autonomic regulation in cancer patients treated with the Neuro Emotional Technique for traumatic stress symptoms.

PURPOSE: A growing number of research studies have implicated the cerebellum in emotional processing and regulation, especially with regard to negative emotional memories. However, there currently are no studies showing functional changes in the cerebellum as a result of treatment for traumatic stress symptoms. The Neuro Emotional Technique (NET) is an intervention designed to help improve symptoms related to traumatic stress using an integrative approach that combines emotional, cognitive, and motor processing, with a particular focus on autonomic nervous system regulation. In this study, we evaluated whether the NET intervention alters functional connectivity in the brain of patients with traumatic stress symptoms associated with a cancer-related event. We hypothesized that the NET intervention would reduce emotional and autonomic reactivity and that this would correlate with connectivity changes between the cerebellum and limbic structures as well as the brain stem. METHODS: We enrolled patients with a prior cancer diagnosis who experienced distressing cancer-related memories associated with traumatic stress symptoms of at least 6 months in duration. Participants were randomized to either the NET intervention or a waitlist control. To evaluate the primary outcome of neurophysiological effects, all participants received resting-state functional blood oxygen level-dependent (BOLD) magnetic resonance imaging (rs-fMRI) before and after the NET intervention. In addition, autonomic reactivity was measured using heart rate response to the traumatic stimulus. Pre/post comparisons were performed between the NET and control groups. RESULTS: The results demonstrated significant changes in the NET group, as compared to the control group, in the functional connectivity between the cerebellum (including the vermis) and the amygdala, parahippocampus, and brain stem. Likewise, participants receiving the NET intervention had significant reductions in autonomic reactivity based on heart rate response to the traumatic stimulus compared to the control group. CONCLUSIONS: This study is an initial step towards establishing a neurological signature of treatment effect for the NET intervention. Specifically, functional connectivity between the cerebellum and the amygdala and prefrontal cortex appear to be associated with a reduction in autonomic reactivity in response to distressing cancer-related memories. IMPLICATIONS FOR CANCER SURVIVORS: This study contributes to the understanding of possible mechanisms by which interventions like NET may help reduce emotional distress in cancer patients who suffer from traumatic stress symptoms.

Reduced Field of View Diffusion Tensor Imaging and Fiber Tractography of the Pediatric Cervical and Thoracic Spinal Cord Injury.

The aim of this study is to assess the utility and effectiveness of diffusion tensor imaging (DTI) and diffusion tensor tractography (DTT) of the entire pediatric cervical and thoracic spinal cord toward discrimination of typically developing (TD) controls and subjects with spinal cord injury (SCI). A total of 43 pediatric subjects, including 23 TD subjects ranging in age from 6 to 16 years old and 20 subjects with SCI ranging in age from 7 to 16 years, were recruited and scanned using a 3.0 Tesla magnetic resonance scanner. Reduced field of view diffusion tensor images were acquired axially to cover the entire spinal cord across two slabs. For DTI analysis, motion correction was performed by coregistration of the diffusion-weighted images to the reference image (b0). Streamline deterministic tractography results were generated from the preprocessed data. DTI and DTT parameters of the whole cord, including fractional anisotropy (FA), mean diffusivity (MD), tract length, and tract density, were calculated, averaged across the whole spinal cord, and compared between the TD and SCI groups. Statistically significant decreases have been shown in FA (TD = 0.46 ± 0.11; SCI = 0.37 ± 0.09; p < 0.0001) and tract density (TD = 405.93 ± 243.84; SCI = 268.90 ± 270.34; p < 0.0001). However, the mean length of tracts and MD did not show significant differences. When investigating differences in DTI and DTT parameters above and below the injury site, it was shown that the FA and tract density in patients with cervical SCI decreased significantly in the thoracic region. An identical trend was observed in the cervical region for patients with thoracic SCI as well. When comparing TD and SCI subjects, FA and tract density were the most sensitive parameters in detecting functional changes of the spinal cord in chronic pediatric SCI. The results show that both DTI and DTT have the potential to be imaging biomarkers in the diagnosis of SCI.

Identification of ghost artifact using texture analysis in pediatric spinal cord diffusion tensor images.

