Neonatal SEP - back to bedside with basic science

Scalp-recorded somatosensory evoked potentials (SEPs) have been successfully used in neonatal assessment for several decades. The current routine SEP paradigm is markedly predictive for future cerebral palsy (CP) or other neurocognitive sequelae in brain-injured babies. Recent advances in basic science have dramatically increased our knowledge about structural–functional development of SEP-related brain mechanisms. It has thereby become apparent that preterm SEP differs from that in more mature counterparts in that it also comprises responses from transient brain structures, and hence being unique to the preterm period. It is now obvious also that several aspects in the current SEP paradigm, ranging from the type of stimulation to the methods of recording and analysis, are suboptimal for preterm babies. Recent progress in recording and analysis techniques have made it possible to combine SEP studies with EEG recordings, as well as to implement advanced analyses (e.g. time–frequency analysis) into routine practice. This review summarizes literature from relevant areas in basic science, and proposes a novel, integrated approach in neonatal SEP studies in order to significantly increase the fidelity of testing somatosensory system.     

High frequency of TTK mutations in microsatellite-unstable colorectal cancer and evaluation of their effect on spindle assembly checkpoint

Frameshift mutations frequently accumulate in microsatellite-unstable colorectal cancers (MSI CRCs) typically leading to downregulation of the target genes due to nonsense-mediated messenger RNA decay. However, frameshift mutations that occur in the 3' end of the coding regions can escape decay, which has largely been ignored in previous works. In this study, we characterized nonsense-mediated decay-escaping frameshift mutations in MSI CRC in an unbiased, genome wide manner. Combining bioinformatic search with expression profiling, we identified genes that were predicted to escape decay after a deletion in a microsatellite repeat. These repeats, located in 258 genes, were initially sequenced in 30 MSI CRC samples. The mitotic checkpoint kinase TTK was found to harbor decay-escaping heterozygous mutations in exon 22 in 59% (105/179) of MSI CRCs, which is notably more than previously reported. Additional novel deletions were found in exon 5, raising the mutation frequency to 66%. The exon 22 of TTK contains an A(9)-G(4)-A(7) locus, in which the most common mutation was a mononucleotide deletion in the A(9) (c.2560delA). When compared with identical non-coding repeats, TTK was found to be mutated significantly more often than expected without selective advantage. Since TTK inhibition is known to induce override of the mitotic spindle assembly checkpoint (SAC), we challenged mutated cancer cells with the microtubule-stabilizing drug paclitaxel. No evidence of checkpoint weakening was observed. As a conclusion, heterozygous TTK mutations occur at a high frequency in MSI CRCs. Unexpectedly, the plausible selective advantage in tumourigenesis does not appear to be related to SAC.     

GAT-1 acts to limit a tonic GABA(A) current in rat CA3 pyramidal neurons at birth

Tonic activation of GABA(A) receptors takes place before the development of functional synapses in cortical structures. We studied whether inefficient GABA uptake might explain the presence of a tonic GABA(A)-mediated current (I(GABA-A)) in early postnatal hippocampal pyramidal neurons. The data show, however, that the tonic I(GABA-A) is enhanced by the specific blocker of GABA transporter-1 (GAT-1), NO-711 (1-[2-[[(Diphenylmethyleneimino]oxy]ethyl]-1,2,5,6-tetrahydro-3-pyridinecarboxylic acid hydrochloride), at birth in rat CA3 pyramidal neurons. NO-711 also prolonged the duration of GABA transients during endogenous hippocampal network events (known as giant depolarizing potentials) at postnatal day 0. The endogenous tonic I(GABA-A) was seen and it was enhanced by NO-711 in the presence of tetrodotoxin, which itself had only a minor effect on the holding current under control conditions. This indicates that the source of interstitial GABA is largely independent of action-potential activity. The tonic I(GABA-A) in neonatal CA3 pyramidal neurons was increased by zolpidem, indicating that at least a proportion of the underlying GABA(A) receptors contain gamma2 and alpha1-alpha3 subunits. The present data point to a significant role for GAT-1 in the control of the excitability of immature hippocampal neurons and networks.     

Determination of critical power using a 3-min all-out cycling test

PURPOSE: We tested the hypothesis that the power output attained at the end of a 3-min all-out cycling test would be equivalent to critical power. METHODS: Ten habitually active subjects performed a ramp test, two 3-min all-out tests against a fixed resistance to establish the end-test power (EP) and the work done above the EP (WEP), and five constant-work rate tests to establish the critical power (CP) and the curvature constant parameter (W') using the work-time and 1/time models. RESULTS: The power output in the 3-min trial declined to a steady level within 135 s. The EP was 287 +/- 55 W, which was not significantly different from, and highly correlated with, CP (287 +/- 56 W; P = 0.37, r = 0.99). The standard error for the estimation of CP using EP was approximately 6 W, and in 8 of 10 cases, EP agreed with CP to within 5 W. Similarly, the WEP derived from the 3-min test (15.0 +/- 4.7 kJ) was not significantly different from, and correlated with, W' (16.0 +/- 3.8 kJ; P = 0.35; r = 0.84). CONCLUSIONS: During a 3-min all-out cycling test, power output declined to a stable value in approximately the last 45 s, and this power output was not significantly different from the independently measured critical power.     

