Mars Perseverance Sol 1515: Right Mastcam-Z Camera Image Credit: NASA/JPL-Caltech/ASU Larger image
A PhD student and his supervisor have developed a simple way for testing for active life on Mars and other planets using equipment already on the Mars Curiosity rover and planned for future use on the ExoMars Rosalind Franklin rover.
There is enormous interest in the possibility of past or present life beyond Earth, with space agencies spending a great deal of time and money exploring suitable extraterrestrial homes and searching for signals of life.
PhD student Solomon Hirsch and his supervisor Professor Mark Sephton, from Imperial College London’s Department of Earth Science & Engineering, have realised that an existing instrument could be used to detect signs of life at a fraction of the cost of developing new missions and instruments.
It has the potential to be used to detect living organisms on other planets or moons. The instrument, called a gas chromatograph-mass spectrometer (GC-MS), has been installed on Martian probes since the mid-1970s with early versions on the Viking I and Viking II landers. Solomon and Mark determined that it could be used to detect a chemical bond within cell membrane molecules that are found in many living, and very recently deceased, organisms.
The research is published in the journal Nature Space Exploration. (Intact polar lipids as organic biomarkers of viable extraterrestrial life)
“Space Agencies such as NASA and ESA don’t know their instruments can already do this,” said Professor Sephton. “Here we have developed an elegant method that rapidly and reliably identifies a chemical bond that shows the presence of viable life,” he says. “The Curiosity rover just turned 13 on Mars, but who says you can’t teach an old dog new tricks.”
The new method detects a unique sequence of atoms that bind the constituent molecules of the external membranes of living bacteria and eukarya cells. These constitute the vast majority of biological matter on Earth and include the kinds of lifeforms the scientists would also expect to find beyond our planet.
The signatures of these bonds found in molecules called intact polar lipids (IPLs) show up as a clear spike in a graph produced by the GC-MS instrument (see figure).
Solomon says: “When we put the intact polar lipid compounds into our GC-MS we didn’t know what to expect because these compounds are usually analysed using other techniques. The characteristic signature we identified provides a clear indicator of viable life using space-ready equipment already in use on many extraterrestrial missions. If we find signs of life beyond Earth, the first question will be: is it living right now? It’s thrilling to think that the technique we developed here could be used to help answer that question.”
Once an organism dies, its IPL bonds disintegrate within hours, after which time they can no longer be detected, and a spike no longer appears in the instrument readout.
The method is not only useful for detecting life elsewhere in the solar system but also for protecting life back here on Earth. Groups of scientists from around the world are planning to spend multiple millions of dollars, to detect signs of active life in samples returned from Mars. Their task will be made easier with a quick and simple method to screen for life.
Professor Sephton says, “Our active life detection method could be deployed on Mars and the plumes of icy moons in the outer solar system from where the data can be sent back to Earth for interpretation, or in samples returned to Earth from potential alien biospheres.”
Solomon says: “Our expectation of finding things alive on the Martian surface is low due to the hostile temperature and radiation conditions. Still, we aren’t ruling out the possibility – life finds amazing ways to survive in extreme circumstances. Furthermore, future missions such as the ExoMars mission plan to drill metres deep into the surface of the planet where the likelihood of finding active life is significantly higher.”
Intact polar lipids as organic biomarkers of viable extraterrestrial life, NPJ Space Exploration (open access)
Neural organoids were differentiated from iPSC-derived Neural Progenitor Cells (NPC) for up to 14 weeks and characterized throughout development (Fig. 1a). Gene expression of synaptic plasticity markers was quantified from week 0 to week 12. Calcium signaling development was analyzed from week 2 to week 14. Finally, electrical activity was characterized by High-Density Microelectrode Arrays (HD-MEAs) over two independent time periods, from weeks 6-to-9 and 10-to-13. In addition, pharmacological modulation of neurotransmission was performed at weeks 8 and 13. Lastly, input-specific Theta Burst Stimulation (TBS) was implemented at week 14 to induce synaptic plasticity. To analyze both spontaneous and evoked electrical activity from the HD-MEA data, functional connectivity and criticality analysis were performed. A schematic overview of the neurocomputational investigations is shown in Fig. 1b. In addition, an example of how evoked activity from pharmacological or electrical stimuli can modulate synaptic transmission to induce synaptic plasticity is shown in Fig. 1c. In long-term potentiation, IEGs play a role in trafficking of glutamatergic receptors into the postsynaptic terminal (Fig. 1c), therefore expression of these IEGs can serve as a molecular marker for long-term memory27,37.
Fig. 1: Schematic overview of the experimental approach.
a Experimental timeline. Created in BioRender. Alam El Din, D. (2025) https://BioRender.com/v4k2lpzb Overview of avalanche and network connectivity analysis for time series electrophysiology data from organoids plated on HD-MEAs. Created in BioRender. Alam El Din, D. (2025) https://BioRender.com/trj7ehfc Schematic representation of synaptic transmission modulation by pharmacological and electrical stimuli to induce synaptic plasticity. Adapted from Kim, S. (2025). Long-Term Potentiation. https://app.biorender.com/biorender-templates/details/t-61006a6924e0d000a40de3a1-long-term-potentiation. Created in BioRender. Alam El Din, D. (2025) https://BioRender.com/eqpwhdw.
Neural organoids develop proper synapse formation and express receptors necessary for synaptic transmission
Neural organoids were differentiated following our in-house established protocol11. The expression of markers for astrocytes (GFAP), oligodendrocytes (MBP, OLIG2) and mature neurons (MAP2) increased in the first 8 weeks of maturation and then plateaued in the following weeks, indicating that the differentiation predominantly occurs rapidly until week 8 and then reaches a more stable, mature cell composition from week 8 to 12 (Supplementary Fig. 1). Hence, two time points were selected for the experiments in this paper.
RNA-sequencing (RNAseq) revealed some interesting trends in gene expression between week 8 and week 12/13 organoids. Firstly, while many GABAergic markers showed no difference in expression (e.g., PVALB, GABRA1, LHX6), we observed a slight trend in downregulation of several GABAergic markers, such as SST, SLC32A1, and GAD 1 and 2 in the week 12/13 organoids compared to the week 8 group, as shown in Supplementary Fig. 2a and b.
By comparing expression levels across main cell lineages and brain regions (Supplementary Figs. 3 and 4), we observe an increase in the expression of astrocyte markers (e.g, AQP4, GFAP, HOPX, S100B, and SLC1A3) in the week 12/13 group compared to week 8. These data suggest that the observed trends in downregulation in expression of GABA and glutamatergic markers (GRIN3A, GRIN3B, and SLC17A7) may be linked to the shift in cellular populations with more astrocytes (based on the higher expression of astrocyte markers in the week 12/13 organoids), (Supplementary Figs. 2 and 3). We also profiled the expression of brain region markers, which showed the presence of genes across the forebrain, hindbrain, and midbrain and no consistent differences in expression between the two age groups (Supplementary Fig. 4).
These findings are further supported by the functional enrichment analysis (Supplementary Fig. 5), which revealed that genes associated with synaptic signaling, neuronal differentiation, and axonogenesis were downregulated and cell cycle genes were upregulated in the week 12/13 group (Supplementary Fig. 5).
We then evaluated the presence of pre- and postsynaptic markers as well as IEGs as the key proteins involved in synaptic plasticity and learning formation. Presence of the presynaptic marker Synaptophysin (SYP) and postsynaptic marker HOMER1 was detected in both week 8 and 12 organoids, displaying typical punctual staining (Fig. 2a). Gephyrin-positive signal was close to background with few positive cells at week 8 and increased at week 12 (Fig. 2b). This indicates that there are more developed inhibitory synapses at the later stage of differentiation. Gene expression of GABRA1, which encodes the inhibitory GABAA receptor, followed the same pattern (Fig. 2c). Gene expression of postsynaptic marker HOMER1 was steady over time (Fig. 2c). Presence of Synapsin1 and MAP2 seemed to be reduced at week 12 vs. week 8, supporting RNAseq data of the lineage shift towards more astrocytes (Fig. 2b), although cell number quantification is needed to drive the final conclusions.
Fig. 2: Expression of glutamatergic and GABAergic receptor and synaptic plasticity-related genes in neural organoids over course of differentiation.
a Representative immunocytochemistry images of organoids showing postsynaptic marker (HOMER1) and presynaptic marker (SYP) in 8- and 12-week cultures. In composite images, HOMER1 is shown in blue, and SYP is shown in yellow. Scale bars are 100 µm, 50 µm, and 10 µm, respectively. b Presence of inhibitory post-synaptic marker (Gephyrin), pre-synaptic marker (SYN1) and dendrites (MAP2) in 8- and 12-week organoids. In composite images, Gephyrin is shown in blue, SYN1 in yellow, and MAP2 in grey. Scale bars are 100 µm and 50 µm, respectively. For a, and b, all images were taken at 20x, 100x, and 100x + 4x zoom and processed with ImageJ for visualization. c Gene expression of Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha1 (GABRA1), Glutamate Ionotropic Receptor NMDA Type Subunit 1 (GRIN1), Glutamate [NMDA] Receptor Subunit Epsilon-1 (GRIN2A), and Glutamate [NMDA] Receptor Subunit Epsilon-2 (GRIN2B), Glutamate Ionotropic Receptor AMPA Type Subunit 1 (GRIA1), homer scaffold protein 1 (HOMER1) in organoids over the course of differentiation. d Representative immunocytochemistry images of weeks 8 and 12 organoids stained for Neuronal Pentraxin 2 (NPTX2), Activity-Regulated Cytoskeleton-associated protein (ARC), cAMP response element-binding protein (CREB), and Brain-Derived Neurotrophic Factor (BDNF). Scale bar is 100 µm. e Gene expression over the course of differentiation of immediate early genes (IEGs) ARC, BDNF, Neuronal PAS Domain Protein 4 (NPAS4), NPTX2, Fos proto-oncogene AP-1 transcription factor subunit (FOS), and Early Growth Response 1 (EGR1). f Gene expression of synaptic slasticity- related genes: CREB, calcium/calmodulin-dependent protein kinase II A (CAMK2A), Synaptic Ras GTPase-activating protein 1 (SYNGAP1). g Gene expression of synaptic plasticity -related miRNAs. For all gene expression plots, data is shown as a box and whisker plot (with the box extending from the 25th to 75th percentiles) and represented as log2(Fold Change) normalized to NPCs from 2-3 independent experiments with 3 technical replicates each. In all qPCR experiments, ACTB was used as a reference gene.
