Functional network modules overlap and are linked to interindividual connectome differences during human brain development.

The modular structure of functional connectomes in the human brain undergoes substantial reorganization during development. However, previous studies have implicitly assumed that each region participates in one single module, ignoring the potential spatial overlap between modules. How the overlapping functional modules develop and whether this development is related to gray and white matter features remain unknown. Using longitudinal multimodal structural, functional, and diffusion MRI data from 305 children (aged 6 to 14 years), we investigated the maturation of overlapping modules of functional networks and further revealed their structural associations. An edge-centric network model was used to identify the overlapping modules, and the nodal overlap in module affiliations was quantified using the entropy measure. We showed a regionally heterogeneous spatial topography of the overlapping extent of brain nodes in module affiliations in children, with higher entropy (i.e., more module involvement) in the ventral attention, somatomotor, and subcortical regions and lower entropy (i.e., less module involvement) in the visual and default-mode regions. The overlapping modules developed in a linear, spatially dissociable manner, with decreased entropy (i.e., decreased module involvement) in the dorsomedial prefrontal cortex, ventral prefrontal cortex, and putamen and increased entropy (i.e., increased module involvement) in the parietal lobules and lateral prefrontal cortex. The overlapping modular patterns captured individual brain maturity as characterized by chronological age and were predicted by integrating gray matter morphology and white matter microstructural properties. Our findings highlight the maturation of overlapping functional modules and their structural substrates, thereby advancing our understanding of the principles of connectome development.

The neural and genetic underpinnings of different developmental trajectories of Attention-Deficit/Hyperactivity Symptoms in children and adolescents.

BACKGROUND: The trajectory of attention-deficit hyperactivity disorder (ADHD) symptoms in children and adolescents, encompassing descending, stable, and ascending patterns, delineates their ADHD status as remission, persistence or late onset. However, the neural and genetic underpinnings governing the trajectory of ADHD remain inadequately elucidated. METHODS: In this study, we employed neuroimaging techniques, behavioral assessments, and genetic analyses on a cohort of 487 children aged 6-15 from the Children School Functions and Brain Development project at baseline and two follow-up tests for 1 year each (interval 1: 1.14 ± 0.32 years; interval 2: 1.14 ± 0.30 years). We applied a Latent class mixed model (LCMM) to identify the developmental trajectory of ADHD symptoms in children and adolescents, while investigating the neural correlates through gray matter volume (GMV) analysis and exploring the genetic underpinnings using polygenic risk scores (PRS). RESULTS: This study identified three distinct trajectories (ascending-high, stable-low, and descending-medium) of ADHD symptoms from childhood through adolescence. Utilizing the linear mixed-effects (LME) model, we discovered that attention hub regions served as the neural basis for these three developmental trajectories. These regions encompassed the left anterior cingulate cortex/medial prefrontal cortex (ACC/mPFC), responsible for inhibitory control; the right inferior parietal lobule (IPL), which facilitated conscious focus on exogenous stimuli; and the bilateral middle frontal gyrus/precentral gyrus (MFG/PCG), accountable for regulating both dorsal and ventral attention networks while playing a crucial role in flexible modulation of endogenous and extrinsic attention. Furthermore, our findings revealed that individuals in the ascending-high group exhibited the highest PRS for ADHD, followed by those in the descending-medium group, with individuals in the stable-low group displaying the lowest PRS. Notably, both ascending-high and descending-medium groups had significantly higher PRS compared to the stable-low group. CONCLUSIONS: The developmental trajectory of ADHD symptoms in the general population throughout childhood and adolescence can be reliably classified into ascending-high, stable-low, and descending-medium groups. The bilateral MFG/PCG, left ACC/mPFC, and right IPL may serve as crucial brain regions involved in attention processing, potentially determining these trajectories. Furthermore, the ascending-high pattern of ADHD symptoms exhibited the highest PRS for ADHD.

Two new sesterterpenoids from Atractylodes japonica Koidz. ex Kitam.

Two new sesterterpenoids, atractylodes japonica terpenoid acid I (1) and atractylodes japonica terpenoid aldehyde I (2), were isolated from the rhizomes of Atractylodes japonica Koidz. ex Kitam together with ten known compounds (3-12). Their structures were elucidated on the basis of comprehensive spectroscopic analysis (1D/2D NMR, HRESIMS and IR). In addition, all of these isolated compounds were evaluated for their cytotoxic activities against human gastric cancer cell MGC-803 and human hepatocellular cancer cell HepG-2. Most of them exhibited moderate to weak inhibitory effects with IC50 values in the range of 25.15-88.85 µM except for 9-12.

