Prognostic value of tumor deposits and positive lymph node ratio in stage III colorectal cancer: a retrospective cohort study.

BACKGROUND: In colorectal cancer (CRC), tumor deposits (TD) have been used to guide the N staging only in node-negative patients. It remains unknown about the prognostic value of TD in combination with positive lymph node ratio (LNR) in stage III CRC. PATIENTS AND METHODS: We analyzed data from 31,139 eligible patients diagnosed with stage III CRC, including 30,230 from the Surveillance, Epidemiology, and End Results (SEER) database as a training set and 909 from two Chinese hospitals as a validation set. The associations of TD and LNR with cancer-specific survival (CSS) and overall survival (OS) were evaluated using the Kaplan-Meier method and Cox regression models. RESULTS: Both TD-positive and high LNR (value≥0.4) were associated with worse CSS in the training (multivariable hazard ratio [HR], 1.50; 95% confidence interval [CI], 1.43-1.58 and HR, 1.74; 95% CI, 1.62-1.86, respectively) and validation sets (HR,1.90; 95%CI, 1.41-2.54 and HR,2.01; 95%CI, 1.29-3.15, respectively). Compared to patients with TD-negative and low LNR (value<0.4), those with TD-positive and high LNR had a 4.09-fold risk of CRC-specific death in the training set (HR, 4.09; 95% CI, 3.54-4.72) and 4.60-fold risk in the validation set (HR, 4.60; 95% CI, 2.88-7.35). Patients with TD-positive/H-LNR CRC on the right side had the worst prognosis (P<0.001). The combined variable of TD and LNR contributed the most to CSS prediction in the training (24.26%) and validation (32.31%) sets. A nomogram including TD and LNR showed satisfactory discriminative ability, and calibration curves indicated favorable consistency in both the training and validation sets. CONCLUSIONS: TD and LNR represent independent prognostic predictors for stage III CRC. A combination of TD and LNR could be used to identify those at high risk of CRC deaths.

Ultrasound treatment alleviates external pericarp browning and improves fruit quality of pomegranate during storage.

BACKGROUND: Ultrasound treatment has a beneficial role in horticultural production from harvest to consumption. The quality traits and microbiological load in pomegranate fruit were explored during 30 days' storage at 20 °C after 10 min and 30 min ultrasound treatments. RESULTS: Ultrasound treatment significantly reduced the microbiological load during storage, providing a relatively clean and suitable storage environment. This was especially true for the 30 min treatment, which also maintained relatively lower weight loss and kept the browning rate below 5% during storage. Meanwhile, the fruit treated with ultrasound had higher ascorbic acid and anthocyanin content, which provided better antibacterial properties and higher nutraceutical properties until the end of storage. The 30 min ultrasound treatment significantly delayed the decrease in catalase (CAT) enzyme activity and the increase in peroxidase (POD) enzyme activity. Combined with weighted gene co-expression network analysis (WGCNA), and correlation analysis, color indicators and antioxidant activity induced by ultrasound treatment were responsible for the relatively higher fruit quality of pomegranate. CONCLUSION: Ultrasound treatment can improve the sensory quality and nutritional characteristics of pomegranate fruits during storage, and reduce the microbiological load. Ultrasound for 30 min was better than 10 min for prolonging the storage life of pomegranate. Our results will provide valuable information for ultrasound application in other horticultural products. © 2023 Society of Chemical Industry.

Study on the Overmolding Process of Carbon-Fiber-Reinforced Poly (Aryl Ether Ketone) (PAEK)/Poly (Ether Ether Ketone) (PEEK) Thermoplastic Composites.