Both AMPA and NMDA receptors play an important role in synaptic plasticity, including STP/LTP24,38,39, therefore showing expression of these receptors was imperative for this study to give insight into the mechanisms of learning and memory in neural organoids. The increase in gene expression over time was the greatest for GRIN1, which plateaued around week 8 to week 12 (Fig. 2c). GRIN2A and GRIN2B both increased over time with a higher increase of GRIN2A expression than GRIN2B, suggesting increasing maturity40(Fig. 2c). GRIA1 expression also increased over time and plateaued after week 8 (Fig. 2c). Thus, plateau in expression of these subunits suggests the organoids reached a mature state between week 8 to 12.
Dynamic expression of immediate early genes associated with synaptic plasticity and cognitive functions over time
IEGs are crucial for cognitive functions as they act directly at the synapse and mediate the cellular processes that are essential for learning and memory consolidation27. To demonstrate that the neural organoids have the necessary cellular components for cognitive processes, we quantified IEG expression during the course of differentiation (Fig. 2d, e, Supplementary Fig. 6). Gene expression of ARC, BDNF, NPAS4, NPTX2, and FOS was significantly increased over time, while EGR1 was already expressed in NPCs and remained at levels close to those in NPCs. Expression of upstream regulators of IEGs (CREB and CAMK2A) also increased over time with the largest increase in expression of CAMK2A (Fig. 2f). In addition, SYNGAP1, which plays a key role in regulating synaptic function and plasticity41, was stably expressed throughout the course of differentiation, starting from NPCs. The levels of IEG proteins (NPTX2, ARC, and BDNF) and upstream IEG transcription factor CREB were comparable between week 8 and 12, confirming the plateau observed in RT-qPCR data (Fig. 2d). These results were consistent with RNAseq data which showed no changes in IEG expression between 8 and 12/13 week old organoids (Supplementary Fig. 6a and Supplementary Fig. 6b).
Finally, we assessed the expression of microRNAs known to be involved in synaptic plasticity (Fig. 2g)42 and observed expected dynamics. A strong increase in expression of mir-124-3p over the course of differentiation was observed. mir-132-3p and mir-134-3p had opposite expression patterns: mir-132-3p was increased over time while expression of mir-134-3p was first strongly induced from NPC to 2 weeks of differentiation and was downregulated thereafter (Fig. 2g).
Evidence of spontaneous electrical activity and highly interconnected neuronal networks in neural organoids
Electrophysiology over the course of organoid development was characterized using calcium imaging and HD-MEAs. In addition to the expression of molecular machinery involved in synaptic plasticity, neural organoids showed highly patterned spontaneous electrical activity (Figs. 3 and 4). Calcium transients were measured using Fluo-4 biweekly from week 2 to 14. Whole organoid change in fluorescence over resting fluorescence intensity (∆F/F) was quantified and compared across age groups (Fig. 3a). From these ∆F/F plots, the average rise time, peak amplitude, firing rate, decay time, burst duration, number of peaks, and percentage of active organoids per time point were calculated (Fig. 3b). Bursts were identified as peaks in calcium transients. Burst firing rate was calculated as the number of burst peaks per second.
Fig. 3: Neural organoid calcium oscillatory dynamics across different time points to show maturation of spontaneous network bursting.
a Representative changes in fluorescence over resting fluorescence (∆F/F) graphs across 360 seconds for each time point from week 2 (W2) to week 14 (W14) of differentiation. b Average rise time, peak amplitude, firing rate, decay time, burst duration, number of peaks, and percentage of active organoids shown across different time points. At least 8 individual organoids across at least 3 independent experiments were imaged and quantified for each time point. Data is shown as box and whisker plots (with the box extending from the 25th to 75th percentiles). Statistics were performed using one-way ANOVA and a Tukey post-hoc test. Changes over time were significant for rise time (p < 0.05), burst firing rate (p < 0.0001), peak amplitude (p < 0.0001), decay time (p < 0.01), burst duration (p < 0.001), and total number of peaks per organoid (p < 0.0001). Pairwise comparisons are shown on the figure: # = Significant difference from week 4, Ŧ = Significant difference from week 6, $ = Significant difference from week 8, ¥ = Significant difference from week 10, ȼ = Significant difference from week 12, • = Significant difference from week 14, * = Significant difference from all weeks. For exact p values see Supplementary Tables 4–9. See also Supplementary Fig. 7 for single neuron calcium imaging analysis.
Fig. 4: Changes in spontaneous electrical activity in neural organoids throughout development.
Representative raster plots and active area plots from HD-MEA recordings showing spontaneous electrical activity over time during a weeks 6-to-9 and b weeks 10-to-13 of differentiation. DOM: Days on MEAs. c Network dynamic metrics from both organoid age groups over time (blue line represents 6-to-9 week organoids, red line – 10-to-13 week organoids. The line shown represents mean and the shaded region represents the standard deviation plotted from 2 independent experiments with 5 to 6 HD-MEA wells per group per experiment with 2–5 organoids per well (n = 11–12 wells per age group). Statistics were performed using a mixed-effects model with matching and a Tukey post-hoc test. p < 0.05 was considered significant. For exact p values from pairwise comparisons, see the Supplementary Data 3 file. ISI: Interspike Interval. IBI: Interburst Interval.
Calcium imaging showed that 2-week-old organoids did not exhibit spontaneous oscillatory calcium dynamics. The first signs of oscillatory calcium activity were detected at week 4, with high-frequency oscillations at weeks 4 and 6, as shown by higher burst firing rates and number of peaks (Fig. 3, Supplementary Video 1, and Supplementary Fig. 7). At week 8, oscillation patterns shifted to lower frequency with fewer calcium peaks, lower burst firing rates, higher amplitudes, longer burst durations, and larger decay times (Fig. 3, Supplementary Video 2, and Supplementary Fig. 7). The plateau shape of the oscillations at week 8 indicated multiple neuronal action potentials contributing to the calcium oscillation (Fig. 3a). The decrease in the number of peaks from weeks 6 to 8 suggested more synchronous calcium transients, indicating a more densely connected mature network. From weeks 10 to 14, burst duration, decay time, and number of peaks did not change significantly, but amplitude and percentage of active organoids decreased, suggesting a plateau in differentiation around week 8.
In addition to whole organoid analysis, ∆F/F was quantified in single neurons for at weeks 4–10 (Supplementary Fig. 7). Maximum intensity z-projections of time course videos showed that neuronal networks at weeks 4 and 6 were less connected compared to weeks 8 and 10 (Supplementary Fig. 7). At weeks 4 and 6, neurons spiked at higher frequencies and with less synchronization (Supplementary Fig. 7a and b). By weeks 8 and 10, larger burst amplitudes and longer burst durations are likely contributed to multiple action potentials across different neurons, which were spiking simultaneously (Supplementary Fig. 7c and d). At week 10, the propagation of an action potential across connected neurons was observed by the slightly delayed peak burst amplitude of region of interest (ROI) 1 compared to ROIs 2 and 3 (Supplementary Fig. 7d).
To measure network activity over time, HD-MEAs were used to obtain high spatial and temporal resolution of organoids’ electrical activity across two different time periods (weeks 6-to-9 and 10-to-13) (Fig. 4). Representative raster plots indicated differences in spontaneous electrical activity in organoids depending on their age (Fig. 4a, b). The 6-to-9-week organoids have a significantly higher burst frequency, number of spikes within burst, and percent active area than those in the later time point group (Fig. 4c). They also had significantly shorter interburst intervals compared to the more mature group, consistent with the calcium imaging data in Fig. 3.
To further assess the organoids’ functionality, neuronal connectivity and criticality were quantified from the same HD-MEA time course data (Figs. 5 and 6). In both age groups, changes in functional connections between electrodes were observed over time on the HD-MEA (Fig. 5a). Denser connections and more active electrodes were observed in the 10-to-13-week group compared to the 6-to-9-week group, as denoted by the thickness of the black lines and red electrodes, respectively in the connectivity graphs (Fig. 5a). However, while both groups showed significant increases in the number of nodes over time, the 10-to-13-week group had a significantly lower number of nodes overall in their functional connectivity matrices compared to the 6-to-9-week group (Fig. 5b). To quantify the shift in the strength of the edges over time, an edge weight distribution was calculated by measuring the fraction of total possible edges that are realized (Fig. 5c). Interestingly, most edges were activated across all samples over time (Fig. 5c). The 10-to-13-week group showed no significant changes over time, while the 6-to-9-week group showed a temporary significant decrease in strength of edges at day on MEA (DOM) 7 (Fig. 5c). Finally, the organoid’s modularity was significantly different across age groups and significantly decreased in both age groups over time, indicating that the networks started with multiple communities but then became more of a single community over time (Fig. 5d). The decrease in modularity may also be due to an increased number of nodes. Despite the similarity in modularity, the 10-to-13-week group maintained a significantly higher modularity over time, indicating that it maintained more communities or network connections (Fig. 5d).
Fig. 5: Neural organoids show highly interconnected neuronal networks and criticality throughout development.
a Representative plots of functional connectivity at day on MEA (DOM) 3, 9, 15, and 21 for the week 6-to-9 and week 10-to-13 old organoids. For clarity of visualization, only the 200 connections (edges) with the highest mutual information are shown. Each red dot represents an electrode, and the lines indicate the connections between electrodes. The thickness of the line indicates the weight of connectivity. b Average number of nodes; c Average fraction of total possible edges; d Average modularity over time in week 6-to-9 and week 10-to-13 organoids. e Deviation from Criticality Coefficient (DCC). f Branching Ratio (BR) g Shape collapse error (SCe) over time in 6-to-9 week and 10-to-13 week old organoids. In b–d the line shown represents the mean and the shaded region represents the standard deviation plotted from 2 independent experiments with 5 to 6 HD-MEA wells per group per experiment with 2–5 organoids per well (n = 11–12 wells per age group). Panels e–g show regression lines (blue line- 6–9 week old organoids, red line – 10-13 week old organoids) with a 95% confidence interval. Data plotted is from 2 independent experiments with 5-6 HD-MEA wells per group per experiment (n = 11–12 wells per age group). Statistics were performed using a two-way ANOVA and a Tukey post-hoc test. p < 0.05 was considered significant. For exact p values from pairwise comparisons, see the Supplementary Data 3 file.