Association of emotional and behavioral problems with the development of the substantia nigra, subthalamic nucleus, and red nucleus volumes and asymmetries from childhood to adolescence: A longitudinal cohort study.

The substantia nigra (SN), subthalamic nucleus (STN), and red nucleus (RN) have been widely studied as important biomarkers of degenerative diseases. However, how they develop in childhood and adolescence and are affected by emotional behavior has not been studied thus far. This population-based longitudinal cohort study used data from a representative sample followed two to five times. Emotional and behavioral problems were assessed with the Strengths and Difficulties Questionnaire (SDQ). Linear mixed models were used to map developmental trajectories and behavioral regulation. Using an innovative automated image segmentation technique, we quantified the volumes and asymmetries of the SN, STN and RN with 1226 MRI scans of a large longitudinal sample of 667 subjects aged 6-15 years and mapped their developmental trajectories. The results showed that the absolute and relative volumes of the bilateral SN and right STN showed linear increases, while the absolute volume of the right RN and relative volume of the bilateral RN decreased linearly, these effects were not affected by gender. Hyperactivity/inattention weakened the increase in SN volume and reduced the absolute volume of the STN, conduct problems impeded the RN volume from decreasing, and emotional symptoms changed the direction of SN lateralization. This longitudinal cohort study mapped the developmental trajectories of SN, STN, and RN volumes and asymmetries from childhood to adolescence, and found the association of emotional symptoms, conduct problems, and hyperactivity/inattention with these trajectories, providing guidance for preventing and intervening in cognitive and emotional behavioral problems.

Functional connectome through the human life span.

The functional connectome of the human brain represents the fundamental network architecture of functional interdependence in brain activity, but its normative growth trajectory across the life course remains unknown. Here, we aggregate the largest, quality-controlled multimodal neuroimaging dataset from 119 global sites, including 33,809 task-free fMRI and structural MRI scans from 32,328 individuals ranging in age from 32 postmenstrual weeks to 80 years. Lifespan growth charts of the connectome are quantified at the whole cortex, system, and regional levels using generalized additive models for location, scale, and shape. We report critical inflection points in the non-linear growth trajectories of the whole-brain functional connectome, particularly peaking in the fourth decade of life. Having established the first fine-grained, lifespan-spanning suite of system-level brain atlases, we generate person-specific parcellation maps and further show distinct maturation timelines for functional segregation within different subsystems. We identify a spatiotemporal gradient axis that governs the life-course growth of regional connectivity, transitioning from primary sensory cortices to higher-order association regions. Using the connectome-based normative model, we demonstrate substantial individual heterogeneities at the network level in patients with autism spectrum disorder and patients with major depressive disorder. Our findings shed light on the life-course evolution of the functional connectome and serve as a normative reference for quantifying individual variation in patients with neurological and psychiatric disorders.

Reduced volume of the left cerebellar lobule VIIb and its increased connectivity within the cerebellum predict more general psychopathology one year later via worse cognitive flexibility in children.

Predicting the risk for general psychopathology (the p factor) requires the examination of multiple factors ranging from brain to cognitive skills. While an increasing number of findings have reported the roles of the cerebral cortex and executive functions, it is much less clear whether and how the cerebellum and cognitive flexibility (a core component of executive function) may be associated with the risk for general psychopathology. Based on the data from more than 400 children aged 6-12 in the Children School Functions and Brain Development (CBD) Project, this study examined whether the left cerebellar lobule VIIb and its connectivity within the cerebellum may prospectively predict the risk for general psychopathology one year later and whether cognitive flexibility may mediate such predictions in school-age children. The reduced gray matter volume in the left cerebellar lobule VIIb and the increased connectivity of this region to the left cerebellar lobule VI prospectively predicted the risk for general psychopathology and was partially mediated by worse cognitive flexibility. Deficits in cognitive flexibility may play an important role in linking cerebellar structure and function to the risk for general psychopathology.

Engineering Nucleotidoproteins for Base-Pairing-Assisted Cytosolic Delivery and Genome Editing.

