Elizabeth W. Pang

Assessment of Genetic Changes in Hepatocellular Carcinoma by Comparative Genomic Hybridization Analysis : Relationship to Disease Stage, Tumor Size, and Cirrhosis

Hepatocellular carcinoma (HCC) is a common and highly malignant tumor that is prevalent in Southeast Asia. Although the etiological factors associated are now well recognized, the interactions between individual factors and the molecular mechanisms by which they lead to cancer remain unclear. Cytogenetic analysis on HCC has been limited because of poor hepatocyte growth in vitro. The recently developed technique of comparative genomic hybridization (CGH), however, permits screening of the entire genome without the need of cell culture. CGH was applied to the study of genomic aberrations in 67 surgically resected samples of HCC, 3 of adenomatous hyperplasia (AH), and 12 of nontumorous cirrhotic liver surrounding the tumors. All samples were from patients of a racially and etiologically homogeneous population in Southern China, where chronic hepatitis B virus infection is the main etiological factor. CGH analysis of the HCC samples revealed frequent copy number gain of 1q (48/67 cases, 72%), 8q (32/67 cases, 48%), 17q (20/67 cases, 30%), and 20q (25/67 cases, 37%) and common losses on 4q (29/67 cases, 43%), 8p (25/67 cases, 37%), 13q (25/67 cases, 37%), and 16q (20/67 cases, 30%). Our finding of a high incidence of 1q gain strongly suggested this aberration was associated with the development of HCC. Genomic abnormalities were detected in 1 of the 3 AH specimens but absent in all 12 cirrhotic tissues surrounding the tumor. Clinical staging classified 3/67 HCC cases as T1, 53 cases as T2, and 11 cases as T3. No significant difference in the pattern of genomic imbalances was detected between stages T2 and T3. A significant copy number loss of 4q11-q23 was, however, identified in those tumors larger than 3 cm in diameter. Of particular interest was the identification of 8q copy number gain in all 12 cases of HCC that arose in a noncirrhotic liver, compared with only 20/55 cases in HCC arising in a cirrhotic liver. We suggest that 8q over-representation is likely associated with a growth advantage and proliferative stimulation that have encouraged malignant changes in the noncirrhotic human liver.

Event-related beamforming: a robust method for presurgical functional mapping using MEG.

OBJECTIVE We describe the application of a new spatial filtering technique--event-related beamforming (ERB)--for presurgical functional mapping of primary sensory areas using MEG. This method provides an alternative to equivalent current dipole (ECD) modeling that potentially eliminates problems of intracranial magnetic artifacts due to movement of ferromagnetic materials (e.g., orthodontic braces) or eye movements. METHODS We compared localization results for ERB and ECD localization of primary somatosensory (M20) and auditory (M100) evoked responses in 12 healthy control subjects and four subjects with metallic dental implants. Data were recorded with a 151-channel CTF MEG system using standard presurgical mapping protocols. RESULTS We found a high level of agreement between the two methods in control subjects (overall localization difference was 5.9+/-2.2 mm for M20 and 10.4+/-5.6 mm for M100). Subjects with dental implants showed severely distorted evoked responses that could not be analyzed using ECD, whereas the ERB method localized sources to expected anatomical locations. CONCLUSIONS MEG functional mapping may be carried out without removal of orthodontic or other metallic implants using event-related beamformer analysis. SIGNIFICANCE Spatial filtering methods can overcome some of the limitations associated with MEG expanding its applicability, particularly in pediatric clinical environments.

Hypomethylation of Chromosome 1 Heterochromatin DNA Correlates with q-Arm Copy Gain in Human Hepatocellular Carcinoma

Using comparative genomic hybridization (CGH) analysis, we, and others, have shown that there is a high and consistent incidence of chromosome 1q copy gain in human hepatocellular carcinoma (HCC). Chromosome 1 rearrangements, that involved peri-centromeric breakpoints, have also been frequently reported in karyotypic studies of HCC. Satellite DNA hypomethylation has been postulated as the mechanism underlying the induction of chromosome 1 peri-centromeric instability in many human cancers and in individuals with the rare recessive disorder ICF (immunodeficiency, centromeric heterochromatin instability, facial anomalies). In this study, we have investigated the role of DNA hypomethylation in 1q copy gain in HCC by examining the methylation status of chromosome 1 heterochromatin DNA (band 1q12). Thirty-six histologically confirmed samples of HCC were studied (24 paired tumor and adjacent nontumorous liver tissues, and 12 tumor only). Hypomethylation of satellite 2 (Sat2) DNA in 1q12 was analyzed by Southern blotting using methyl-sensitive enzyme digestion. In parallel, all cases were analyzed by CGH. A strong correlation between hypomethylated Sat2 sequences and 1q copy gain with a 1q12 breakpoint was found (P < 0.001). We postulate that such hypomethylation alters the interaction between the CpG-rich satellite DNA and chromatin proteins, resulting in heterochromatin decondensation, breakage and aberrant 1q formation. Spectral karyotyping further supported the presence of fragile 1q12 in HCC. Of particular interest was the finding of Sat2 DNA hypomethylation in 5 of 24 adjacent nontumorous liver tissues examined. These tissues showed no evidence of malignancy on histological examination nor did they display any CGH abnormalities. Our findings suggest a role for Sat2 demethylation in the early stages of the stepwise progression of liver carcinogenesis.