Fig. 6: Pharmacological characterization of synaptic transmission changes of neuronal spiking and bursting activity and Immediate Early Gene expression.
Expression of ARC, NPAS4, FOS, and EGR1 after 2 hours of exposure to 20 µM AP5 + 20 µM NBQX (pink box), 10 µM bicuculline (green box) and 100 µM 4-AP (purple box) in a 8-week and b 13-week-old organoids, represented as box and whisker plots (25th to 75th percentiles) and as log2(Fold Change) normalized to negative control (organoids with no chemical treatment = 2 h control). ACTB was used as a reference gene. The data represents 3 independent experiments with 2 technical replicates each for 8 weeks and 4-5 independent experiments with 2 technical replicates each for the 13-week time point. Statistics were calculated based on the replicate average from each independent experiment, with one-way ANOVA and post-hoc Dunnett’s tests *p < 0.05, ***p < 0.001, ****p < 0.0001 c Representative raster plots from MEA recordings in 13-week-old organoids (from 6 wells per condition) before and after treatment with bicuculline, 4-AP, and NBQX + AP5. d Burst frequency, Interburst interval coefficient of variation, burst duration, and percentage of spikes within bursts plotted as box and whisker plot (with the box extending from the 25th to 75th percentiles) for bicuculline, 4-AP, and NBXQ + AP5 treated wells prior to (baseline), 0 mins, 2 hours, and 4 hours after exposure. The data represents 3 independent experiments with 2 HD-MEA wells per experiment per chemical (n = 6). Statistical significance was calculated with repeated measures ANOVA with post-hoc Dunnett tests. p < 0.05 was considered significant. Pairwise comparisons can be seen in the Supplementary Tables 10-21 and significant groups are shown in the figure. ARC – Activity-Regulated Cytoskeleton-Associated Protein; NPAS4 – Neuronal PAS Domain Protein 4; FOS – Fos proto-oncogene AP-1 transcription factor subunit; EGR1 – Early Growth Response Protein 1; AP5 – 2-Amino-5-phosphonopentanoic acid (an NMDA receptor antagonist); NBQX – 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f]quinoxaline (an AMPA receptor antagonist).
Criticality is a state in which complex systems such as a brain operates at the critical point between organization and randomness, demonstrating how neuronal networks may navigate between the two stages of chaos and order43. The critical point state is key for brain functionality, as it operates at its optimal and most efficient computational capacity and is highly sensitive to external stimuli during this stage. Organoids exhibited properties of criticality over the course of differentiation (Fig. 5e–g). The more mature 10-to-13-week group showed a consistently lower and more tightly regulated Deviation from Criticality Coefficient (DCC) value and higher Branching Ratio (BR), approaching 1, compared to the 6-to-9-week group (Fig. 5e). While the BR in the 10-to-13-week group decreased non significantly over the period of 3 weeks on the HD-MEAs, the 6-to-9-week group gradually increased significantly, demonstrating maturation and pursuit of criticality and thus a stable state (Fig. 5f). Additionally, the Shape Collapse error (SCe) for the 10-to-13-week group was significantly lower than that of the 6-to-9-week group, indicating a more accurate scaling of avalanches of varying durations to an universal shape in the 10-to-13-week group (Fig. 5g). This analysis suggests that the 10-to-13-week group was in a more critical state compared to the 6-to-9-week group. However, over time, both the BR and SCe appeared to converge for both groups, suggesting that the 6-to-9-week group exhibited increasingly critical dynamics, while the 10-to-13-week group showed diminishing critical dynamics on the MEA over time.
Pharmacological characterization of synaptic transmission changes neuronal bursting activity and immediate early gene expression
To validate reactiveness to network modulations, pharmacological agents were used to cause neuronal depolarization and disrupt excitatory glutamatergic synaptic transmission. Expression of IEGs and synaptic plasticity-related genes was measured 2 hours after exposure to pharmacological agents and compared to the corresponding untreated control in two age groups (8 weeks and 13 weeks) (Fig. 6). To disrupt excitatory glutamatergic synaptic transmission, organoids were treated with 2,3-dioxo-6-nitro-7-sulfamoyl-benzo[f]quinoxaline (NBQX), an AMPA receptor antagonist, D-2-amino-5-phosphonovalerate (AP5), a NMDA receptor antagonist, 4-Aminopyridine (4-AP), a voltage-gated potassium (Kv) channel antagonist, and bicuculline, a GABA receptor antagonist, were used to enhance neuronal depolarization and synaptic transmission (Fig. 6).
Bicuculline induced a slight increasing trend in gene expression across both age groups (Fig. 6a), while exposure to 4-AP led to significant changes in NPAS4 and FOS expression at both age groups. Expression of ERG1 was significantly induced only at week 13. Lastly, ARC expression showed an increased trend in expression after 4-AP exposure (Fig. 6a). No significant changes in gene expression were seen after exposure to NBQX and AP5 individually or combined (Fig. 6 and Supplementary Fig. 8a–d). Since IEGs were more strongly perturbed at week 13, the effects of these chemicals on electrophysiological activity were assessed in this age group. Organoids were exposed to the pharmacological agents directly on the HD-MEA at DOM 29. Network recordings were taken before the addition of the chemicals as a baseline. Network activity was then recorded immediately after exposure, followed by 2 and 4 hours thereafter, after which the recorded parameters were compared to baseline activity (Fig. 6c and d). 4-AP and bicuculline increased network activity while NBQX + AP5 decreased network activity over time (Fig. 6c). More specifically, bicuculline caused an insignificant increasing trend in mean burst frequency and interburst interval coefficient of variation (CV) over time, a significant increase in percent of spikes within bursts 0 minutes after and an increasing trend in percent of spikes within bursts 2 and 4 hours after exposure. In addition, bicuculline caused no significant changes or trends in burst duration over time. 4-AP exposure caused a significant increase in mean burst frequency and an increasing trend in mean percent of spikes within bursts 0 minutes after. In addition, the percentage of spikes within bursts maintained an increasing trend within 2 and 4 hours after exposure. 4-AP also caused a decreasing trend in burst duration that was maintained over time. Finally, 4-AP caused no significant changes or trends in interburst interval CV over time. Additionally, NBQX + AP5 exposure completely abolished network bursting activity (Figs. 6c and d, Supplementary Fig. 8). Overall, NBQX + AP5 significantly decreased mean burst frequency, interburst interval CV, burst duration, and percentage of spikes within bursts from 0 minutes to 4 hours. Interestingly, we found that NMDA receptors are largely responsible for neuronal network bursting, as exposure to only AP5 was enough to abolish the bursting, while blocking only AMPA receptors with NBQX only partially reduced the bursting (Supplementary Fig. 8). These results agreed with previous reports showing that ketamine and xenon, which act on NMDA receptors, lead to burst silencing and reduction in vitro44,45. No changes in firing rate, spikes per burst, and burst duration were seen after NBQX application alone, but when AP5 or NBQX + AP5 was applied, no bursts were observed; therefore, firing rate, spikes per burst, and burst duration were not quantifiable (Supplementary Fig. 8).
While NBQX + AP5 reduced the network bursting, some spiking activity was still seen (Fig. 6c and Supplementary Fig. 8e). To elucidate which type of neurons contributed to the remaining activity, different modulators of the glutamatergic, GABAergic, dopaminergic, noradrenergic and cholinergic neurons were tested in different combinations (Supplementary Fig. 9). The first set of experiments further blocked NMDA and AMPA receptors, by increasing concentrations of NBQX + AP5 to 40 µM. As for the previous 20 µM exposure (Fig. 6c and Supplementary Fig. 8e), all bursting was abolished and spiking was diminished, but increased concentration did not remove the activity completely. Therefore, next, we blocked different types of neurons. First, 10 µM bicuculline and 10 µM CPG 55845 hydrochloride (CPG 55845) were added to block GABA(A) and GABA(B) receptors in addition to NMDA and AMPA receptors. Upon addition of bicuculline and CPG 55845, a further reduction in spiking activity was observed (Supplementary Fig. 9a, last panel, 10–20 min). The quantification of the effect after blocking excitatory receptors and the subsequent blocking of inhibitory neurons is shown in Supplementary Fig. 9d.
Next, inhibitory and excitatory receptors were blocked simultaneously by adding 40 µM NBQX, 40 µM AP5, 10 µM bicuculline, and 10 µM CPG 55845 at the same time with similar effects as before (Supplementary Fig. 9b). Subsequently, 5 µM 6-hydroxydopamine (6-OHDA) and 10 µM haloperidol were added, which induce lesions in dopaminergic (and noradrenergic) neurons and block dopamine D2 receptors, respectively. This resulted in an even more pronounced reduction in spiking activity (Supplementary Fig. 9b, the last panel). Supplementary Fig. 9e quantifies the corresponding changes in bursting and spiking metrics. These results suggest that most of the remaining activity originated from dopaminergic and noradrenergic neurons.
The last population of neurons examined for chemical modulation effects was cholinergic neurons. To achieve this, 40 µM NBQX, 40 µM AP5, 10 µM bicuculline, 10 µM CPG 55845, 5 µM 6-OHDA, and 10 µM haloperidol were added simultaneously to block excitatory, inhibitory, dopaminergic and noradrenergic neurons. The same effect as shown in Supplementary Fig. 9b was also observed in Supplementary Fig. 9c, demonstrating a drastic reduction in bursting and spiking, as quantified in Supplementary Fig. 9f. Finally, 1 nM chlorpyrifos (CPF)-Oxone was added to that same well to attempt to increase activity in the network by activating cholinergic neurons. However, no changes in bursting or spiking were observed, as shown in Supplementary Fig. 9c and quantified in Supplementary Fig. 9f. Therefore, findings suggest that the remaining activity likely originated from partially blocked AMPA and NMDA receptors, as well dopaminergic and noradrenergic neurons.