Protein therapeutics targeting intracellular machineries hold profound potential for disease treatment, and hence robust cytosolic protein delivery technologies are imperatively demanded. Inspired by the super-negatively charged, nucleotide-enriched structure of nucleic acids, adenylated pro-proteins (A-proteins) with dramatically enhanced negative surface charges have been engineered for the first time via facile green synthesis. Then, thymidine-modified polyethyleneimine is developed, which exhibits strong electrostatic attraction, complementary base pairing, and hydrophobic interaction with A-proteins to form salt-resistant nanocomplexes with robust cytosolic delivery efficiencies. The acidic endolysosomal environment enables traceless restoration of the A-proteins and consequently promotes the intracellular release of the native proteins. This strategy shows high efficiency and universality for a variety of proteins with different molecular weights and isoelectric points in mammalian cells. Moreover, it enables highly efficient delivery of CRISPR-Cas9 ribonucleoproteins targeting fusion oncogene EWSR1-FLI1, leading to pronounced anti-tumor efficacy against Ewing sarcoma. This study provides a potent and versatile platform for cytosolic protein delivery and gene editing, and may benefit the development of protein pharmaceuticals.

Emotional and behavioral problems change the development of cerebellar gray matter volume, thickness, and surface area from childhood to adolescence: A longitudinal cohort study.

AIMS: Increasing evidence indicates that major neurodevelopmental disorders have potential links to abnormal cerebellar development. However, the developmental trajectories of cerebellar subregions from childhood to adolescence are lacking, and it is not clear how emotional and behavioral problems affect them. We aim to map the developmental trajectories of gray matter volume (GMV), cortical thickness (CT), and surface area (SA) in cerebellar subregions from childhood to adolescence and examine how emotional and behavioral problems change the cerebellar development trajectory in a longitudinal cohort study. METHOD: This population-based longitudinal cohort study used data on a representative sample of 695 children. Emotional and behavioral problems were assessed at baseline and at three annual follow-ups with the Strengths and Difficulties Questionnaire (SDQ). RESULTS: Using an innovative automated image segmentation technique, we quantified the GMV, CT, and SA of the whole cerebellum and 24 subdivisions (lobules I-VI, VIIB, VIIIA&B, and IX-X plus crus I-II) with 1319 MRI scans from a large longitudinal sample of 695 subjects aged 6-15 years and mapped their developmental trajectories. We also examined sex differences and found that boys showed more linear growth, while girls showed more nonlinear growth. Boys and girls showed nonlinear growth in the cerebellar subregions; however, girls reached the peak earlier than boys. Further analysis found that emotional and behavioral problems modulated cerebellar development. Specifically, emotional symptoms impede the expansion of the SA of the cerebellar cortex, and no gender differences; conduct problems lead to inadequate cerebellar GMV development only in girls, but not boys; hyperactivity/inattention delays the development of cerebellar GMV and SA, with left cerebellar GMV, right VIIIA GMV and SA in boys and left V GMV and SA in girls; peer problems disrupt CT growth and SA expansion, resulting in delayed GMV development, with bilateral IV, right X CT in boys and right Crus I GMV, left V SA in girls; and prosocial behavior problems impede the expansion of the SA and lead to excessive CT growth, with bilateral IV, V, right VI CT, left cerebellum SA in boys and right Crus I GMV in girls. CONCLUSIONS: This study maps the developmental trajectories of GMV, CT, and SA in cerebellar subregions from childhood to adolescence. In addition, we provide the first evidence for how emotional and behavioral problems affect the dynamic development of GMV, CT, and SA in the cerebellum, which provides an important basis and guidance for the prevention and intervention of cognitive and emotional behavioral problems in the future.

Vertically Aligned Boron Nitride Nanosheets Films for Superior Electronic Cooling.

Thermally conductive and electrically insulating thermal interface materials (TIMs) are highly desired for electronic cooling. To improve heat transfer efficiency, thermally conductive fillers with a high loading content have been incorporated into the polymer-based TIMs. However, this is usually at the expense of the interfacial thermal resistance reduction and reliability. In this study, vertically aligned boron nitride nanosheet films (VBNFs) have been prepared by a scalable microfluidic spinning process and template-assisted chemical vapor deposition conversion method. A further high-temperature annealing was applied to achieve high crystallinity. VBNFs have been applied as fillers to fabricate TIMs and achieve a superior through-plane thermal conductivity of 6.4 W m-1 K-1 and low modulus of 2.2 MPa at low BN loading of 9.85 vol %, benefitting from the well-aligned vertical sheet structure and high crystallinity. In addition, the fabricated TIMs present high-volume resistivity and breakdown strength, satisfying the electrical insulation demands. The high thermal conductivity and low modulus contribute an outstanding cooling performance to the TIMs in the heat dissipation application for high-power LEDs. This template-assisted conversion technology for the fabrication of orientated BN nanosheets structure and the prepared high-performance TIMs pave the way for efficient thermal management of high-power electronics.