Epileptogenic neocortical networks are revealed by abnormal temporal dynamics in seizure-free subdural EEG

Long-term video electroencephalographic (EEG) recording is currently a routine procedure in the presurgical evaluation of localization-related epilepsies. Cortical epileptogenic zone is usually localized from ictal recordings with intracranial electrodes, causing a significant burden to patients and health care. Growing literature suggests that epileptogenic networks exhibit aberrant dynamics also during seizure-free periods. We examined if neocortical epileptogenic regions can be circumscribed by quantifying local long-range temporal (auto-)correlations (LRTC) with detrended fluctuation analysis of seizure-free ongoing subdural EEG activity in 4 frequency bands in 5 patients. We show here with subdural EEG recordings that the LRTC are abnormally strong near the seizure onset area. This effect was most salient in neocortical oscillations in the beta frequency band (14-30 Hz). Moreover, lorazepam, a widely used antiepileptic drug, exerted contrasting effects on LRTC (n = 2): lorazepam attenuated beta-band LRTC near the epileptic focus, whereas it strengthened LRTC in other cortical areas. Our findings demonstrate that interictal neuronal network activity near the focus of seizure onset has pathologically strong intrinsic temporal correlations. The observed effect by lorazepam on beta-band activity suggests that the antiepileptic mechanism of benzodiazepines may be related to the normalization of LRTC within the epileptic focus. We propose that this method may become a promising candidate for routine invasive and noninvasive presurgical localization of epileptic foci.     

Hierarchical Bayesian inference for the EEG inverse problem using realistic FE head models: depth localization and source separation for focal primary currents

The estimation of the activity-related ion currents by measuring the induced electromagnetic fields at the head surface is a challenging and severely ill-posed inverse problem. This is especially true in the recovery of brain networks involving deep-lying sources by means of EEG/MEG recordings which is still a challenging task for any inverse method. Recently, hierarchical Bayesian modeling (HBM) emerged as a unifying framework for current density reconstruction (CDR) approaches comprising most established methods as well as offering promising new methods. Our work examines the performance of fully-Bayesian inference methods for HBM for source configurations consisting of few, focal sources when used with realistic, high-resolution finite element (FE) head models. The main foci of interest are the correct depth localization, a well-known source of systematic error of many CDR methods, and the separation of single sources in multiple-source scenarios. Both aspects are very important in the analysis of neurophysiological data and in clinical applications. For these tasks, HBM provides a promising framework and is able to improve upon established CDR methods such as minimum norm estimation (MNE) or sLORETA in many aspects. For challenging multiple-source scenarios where the established methods show crucial errors, promising results are attained. Additionally, we introduce Wasserstein distances as performance measures for the validation of inverse methods in complex source scenarios.     

Netrin-4 Promotes Glioblastoma Cell Proliferation through Integrin ��4 Signaling

Netrin-4 is a laminin-related secreted molecule originally found to have roles in neuronal axon migration. Recent studies have indicated that netrin-4 also participates in the development of nonneural tissues and modulates tumor cell proliferation and tumor metastasis. Here we have explored the functions and molecular mechanisms of netrin-4 in glioblastoma multiforme. The suppression of netrin-4 expression in glioblastoma cell lines significantly reduced cell proliferation and motility and increased serum deprivation-induced apoptosis. Using tandem affinity purification combined with protein identification by mass spectrometry, we found that integrin β₄ interacts with netrin-4 and that it mediates mitogenic effects as well as AKT and mammalian target of rapamycin phosphorylation induced by netrin-4. Interestingly, netrin-4 acted as an inhibitor of cell proliferation in integrin β₄-silenced glioblastoma cells, and high concentrations of netrin-4 reduced cell proliferation. The negative effects of netrin-4 on proliferation were mediated by UNC5B. Analysis of more than 400 primary tumors from The Cancer Genome Atlas repository revealed that the expression of netrin-4 is significantly downregulated in glioblastoma and that the reduced expression is linked to poor patient survival time. The expression of integrin β₄ is increased in glioblastoma, and it predicts poor patient survival time. Current results illustrate a novel mechanism for glioma progression, where glioma cells reduce netrin-4 expression to decrease its inhibitory effects. In parallel, the expression of integrin β₄ is upregulated to sensitize the cells to low concentrations of netrin-4 for maintaining cell proliferation.