To generate input-specific evoked activity from electrical stimulation, Theta Burst Stimulation (TBS) was delivered to 14-week-old organoids 4 times with 13-minute intervals between TBS (Fig. 7a) on the HD-MEA. Two independent experiments (referred to as A and B in the figures and below) were conducted. In both experiments, four to five organoids were seeded on each well at week 9.5 of differentiation and grown on the MEA until 33 ± 1 DOM before stimulation (Supplementary Fig. 10a and Supplementary Fig. 11a). First, we recorded the baseline network activity across all wells and found that basal activity was lower in wells 1A-3A than wells 4A-6A and 1B-4B (Supplementary Figs. 10b and 11b). The MaxWell HD-MEA has an electrode size of 8.75 × 12.50 µm², and the electrode center-to-center distance is 17.5 µm, allowing one neuron to be recorded by multiple electrodes. For input-specific synaptic plasticity, one neuron from each well was identified based on its footprint (Supplementary Figs. 12a and 13a) and spike-sorted neuron traces using the Axon Tracking assay in the MaxLab Live Software (Supplementary Figs. 12b and 13b). Then, 32 electrodes focusing on a single neuron in each well were stimulated using a modified version of previously described LTP induction protocols46,47,48 (Fig. 7a). To optimize the stimulation of each neuron, electrodes along the entire neuron including the soma and axon were targeted for stimulation.
Fig. 7: Theta-burst stimulation modulated short-term plasticity for Experiment A.
a Graphical summary of TBS protocol. i-The TBS was performed four times spaced by 13 minutes. ii-Within each TBS there are 10 trials with four spikes per trial. iii-The schematic of each trial. b Percent active area before and after stimulation across all 6 wells. Wells 4A–6A show consistent increase or decrease in active area in response to stimulation while wells 1A–3A show little change. c Representative heat map evoked activity response for wells 4A–6A. Bin size is equal to 10 ms. The stimulation pulses are the light grey vertical lines, and the dashed orange lines indicate the start/stop time of the analysis window for calculating evoked activity. d percentage of active electrodes, total spikes, and evoked activity for wells 1A-3A and then 4A-6A. Purple circle represents well 1A, pink square – well 2 A, turquoise triangle – well 3A, green circle – well 4A, blue square – well 5A, and yellow triangle – well 6A. The data represents the mean with 10th to 90th percentile for each well. The 90th percentile response of a well treated with NBQX/AP5 before and during stimulation is shown with a blue dashed line overlayed on all graphs. The mean response of a well-treated with NBQX/AP5 before and during stimulation is shown in a black dashed line overlayed on all graphs. The 10th percentile response of a well treated with NBQX/AP5 before and during stimulation is shown in a red dashed line overlayed on all graphs. Responses above this NBQX/AP5 region indicate responses generated by glutamatergic receptors. e Histograms of total evoked activity per bin (bin size of 10 ms), total spikes, and total active area. The top three graphs show data aggregated across all electrodes for all 4 TBS for wells 1A–3A, and the bottom three graphs show data aggregated across all electrodes for all 4 TBS for wells 4A-6A. Wells 1A-3A show little to no response while wells 4A-6A indicate evoked responses on the millisecond timescale.
To investigate short-term changes in evoked activity, total evoked activity per bin (10 ms), total spikes, and total active area were measured. Active area before and after each stimulation are shown for all wells in both experiments (Fig. 7b and Supplementary Fig. 14a). Wells 4A-6A, 1B-4B showed substantial changes in active area in response to the stimulus while wells 1A-3A showed little changes (Fig. 7b, Supplementary Fig. 14a).
Representative evoked activity heatmaps from wells 4A-6A and 1B-4B demonstrated strong short-term responses within milliseconds following stimulation (Fig. 7c, Supplementary Figs. 15, and 16). Wells with lower baseline activity (e.g., 1A-3A) did not show any response.
To determine an activity threshold, we treated one well with NBQX/AP5 to block glutamatergic receptors-dependent synaptic plasticity. The 90th, mean, and 10th percentile responses from the NBQX/AP5-treated well is shown overlayed on the plots as the dotted blue, black, and red lines, respectively (Fig. 7d, Supplementary Fig. 14c). Wells 1A-3A did not exceed this threshold, while wells 4A-6A and 1B-4B consistently did across all four TBS sets (Fig. 7d, Supplementary Fig. 14c).
Aggregated data for active area, total spikes, and evoked activity showed that wells 4A-6A and 1B-4B had a distribution skewed to the right of 0, while wells 1A-3A only exhibited a mode around 0 (Fig. 7e, Supplementary Fig. 14d). Wells 1A-3A, with lower baseline activity and connectivity compared to wells 4A-6A and 1B-4B, did not respond above threshold, whereas the shift to the right of 0 in wells 4A-6A and 1B-4B suggests short-term potentiation, as stimulation led to short-term increases in activity.
To further confirm STP, connectivity and criticality was quantified across each TBS for well that exhibited STP (Well 4A, 5A, 6A, 1B, 2B, 3B, 4B) across both experiments (Fig. 8a–c). We observed a significant increase in the number of nodes and edges while modularity significantly decreased after every stimulation indicating a more connected network of neurons after stimulation (Fig. 8a and b). These results suggest that wells that exhibit evoked electrical activity also became more connected, more of a single community. While metrics of connectivity were extremely consistent across stimulations, metrics of criticality varied across the stimulations including the deviation of criticality coefficient and shape collapse error (Fig. 8c). There were no significant differences in either metric (Fig. 8c). The branching ratio, however, showed a significant increase following stimulation (Fig. 8c), suggesting that the neural organoids are becoming more critical after stimulation.
Fig. 8: Theta-burst stimulation drives short-term changes in connectivity and criticality and long-term potentiation and depression of neuronal units.
a Connectivity metrics for all wells that demonstrated STP. b Representative connectivity graph before and immediately following TBS #1 for well 1B. c Criticality metrics for all wells that demonstrated STP. The data represents the mean of seven biological replicates from two independent experiments. A Wilcoxon matched-pairs signed rank test was performed to determine statistical significance for a and c, **p < 0.01, ***p < 0.001, ****p < 0.0001. Exact p-values are listed in Supplementary Tables 22–27. For a and c green circle represents well 4A, blue square – well 5 A, yellow triangle -well 6A, red line – well 1B, purple circle – 2B, open blue circle – 3B, open pink square – 4B. d) Quantification of input-specific long-term potentiation (LTP) and depression (LTD) by measuring firing rate over time in neuronal units. Two example units demonstrating either LTP (blue line with 95% confidence intervals depicted with dashed lines) or LTD (red line, with 95% confidence intervals depicted with dashed lines) are shown on the left. The proportion of neuronal units that demonstrated LTP (red) or LTD (blue) across wells is demonstrated and quantified on the right.
Long-term effects of TBS on organoids were assessed by quantifying criticality, connectivity, network-level dynamics (including interspike interval, interspike interval CV, and firing rate), and spike sorted unit (aka neuronal) level changes in firing rate in wells that demonstrated input-specific STP (Supplementary Figs. 17 and 18). These metrics were quantified before and at 60 – 180 minutes post-TBS. There were no significant differences before and after stimulation (after stimulation includes grouped data from 60-180 minutes post stimulation) for all connectivity and criticality metrics (Supplementary Figs. 17 and 18). In addition, there were no consistent trends over time for network level interspike interval, interspike interval CV, or firing rate for wells exhibiting STP (Supplementary Figs. 17 and 18). Interspike Interval (ISI) was calculated with a 4 Hz threshold (up to 250 ms) to account for changes in theta entrainment/phase locking. Well 4A, 5A, 1B, 2B, 3B, and 4B showed no long-term changes in ISI after stimulation (Supplementary Figs. 17c and 18c). While well 6A showed a significant increase in ISI 60–180 min post stimulation, apart from at 90 minutes (Supplementary Fig. 17c). The CV was used to measure ISI variability across timepoints7. A CV of 2.5 indicates a perfect Poisson process7,49, while a CV near zero indicates a perfectly periodic spike train. All wells analyzed showed no significant change in ISI CV after stimulation (Supplementary Figs. 17c and 18c). In addition, none of the wells showed any changes in firing rate over time compared to the baseline (Supplementary Figs. 17d and 18d). Since network level ISI, ISI CV, and firing rate graphs were analyzed based on overall trends across grouped units for each well and not on a unit level, the results indicate that there are no network level changes in interspike interval or firing rate. Together, these results suggest that there are no long-term changes to the network dynamics after TBS to an individual neuron. This is expected as the TBS was input-specific; therefore, we focused on unit-level changes rather than network-level changes. Upon further investigation of unit-level firing rates over time, linear regression analysis showed that all wells had units that were either potentiated or depressed and were maintained for 180 minutes post-stimulation (Fig. 8d). To account for random changes in firing rate, the same analysis was performed on a time series recording without stimulation. In this recording, only one out of 40 units showed potentiated firing rate over time, therefore, this can be considered the noise level of changes due to basal synaptic plasticity within the organoids. Wells 4A-6A and 2B-4B show potentiated and depressed units above this level (Fig. 8d). Representative LTP and LTD units are shown in Fig. 8d. Together, these data indicate there are input-specific TBS-induced changes in connected neurons (aka units) over hours but not the overall network, supporting the use of this model to modulate input-specific short- and long-term synaptic plasticity and detect changes in synaptic plasticity in connected neurons. In addition, due to the nature of input-specific synaptic plasticity on a HD-MEA, it is difficult to control what kind of neuronal circuits are being modulated, whether they are increased or decreased, which could explain why the population of LTP compared to LTD is different in each well.
SPOILER ALERT:This interview contains major spoilers from “Mirror, Mirror,” the finale of Apple TV+’s “Smoke.”
