Network connectivity underlying information processing speed in children: Application of a pediatric brain tumor survivor injury model.

Elucidating how adaptive and maladaptive changes to the structural connectivity of brain networks influences neural synchrony, and how this structure-function coupling impacts cognition is an important question in human neuroscience. This study assesses these links in the default mode and executive control networks during resting state, a visual-motor task, and through computational modeling in the developing brain and in acquired brain injuries. Pediatric brain tumor survivors were used as an injury model as they are known to exhibit cognitive deficits, structural connectivity compromise, and perturbations in neural communication. Focusing on information processing speed to assess cognitive performance, we demonstrate that during the presence and absence of specific task demands, structural connectivity of these critical brain networks directly influences neural communication and information processing speed, and white matter compromise has an indirect adverse impact on reaction time via perturbed neural synchrony. Further, when our experimentally acquired structural connectomes simulated neural activity, the resulting functional simulations aligned with our empirical results and accurately predicted cognitive group differences. Overall, our synergistic findings further our understanding of the neural underpinnings of cognition and when it is perturbed. Further establishing alterations in structural-functional coupling as biomarkers of cognitive impairments could facilitate early intervention and monitoring of these deficits.

A Versatile Magnetic Nanoplatform for Plug-and-Play Functionalization: Genetically Programmable Cargo Loading to Bacterial Magnetosomes by SpyCatcher "Click Biology".

Bacterial magnetosomes ("MAGs") represent a promising class of magnetic iron oxide nanoparticles with exceptional material characteristics and high application potential in the biomedical and biotechnological field. For the surface functionalization of MAGs with different protein cargos, their enveloping membrane can be addressed by genetic means. However, the expression of foreign polypeptides as translational fusion to magnetosome membrane proteins is still laborious and lacks versatility as the generated particles are monospecific and thus restricted to predetermined functions. Utilizing the SpyTag-SpyCatcher (ST-SC) bioconjugate system, we here establish a flexible platform for the targeted nanoassembly of multifunctional MAGs that combines the rapidity of chemical coupling (e.g., by cross-linking reactions) and the unmatched selectivity and controllability of in vivo functionalization. MAGs genetically engineered to display either SC- or ST-connectors are shown to efficiently bind a variety of complementary tagged (protein) cargo. Specifically, we cover a broad spectrum of representative functional moieties and foreign cargo (such as enzymes, antibodies, fluorophores, and silica beads) with relevance in biotechnology and biomedicine and demonstrate the interchangeability of the MAGs-adapted ST-SC system. For the controlled generation of artificial shells surrounding the particles, SC-MAGs are effectively coated by protein-corona proteins. The potential of the here-provided toolkit is even more enhanced by using SC-MAGs as an affinity tool for selective protein pulldown in vitro and in vivo. Overall, this innovative technology turns bacterial MAGs into a flexible magnetic nanoscaffold for the targeted plug-and-play display of virtually unlimited additional functionalities, thereby generating a multitude of magnetic hybrid materials that can be used in many applications.

Predictive value of the left atrioventricular coupling index for recurrence after radiofrequency ablation of paroxysmal atrial fibrillation.

BACKGROUND: Although patients with paroxysmal atrial fibrillation (PAF) are preferred to undergo catheter ablation (CA), the high possibility of recurrence following surgery is still concerning. We aimed to evaluate the ability of the left atrioventricular coupling index (LACI), which is the ratio of the left atrium end-diastolic volume to the left ventricle end-diastolic volume, to predict PAF recurrence after CA. METHODS: Patients with PAF undergoing CA for the first time between January 2018 and June 2021 were admitted and grouped by recurrence within a year. LACI was measured before CA using ultrasonography. Risk factors identified by multivariable logistic regression analysis, and the area under the receiver operating characteristic (ROC) curve was used to assess the ability of LACI to predict PAF recurrence after CAP. RESULTS: Among the 204 patients treated at our hospital, 164 patients were included in the research after eliminating those who were lost to follow-up. Among them, 56 individuals had recurrence following a 90-day blanking period. Recurrence is more likely in elderly patients with high blood pressure. Patients who suffered recurrence exhibited lower left atrial ejection fraction and increased LACI, left atrial volume minimum, and left atrium volume index maximum. LACI was an independent risk factor for postoperative recurrence (OR: 1.526, 95% CI: 1.325-1.757, P < 0.001), and ROC displayed remarkable predictive value [area under the curve (AUC) = 0.868]. CONCLUSIONS: High LACI is significantly associated with postoperative recurrence in PAF patients, and LACI has incremental prognostic value to predict recurrence.