Scalable microfluidic fabrication of vertically aligned two-dimensional nanosheets for superior thermal management.

Two-dimensional (2D) nanosheets have been assembled into various macroscopic structures for wide engineering applications. To fully explore their exceptional thermal, mechanical, and electrical properties, 2D nanosheets must be aligned into highly ordered structures due to their strong structural anisotropy. Structures stacked layer by layer such as films and fibers have been readily assembled from 2D nanosheets due to their planar geometry. However, scalable manufacturing of macroscopic structures with vertically aligned 2D nanosheets remains challenging, given their large lateral size with a thickness of only a few nanometers. Herein, we report a scalable and efficient microfluidics-enabled sheet-aligning process to assemble 2D nanosheets into a large-area film with a highly ordered vertical alignment. By applying microchannels with a high aspect ratio, 2D nanosheets were well aligned vertically under strong channel size confinement and high flow shear stress. A vertically aligned graphene sheet film was obtained and applied to effectively improve the heat transfer of thermal interfacial materials (TIMs). Superior through-plane thermal conductivity of 82.7 W m-1 K-1 at a low graphene content of 11.8 vol% was measured for vertically aligned TIMs. Thus, they demonstrate exceptional thermal management performance for switching power supplies with high reliability.

Two new diarylheptanoids from the green walnut husks of Juglans mandshurica Maxim.

Chemical investigation on the 95% ethanol extract of green walnut husks of Juglans mandshurica Maxim. led to the isolation of two new diarylheptanoid compounds, including Juglanin K (1) and Myricananin I (2), together with ten known compounds (3-13). Their structures were elucidated by extensive analyses of comprehensive spectroscopic methods. The isolated compounds were evaluated for their antibacterial activities against Escherichia coli and Staphylococcus aureus. The results showed that compound 2 had moderate antibacterial activity with the MIC values of 0.313 and 0.156 mg/mL, respectively. compounds 1, 3-7 and 13 showed weakly antibacterial activities against Escherichia coli and Staphylococcus aureus with the MIC values of 0.625-2.5 mg/mL.

Serial and discrete naming and reading in Chinese first graders: Testing predictions from the cascaded processing hypothesis.

Recent studies have suggested that-beyond automaticity and prosody-reading fluency involves parallel processing of adjacent items presented in a sequence, termed "cascaded processing." To date, most studies examining cascaded processing have been conducted in alphabetic orthographies. Thus, the purpose of this study was to examine the cascaded processing hypothesis in Chinese. A total of 119 Grade 1 Chinese children (61 boys and 58 girls; Mage = 7.30 years, SD = 0.31) were assessed on serial and discrete naming of digits as well as on serial and discrete naming of high-frequency one- and two-character words and low-frequency one-character words presented with pinyin. Results of hierarchical regression analyses showed, first, that serial digit naming was a unique predictor of discrete naming of low-frequency one-character words and two-character words, but not of high-frequency one-character words. Second, serial digit naming was a unique predictor of reading of high-frequency one- and two-character word reading after controlling for discrete word reading. These findings suggest that Chinese first graders process high-frequency characters holistically (similar to simple digits), which then facilitates parallel processing of multiple stimuli when they are presented in a sequence.

Drought stress delays photosynthetic induction and accelerates photoinhibition under short-term fluctuating light in tomato.

