Human organotypic brain slice cultures: a detailed and improved protocol for preparation and long-term maintenance.

The investigation of the human brain at cellular and microcircuit level remains challenging due to the fragile viability of neuronal tissue, inter- and intra-variability of the samples and limited availability of human brain material. Especially brain slices have proven to be an excellent source to investigate brain physiology and disease at cellular and small network level, overcoming the temporal limits of acute slices. Here we provide a revised, detailed protocol of the production and in-depth knowledge on long-term culturing of such human organotypic brain slice cultures for research purposes. We highlight the critical pitfalls of the culturing process of the human brain tissue and present exemplary results on viral expression, single-cell Patch-Clamp recordings, as well as multi-electrode array recordings as readouts for culture viability, enabling the use of organotypic brain slice cultures of these valuable tissue samples for basic neuroscience and disease modeling (Fig. 1).

Dimensionality Reduction: Foundations and Applications in Clinical Neuroscience.

Advancements in population neuroscience are spurred by the availability of large scale, open datasets, such as the Human Connectome Project or recently introduced UK Biobank. With the increasing data availability, analyses of brain imaging data employ more and more sophisticated machine learning algorithms. However, all machine learning algorithms must balance generalization and complexity. As the detail of neuroimaging data leads to high-dimensional data spaces, model complexity and hence the chance of overfitting increases. Different methodological approaches can be applied to alleviate the problems that arise in high-dimensional settings by reducing the original information into meaningful and concise features. One popular approach is dimensionality reduction, which allows to summarize high-dimensional data into low-dimensional representations while retaining relevant trends and patterns. In this paper, principal component analysis (PCA) is discussed as widely used dimensionality reduction method based on current examples of population-based neuroimaging analyses.

Introduction to Machine Learning in Neuroimaging.

Advancements in neuroimaging and the availability of large-scale datasets enable the use of more sophisticated machine learning algorithms. In this chapter, we non-exhaustively discuss relevant analytical steps for the analysis of neuroimaging data using machine learning (ML), while the field of radiomics will be addressed separately (c.f., Chap. 18 -Radiomics). Broadly classified into supervised and unsupervised approaches, we discuss the encoding/decoding framework, which is often applied in cognitive neuroscience, and the use of ML for the analysis of unlabeled data using clustering.

Foundations of Time Series Analysis.

For almost a century, classical statistical methods including exponential smoothing and autoregression integrated moving averages (ARIMA) have been predominant in the analysis of time series (TS) and in the pursuit of forecasting future events from historical data. TS are chronological sequences of observations, and TS data are therefore prevalent in many aspects of clinical medicine and academic neuroscience. With the rise of highly complex and nonlinear datasets, machine learning (ML) methods have become increasingly popular for prediction or pattern detection and within neurosciences, including neurosurgery. ML methods regularly outperform classical methods and have been successfully applied to, inter alia, predict physiological responses in intracranial pressure monitoring or to identify seizures in EEGs. Implementing nonparametric methods for TS analysis in clinical practice can benefit clinical decision making and sharpen our diagnostic armory.

The Artificial Intelligence Doctor: Considerations for the Clinical Implementation of Ethical AI.

The applications of artificial intelligence (AI) and machine learning (ML) in modern medicine are growing exponentially, and new developments are fast-paced. However, the lack of trust and appropriate legislation hinder its clinical implementation. Recently, there is a clear increase of directives and considerations on Ethical AI. However, most literature broadly deals with ethical tensions on a meta-level without offering hands-on advice in practice. In this article, we non-exhaustively cover basic practical guidelines regarding AI-specific ethical aspects, including transparency and explicability, equity and mitigation of biases, and lastly, liability.

18F-FET-PET-guided gross total resection improves overall survival in patients with WHO grade III/IV glioma: moving towards a multimodal imaging-guided resection.

PURPOSE: PET using radiolabeled amino acid [18F]-fluoro-ethyl-L-tyrosine (FET-PET) is a well-established imaging modality for glioma diagnostics. The biological tumor volume (BTV) as depicted by FET-PET often differs in volume and location from tumor volume of contrast enhancement (CE) in MRI. Our aim was to investigate whether a gross total resection of BTVs defined as < 1 cm3 of residual BTV (PET GTR) correlates with better oncological outcome. METHODS: We retrospectively analyzed imaging and survival data from patients with primary and recurrent WHO grade III or IV gliomas who underwent FET-PET before surgical resection. Tumor overlap between FET-PET and CE was evaluated. Completeness of FET-PET resection (PET GTR) was calculated after superimposition and semi-automated segmentation of pre-operative FET-PET and postoperative MRI imaging. Survival analysis was performed using the Kaplan-Meier method and the log-rank test. RESULTS: From 30 included patients, PET GTR was achieved in 20 patients. Patients with PET GTR showed improved median OS with 19.3 compared to 13.7 months for patients with residual FET uptake (p = 0.007; HR 0.3; 95% CI 0.12-0.76). This finding remained as independent prognostic factor after performing multivariate analysis (HR 0.19, 95% CI 0.06-0.62, p = 0.006). Other survival influencing factors such as age, IDH-mutation, MGMT promotor status, and adjuvant treatment modalities were equally distributed between both groups. CONCLUSION: Our results suggest that PET GTR improves the OS in patients with WHO grade III or IV gliomas. A multimodal imaging approach including FET-PET for surgical planning in newly diagnosed and recurrent tumors may improve the oncological outcome in glioma patients.

Failed Neuroprotection of Combined Inhibition of L-Type and ASIC1a Calcium Channels with Nimodipine and Amiloride.

Effective pharmacological neuroprotection is one of the most desired aims in modern medicine. We postulated that a combination of two clinically used drugs-nimodipine (L-Type voltage-gated calcium channel blocker) and amiloride (acid-sensing ion channel inhibitor)-might act synergistically in an experimental model of ischaemia, targeting the intracellular rise in calcium as a pathway in neuronal cell death. We used organotypic hippocampal slices of mice pups and a well-established regimen of oxygen-glucose deprivation (OGD) to assess a possible neuroprotective effect. Neither nimodipine (at 10 or 20 µM) alone or in combination with amiloride (at 100 µM) showed any amelioration. Dissolved at 2.0 Vol.% dimethyl-sulfoxide (DMSO), the combination of both components even increased cell damage (p = 0.0001), an effect not observed with amiloride alone. We conclude that neither amiloride nor nimodipine do offer neuroprotection in an in vitro ischaemia model. On a technical note, the use of DMSO should be carefully evaluated in neuroprotective experiments, since it possibly alters cell damage.