Cardiovascular magnetic resonance-derived left atrioventricular coupling index as a novel prognostic marker for light-chain amyloidosis.

BACKGROUND: Left atrioventricular coupling index (LACI) is a novel biomarker, and the prognostic value of LACI to predict cardiovascular events has been validated. The present study aimed to explore the prognostic value of LACI in patients with light-chain (AL) amyloidosis. METHODS: We prospectively enrolled 179 patients with AL amyloidosis who underwent cardiovascular magnetic resonance imaging between December 2011 and January 2020. LACI was defined as the ratio between the left atrial volume and the left ventricular volume at end-diastole. The primary endpoint was all-cause mortality. Receiver operating characteristic curve was used to identify the optimal cut-off of LACI in predicting all-cause mortality. Univariable and multivariable Cox proportional hazard models were used to assess the association of LACI and primary endpoint. RESULTS: During a median follow-up of 30 months, 118 (65.9 %) patients with all-cause mortality were documented. LACI was significantly higher in patients with primary endpoint compared to those without primary endpoint (55.4 %, interquartile range: 31.6 %-71.5 % vs. 39.4 %, interquartile range: 24.1 %-54.7 %, p = 0.001). The optimal cut-off for LACI to predict mortality was 49.3 %. In multivariate Cox regression analysis, LACI≥49.3 % (HR 1.907, 95 % CI 1.273-2.857, p = 0.002) was an independent predictor of all-cause mortality. On Kaplan-Meier analysis, patients at advanced Mayo stage (IIIa and IIIb) can be further risk stratified using LACI≥49.3 % (log-rank p = 0.035, p = 0.025). CONCLUSION: The LACI provides powerful independent prognostic value in AL amyloidosis. The LACI has incremental prognostic value to predict all-cause mortality over the Mayo stage in patients at the advanced Mayo stage.

Drought-dependent regulation of cell coupling in Arabidopsis leaf epidermis requires plasmodesmata protein NHL12.

The cytoplasms of most plant cells are connected by membrane-lined cell wall channels, the plasmodesmata (PD). Dynamic regulation of sugar, hormone and protein diffusion through PD is essential for plant development and stress responses. Understanding this regulation requires knowledge of factors and mechanisms that control PD permeability through the modulation of callose levels in the cell wall around PD openings. We investigated PD regulation in leaf epidermis cells in relation to drought stress in Arabidopsis thaliana. Upon finding PD-mediated cell wall permeability decreased by drought stress and the hormone ABA, we tested several PD-associated genes with drought-responsive expression for their involvement in this response. Mutants of NHL12 showed relatively low PD permeability that was unaffected by drought or ABA treatment. Overexpression of NHL12 in Nicotiana benthamiana epidermis cells increased PD permeability. Moreover, we show that NHL12 can potentially interact with the callose synthase-regulator NHL3 and we explored the effect of NHL12 abundance and/or lower interface permeability on ABA signaling genes. Our results indicate that NHL12 is a drought-responsive negative regulator of PD callose levels and, thereby, interface permeability. Results are discussed with regard to PD function during drought stress and the regulation of intercellular transport.

Peroxymonosulfate-assisted photocatalysis system enhanced magnetic Fe3O4@P-C3N4 treatment of tetracycline wastewater: Multi-pathways mediated electrons migration to generate reactive species.

Photocatalytic wastewater purification is essential for environmental remediation, but rapid carrier recombination and limited oxidative capacity hinder progress. This study proposes an innovative strategy by integrating homogeneous and heterogeneous electron acceptors into a g-C3N4-based photocatalytic system, significantly enhancing the multipath utilization of photogenerated electrons. A novel Fe3O4@P-C3N4 was developed to activate an advanced peroxymonosulfate-assisted photocatalysis (PAP) system, achieving complete degradation and significant mineralization of tetracycline (TC) in real water environments, outperforming others reported in the last five years. Phytic acid, as a key precursor, modifies the hollow tubular morphology and introduces phosphorus (P) heteroatoms as electronic trapping centers, enhancing the visible light response and carrier separation, thereby promoting the Fe2+/Fe3+ cycle and the formation of reactive species. Density functional theory (DFT) calculations pinpointed TC's vulnerable sites and synergically identified reactive species, revealing almost non-toxic degradation processes. Moreover, the recyclable magnetic Fe3O4@P-C3N4/PAP system demonstrates practical application potential and leaching stability in cyclic and continuous testing. This study offers unique insights into the strategic design of photocatalysts and catalytic environments, potentially advancing practical wastewater remediation.

