Checking a set of hypotheses

Mechanism. ATP-dependent P2X7 may link damage and inflammatory signaling to hyperexcitability. The effect is proposed to depend on network state; the contributions of neurons and glia are not considered established in advance.

Prediction. Support requires the inflammatory phenotype to be associated with a greater response to AFC after accounting for baseline severity, with an action consistent with P2X7 blockade. If the advantage disappears at comparable baseline activity, inflammation is not confirmed as an independent predictor. If AFC does not block P2X7 in the selected species, the network test is uninformative.

Mechanism. GSK2795039 inhibits NOX2, one source of reactive oxygen species. Feedback between oxidative changes and excitability is proposed. Stopping discharges and protecting against their consequences are treated as distinct possible effects.

Prediction. Support requires reduced electrical burden following late intervention with confirmed action on NOX2. Replication using an independent tool distinguishes the mechanism from an off-target effect of the compound. If ROS and damage decrease but discharges persist, the role of NOX2 in sustaining activity in this model is rejected, while neuroprotection may remain a separate result.

Mechanism. LY487379 is a positive allosteric modulator of mGlu2. In characterized systems, it enhanced the action of glutamate without an independent agonist effect. Dependence on the endogenous signal may provide selectivity for the activity pattern, but does not guarantee it.

Prediction. Support requires confirmed functional mGlu2, better preservation of normal responses than with the comparator at an equal network effect, and dependence on GRM2 without impairment of inhibitory transmission. The proposed advantage is rejected if disinhibition appears, there is no benefit at an equal network effect, or the action does not depend on GRM2.

Mechanism. KCC2 promotes chloride extrusion from neurons. Restoration of the GABA response reversal potential may restore the effectiveness of GABAergic inhibition.

Prediction. 350 corrects the functional Cl⁻ defect, followed by restoration of diazepam action and reduction of pathological electrical discharges. An independent intervention in KCC2 with its own control is added for a causal conclusion. If the gradient is corrected but diazepam remains ineffective, the specific hypothesis is not supported. If the defect is not reproduced, a negative network result does not yet test the hypothesis.

Mechanism. Activation of Kv7 potassium channels limits excitability, while blockade of AMPA receptors reduces the spread of excitation. The possibility of reducing the burden on each mechanism is being tested.

Prediction. The combination preserves evoked responses better at a comparable reduction in discharges and without loss of viability, with the advantage confirmed in a new independent series. The claimed advantage is rejected if the effect is explained by deeper network suppression or if the difference disappears after matching the antiepileptiform effect.

Mechanism. The proposed chain is albumin exposure → astrocytic TGF-β signaling → impaired potassium buffering → persistent hyperexcitability. SB-431542 is used as a tool, but blocks ALK4/5/7 rather than only ALK5.

Prediction. Support requires early intervention to prevent persistent hyperexcitability after compound removal and preserve functional K⁺ clearance; late intervention separately assesses the reversibility of the changes. If the effect disappears after complete washout, it does not represent prevention of persistent remodeling. If hyperexcitability decreases without restoration of K⁺ control, the potassium explanation specifically is rejected.

Mechanism. LDN/OSU-0212320 has been reported to increase EAAT2 translation with involvement of PKC–YB-1. The functional target is glutamate transport; this does not imply direct binding of the compound to EAAT2.

Prediction. Glutamate transport and network function improve together; whether the effect is stronger in cultures with a baseline transport deficit is assessed. If uptake increases but discharges remain unchanged, the explanation based on insufficient glutamate clearance in this model is rejected. If discharges decrease without increased uptake, the EAAT2 mechanism is not established.

Mechanism. SK channels couple increased intracellular calcium to a potassium current and subsequent hyperpolarization. CyPPA increases their sensitivity to calcium. Discharge suppression and disruption of calcium homeostasis may be separable in time.

Prediction. Support requires reproducibly identifying an interval in which electrical discharges decrease, normal responses are preserved, and basal Ca²⁺ and delayed damage do not worsen. The useful-window hypothesis is rejected if every effective regimen simultaneously increases basal Ca²⁺, damages cells, or suppresses normal responses equally.

Mechanism. ADK inhibition may increase adenosine availability. Subsequent activation of A₁ receptors limits excitatory transmission. Selectivity according to network state is an assumption to be tested.

Prediction. ABT reduces hyperactivity while relatively preserving ordinary responses, and the effect is consistent with A1 dependence and confirmed action on ADK/adenosine. Loss of the effect under A1 blockade supports the pathway but does not directly measure ADK inhibition. Selective inhibition is rejected if the ordinary network is suppressed at least as much as the pathological network; absence of A1 dependence requires reconsidering the proposed mechanism.

Mechanism. NBI-921352 preferentially blocks inactivated Naᵥ1.6. Relative sparing of Naᵥ1.1 may reduce suppression of some classes of interneurons.

Prediction. At an equal network effect, NBI better preserves the firing capacity of identified inhibitory cells and functional inhibition. The advantage is rejected if both drugs suppress inhibitory cells equally or if the advantage disappears at comparable network suppression.