This paper used poly (aryl ether ketone) (PAEK) resin with a low melting temperature to prepare laminate via the compression-molding process for continuous-carbon-fiber-reinforced composites (CCF-PAEK). Then, poly (ether ether ketone) (PEEK), or a short-carbon-fiber-reinforced poly (ether ether ketone) (SCF-PEEK) with a high melting temperature, was injected to prepare the overmolding composites. The shear strength of short beams was used to characterize the interface bonding strength of composites. The results showed that the interface properties of the composite were affected by the interface temperature, which was adjusted by mold temperature. PAEK and PEEK formed a better interfacial bonding at higher interface temperatures. The shear strength of the SCF-PEEK/CCF-PAEK short beam was 77 MPa when the mold temperature was 220 °C and 85 MPa when the mold temperature was raised to 260 °C. The melting temperature did not significantly affect the shear strength of SCF-PEEK/CCF-PAEK short beams. For the melting temperature increasing from 380 °C to 420 °C, the shear strength of the SCF-PEEK/CCF-PAEK short beam ranged from 83 MPa to 87 MPa. The microstructure and failure morphology of the composite was observed using an optical microscope. A molecular dynamics model was established to simulate the adhesion of PAEK and PEEK at different mold temperatures. The interfacial bonding energy and diffusion coefficient agreed with the experimental results.

Growth arrest-specific protein 2 (GAS2) interacts with CXCR4 to promote T-cell leukemogenesis partially via c-MYC.

Although growth arrest-specific protein 2 (GAS2) promotes the growth of T-cell acute lymphoblastic leukemia (T-ALL) cells in culture, the effect of GAS2 on T-cell leukemogenesis has not been studied, and the mechanism remains unclear. In the present study, xenograft studies showed that GAS2 silencing impaired T-cell leukemogenesis and decreased leukemic cell infiltration. Mechanistically, GAS2 regulated the protein expression of C-X-C chemokine receptor type 4 (CXCR4) rather than its transcript expression. Immunoprecipitation revealed that GAS2 interacted with CXCR4, and confocal analysis showed that GAS2 was partially co-expressed with CXCR4, which provided a strong molecular basis for GAS2 to regulate CXCR4 expression. Importantly, CXCR4 overexpression alleviated the inhibitory effect of GAS2 silencing on the growth and migration of T-ALL cells. Moreover, GAS2 or CXCR4 silencing inhibited the expression of NOTCH1 and c-MYC. Forced expression of c-MYC rescued the growth suppression induced by GAS2 or CXCR4 silencing. Meanwhile, GAS2 deficiency, specifically in blood cells, had a mild effect on normal hematopoiesis, including T-cell development, and GAS2 silencing did not affect the growth of normal human CD3+ or CD34+ cells. Overall, our data indicate that GAS2 promotes T-cell leukemogenesis through its interaction with CXCR4 to activate NOTCH1/c-MYC, whereas impaired GAS2 expression has a mild effect on normal hematopoiesis. Therefore, our study suggests that targeting the GAS2/CXCR4 axis is a potential therapeutic strategy for T-ALL.

Overexpression of PgCBF3 and PgCBF7 Transcription Factors from Pomegranate Enhances Freezing Tolerance in Arabidopsis under the Promoter Activity Positively Regulated by PgICE1.

Cold stress limits plant growth, development and yields, and the C-repeat binding factors (CBFs) function in the cold resistance in plants. However, how pomegranate CBF transcription factors respond to cold signal remains unclear. Considering the significantly up-regulated expression of PgCBF3 and PgCBF7 in cold-tolerant Punica granatum 'Yudazi' in comparison with cold-sensitive 'Tunisia' under 4 °C, the present study focused on the two CBF genes. PgCBF3 was localized in the nucleus, while PgCBF7 was localized in the cell membrane, cytoplasm, and nucleus, both owning transcriptional activation activity in yeast. Yeast one-hybrid and dual-luciferase reporter assay further confirmed that PgICE1 could specifically bind to and significantly enhance the activation activity of the promoters of PgCBF3 and PgCBF7. Compared with the wild-type plants, the PgCBF3 and PgCBF7 transgenic Arabidopsis thaliana lines had the higher survival rate after cold treatment; exhibited increased the contents of soluble sugar and proline, while lower electrolyte leakage, malondialdehyde content, and reactive oxygen species production, accompanying with elevated enzyme activity of catalase, peroxidase, and superoxide dismutase; and upregulated the expression of AtCOR15A, AtCOR47, AtRD29A, and AtKIN1. Collectively, PgCBFs were positively regulated by the upstream PgICE1 and mediated the downstream COR genes expression, thereby enhancing freezing tolerance.