Effective Alkyl-Alkyl Cross-Coupling with an Iron-Xantphos Catalyst: Mechanistic and Structural Insights.

While iron-catalyzed C(sp2)-C(sp3) cross-couplings have been widely studied and developed in the last decade, alkyl-alkyl cross-coupling systems with iron remain underdeveloped despite the importance of C(sp3)-C(sp3) bonds in organic synthesis. A major challenge to the development of these reactions is the current lack of fundamental insight into ligand effects and organoiron intermediates that enable effective alkyl-alkyl couplings. The current study addresses this longstanding limitation using a combination of 57Fe Mössbauer spectroscopy, SC-XRD (single-crystal X-ray diffraction) and reactivity studies of alkyl-alkyl coupling with iron-Xantphos to define the in situ formed iron-Xantphos intermediates in catalysis. Combined with detailed reactivity studies, the nature of the key mechanistic pathways in catalysis and ligands effects to achieve effective alkyl-alkyl cross-coupling over competing b-H elimination pathways are probed. Overall, these foundational studies provide a platform for future bespoke ligand and pre-catalyst design for alkyl-alkyl cross-coupling methods development with sustainable iron catalysis.

Fire-Resistant and Thermal Stability Properties of Fluorosilicone Adhesives by Incorporation of Surface-Modified Aluminum Trihydrate.

Fluorosilicone was combined with aluminum trihydrate (ATH) to induce synergistic flame-retardant and thermal-resistant properties. The surface of ATH was modified with four different silane coupling agents. The flammability and mechanical properties of the fluorosilicone/ATH composites were assessed using an UL94 vertical test and a die shear strength test. The change in shear strength was investigated under aging for 1000 h at -55 °C and 150 °C. Pure fluorosilicone had inherent fire resistance and thus achieved a V-0 rating even at 20 wt.% ATH loading. Upon addition of ATH treated with 3-glycidoxypropyl trimethoxysilane, the composites exhibited the highest shear strength of 3.9 MPa at 23 °C because of the additional crosslinking reaction of fluorosilicone resin with the epoxide functional group of the coupling agent. Regardless of the types of coupling agents, the composites exhibited similar flame retardancy at the same ATH content, with a slight reduction in shear strength at 180 °C and 250 °C. The shear strength of the adhesives gradually decreased with aging time at -55 °C, but increased noticeably from 3.9 MPa to 11.5 MPa when aged at 150 °C due to the occurrence of the additional crosslinking reaction of fluorosilicone.

Gas-Phase Studies of NMR Shielding and Indirect Spin-Spin Coupling in 13C-Enriched Ethane and Ethylene.

13C and 1H NMR spectra were observed as the function of density in 1,2-13C-enriched ethane and ethylene for the pure gaseous compounds and their binary mixtures with xenon and carbon dioxide gases as the solvents. All the chemical shifts and indirect spin-spin couplings were linearly dependent on the solvent density. The appropriate NMR parameters (σ and nJ) in isolated 13C2H6 and 13C2H4 molecules and the coefficients responsible for the binary molecular interactions were determined and compared with previous similar measurements and selected calculated shielding data. The newly obtained 13C shielding values in the isolated ethane and ethylene molecules suggest visible secondary isotope effects due to the additional carbon-13 atom. All the investigated shielding parameters depend on intermolecular interactions, and the dependence of 13C shielding is much more marked. In contrast, the indirect spin-spin couplings in 13C2H6 and 13C2H4 molecules are almost independent of solvent molecules. Their nJ values determined in liquids over sixty years ago are generally consistent with the same nJ parameters in isolated 13C2H6 and 13C2H4 molecules.