Dave Gudsen (Taron Egerton) was never the man he said he was. He may have been an arson investigator by day, but he was one of the two serial arsonists who would terrorize his Pacific Northwest town by night. He may have had aspirations of becoming a novelist, but he was practically laughed out of a publisher’s office for his bad writing. He may have seen himself as a loving husband and stepfather, but his “nice guy” persona belied his abusive nature.
And in the final minutes of the Apple TV+ drama “Smoke,” viewers come to learn that the man they’ve watched for nine episodes doesn’t actually look like Egerton (who also serves as an executive producer). While sitting opposite his former partner, Det. Michelle Calderone (Jurnee Smollett) — who had finally been able to apprehend him after spending most of the nine-part series gathering evidence of his fiery misdeeds — Gudsen’s mask finally drops in the interrogation room, revealing a man who looks markedly older and heavier staring back at himself in the mirror.
It’s an unsettling final twist that creator and showrunner Dennis Lehane fought to include in the final chapter of his fiery saga, which was inspired by the real-life case of convicted serial arsonist John Leonard Orr.
Courtesy of Apple TV+
“We hid it from Apple because we knew they’d hate it — and they did. And everybody was like, ‘What?!’ And I was like, “No, he’s projecting! That storyline is about his projection. He’s projecting who he is,’” Lehane tells Variety. “This is a guy who walked around thinking of himself as a hero, who was simultaneously running out and doing some really terrible things. To me, we’ve become victims of a type of performative persona, performative masculinity, whatever you want to call it. People are losing complete grasp with who they actually are, and Orr represented that to me.”
In a wide-ranging chat, Lehane opens up about how his personal experiences with men like Gudsen inspired his and Egerton’s latest foray into TV after their acclaimed 2022 miniseries “Black Bird,” why he was not surprised by Trump’s return to power (and why he thinks Democrats are partially to blame), the harrowing finale scene that he had to cut down in the edit — and why he believes that finale could represent either an end or a new beginning for Gudsen and Calderone.
How did you figure out what motivates Dave’s actions? What did you settle on as the root cause of his behavior?
I grew up in a hyper-masculine culture. I knew a lot of people who were incredibly full of shit and who would make incredible justifications for their own failure, so I’d had a front row seat to a lot of that for years. I hung out in a lot of the bars where guys talked about how they got shafted out of jobs because they were white. And I then moved into literary circles where you would come across the same type of guy. I remember a guy saying he lost out on a fellowship because the other person applying for the fellowship was gay, and I was like, “Did he put that on his CV? You really have gone that far down the rabbit hole of bullshit?”
I guess what I’m saying is, I’ve had a front row seat for a lot of toxic masculinity for six decades, and Dave, to me, was that guy. There’s always an excuse with him. Michelle plays a card late in the show where she says, “Huh, so you failed the psych eval,” and that was true of Orr. Orr didn’t get into the police department — that was his big axe to grind — not because he was single and white, but because he failed the psych eval. So I thought [toxic masculinity] was something to examine further as we live it now.
You have insisted in past interviews that you are not a political writer by any means, but many of the social issues you tackle in your work — including race, wealth, privilege, masculinity — can be construed in a way to make a political statement. Do you think this show has landed with viewers any differently in the current political climate with the Trump administration?
None of this goes away; it hadn’t gone away. It wasn’t like in 2020, everybody waved a wand and we’re like, “Hey, we’re sane again!” There were still a million people running around talking about January 6th as if it wasn’t an insurrection. There’s still plenty of ugly shit going on. And then you had all the anti-woke stuff. I’m with the guys on “South Park” — the woke stuff was getting a little silly when you were taking it to the extreme, but it’s a hell of a lot better than the alternative, which is where we’re going now.
And now you look at what’s happening in Washington D.C. today — it is a test run; it is a dry run. He did one in California, now he’s doing one in D.C. What’s going to happen when he decides to run for a third term? He’s already established he can send troops into American cities. That’s the reality we’re living in. So I don’t think any of this went away. I think it’s been underground for a very long time, and I think liberals and Democrats, certainly, by being out of touch with the white working class, they lost their finger on the pulse. I was not surprised by any of it. I wasn’t surprised by 2016. I wasn’t surprised by 2024. I won’t be surprised by what’s coming, because I live in those bars; I hang out with those people; I’ve spent a lot of time in their kitchens. I know how they think, and there’s a lot of rage out there.
Courtesy of Robert Falconer/Apple TV+
On some fundamental level, Dave and Michelle are very much the same person. They both share self-destructive tendencies, and they each have to find ways to justify their own behavior to live with themselves. How did you think about evolving that central relationship over the course of this season?
We knew Michelle had heroic qualities, but was very much not a hero. That makes her like Dave in some ways, although Dave’s the real extreme version of it. So we started to look at Michelle in terms of her own damage, her own dysfunction, and her own denial, which is pretty prodigious. So as we progressed in the writers’ room, we kept going, “How much more extreme can we take this?” Because we are living in extreme times.
I remember the day I came in and said, “Dave is not Dave. Dave does not look like Taron Egerton.” It was pretty early, and we hid it from Apple because we knew they’d hate it — and they did. And everybody was like, “What?!” And I was like, “No, he’s projecting! That storyline is about his projection. He’s projecting who he is.”
The Freddy storyline is clearly the storyline you’ve seen from me before. My heart and soul lies in those types of stories [about] the people who are truly disadvantaged, the people who are true victims of society, who then lash out in very unfortunate and terrible ways. Once we wrapped that, we were like, “OK, the last two episodes, now that’s going to be the Michelle and Dave show. We’re going to really see how nuts these two people are and let it ring out to the very last scene.”
You mentioned Freddy — the fast-food worker arsonist played by Ntare Guma Mbaho Mwine — who uses a milk jug filled with oil as his incendiary device of choice, and who dies by suicide in the seventh episode. Dave and Freddy were both going to burn down the world for not giving them what they thought they deserved. But while they are both criminals, their circumstances could not be more different. How did you think about juxtaposing Dave and Freddy’s arcs, and how conscious were you of the role that race plays in this story?
I was 100% aware of that at all times. But I never want to write polemics. I never want to make you feel like you’re doing your homework politically or socially. I just want to tell a story about people. But I think it completely goes without saying that Freddy is the true disadvantaged one because of the color of his skin, because of his education, because of his background. It’s not like he has mommy issues. He got screwed on every single possible level as a person.
He was inspired a bit by a guy named Thomas Sweatt, who was an arsonist in Washington D.C. and shows up at the very end of the “Firebug” podcast, which is what inspired the show. I thought, “What if Thomas Sweatt was working at the same time as John Orr in the same area?” So there’s not a single misinterpretation you could make of the Freddy storyline that’s probably mistaken. We meant everything, and Ntare was game.
We were shocked that nobody brought up more the way he’s vanquished by Dave [at the end of Episode 6, when Dave gets some sick pleasure out of stopping Freddy from lighting himself and Brenda, the kind hairdresser who gave him a makeover, on fire]. In the end, it’s almost like Dave ejaculates on him. Dave gets a hard-on from it! I mean, that’s white power at its essence, and then he turns to the African American woman and says, “Don’t worry. You’re safe now.” It’s pretty brutal. All of us — me, Taron, Ntare, Adina [Porter, who played Brenda] — went to the wall on that storyline.
We knew we were telling two stories. One was the crazy story that’s kind of fun. One was the very dark and very sad story that was going to end tragically. I think that dark, tragic story is certainly a wheelhouse for me. That’s something that I obsess over. You see it in all my work — in Larry Hall in “Black Bird,” in “Small Mercies,” in “Mystic River.” But the other [storyline] was to balance that, because I don’t want to show this just like, “Oh, wow, man, life really sucks.” But for Freddy, it sure does.
Courtesy of Robert Falconer/Apple TV+
In the penultimate episode, Michelle planted a discarded glove that Dave had left in her car at the rental house of police captain Steven Burke (Rafe Spall), whom she had accidentally killed during a heated confrontation about their ill-advised affair, before torching the place to remove any trace of her being there. How did you arrive at that final twist, and what were the most important considerations for you as you were writing the finale?
The idea was, we live in this world of non-truth — she’s not going to get him clean. She’s getting him for something that she did, not that he did. And once we came up with that, we were like, “That’s the show! That’s the story we’ve been telling since the beginning.” Dave’s looking at the TV in Episode 9, and he goes, “I’m being framed.” And then his ex-wife says to him, “Nobody says that. That doesn’t happen.” And he’s like, “No, I’m being framed!” And he’s right! We really thought, “Wouldn’t it be interesting if Dave had to try and figure out a way to dig himself out of this one?” Because he left such little physical evidence behind — and that’s the truth of arsons. They’re extremely hard to prosecute, because you burn the evidence. So we just thought it would be highly ironic.
We had to give the audience a huge fire. That’s something that we’ve been promising. The director, Joe Chappelle, and I are both Sergio Leone fans, we’re both George Miller fans, and we thought: What if we mashed up Sergio Leone and George Miller and had [the characters] driving through a fire, and then had her be catapulted from the car, turn around and come out like a true Sergio Leone hero? And instead of reaching for a gun, she reaches for a scrunchie.
I wrote the notes for this episode in a kind of fever. My assistant viewed some of it, and it’s one of the craziest things she ever saw. I went into a room and turned on the “Oppenheimer” soundtrack on a complete loop, and then I just began to envision everything and threw it up on a board. I felt like a mad scientist, but that’s where all this came from. I knew what [writer] Molly [Miller] had done in Episode 8, so I was like, “Oh, man, the gauntlet is thrown. I gotta top that.” Go big or go home was the whole point of that episode.
In the finale, when he knows Michelle is on to him, Dave tries to kill her by driving both of them straight into a wildfire, and after she survives the inevitable car accident, she is able to bring him into custody — but not before she puts a gun in his mouth and threatens to shoot him. Was there ever a version of this story where Michelle would have killed Dave or vice versa?