PURPOSE: Ghost artifacts are a major contributor to degradation of spinal cord diffusion tensor images. A multi-stage post-processing pipeline was designed, implemented and validated to automatically remove ghost artifacts arising from reduced field of view diffusion tensor imaging (DTI) of the pediatric spinal cord. METHOD: A total of 12 pediatric subjects including 7 healthy subjects (mean age=11.34years) with no evidence of spinal cord injury or pathology and 5 patients (mean age=10.96years) with cervical spinal cord injury were studied. Ghost/true cords, labeled as region of interests (ROIs), in non-diffusion weighted b0 images were segmented automatically using mathematical morphological processing. Initially, 21 texture features were extracted from each segmented ROI including 5 first-order features based on the histogram of the image (mean, variance, skewness, kurtosis and entropy) and 16s-order feature vector elements, incorporating four statistical measures (contrast, correlation, homogeneity and energy) calculated from co-occurrence matrices in directions of 0°, 45°, 90° and 135°. Next, ten features with a high value of mutual information (MI) relative to the pre-defined target class and within the features were selected as final features which were input to a trained classifier (adaptive neuro-fuzzy interface system) to separate the true cord from the ghost cord. RESULTS: The implemented pipeline was successfully able to separate the ghost artifacts from true cord structures. The results obtained from the classifier showed a sensitivity of 91%, specificity of 79%, and accuracy of 84% in separating the true cord from ghost artifacts. CONCLUSION: The results show that the proposed method is promising for the automatic detection of ghost cords present in DTI images of the spinal cord. This step is crucial towards development of accurate, automatic DTI spinal cord post processing pipelines.

Genetic and neural predictors of behavioral weight loss treatment: A preliminary study.

OBJECTIVE: To examine neural mechanisms of action in behavioral weight loss treatment (BWL) and explore neural and genetic predictors of BWL. METHODS: Neural activation to milkshake receipt and genetics were compared in 17 women with obesity who received 12 weeks of BWL and 17 women who received no intervention. Participants were scanned twice using functional magnetic resonance imaging at baseline and 12 weeks. Weight was assessed at baseline, 12, 36, and 60 weeks. RESULTS: BWL participants lost more weight than controls at 12 weeks (-4.82% versus -0.70%). After 12 weeks, BWL had greater reduction in right caudate activation response to milk shake receipt than did controls. Among BWL participants, baseline to 12-week reduction in frontostriatal activation to milk shake predicted greater weight loss at 12, 36, and 60 weeks. Possessing the A/A or T/A genotype of the fat mass and obesity-associated (FTO) variant rs9939609 predicted greater weight loss at 12 and 36 weeks. CONCLUSIONS: These preliminary data reveal that reduction in right caudate activation may be a neural mechanism of weight loss in BWL, and baseline FTO variant and reduction in frontostriatal activation during BWL predict short- and long-term weight loss. These findings require replication in larger samples.

Spatially selective 2D RF inner field of view (iFOV) diffusion kurtosis imaging (DKI) of the pediatric spinal cord.

Magnetic resonance based diffusion imaging has been gaining more utility and clinical relevance over the past decade. Using conventional echo planar techniques, it is possible to acquire and characterize water diffusion within the central nervous system (CNS); namely in the form of Diffusion Weighted Imaging (DWI) and Diffusion Tensor Imaging (DTI). While each modality provides valuable clinical information in terms of the presence of diffusion and its directionality, both techniques are limited to assuming an ideal Gaussian distribution for water displacement with no intermolecular interactions. This assumption neglects pathological processes that are not Gaussian therefore reducing the amount of potentially clinically relevant information. Additions to the Gaussian distribution measured by the excess kurtosis, or peakedness, of the probabilistic model provide a better understanding of the underlying cellular structure. The objective of this work is to provide mathematical and experimental evidence that Diffusion Kurtosis Imaging (DKI) can offer additional information about the micromolecular environment of the pediatric spinal cord. This is accomplished by a more thorough characterization of the nature of random water displacement within the cord. A novel DKI imaging sequence based on a tilted 2D spatially selective radio frequency pulse providing reduced field of view (FOV) imaging was developed, implemented, and optimized on a 3 Tesla MRI scanner, and tested on pediatric subjects (healthy subjects: 15; patients with spinal cord injury (SCI):5). Software was developed and validated for post processing of the DKI images and estimation of the tensor parameters. The results show statistically significant differences in mean kurtosis (p < 0.01) and radial kurtosis (p < 0.01) between healthy subjects and subjects with SCI. DKI provides incremental and novel information over conventional diffusion acquisitions when coupled with higher order estimation algorithms.

Technical considerations for functional magnetic resonance imaging analysis.

Clinical application of functional magnetic resonance imaging (fMRI) based on blood oxygenation level-dependent (BOLD) effect has increased over the past decade because of its ability to map regional blood flow in response to brain stimulation. This mapping is primarily achieved by exploiting the BOLD effect precipitated by changes in the magnetic properties of hemoglobin. BOLD fMRI has utility in neurosurgical planning and mapping neuronal functional connectivity. Conventional echo planar imaging techniques are used to acquire stimulus-driven fMR imaging BOLD data. This article highlights technical aspects of fMRI data analysis to make it more accessible in clinical settings.

Immunohistochemistry is a reliable screening tool for identification of ALK rearrangement in non-small-cell lung carcinoma and is antibody dependent.