3D-Cultured MC3T3-E1-Derived Exosomes Promote Endothelial Cell Biological Function under the Effect of LIPUS.

Porous Ti-6Al-4V scaffold materials can be used to heal massive bone defects because they can provide space for vascularisation and bone formation. During new bone tissue development, rapid vascular ingrowth into scaffold materials is very important. Osteoblast-derived exosomes are capable of facilitating angiogenesis-osteogenesis coupling. Low-intensity pulsed ultrasound (LIPUS) is a physical therapy modality widely utilised in the field of bone regeneration and has been proven to enhance the production and functionality of exosomes on two-dimensional surfaces. The impact of LIPUS on exosomes derived from osteoblasts cultured in three dimensions remains to be elucidated. In this study, exosomes produced by osteoblasts on porous Ti-6Al-4V scaffold materials under LIPUS and non-ultrasound stimulated conditions were co-cultured with endothelial cells. The findings indicated that the exosomes were consistently and stably taken up by the endothelial cells. Compared to the non-ultrasound group, the LIPUS group facilitated endothelial cell proliferation and angiogenesis. After 24 h of co-culture, the migration ability of endothelial cells in the LIPUS group was 17.30% higher relative to the non-ultrasound group. LIPUS may represent a potentially viable strategy to promote the efficacy of osteoblast-derived exosomes to enhance the angiogenesis of porous Ti-6Al-4V scaffold materials.

Right Ventricular to Pulmonary Artery Uncoupling Is Associated With Impaired Exercise Capacity in Patients With Transthyretin Cardiac Amyloidosis.

BACKGROUND: Exercise capacity is related to mortality and morbidity in heart failure (HF) patients. Determinants of exercise capacity in transthyretin cardiac amyloidosis (ATTR-CA) have not been established. METHODS AND RESULTS: This single-center study retrospectively evaluated ATTR-CA patients and patients with non-amyloidosis HF with preserved/mildly reduced ejection fraction (HFpEF/HFmrEF) (n=32 and n=51, respectively). In the ATTR-CA group, the median age was 75.5 years (interquartile range [IQR] 71.3-78.8 years), 90.6% were male, and the median left ventricular (LV) ejection fraction was 53.5% (IQR 41.4-65.6%). Cardiopulmonary exercise tests revealed a median peak oxygen consumption and anaerobic threshold of 15.9 (IQR 11.6-17.4) and 10.6 (IQR 8.5-12.0] mL/min/kg, respectively, and ventilatory efficiency (minute ventilation/carbon dioxide production [V̇E/V̇CO2] slope) of 35.5 (IQR 32.0-42.5). Among exercise variables, V̇E/V̇CO2slope has the greatest prognostic value. Univariate analysis revealed a significant correlation between V̇E/V̇CO2slope and age, LV global longitudinal strain, tricuspid annular plain systolic excursion/pulmonary arterial systolic pressure (TAPSE/PASP) ratio, and mixed venous oxygen saturation. In multivariate analyses, the TAPSE/PASP ratio was an independent predictor of V̇E/V̇CO2slope (95% confidence interval -44.5, -10.8; P=0.0067). In non-amyloidosis HFpEF/HFmrEF patients, the TAPSE/PASP ratio was not independently correlated with V̇E/V̇CO2slope. CONCLUSIONS: Right ventricular-pulmonary artery coupling estimated by the TAPSE/PASP ratio determines exercise capacity in ATTR-CA patients. This highlights the importance of early therapeutic intervention against underappreciated right ventricular dysfunction associated with ATTR-CA.

Adhesion Strength Enhancement of Butyl Rubber and Aluminum Using Nanoscale Self-Assembled Monolayers of Various Silane Coupling Agents for Vibration Damping Plates.

In this paper, we enhance the adhesion strength of butyl rubber-based vibrational damping plates using nanoscale self-assembled monolayers of various silane coupling agents. The silane coupling agents used to chemically modify the plate's aluminum surface include 3-aminopropyltriethoxysilane (APTES), (3-glycidyloxypropyl) triethoxysilane (GPTES), 3-mercaptopropyltrimethoxysilane (MPTMS), and 3-(triethoxysilyl)propyl isocyanate (ICPTES). The modified surfaces were analyzed using Fourier transform infrared spectroscopy (FT-IR) and X-ray photoelectron spectroscopy (XPS), and the enhancement in adhesion strength between the rubber and aluminum was estimated through T-Peel tests. As a result, MPTMS showed the highest enhancement in adhesion strength, of approximately 220% compared to the untreated sample, while GPTES, ICPTES, and APTES resulted in adhesion strength enhancements of approximately 200%, 150%, and 130%, respectively.