Fluctuating light (FL) and drought stress usually occur concomitantly. However, whether drought stress affects photosynthetic performance under FL remains unknown. Here, we measured gas exchange, chlorophyll fluorescence, and P700 redox state under FL in drought-stressed tomato (Solanum lycopersicum) seedlings. Drought stress significantly delayed the induction kinetics of stomatal and mesophyll conductances after transition from low to high light and thus delayed photosynthetic induction under FL. Therefore, drought stress exacerbated the loss of carbon gain under FL. Furthermore, restriction of CO2 fixation under drought stress aggravated the over-reduction of photosystem I (PSI) upon transition from low to high light. The resulting stronger FL-induced PSI photoinhibition significantly suppressed linear electron flow and PSI photoprotection. These results indicated that drought stress not only caused a larger loss of carbon gain under FL but also accelerated FL-induced photoinhibition of PSI. Furthermore, drought stress enhanced relative cyclic electron flow in FL, which partially compensated for restricted CO2 fixation and thus favored PSI photoprotection under FL. To our knowledge, we here show new insight into how drought stress affects photosynthetic performance under FL.

Inflammatory Cell-Inspired Cascade Nanozyme Induces Intracellular Radical Storm for Enhanced Anticancer Therapy.

Manipulating intracellular levels of reactive oxygen and nitrogen species (RONS) is of great potential for cancer treatment. Inspired by the natural mechanism of a radical storm in inflammatory cells via activated and regulatable biocatalysis, the authors herein report a self-powered nanozyme that can enable RONS production in tumor cells via cascade reactions. The nanozymes are constructed via glucose oxidase (GOx)-templated inverse microemulsion polymerization from acrylamide, arginine-acrylamide, ferrocene-acrylate, and N,N'-bis(acryloyl)cystamine, followed by surface coating with hyaluronic acid. After targeted delivery into cancer cells, the nanozymes are dissociated by intracellular glutathione to release GOx, which decomposed glucose to generate gluconic acid and H2 O2 . Under such acidified conditions, H2 O2 efficiently oxidized pendant arginine residues to produce nitric oxide , transformed into a highly toxic hydroxyl radical and superoxide anion via ferrocene-mediated Fenton reaction and Haber-Weiss cycle, and simultaneously generated peroxynitrite anion via reaction between NO and ·O2 - , thus provoking the RONS radical storm to effectively eradicate A549 tumor cells both in vitro and in vivo. This nature-inspired enzyme-chemical dynamic therapy may provide a promising modality for anti-cancer treatment.

Enantioselective Single and Dual α-C-H Bond Functionalization of Cyclic Amines via Enzymatic Carbene Transfer.

Cyclic amines are ubiquitous structural motifs found in pharmaceuticals and biologically active natural products, making methods for their elaboration via direct C-H functionalization of considerable synthetic value. Herein, we report the development of an iron-based biocatalytic strategy for enantioselective α-C-H functionalization of pyrrolidines and other saturated N-heterocycles via a carbene transfer reaction with diazoacetone. Currently unreported for organometallic catalysts, this transformation can be accomplished in high yields, high catalytic activity, and high stereoselectivity (up to 99:1 e.r. and 20,350 TON) using engineered variants of cytochrome P450 CYP119 from Sulfolobus solfataricus. This methodology was further extended to enable enantioselective α-C-H functionalization in the presence of ethyl diazoacetate as carbene donor (up to 96:4 e.r. and 18,270 TON), and the two strategies were combined to achieve a one-pot as well as a tandem dual C-H functionalization of a cyclic amine substrate with enzyme-controlled diastereo- and enantiodivergent selectivity. This biocatalytic approach is amenable to gram-scale synthesis and can be applied to drug scaffolds for late-stage C-H functionalization. This work provides an efficient and tunable method for direct asymmetric α-C-H functionalization of saturated N-heterocycles, which should offer new opportunities for the synthesis, discovery, and optimization of bioactive molecules.

Upward propagation of gigantic jets revealed by 3D radio and optical mapping.

Occasionally, lightning will exit the top of a thunderstorm and connect to the lower edge of space, forming a gigantic jet. Here, we report on observations of a negative gigantic jet that transferred an extraordinary amount of charge between the troposphere and ionosphere (∼300 C). It occurred in unusual circumstances, emerging from an area of weak convection. As the discharge ascended from the cloud top, tens of very high frequency (VHF) radio sources were detected from 22 to 45 km altitude, while simultaneous optical emissions (777.4 nm OI emitted from lightning leaders) remained near cloud top (15 to 20 km altitude). This implies that the high-altitude VHF sources were produced by streamers and the streamer discharge activity can extend all the way from near cloud top to the ionosphere. The simultaneous three-dimensional radio and optical data indicate that VHF lightning networks detect emissions from streamer corona rather than the leader channel, which has broad implications to lightning physics beyond that of gigantic jets.