Laser Ablation Mechanism and Performance of Carbon Fiber-Reinforced Poly Aryl Ether Ketone (PAEK) Composites.

The ablation mechanism and performance of carbon fiber (CF)-reinforced poly aryl ether ketone (PAEK) thermoplastic composites were studied in this paper. The results show that the ablation damaged area is controlled by the irradiation energy, while the mass loss rate is controlled by the irradiation power density. In the ablation center, the PAEK resin and CFs underwent decomposition and sublimation in an anaerobic environment. In the transition zone, the resin experienced decomposition and remelting in an aerobic environment, and massive char leaves were present in the cross section. In the heat-affected zone, only remelting of the resin was observed. The fusion and decomposition of the resin caused delamination and pores in the composites. Moreover, oxygen appeared crucial to the ablation morphology of CFs. In an aerobic environment, a regular cross section formed, while in an anaerobic environment, a cortex-core structure formed. The cortex-core structure of CF inside the ablation pit was caused by the inhomogeneity of fibers along the radial direction and the residual carbon layer generated by resin decomposition in an anoxic environment. The description of the ablation mechanism presented in this study broadens our understanding of damage evolution in thermoplastic composites subjected to high-energy CW laser irradiation.

The spirobichroman-based polyimides with different side groups: from structure-property relationships to chain packing and gas transport performance.

Spirobichroman-based polymers with high gas permeability and selectivity are promising for their applications as membranes in gas separation. In this study, three spirobichroman-based polyimides (PIs; 6FDA-FH, 6FDA-DH, and 6FDA-MH) were synthesised by the polyreaction between diamines containing different substituents (benzene ring, pyridine ring, and methyl group) and 4,4'-(hexafluoroisopropylidene)-diphthalic anhydride (6FDA). The physical properties, gas transport behaviour, d-spacing, dihedral angle of molecules, and fractional free volume of the PIs were investigated through experiments and molecular simulations. The PIs exhibited excellent thermal stability and good solubility in common organic solvents. The gas permeability of the PIs was investigated; the results highlighted the critical role of the substituents in the enhancement of the gas separation performance of polymer membranes. Detailed analysis of the PIs showed that 6FDA-FH exhibits the highest gas permeability. This can be ascribed to the loose packing of the polymer chain owing to the increased dihedral angle between the two planes. However, the methyl substituent in 6FDA-MH disrupts the polymer chain packing rather than changing the dihedral angle between the two planes, thus enhancing the gas permeability of 6FDA-MH. Furthermore, 6FDA-DH exhibited the highest CO2/CH4 selectivity, which is attributed to the CO2 affinity of the polymer containing the pyridine unit.

Dual-Switching Electrochromism and Electrofluorochromism Derived from Diphenylamine-Based Polyamides with Spirobifluorene/Pyrene as Bridged Fluorescence Units.

Electrochromic (EC)/electrofluorochromic (EFC) bifunctional materials are receiving great attention because of their promising applications in optoelectronic devices. However, the development of ideal EC/EFC bifunctional materials is still a great challenge because of the poor integration of EC/EFC performances (optical contrast, response speed, and switching stability). Herein, we reported two novel diphenylamine-based mixed valence (MV) polyamides (S-HPA and P-HPA) with spirobifluorene (2,7-positions) and pyrene (1,6-positions) as bridged fluorescence units, respectively, showing impressive cyclability and fluorescence contrast with rapid switching. Through the formation of an effective electronic coupling between the two nitrogen centers using spirobifluorene/pyrene bridges, we demonstrated that different bridges have significant effects on the thermal and electrooptical characteristics of polyamides. In addition to lower fluorescence quantum yield and glass transition temperature, the S-HPA exhibited superior cyclability (contrast change <3.4%/14% over 500/300 cycles for EC/EFC switching), higher color/fluorescence contrast (64%/304%), and faster switching time (<2.6 s), mainly owing to the shorter conjugated length and more twisted configuration of the spirobifluorene bridge. The design principle of MV polymers with fluorophore bridges proposed here will be a promising way to realize high-performance EC/EFC devices and will also provide new insights into their future development and applications.