The Impact of Chemotherapy on Arterial Stiffness and Ventricular-Arterial Coupling in Women with Breast Cancer.

BACKGROUND: The cardiac toxicity of chemotherapy for breast cancer is not uncommon and has been associated with elevated morbidity and mortality. In the present study, we assessed the impact of chemotherapy on cardiovascular function by assessing the cardio-ankle vascular index (CAVI), global longitudinal strain (GLS) and ventricular-arterial coupling (VAC: CAVI/GLS ratio) in chemotherapy-treated women. METHODS: This prospective study enrolled 78 women with breast cancer who were receiving anthracycline-based chemotherapy +/- anti-HER2 therapy (trastuzumab +/- pertuzumab). Forty-one age-matched healthy women served as controls. We comparatively evaluated left ventricular ejection fraction (LVEF), CAVI, GLS and VAC, between the chemotherapy and control groups. We also assessed their changes over time (baseline, 3-month and 6-month time point) and their independent association with the incidence of cancer therapy-related cardiovascular dysfunction (CTRCD) in the chemotherapy group. RESULTS: In comparison to healthy controls, women receiving chemotherapy presented with significantly higher GLS (from -21.02 ± 2.09% to -19.01 ± 2.81%, p < 0.001) and VAC (-0.36 ± 0.06 to -0.41 ± 0.11, p < 0.001). The presence of CTRCD was associated with a further increase in GLS and CAVI and a significant decline in LVEF and VAC compared to CTRCD-free women (p < 0.001). Baseline, CAVI, GLS and VAC were independently associated with CTRCD development during follow-up. CONCLUSION: Women with breast cancer undergoing chemotherapy displayed abnormal levels of CAVI, VAC and GLS, compared to healthy individuals. Those effects on VAC and CAVI were more exaggerated among women with CTRCD, implicating their potential use to refine screening and therapeutic strategies for this specific population.

Neuroprotective effects of salvianolic acids combined with Panax notoginseng saponins in cerebral ischemia/reperfusion rats concerning the neurovascular unit and trophic coupling.

BACKGROUND: The neurovascular unit (NVU) and neurovascular trophic coupling (NVTC) play a key regulatory role in brain injury caused by ischemic stroke. Salvianolic acids (SAL) and Panax notoginseng saponins (PNS) are widely used in China to manage ischemic stroke. Neuroprotective effects of SAL and PNS, either taken alone or in combination, were examined in this research. METHODS: Wistar rats were randomly divided into the following groups: Sham group (Sham), cerebral ischemia/reperfusion group (I/R), I/R with SAL group (SAL), I/R with PNS group (PNS), I/R with SAL combined with PNS (SAL + PNS), and I/R with edaravone group (EDA). Treatment was administered once daily for two days after modeling of middle cerebral artery occlusion/reperfusion (MCAO/R). RESULTS: Compared with the I/R group, SAL, PNS, or SAL + PNS treatment reduced infarct size, improved neurological deficit score, reduced Evans blue extravasation, increased expression of CD31 and tight junction proteins (TJs), including zonula occludens-1 (ZO-1), zonula occludens-2 (ZO-2), and junctional adhesion molecule-1 (JAM-1). Furthermore, SAL, PNS, or SAL + PNS suppressed the activations of microglia and astrocyte and led to the amelioration of neuron and pericyte injury. Treatment also inhibited NVU dissociation of GFAP/PDGFRß and Collagen IV/GFAP while upregulated the expression level of BDNF/TrkB and BDNF/NeuN. CONCLUSIONS: SAL and PNS have significantly remedied structural and functional disorders of NVU and NVTC in I/R injury. These effects were more pronounced when SAL and PNS were combined than when used separately.

Evaluation of the Polypyrrole Coupling Mode for High-Performance Dual-Ion Batteries.