No. We always knew we were driving towards [that interrogation room scene]. In case it’s not brutally obvious by now, I love nothing more than two people in a room facing off against each other. So I always knew there was going to be a camera move behind Dave in the final scene that was going to reveal that he’s not the Dave you’ve been looking at for nine episodes. I always knew that was there, so it was all about just getting them in that room.
I liked the idea of her beating the ever-living shit out of him, and then his life being saved by the rain. Right up until that moment, she could kill him, and then the rain comes and washes her clean, and she says, “I’m going to walk him in. I’m going to bust him.” She probably should have killed him. If she killed him, then all questions go away.
The stuff with the gun — we trimmed that and trimmed that, because we all went so over the top with that. It was pretty brutal. That scene was so clearly a type of rape of Dave, and she was shoving that gun in and out, in and out, in and out [of his mouth]. She was showing him what it feels like to be a woman, basically, and to have no power. So we just said, “I think less equals more here.” I’d say we cut 45 seconds out of that bit. It was so long! Joe had all these incredible closeups. And I remember when I wrote it and I turned it in, my writers were like, “Uh … what’s with the face-fucking?” I was like, “Yeah, I know.” We dialed it back, but all of Michelle’s rage at every single man [in her life] — Arch Stanton, Burke, her long-gone father, her brother — comes out in that scene.
Courtesy of Apple TV+
How would you characterize that charged final look between Dave and Michelle in the interrogation room?
At first — and this is Taron’s performance — when he says, “I know who I am,” he’s shook. His persona drops. It falls away. He’s lost for a second. And that’s why you get a glimpse of [the real] him. It’s the same after his car accident. After his car accident, his wife blows him up, she breaks him down for a second. He stands up, he walks to the mirror, and he sees himself for just a second, and then he goes back into his bullshit. So when he says, “I know who I am,” at first, he says it almost hopefully. Taron lets his face settle, and then he gets this predatory look in his eyes again, and he’s Dave again, and he just says, “I know who I am.” And then Michelle says, “So do I.” And it’s the two of them looking at each other: “Now what do we do? What happens next?” Because this dance is what has been fueling them, and it ends the moment she walks out of that room, potentially.
What do you imagine happens to Dave and Michelle after the screen fades to black?
I think Dave becomes a media darling, a hero of the right, a victim. I think Dave becomes the Karen Reed of his time, and he’s going to have a whole contingent of the population who says, “This guy’s innocent. This guy was framed.” Which, in this case, they’re right! And then Michelle, what Burke’s ghost says to her is very true: “This is a flimsy story. All it takes is one person to put us together, and your whole story is going to fall apart.” So there’s a lot of grist for the mill for it to go on.
But “Smoke” was billed as a limited series. Does this mean that you are actively looking for ways to continue telling this story, or have you already let the story go?
No, we haven’t let the story go. We have plans if it were to go forward, but we don’t know if it will or won’t. So I don’t know if it’s quite … lit the world on fire. We always knew we were taking a very calculated risk with this show, because it’s not like too many things. We wanted it to be audacious, and we wanted the show to have a tonal shift within the heart of every episode. That’s a very risky thing to do in the current climate — and now I think even more so. Because I’m like everybody else, man. I just want a break from the really dark things I see in the news every day. So we always knew the show could be a tough sell. I’ve been connected to a smash hit or two — I know what it feels like — and this doesn’t feel like that. This feels like a show that’s going to need to grow its audience, and we’ll see what happens. If it goes someplace, we have a plan; we know where it can go. If it doesn’t, I’m moving on to the next thing. I did the show I wanted to do.
Taron Egerton and Dennis Lehane on the set of “Smoke.”
Courtesy of Robert Falconer/Apple TV+
What is next for you then? Have you and Taron discussed what your next collaboration could look like?
I think we’re both going to take a break for the next project. I don’t see a part necessarily for him in there, and he’s mulling his own possibilities. He just did this magnificent film called “She Rides Shotgun,” and the studio just dumped it, man. It’s so sad. It’s a great film, and it’s gotten great reviews. So I think Taron’s in a space where he’s just trying to figure out what his next move is. We are close friends, and we’ll get together on something else at some point for sure. He likes to do one for somebody else, and he likes to do one for me.
As a novelist, I just don’t have anything in me right now. I’m not thinking that way. If another “Small Mercies” wants to come bursting out of me, that’d be awesome. But if it doesn’t, then I’ll just keep doing [TV] because I take so much joy in the collaborative nature of this , and you don’t have that when you write a novel. You’re just alone all the time, and I’m really sick of being alone all the time.
We’ve already developed [the TV adaptation of] “Small Mercies.” There’s scripts, there’s a cast. I can’t speak to it yet. There’s a lot of questions swirling around it right now that are just way past my pay grade, and then I’ll get to it. So I have “Small Mercies” ready to go, but it’s just a matter of when. I’m waiting to see what Apple wants to send me next, because I’m under contract with them. And if they want to throw something my way, then I’m happy to work. But I have nothing to pitch right now. I really burned myself — God, these fire metaphors! — when I did “Smoke.” There was nothing left, so we’ll see what happens.
“We were really awful, and they were very good, but we made them better,” said Erasmus after the 38-22 defeat.
“We can find excuses but they gutsed it out and, yes, we gave them one or two soft tries with an intercept here and a loose pass there, but overall, they were just better than us on the day.
“They beat us in most departments. We didn’t scrum them, they beat us in the lineouts, and they bullied us at the breakdown after Siya Kolisi went off and Marco van Staden had to go off for an HIA.
“We as coaches got it terribly wrong and we have to look at ourselves before we point fingers at the players.
“From now until next Saturday, we’re going to take a lot of flak, but we take credit when we do well, and we have to take the flak when we do badly.
“I’m saying it with a smile not because I’m happy but I’m saying it with a smile because that’s the reality of rugby and we’re really disappointed and we’re feeling bad for our players and we’re feeling bad for our supporters and for overall what we produced on the field.”
Erasmus admitted that the team for next week’s return match at DHL Stadium in Cape Town had already been picked but that he may now reconsider.
“We have always said that if we don’t play well and we lose momentum – and we did both, we played badly and lost momentum – we might change our thinking,” he said.
“We had a chat now in the change room and that may now change, but we want to get Ethan Hooker, Canan Moodie and Morne van den Berg and a few others a start but we’ll have to rethink.”
Erasmus also admitted that the physicality of the Wallabies had caused problems that the Boks failed to overcome: “I don’t they tactically outsmarted us, but they physically dominated us, and the interesting thing is that the longer the game went on they were supposed to struggle but it just shows what Wallaby coach Joe Schmidt is building there.
“The saddest thing is that they took five points, and we didn’t fight back to take a bonus point. I can butter this up and make excuses, but we were really terrible on the day.”
Erasmus rued missed chances and said he would have to review the messaging the players were being given.
“Grant Williams had so many breaks where he almost got away, and Manie had so many breaks where he almost got away the same with Edwill and that changes games,” said Erasmus.
“When you’re 22-0 up and there’s lots of space and the player thinks ‘let’s take a chance’, and that’s coaching; that’s us. Telling the guys let’s build an innings, 22-0 is not winning the game.
“It was a bad loss in a bad way – not against a bad team – and we didn’t have the fight until the end; there was a stage where I felt our heads were dropping and our shoulders were slumping and that bothered.
“But we had a quick chat, and everyone is very disappointed and how you see the game and how our supporters will feel but tactically we totally overplayed every time we thought something was on.”
The second match against Australia takes place at DHL Stadium in Cape Town on Saturday, kick off at 17h10.
The supernova known as SN 2023zkd was first spotted in July 2023 by the Zwicky Transient Facility, but what made it remarkable was how quickly astronomers were able to study it. A newly developed AI algorithm, designed to flag unusual stellar explosions in real time, raised the initial alert.
That early detection proved crucial, giving scientists the chance to launch immediate follow-up observations with an array of telescopes on Earth and in space. Leading the effort were researchers from the Center for Astrophysics at Harvard & Smithsonian (CfA) and MIT, working under the Young Supernova Experiment.
Their analysis suggests that the most likely explanation is a massive star caught in a fatal orbit with a black hole. As the two spiraled closer together, gravitational stress reached a breaking point, triggering the star’s collapse in a supernova while it partially engulfed its black hole companion.
Machine learning system flags rare stellar explosion months in advance
According to Alexander Gagliano, lead author of the study and fellow at the NSF Institute for Artificial Intelligence and Fundamental Interactions, the blast was most likely sparked by a catastrophic encounter with a black hole companion, providing the strongest evidence so far that such close interactions can actually detonate a star.
The team’s machine learning system flagged SN 2023zkd months before its most unusual behavior, giving astronomers enough time to secure the critical observations needed to decode the explosion. An alternative interpretation is that the black hole completely tore the star apart before it could explode on its own. In this scenario, the black hole rapidly consumed the star’s debris, and the observed emission came from the debris slamming into surrounding gas.
SN 2023zkd first appeared to astronomers as a fairly ordinary supernova, marked by a single burst of light roughly 730 million light-years from Earth. Yet as they monitored it over the following months, the event took an unexpected turn when the fading star brightened again.
Strange light pattern points to hidden black hole companion
To investigate this unusual behavior, the team dug into archival observations and uncovered an even stranger clue: the system had been gradually brightening for more than four years prior to the explosion. Such long-term pre-explosion activity is extremely rare in supernovas, making SN 2023zkd stand out as a cosmic anomaly, the study notes.
Analysis showed that the explosion’s unusual light pattern was shaped by gas the star had shed in its final years. The first brightening came as the blast wave hit low-density material, while the second peak arose from a slower collision with a dense, disk-like cloud. This structure, along with the star’s erratic pre-explosion activity, suggests it was under intense gravitational stress – likely caused by a nearby compact companion such as a black hole.
Gagliano further pointed out that the discovery highlights the importance of studying how massive stars interact with their companions in their final stages of life. While it has long been known that most massive stars exist in binary systems, direct evidence of one exchanging mass shortly before exploding is exceptionally rare. Now, SN 2023zkd offers a rare glimpse into this process, giving astronomers valuable insight into how such close interactions can shape the violent deaths of stars.