Mismatch negativity to speech stimuli in 8-month-old infants and adults

The mismatch negativity (MMN) was measured in 15 normal awake 8-month-old infants and 10 adults to the speech consonants /da/ and /ta/. ERPs were analyzed at 11 electrodes (Fz, Cz, Pz, C3, C4, T3, T4, T5, T6, P3, P4). Four-hundred trials were presented: the /da/ standards with 80% probability and the /ta/ deviants with 20% probability. The ISI was 600 ms. An MMN was observed for both adults and infants but with different scalp distributions. A clear infant MMN was observed only at C3 and T3 electrodes, whereas the adult MMN was present at Fz, Cz, C3, C4 and Pz. A repeated-measures ANOVA on the normalized summed area between 200 and 250 ms revealed an age (adult vs. infant) x electrode interaction. Paired t-tests indicated that adults and infants showed significant differences at the C3, Cz, T3, Pz and T6 electrodes. The adult MMN was largest at Cz and C3 whereas the infant MMN was largest at T3. These data are discussed in terms of possible maturational changes in the MMN.

Sound motion evoked magnetic fields

OBJECTIVE The aim of present study was to determine which brain regions are involved in the conscious perception of sound motion in humans. METHODS Six kinds of sound stimuli were studied. Two static sound stimuli with durations of 100 or 1000 ms remained at a fixed position during the stimulation period. Four moving sound stimuli with duration of 100 or 1000 ms were moving from left to right, or right to left, during the stimulation period. Evoked magnetic fields were recorded using a 151-channel whole cortex magnetoencephalographic system. RESULTS The response identified in all sound stimuli was M100. Responses identified only in moving sound stimuli were M180, M280 and M680. Contour maps and dipoles overlapped on magnetic resonance imaging indicated that both the M100 and M680 responses were generated in the superior temporal cortex (left and right), while M180 and M280 were generated in the parietal cortex (right). CONCLUSIONS The results of this MEG study indicated that the right parietal cortex was involved in sound motion processing. We hypothesize that the right parietal cortex, in association with the left and right superior temporal cortex, forms a network to process sound motion information.

Three-dimensionally reconstructed magnetic source imaging and neuronavigation in pediatric epilepsy: technical note.

OBJECTIVE: To determine the role of reconstructing three-dimensional magnetic source imaging (MSI) data on cortical resections for children undergoing epilepsy surgery using neuronavigation. METHODS: Magnetoencephalographic recordings were analyzed in 16 children under 18 years of age with intractable epilepsy. The data were transferred to the neuronavigation workstation for intraoperative localization of MSI spike sources in selected patients. With the aid of neuronavigation, the MSI spike sources were resected. Intraoperative electrocorticography was then used to survey the surrounding field for residual epileptiform activity. RESULTS: MSI spike sources were obtained in 13 of 16 patients. MSI spike sources localized the cortical and subcortical discharges before intraoperative electrocorticography in nine patients and before extraoperative subdural grid electroencephalographic monitoring in four patients. The localization of MSI spikes sources was characterized by clustered spike sources in 10 patients. By use of neuronavigation, the clustered spike sources were correlated to the interictal zone indicated by intraoperative electrocorticography in six patients and to the ictal onset zone shown on extraoperative subdural grid electroencephalography in three patients. Cortical excision of the spike cluster focus was then performed in these six patients. The technique used here to resect MSI spike source clusters that correlate with the ictal onset zone by invasive subdural grid monitoring is illustrated in one patient who underwent cortical resection for epilepsy surgery. CONCLUSION: Three-dimensional reconstruction of MSI data linked to neuronavigation is a promising technique to facilitate resections around eloquent cortex in children with epilepsy.

The role of the insula in speech and language processing

Lesion and neuroimaging studies indicate that the insula mediates motor aspects of speech production, specifically, articulatory control. Although it has direct connections to Brocas area, the canonical speech production region, the insula is also broadly connected with other speech and language centres, and may play a role in coordinating higher-order cognitive aspects of speech and language production. The extent of the insulas involvement in speech and language processing was assessed using the Activation Likelihood Estimation (ALE) method. Meta-analyses of 42 fMRI studies with healthy adults were performed, comparing insula activation during performance of language (expressive and receptive) and speech (production and perception) tasks. Both tasks activated bilateral anterior insulae. However, speech perception tasks preferentially activated the left dorsal mid-insula, whereas expressive language tasks activated left ventral mid-insula. Results suggest distinct regions of the mid-insula play different roles in speech and language processing.