Due to the advantages of large interstitial sites, antisolubility, and reversible insertion and extraction of anions, polypyrrole (PPy) has become an excellent P-type electrode material for dual-ion batteries. Unfortunately, PPy electrodes inevitably suffer from low specific capacity and poor cycle stability because of structural disintegration during repeated cycling as well as poor doping ability brought on by aggregation or cross-linking within the PPy chain. In this work, PPy with different proportions of coupling mode (α-α, α-ß, or ß-ß coupling) was derived from different preparation methods. Among them, PPy derived from the interfacial frozen polymerization method (I-PPy) is dominated by the α-α coupling mode and possesses the best anion doping ability and the highest specific capacity of 119 mAh g-1 at 100 mA g-1 compared with PPy derived from electrochemical deposition (E-PPy) and chemical oxidation method (C-PPy) (both less than 40 mAh g-1). This work verifies that increasing the proportion of the α-α coupling mode in the PPy electrode is a useful strategy to enhance the anion doping ability and capacity of dual-ion batteries.

Solid-Liquid-Gas Three-Phase Indirect Electrolysis Enabled by Affinity Auxiliary Imparted Covalent Organic Frameworks.

The design of efficient heterogenous redox mediators with favorable affinity to substrate and electrolyte are much desired yet still challenging for the development of indirect electrolysis system. Herein, for the first time, we have developed a solid-liquid-gas three-phase indirect electrolysis system based on a covalent organic framework (Dha-COF-Cu) as heterogenous redox mediator for S-S coupling reaction. Dha-COF-Cu with the integration of high porosity, nanorod morphology, abundant hydroxyl groups and active Cu sites is much beneficial for the adsorption/activation of thiols, uniform dispersion and high wettability in electrolyte, and efficient interfacial electron transfer. Notably, Dha-COF-Cu as solid-phase redox mediator exhibits excellent electrocatalytic efficiency for the formation of value-added liquid-phase S-S bond product (yields up to 99%) coupling with the generation of gas-phase product of H2 (~1.40 mmol g-1 h-1), resulting in a powerful three-phase indirect electrolysis system. This is the first work about COFs that can be applied in three-phase indirect electrolysis system, which might promote the development of porous crystalline materials in this field.

Ferromagnetic-Antiferromagnetic Coupling in Gas-Phase Synthesized M(Fe, Co, and Ni)-Cr Nanoparticles for Next-Generation Magnetic Applications.

Combining ferromagnetic-antiferromagnetic materials in nanoalloys (i.e., nanoparticles, NPs, containing more than one element) can create a diverse landscape of potential electronic structures. As a result, a number of their magnetic properties can be manipulated, such as the exchange bias between NP core and shell, the Curie temperature of nanoparticulated samples, or their magnetocaloric effect. In this work, such a family of materials (namely M-Cr NPs where M is Fe, Co, Ni, or some combination of them) is reviewed with respect to the tunability of their magnetic properties via optimized doping with Cr up to its solubility limit. To this end, gas-phase synthesis has proven a most effective method, allowing excellent control over the physical structure, composition, and chemical ordering of fabricated NPs by appropriately selecting various deposition parameters. Recent advances in this field (both experimental and computational) are distilled to provide a better understanding of the underlying physical laws and point toward new directions for cutting-edge technological applications. For each property, a relevant potential application is associated, such as memory cells and recording heads, induced hyperthermia treatment, and magnetic cooling, respectively, aspiring to help connect the output of fundamental and applied research with current real-world challenges.

The Progress of Reductive Coupling Reaction by Iron Catalysis.

The transition metal catalyzed coupling reaction has revolutionized the strategies for forging the carbon-carbon bonds. In contrast to traditional cross-coupling methods using pre-prepared nucleophilic organometallic reagents, reductive coupling reactions for the C-C bonds formation provide some advantages. Because both coupling partners are reduced in the final products using a stoichiometric amount of a reductant, this approach not only avoids the need to use sensitive organometallic species, but also provides an orthogonal and complementary access to classical coupling reaction. Notably, the reductive coupling reactions feature readily available fragments, promote good step economy, exhibit high functional group tolerance and unique chemoselectivity, which have propelled their increasingly popular in the organic synthesis. In recent years, due to the low price, minimal toxicity, and environmentally benign character, iron-catalyzed carbon-carbon coupling reactions have garnered significant attention from the organic synthetic chemists and pharmacologists, especially the iron-catalyzed reductive coupling. This review aims to provide an insightful overview of recent advances in iron-catalyzed reductive coupling reactions, and to illustrate their possible reaction mechanisms.

