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.

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.

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.