Alex’s family says they want his legacy to result in kickboxing being safer for others
The family of a 15-year-old champion kickboxer who died after a bout have said while his death has left them “broken”, they hoped it wouldn’t be “in vain”.
Alex Eastwood, from Fazakerley, Liverpool, collapsed after the third and final two-minute round of a “light contact” kickboxing match against a 17-year-old opponent in Wigan in June 2024. He was taken to hospital but had suffered a serious head injury and died three days later.
Alex’s father Stephen Eastwood and step-mother Nikita vowed to make the sport safer after it came to light he had been competing in an unsanctioned fight.
His father Stephen Eastwood said: “As a family, we’re still broken. I don’t think that will ever go away.
“We’re just trying to do thing right and make change for the people involved in that sport.”
Family handout
Alex’s family have said they want change to be the 15-year-old’s legacy
Mr Eastwood said they were hopeful that a recent meeting with Culture, Media and Sport Secretary Lisa Nandy, who is also the MP for Wigan, would result in changes being made.
He said: “She was quite positive that something will happen to prevent this from happening again.
“So as a parent, his death isn’t in vain – there’s a lasting legacy for Alex and to protect other people in the sport.”
‘Massive regret’
A recent inquest into Alex’s death found there was no minimum standard of medical aid or proper risk assessments done at events like the one that resulted in his death.
Coroner Michael Pemberton said neither Alex, nor his parents, appreciated the dangers of the “chaotic and somewhat disjointed” approach to children involved in combat sports.
Mrs Eastwood said Alex was “massively failed” by the lack of safety protocols in the organisations he competed and trained with.
“He was just doing his dream, doing what he wanted and pursuing his career,” she said.
“There was nothing to keep that dream fulfilled and sustained within the safety that should have been there for him.”
She said assuming that there were safety protocols in place was “a massive regret” she had.
Mrs Eastwood said: “I encourage other parents to ask the questions – what is my child going into here?”
The death toll from recent flash floods in Khyber Pakhtunkhwa surged to 332 on Saturday as various districts, especially hard-hit Buner, reported more loss of lives, the Provincial Disaster Management Authority (PDMA) said on Saturday.
This adds to the casualties in Gilgit-Baltistan and Azad Jammu and Kashmir, where the unprecedented floods have claimed at least 12 and nine lives, respectively, besides causing widespread destruction.
KP witnessed devastating scenes yesterday as flash floods caused by heavy rainfall and cloudbursts in multiple districts claimed over 200 lives in a day, including five crew members of a provincial government chopper that crashed in Mohmand during relief and rescue efforts.
Buner was the worst-affected district in the province with 208 lives lost in the past 48 hours, according to a PDMA situation report. It added that 120 people were injured, while Deputy Commissioner Kashif Qayum Khan’s office reported that 50 were still missing.
According to a PDMA report seen by Dawn.com earlier today, Shangla reported 37 deaths, Mansehra 23, Swat 22, Bajaur 21, Battagram 15, Lower Dir five and a child drowned in Abbottabad.
Detailing infrastructure damage, the report said 11 houses were destroyed while 63 were partially damaged due to the floods. Two schools in Swat and another in Shangla were also affected.
This screengrab of a PDMA report issued on Aug 16, 2025 shows a breakdown of the rain-related incidents across KP and the resulting casualties in the past 48 hours. — PDMA report
The KP government declared an emergency in the severely affected districts of Buner, Bajaur, Swat, Shangla, Mansehra, Torghar, Upper and Lower Dir and Battagram.
According to a notification available with Dawn.com, the state of emergency will remain in place until August 31 and local administrations in these districts have been authorised to mobilise all resources for relief activities.
“The Provincial Disaster Management Authority shall provide from its central storage … a reasonable quantity of tents and NFIs (non-food items), and/or any other article available if required for the subject purpose,” the notification read.
The KP government released Rs1 billion in funds to the PDMA for “timely compensation/preparedness and response to cope with any untoward situation”, a notification shared by it on X said.
It also allocated over Rs1.55bn worth of funds for its Communication and Works Department for the restoration of highways and bridges in the flood-affected districts.
Additionally, the Pakistan Telecommunication Authority (PTA) said it was closely monitoring the situation in flood-affected areas, noting that flash floods damaged several cellular and fixed-line sites, causing service disruptions.
“PTA teams are on the ground, coordinating with federal [and] provincial authorities, district administrations and telecom operators for swift restoration,” the PTA said in a statement.
In Buner, the PTA held meetings with Federal Minister for Kashmir Affairs and Gilgit-Baltistan Amir Muqam, PTI Chairman Barrister Gohar Ali Khan, Buner Deputy Commissioner Kashif Qayum and senior officials of PTCL and CMOs (cellular mobile operators) to review recovery efforts.
“PTA, together with operators, is making all-out efforts to restore communication services at the earliest, recognising their vital role in rescue and relief operations,” the statement added.
Additionally, mobile operator Jazz announced in a press release that it would provide free on-net and PTCL calls for all subscribers in KP.
“The Government of Pakistan has also launched the nationwide 911 unified emergency helpline (PEHL), offering citizens free-of-charge access to emergency services across the country,” the operator added.
On the directives of Governor Faisal Karim Kundi, the Pakistan Red Crescent in KP set up an Emergency Relief Centre to assist flood-affected districts. It can be contacted on 0300-5849255 (secretary), 0334-9086169 (admin officer), 091-9333666, or 091-2590846.
Meanwhile, state broadcaster PTV News provided a weather advisory on X, warning people of “a severe risk of floods and landslides on Torghar, Battagram, Shangla, Lower Kohistan, Tatta Pani, Gilgit, Hunza and Swat roads”, discouraging unnecessary travel in these areas.
According to the post, GB’s Somro Bridge, Ghanche, Saltoro Bridge and Baghicha (Skardu) Bridge were severely damaged and citizens were urged to completely avoid travel.
It added that traffic was suspended on the Jaglot-Skardu road, while routes in Ghizer district, Naltar Road, Babusar Top Road and around Hunza were closed, with tourists discouraged from travelling.
Meanwhile, the provincial management service (PMS) chairman requested in a letter that the KP finance secretary deduct a day’s salary from all PMS officers and redirect the money for relief work, further requesting that the secretary redirect his “whole salary”.
In a statement on X, Prime Minister Shehbaz Sharif said he was “deeply anguished by the devastation caused by cloudbursts and flash floods in KP and northern Pakistan”.
Extending his heartfelt condolences and expressing solidarity, he affirmed: “The government is mobilising all resources for rescue and relief operations.”
The premier added that he met National Disaster Management Authority (NDMA) Chairman Lt Gen Inam Haider Malik and directed him to “expedite rescue operations in the nine affected districts of Swat, Buner, Bajaur, Torghar, Mansehra, Shangla, and Battagram, with urgent focus on Bajaur and Battagram”.
“Immediate relief is being provided to stranded residents, medical aid to the injured, and heavy machinery has been deployed to clear roads and restore connectivity,” PM Shehbaz said.
Buner Police’s public relations officer Israr Khan, put the district’s death toll at 207, compared to the 204 stated by the PDMA.
DC Qayum, in a statement, ordered a ban on the entry of private vehicles into Buner, stating that it was hindering rescue and relief work.
PTI’s Barrister Gohar, who is an MNA from Buner, and MPA Riaz Khan visited the flood-hit area of Beshonai and met the bereaved families who lost their loved ones in the floods, the KP government said.
Later, Gohar chaired a meeting at the Buner DC’s office, which was attended by CM Gandapur, the KP chief secretary, provincial ministers and other senior officials.
“Instructions were given to the relevant departments and district administration to expedite the search for missing persons, rehabilitation, and road clearance operations in the affected areas,” a post on Gohar’s X account read.
As the nation observed a day of mourning for the five crew members who died in a helicopter crash during rescue efforts, the national flag flew at half-mast at Governor House in Peshawar.
KP CM orders expedited relief efforts in flood-hit areas
CM Gandapur chaired an emergency meeting in Peshawar to review the damage caused by cloudbursts, heavy rains and flash floods across the province, which was attended by the chief secretary, divisional commissioners, deputy commissioners, senior officials and PDMA representatives, the Associated Press of Pakistan reported.
Officials said rescue operations were largely completed and the focus was now on relief and rehabilitation.
The meeting’s participants were informed that 309 people were killed and 23 were injured in various rain and flood-related incidents, while 63 houses were partially damaged. Surveys of damage to roads and other infrastructure are still underway.
A view of a house in Buner’s Beshonai village on August 16 after it was destroyed by flooding. — Arif Hayat
“Work is being carried out to restore road links to cut-off areas, and medical teams, food, medicines, and essential supplies are being sent to the affected districts,” the report read.
The meeting was informed that the provincial government has released Rs 1.5 billion to the PDMA for rescue, relief, and compensation, while releasing another Rs 1.5bn to the communication and works department for the repair of roads and infrastructure.
In addition, Rs500 million was allocated to deputy commissioners to compensate the families of those killed in the flooding.
“The provincial government stands firmly with the people in this difficult time and will use all available resources for their rehabilitation,” the chief minister said at the meeting.
Expressing solidarity with the bereaved families, CM Gandapur praised the provincial and district administrations for their swift response, saying their performance in the crisis had been commendable and urged them to maintain their commitment during the relief and rehabilitation phase.
“Road connectivity must be restored as quickly as possible, with helicopters used to deliver aid to inaccessible areas,” the CM ordered. “Compensation payments must be completed within two days and additional medical staff need to be deployed from neighbouring districts.
“Food supplies must be ensured so that no affected family faces shortages.”
The chief minister also instructed the chief secretary and the PDMA to strengthen monitoring and coordination between federal and provincial agencies, mobilise heavy machinery for reconstruction and immediately dispatch all available food and non-food items to flood-hit districts.
Earlier, the CM visited Buner to review the damage caused by flooding and ongoing relief operations, according to a post on X by the KP government.
“The divisional and district administration will brief the chief minister on human and financial losses and relief activities,” the government wrote. “The chief minister will also visit the affected areas and meet with the affected families.”
The post added that members of the provincial cabinet, the chief secretary, and other officials would accompany CM Gandapur.