Preventing Site-Specific Calpain Proteolysis of Junctophilin-2 Protects Against Stress-Induced Excitation-Contraction Uncoupling and Heart Failure Development.

BACKGROUND: Excitation-contraction (E-C) coupling processes become disrupted in heart failure (HF), resulting in abnormal Ca2+ homeostasis, maladaptive structural and transcriptional remodeling, and cardiac dysfunction. Junctophilin-2 (JP2) is an essential component of the E-C coupling apparatus but becomes site-specifically cleaved by calpain, leading to disruption of E-C coupling, plasmalemmal transverse tubule degeneration, abnormal Ca2+ homeostasis, and HF. However, it is not clear whether preventing site-specific calpain cleavage of JP2 is sufficient to protect the heart against stress-induced pathological cardiac remodeling in vivo. METHODS: Calpain-resistant JP2 knock-in mice (JP2CR) were generated by deleting the primary JP2 calpain cleavage site. Stress-dependent JP2 cleavage was assessed through in vitro cleavage assays and in isolated cardiomyocytes treated with 1 µmol/L isoproterenol by immunofluorescence. Cardiac outcomes were assessed in wild-type and JP2CR mice 5 weeks after transverse aortic constriction compared with sham surgery using echocardiography, histology, and RNA-sequencing methods. E-C coupling efficiency was measured by in situ confocal microscopy. E-C coupling proteins were evaluated by calpain assays and Western blotting. The effectiveness of adeno-associated virus gene therapy with JP2CR, JP2, or green fluorescent protein to slow HF progression was evaluated in mice with established cardiac dysfunction. RESULTS: JP2 proteolysis by calpain and in response to transverse aortic constriction and isoproterenol was blocked in JP2CR cardiomyocytes. JP2CR hearts are more resistant to pressure-overload stress, having significantly improved Ca2+ homeostasis and transverse tubule organization with significantly attenuated cardiac dysfunction, hypertrophy, lung edema, fibrosis, and gene expression changes relative to wild-type mice. JP2CR preserves the integrity of calpain-sensitive E-C coupling-related proteins, including ryanodine receptor 2, CaV1.2, and sarcoplasmic reticulum calcium ATPase 2a, by attenuating transverse aortic constriction-induced increases in calpain activity. Furthermore, JP2CR gene therapy after the onset of cardiac dysfunction was found to be effective at slowing the progression of HF and superior to wild-type JP2. CONCLUSIONS: The data presented here demonstrate that preserving JP2-dependent E-C coupling by prohibiting the site-specific calpain cleavage of JP2 offers multifaceted beneficial effects, conferring cardiac protection against stress-induced proteolysis, hypertrophy, and HF. Our data also indicate that specifically targeting the primary calpain cleavage site of JP2 by gene therapy approaches holds great therapeutic potential as a novel precision medicine for treating HF.

Mapping Electrophile Chemoselectivity in DalPhos/Nickel N-Arylation Catalysis: The Unusual Influence of Remote Sterics.

We disclose herein our evaluation of competitive (hetero)aryl-X (X: Br > Cl > OTf) reactivity preferences in bisphosphine/Ni-catalyzed C-N cross-coupling catalysis, using furfurylamine as a prototypical nucleophile, and employing DalPhos and DPPF as representative ancillary ligands with established efficacy. Beyond this general (pseudo)halide ranking, other intriguing structure-reactivity trends were noted experimentally, including the unexpected observation that bulky alkyl (e.g., R = tBu) substitution in para-R-aryl-X electrophiles strongly discourages (pseudo)halide reactivity relative to smaller substituents (e.g., nBu, Et, Me), despite being both remote from, and having a similar electronic influence on, the reacting C-X bond; such effects on nickel oxidative addition have not been documented previously and were not observed in our comparator reactions presented herein involving palladium. Density functional theory modeling of such PhPAd-DalPhos/Ni-catalyzed C-N cross-couplings revealed the origins of competitive turnover of C-Br over C-Cl, and possible ways in which bulky para-alkyl substitution might discourage net electrophile uptake/turnover, leading to inversion of halide selectivity.