Neurophysiologic findings of neuronal migration disorders : Intrinsic epileptogenicity of focal cortical dysplasia on electroencephalography, electrocorticography, and magnetoencephalography

We define specific neurophysiologic characteristics for focal cortical dysplasia, a neuronal migration disorder. We reviewed data from published reports and our patients with focal cortical dysplasia. Our patients underwent preoperative scalp video-electroencephalography (EEG), magnetic resonance imaging (MRI), magnetoencephalography, and intraoperative or extraoperative electrocorticography monitoring. Scalp EEG showed trains of rhythmic epileptiform spike or sharp waves. Positive spikes correlated with early seizure onset, MRI lesion around the rolandic fissure, hemiparesis, and a less favorable outcome. Interictal electrocorticography showed continuous epileptogenic discharges: repetitive electrographic seizures and bursting discharges or continuous or quasicontinuous rhythmic spiking. Ictal electrocorticography showed paroxysmal fast and/or repetitive spiking. Magnetoencephalography showed clustered spike sources within and extending from the lesion. Cortical stimulation gave more frequent, lower-threshold afterdischarges and higher-threshold primary motor function. Focal cortical dysplasias are highly and intrinsically epileptogenic. For surgical seizure control, EEG, electrocorticography, and magnetoencephalography must delineate the intrinsic epileptogenic zone within and extending from the focal cortical dysplasia identified by MRI. (J Child Neurol 2005;20:357—363).

Resting-state hippocampal connectivity correlates with symptom severity in post-traumatic stress disorder

Post-traumatic stress disorder (PTSD) is a serious mental health injury which can manifest after experiencing a traumatic life event. The disorder is characterized by symptoms of re-experiencing, avoidance, emotional numbing and hyper-arousal. Whilst its aetiology and resultant symptomology are better understood, relatively little is known about the underlying cortical pathophysiology, and in particular whether changes in functional connectivity may be linked to the disorder. Here, we used non-invasive neuroimaging with magnetoencephalography to examine functional connectivity in a resting-state protocol in the combat-related PTSD group (n = 23), and a military control group (n = 21). We identify atypical long-range hyperconnectivity in the high-gamma-band resting-state networks in a combat-related PTSD population compared to soldiers who underwent comparable environmental exposure but did not develop PTSD. Using graph analysis, we demonstrate that apparent network connectivity of relevant brain regions is associated with cognitive-behavioural outcomes. We also show that left hippocampal connectivity in the PTSD group correlates with scores on the well-established PTSD Checklist (PCL). These findings indicate that atypical synchronous neural interactions may underlie the psychological symptoms of PTSD, whilst also having utility as a potential biomarker to aid in the diagnosis and monitoring of the disorder.

Molecular cytogenetic characterization of nasopharyngeal carcinoma cell lines and xenografts by comparative genomic hybridization and spectral karyotyping.

Nasopharyngeal carcinoma (NPC) cell lines and xenografts represent valuable models for functional and therapeutic studies on this common malignancy in Southeast Asia. The karyotypic information in most NPC cell lines and xenografts, however, remains largely unclear to date. We have characterized the chromosomal aberrations in six commonly used human NPC cell lines and xenografts using the molecular cytogenetic technique of comparative genomic hybridization (CGH). Genomic imbalances identified in cell lines were further correlated with structural abnormalities indicated from spectral karyotyping (SKY) analysis. CGH revealed consistent overrepresentations of 8q (six out of six cases) with a smallest overlapping region identified on 8q21.1 approximately q22. Other common gains included 7p (4/6 cases), 7q (4/6 cases), 12q (4/6), and 20q (4/6 cases), where minimal overlapping regions were suggested on 7p15 approximately p14, 7q11.2 approximately q21, and 12q22 approximately q24.1. Common losses were detected on 3p12 approximately p21 (4/6 cases) and 11q14 approximately qter (4/6 cases). Although SKY analysis on cell lines revealed predominantly unbalanced rearrangements, reciprocal translocations that involved chromosome 2 [i.e., t(1;2), t(2;3), and t(2;4)] were suggested. Furthermore, SKY examination illustrated additional breakpoints on a number of apparently balanced chromosomes. These breakpoints included 3p21, 3q26, 5q31, 6p21.1 approximately p25, 7p14 approximately p22, and 8q22. Our finding of regional gains and losses and breakpoints represents information that may contribute to NPC studies in vitro.