While speaking to reporters in the area, the chief minister said that rescue operations have started and the government is working to clear roads blocked by the flooding.
“Routes in some areas have been restored; however, some have not,” he said. “We have also requested helicopters. Our goal right now is to restore the routes.
“I want to let people know that the provincial government compensate citizens fully for damages to their homes,” the chief minister added. “We are in touch with the NDMA and they will provide us with whatever we need.”
PM to visit affected areas: Amir Muqam
Federal Minister Muqam, visited areas in Buner affected by floods today to assess the damage and meet with victims, the Associated Press of Pakistan reported.
The minister, who is also the president of the PML-N in KP, was accompanied by the Malakand commissioner and other senior officials.
Muqam stated that he was visiting on the instructions of PM Shehbaz, who is personally overseeing the relief operation. “This is not the time for politics; the entire nation stands with victims,” he said.
The minister added that the prime minister would also be visiting the affected districts soon, but did not say specifically when.
During his visit, Muqam directed officials to expedite relief efforts, which are being supported by the local police and district administration. He met with affected families, offering condolences and reviewing ongoing relief activities.
The minister noted that the local administration, the Pakistan Army, and Frontier Corps were all participating in the rescue operation.
“Many villages have been completely destroyed and prayers are being offered for those still missing,” he said. “Medical camps have been established in affected areas and the federal government will provide all necessary resources.”
11 dead, 318 homes destroyed in GB: disaster authority
According to the Gilgit-Baltistan Disaster Management Authority (GBDMA), 11 people were killed, 25 were injured and four were reported missing during the past 48 hours.
The authority added that 92 houses were partially damaged, while 99 were fully destroyed, citing initial reports. Detailed assessments were still ongoing
Interior Minister Shams Lone said that the bridge on the Baltistan Highway was swept away in the flood, cutting off four districts of Baltistan from Gilgit for the second day.
He said that since July 22, “thirty-five people died, 35 were injured and four are missing in the flood waves. Similarly, 318 houses have been completely demolished and 674 have been partially damaged.”
Lone said that the GB government does not have the resources, so the Army has taken steps to help in relief efforts.
“The Pakistan Army played an important role in the rehabilitation after the massive destruction caused by the Babusar flood and provided food and other essential items to the victims of Ghizer today,” he said. “The Army has played an important role and is still at the forefront in this difficult phase.”
Later, Rescue Information Officer Shakriz said in a statement that the body of a person previously reported missing had been recovered, bringing the overall death toll in the region to 36.
Tourists trapped, power plants damaged in GB’s Naltar Valley: govt
Meanwhile, GB government spokesperson Faizullah Faraq said a “great number of tourists” were trapped in the Naltar Valley as the land route was disconnected after floodwaters washed away a huge chunk of the Naltar Expressway.
In his statement, the official said the power supply in the area had been suspended as the three power plants situated there were shut down due to the floods.
According to Faraq, river waters from the Jaglot Goro stream entered several houses and restaurants in low-lying areas.
He added that floods across the province caused severe damage to public property and infrastructure.
“Flooding in four locations in Gilgit district today severely damaged four power plants, a bridge and an expressway, while a large number of buildings were submerged in Jaglot Guru.”
Faraq said that the floods in Naltar Valley damaged three power plants, cutting off power supply to thousands of people.
“A power plant and a bridge were also damaged in Bagrot, cutting off power, while land communication was suspended due to the bridge being washed away,” Faraq said.
“A flood relay containing heavy stones swept away a house within the limits of Allahwali Lake in Astore in the afternoon,” he added. “Local people ran to save their lives.”
A house in Gilgit-Baltistan’s Astore district is washed away by flash flooding on August 16. — Gilgit-Baltistan Government
Rescue efforts continue but face challenges
The KP PDMA told AFP that around 2,000 rescue workers were engaged in recovering bodies from the debris and carrying out relief operations in nine affected districts.
Rescue 1122 Director General Muhammad Tayyab Abdullah told Dawn.com that an elite rescue squad of 80 individuals has been formed for areas where operations are facing “intense difficulty”.
“In the past 12 hours, 3,542 emergencies have been responded to, while 76 vehicles are taking part in the rescue operations,” he detailed.
DG Abdullah noted that the teams were facing challenges in their activities because of ruined roads, while people were also not able to contact helplines due to damaged mobile towers.
Similarly, KP Rescue 1122 spokesperson Bilal Ahmed Faizi told AFP: “Heavy rainfall, landslides in several areas, and washed-out roads are causing significant challenges in delivering aid, particularly in transporting heavy machinery and ambulances.
“Due to road closures in most areas, rescue workers are travelling on foot to conduct operations in remote regions,” he highlighted.
“They are trying to evacuate survivors, but very few people are relocating due to the deaths of their relatives or loved ones being trapped in the debris.”
Meanwhile, PTV News reported that Pakistan Army personnel have arrived in Buner as part of relief efforts. The broadcaster said in a post on X that special equipment from the Army Corps of Engineers will be used to recover bodies and those injured trapped underneath layers of mud.
“Rations and other supplies are being provided through Pakistan Army helicopters, and people are being evacuated from flood-hit areas to safe locations,” PTV News wrote.
A statement issued by Buner’s Rescue 1122 said DG Abdullah, Buner Operations Director Mir Alam, North Region Operations Director Arshad Iqbal and the district emergency officer were supervising the operation themselves.
The Rescue DG urged the public not to go near floodwaters, rainwater drains or bridges, and contact the 1122 helpline in the case of an emergency.
In Battagram, two more bodies were recovered at the Malkal Gali-Neel Band village, which had been impacted by a cloudburst on early Thursday, according to Assistant Commissioner Muhammad Saleem Khan.
The rescue operations had resumed at 6am after being suspended at 7:30pm yesterday due to nightfall and the challenging terrain.
Flash floods also severely impacted the power infrastructure in Swat, as floodwaters entered the 132KV Swat grid station, causing the tripping of 41 feeders and the complete suspension of electric supply across the region.
The KP PDMA has said rainfall would continue until August 21 in different parts of the province.
The Pakistan Meteorological Department (PMD) has also warned the Balochistan government to take emergency measures to prepare for a new spell of monsoon rains in the province, which will start from Aug 18 and continue until Aug 22.
Villagers in Buner deal with the aftermath
Beshonai village in Buner, a settlement of more than 80 households, was left in ruins a day after flash flooding tore through the village centre.
According to a Dawn.com correspondent on the ground, a steady stream was calmly running beneath the large stones the flood brought in, and though there are some remaining houses along the bank of the stream, most others were completely washed away, along with their inhabitants.
Hakeem Jan, a resident of a nearby village, told Dawn.com that he heard “loud bursts” in the mountains, then “a huge rally of water and big stones colliding and raging downward”.
“Multiple cloudbursts in the upper portion of the Elum valley caught the villagers unguarded, leaving them with no time to escape,” Jan said. “The survivors are the children who were at school away from the village or the ones who are on jobs outside Buner.”
Onlookers navigate through stones and rocks left behind after a flash flood ran through Beshonai village, in KP’s Buner district, in this photo from August 16. — Arif Hayat
One survivor, Gul Bacha, told Dawn.com, “Most people working in the relief operations are outsiders and there are no confirmed figures on the death toll here. I was upstream when I saw a big wave of black water coming in the direction of our village.
“I ran like crazy and was screaming to the villagers … some people heard me and ran, but most people downstream were caught unawares.”
Javed Khan, a schoolteacher who lost several relatives, told Dawn.com that while no bodies were recovered in Beshornarry, “seventy corpses were swept away and recovered in Peer Baba and Dagar, later laid to rest by locals”.
An image from August 16 shows the collapsed roof of a house destroyed by flash flooding in Beshonai village, in Buner district. — Arif Hayat
Peer Baba Union Council Nazim Abbas Shaheen told Dawn.com that accessibility was a major hurdle. “It is almost impossible to bring heavy machinery into the affected areas,” he said.
In Qadar Nagar, another valley in the area affected by the floods, a collective funeral for 56 people was offered, where one family lost 40 people. The two survivors among them were a father and his son, who were at Islamabad airport to collect another son who had returned home from abroad.
After hitting Beshonai village and Qadar Nagar, the floodwater left behind wrecked houses and destroyed vehicles in its path, affecting major towns and villages. Images taken by a correspondent on the ground showed cars swept away and lodged in buildings, as well as flooded school yards.
A car can be seen lodged into a building by floodwater in Beshornarry village in Buner, in this photo from August 16. — Arif Hayat
Hurricane Erin has rapidly intensified to become a Category 5 storm on Saturday, one day after being upgraded from a tropical storm, as forecasters warned of flooding and possible landslides in the Caribbean through the weekend.
The Atlantic’s first hurricane of the season had maximum sustained winds of 160 miles per hour, forecasters said.
Associated Press
Key Facts
Hurricane Erin intensified into a Category 5 storm Saturday with maximum sustained winds of 160 miles per hour, according to the National Hurricane Center’s latest forecast.
‘Young Sheldon’ star Iain Armitage reveals heartfelt gift from Warner Bros.
Iain Armitage delivered his heartfelt gratitude to the Warner Bros. Studio Tour Hollywood team for their lovely gesture.
Amitage played young Sheldon Cooper in the spin-off series of hit The Big Bang Theory comedy show.
The Big Little Lies star took to his Instagram account sharing a carousel post which featured a special package from the studio.
“A huge thank you to all my friends at @wbtourhollywood !! Last month, just when I was feeling nostalgic and missing things in California, I got this incredible giant envelope from the Warner Brothers Studio Tour Department,” he wrote in the caption.
He went on to voice his admiration for the TV series and Studio Tour Family.
“You all really know how to make me feel good- your package brightened my day, my week, my month, my year! Thank you for your beautiful artwork, and your thoughtful notes,” Iain said.
He continued, “I miss you all and will see you soon! If anyone wants to get a real glimpse into the passion of the art of film making, please go visit my studio guide friends and please tell them that Iain sends his love.”
As of now, Armitage is set to star in The Adam Trials, a sci-fi psychological triller film directed by Ben Ketai.