INTRODUCTION: Fluorescence in situ hybridization (FISH) is the standard procedure for the detection of anaplastic lymphoma receptor tyrosine kinase (ALK) rearrangement in non-small-cell lung carcinoma (NSCLC) but is expensive and time consuming. We tested three antibodies to ALK, using various detection systems, and hypothesized that ALK immunohistochemistry (IHC) may represent a cost-effective and efficient means of screening for ALK rearrangement in NSCLC. METHODS: We screened 377 stage I or II NSCLC cases in a tissue microarray by FISH and IHC (5A4 [Leica Biosystems Newcastle Ltd, Newcastle upon Tyne, UYnited Kingdom] by Nichirei's N-Histofine ALK detection kit [Nichirei Biosciences inc., Tokyo, Japan], 5A4 by Novocastra with ADVANCE [Dako Canada inc., Burlington, Ontario, Canada], D5F3 by Cell Signaling Technology with ADVANCE [Cell Signalling Technologies inc., Danvers, MA], and DAKO clone ALK1 with FLEX [Dako Canada inc., Burlington, Ontario, Canada] and ADVANCE). IHC was scored as 0, 1+, 2+, or 3+. Possibly positive or positive cases were further analyzed by IHC and FISH on whole section. RESULTS: Tissue microarray results were available on 377 cases by IHC and 273 cases by FISH. Eleven cases were positive or possibly positive by either IHC or FISH, and three cases were positive or possibly positive by both methods. Three cases were ALK-positive by FISH on whole section validation. There was no correlation between semiquantitative IHC score (1+, 2+, 3+) and ALK rearrangement by FISH. D5F3 (Cell Signaling by ADVANCE) and 5A4 (Novocastra by ADVANCE) showed the greatest combination of sensitivity (100%) and specificity (87.5% for 5A4 by Novocastra and 75% for D5F3 by Cell Signaling), and produced no false-negative results. CONCLUSIONS: IHC is a reliable screening tool for identification of ALK rearrangement in NSCLC and is antibody dependent. D5F3 (Cell Signaling) and 5A4 (Novocastra) can be used with FISH for identification of IHC-positive cases to reduce screening costs.

Tissue microarray analysis of connexin expression and its prognostic significance in human breast cancer.

Breast cancer accounts for approximately 15% of all cancer deaths. Currently, axillary nodal status is the most reliable prognostic indicator for breast cancer. Tumor size and histological grade are used to stage breast cancer. Estrogen receptor/progesterone receptor (ER/PR) and HER-2/neu status are useful in predicting patient survival and relapse. Ki67, an indicator of proliferative activity, also correlates well with prognosis. Connexin proteins form gap junction channels, permitting intercellular exchange of ions and small molecules. Reduced connexin protein levels and impaired gap junctional intercellular communication are associated with tumor phenotypes. This study investigated the prognostic value of connexin proteins as breast cancer markers. Tissue microarrays, containing 438 cases of invasive breast carcinoma, were stained with Cx26, Cx32, and Cx43 antibodies. The degree of connexin immunoreactivity was determined and then correlated with patient outcome, tumor grade, tumor size, lymph node status, and immunohistochemical markers, such as p53, ER/PR status, Ki67 and c-erbB-2 expression. Cx26, Cx32, or Cx43 did not correlate well with tumor grade, tumor size, p53 or c-erbB-2 status. There was an inverse correlation between Cx32 and lymph node status (P <0.05) and a positive correlation between Cx43 and PR status (P <0.01). Cx32 and Cx43 correlated positively with ER status (P <0.01). Cx43 correlated negatively with Ki67 expression (P <0.01). Cx26, Cx32, and Cx43 did not correlate with patient outcome. Based on our observations in this study, connexin proteins do not appear to be reliable indicators of breast cancer prognosis.

Genistein and quercetin increase connexin43 and suppress growth of breast cancer cells.

Connexin proteins form gap junctions, which permit direct exchange of cytoplasmic contents between neighboring cells. Evidence indicates that gap junctional intercellular communication (GJIC) is important for maintaining homeostasis and preventing cell transformation. Furthermore, connexins may have independent functions including tumor growth suppression. Most tumors express less connexins, have reduced GJIC and have increased growth rates compared with non-tumorigenic cells. The purpose of this study was to determine whether common flavonoids, genistein and quercetin, increase connexin43 (Cx43) levels, improve GJIC and suppress growth of a metastatic human breast tumor cell line (MDA-MB-231). Quercetin (2.5, 5 microg/ml) and genistein (0.5, 2.5, 15 microg/ml) upregulated Cx43 but failed to increase GJIC. Cx43 localized to the plasma membrane following genistein treatment (2.5, 15 mug/ml). In contrast, Cx43 aggregated in the perinuclear region following quercetin treatment (0.5, 2.5, 5, 15 microg/ml). Both genistein (15 microg/ml) and quercetin (2.5, 5, 15 microg/ml) significantly reduced MDA-MB-231 cell proliferation. In summary, genistein and quercetin increase Cx43 and suppress MDA-MB-231 cell proliferation at physiologically relevant concentrations. These results demonstrate that genistein and quercetin are potential anti-breast cancer agents.