Different predictive roles of phonological awareness and visual attention span for early character reading fluency in Chinese.

Previous studies have usually explored the possible mechanisms underlying reading fluency separately from either linguistic (e.g., phonological awareness, PA) or basic cognitive (e.g., visual attention span, VAS) levels, but relatively little is known about the systematic relationship between multiple levels of cognitive factors and reading fluency development. Thus, the present study, from a combinative point of view, investigated the relationship between VAS at a basic cognitive level and PA at a high linguistic level and explored their predictive power for reading fluency development a year later through a longitudinal method. PA and VAS were measured in sixty-five Chinese children who had just entered Grade 1 of primary school (Time 1); PA tests included syllabic and phonemic aspects, and VAS was reflected by performance in the visual 1-back task with nonverbal stimuli and no verbal response. Reading fluency was tested at the single-character level at the beginning of Grade 2 (Time 2). The results showed that phoneme awareness could not predict variation in reading fluency, while both syllable awareness and VAS showed significant predictive powers for single-character reading fluency. Additionally, further analysis revealed that the relationship between VAS and later single-character reading fluency was partially mediated by syllable awareness. These findings indicated Chinese language characteristics in the relationship among VAS, PA, and reading fluency. The possible roles of multiple levels of cognitive skills in fluent reading have been further discussed and have implications for the early diagnosis and remediation of reading dysfluency.

Electromagnetic Radiation Spectrum of a Composite System.

In many physics fields, the radio emission of a composite system composed of a large number of randomly occurring but similar emission sources is encountered. In general, the composite system lasts longer than each individual component and individual source currents vary much more rapidly. This Letter presents a theory to understand the electromagnetic radiation spectrum of such a system. If the temporal distribution of the random occurrence of the component and the distribution to describe the relevant emission source properties are known, the spectrum of the composite system can be readily found from this theory. There are two main terms that define the spectrum: one term results from the coherent summation of the contributions from individual sources and is proportional to the square of the total number of the components in the system; the other term results from an incoherent summation and is proportional to the first power of that number. This can lead to drastically different spectral magnitudes in different spectral regions, typically with the spectral magnitude in the lower frequency region many orders of magnitude stronger than that in the higher frequency region.

An Enzymatic Platform for the Highly Enantioselective and Stereodivergent Construction of Cyclopropyl-δ-lactones.

Abiological enzymes offers new opportunities for sustainable chemistry. Herein, we report the development of biological catalysts derived from sperm whale myoglobin that exploit a carbene transfer mechanism for the asymmetric synthesis of cyclopropane-fused-δ-lactones, which are key structural motifs found in many biologically active natural products. While hemin, wild-type myoglobin, and other hemoproteins are unable to catalyze this reaction, the myoglobin scaffold could be remodeled by protein engineering to permit the intramolecular cyclopropanation of a broad spectrum of homoallylic diazoacetate substrates in high yields and with up to 99 % enantiomeric excess. Via an alternate evolutionary trajectory, a stereodivergent biocatalyst was also obtained for affording mirror-image forms of the desired bicyclic products. In combination with whole-cell transformations, the myoglobin-based biocatalyst was used for the asymmetric construction of a cyclopropyl-δ-lactone scaffold at a gram scale, which could be further elaborated to furnish a variety of enantiopure trisubstituted cyclopropanes.

Fast negative breakdown in thunderstorms.

Thunderstorms are natural laboratories for studying electrical discharges in air, where the vast temporal, spatial, and energy scales available can spawn surprising phenomena that reveal deficiencies in our understanding of dielectric breakdown. Recent discoveries, such as sprites, jets, terrestrial gamma ray flashes, and fast positive breakdown, highlight the diversity of complex phenomena that thunderstorms can produce, and point to the possibility for electrical breakdown/discharge mechanisms beyond dielectric breakdown theory based mainly on laboratory experiments. Here we present one such confounding discovery, termed fast negative breakdown, that does not fit with our current understanding of dielectric breakdown. Our adaptation of radio astronomy imaging techniques to study extremely transient lightning-associated events confirms that electrical breakdown in thunderstorms can begin with oppositely-directed fast breakdown of negative polarity, similar and in addition to fast positive breakdown expected from conventional dielectric theory and recent observations. The discovery of fast negative breakdown calls for an addendum to the physical description of electrical discharges in air.