Phase Behavior and Phase Diagram of Polystyrene-b-Poly(Perfluorooctylethyl Acrylates).

Fluorocontaining polymers bearing special properties are unique and important materials in modern society. In this work, we focused on the phase behavior and phase diagram of poly(styrene-block-perfluorooctylethyl acrylate) with a volume fraction varying from 0.2 to 0.8. Small-angle X-ray scattering and transmission electron microscopy showed the phase formation in the sequence of hexagonally packed cylinders (HEX) to lamellar layers (LAM) to inverse hexagonally packed cylinders (iHEX) in this series of block polymers. Wide-angle X-ray diffraction experiments proved that the fluorodomains of the LAM phases and the matrix of iHEX phases contained layered structures formed by the crystallization of fluorosegments. During heating, the self-assembled lattice remained intact even after the melting of fluorodomain, with barely changed lattice parameters. Such hierarchical structural formation was understood by chain conformation and domain interaction, which may provide new insight into the molecular design of advanced materials.

Synthesis of Superheat-Resistant Polyimides with Enhanced Dielectric Constant by Introduction of Cu(ΙΙ)-Coordination.

To achieve polyimide-metal complexes with enhanced properties, 5-amine-2-(5-aminopyridin-2-yl)-1-methyl-benzimidazole (PyMePABZ) that contains stiff 2-(2'-pyridyl)benzimidazole (PyBI) was synthesized and exploited to construct the Cu(ΙΙ)-crosslinked polyimides (Cu-PIs). These Cu-PIs exhibited higher dielectric, thermal, and mechanical properties with an increase in Cu2+ content. Among them, their dielectric constants (εrS) were up to 43% superior to that of the neat PI, glass transition temperatures (Tgs) were all over 400 °C, and 5% weight loss temperature (T5%) maintained beyond 500 °C. These data indicate that the metal coordination crosslinking provided a useful guide to develop high performance PIs which possess potential application as useful high temperature capacitors.

Measurement of Quasi-Static 3-D Knee Joint Movement Based on the Registration From CT to US.

The measurement of quasi-static 3-D knee joint movement is an important basis for studying the mechanism of knee joint injury. Most of the existing measurement methods make use of computed tomography (CT) and nuclear magnetic resonance (MR) imaging technology and hence have the disadvantages of invasiveness, ionizing radiation, low accuracy, and high cost. To overcome those drawbacks, this article innovatively proposes a 3-D motion measurement system for the knee joint based on the registration of CT images to ultrasound (US) images. More specifically, the lower limbs of a subject were first scanned once to acquire the CT images. A portable handheld device was designed to control a US probe for mechanically scanning the subject's lower limbs in a linear trajectory. During the movement of the subject's lower limbs, the US scanning was performed quasi-statically. The acquired US images were then registered to the CT images, and the 3-D motions of the lower limb bones could be recreated with the bones scanned in CT images. To guarantee the registration accuracy and efficiency, we used the H-shaped multiview slice assembly as the structural image content for the registration process. The experimental results show that our approach can accurately measure the 3-D motion of the knee joint and meet the needs of 3-D motion analysis of knee joint in practice.

Highly optical transparency and thermally stable polyimides containing pyridine and phenyl pendant.

In order to obtain highly optical transparency polyimides, two novel aromatic diamine monomers containing pyridine and kinky structures, 1,1-bis[4-(5-amino-2-pyridinoxy)phenyl]diphenylmethane (BAPDBP) and 1,1-bis[4-(5-amino-2-pyridinoxy)phenyl]-1-phenylethane (BAPDAP), were designed and synthesized. Polyimides based on BAPDBP, BAPDAP, 2,2-bis[4-(5-amino-2-pyridinoxy)phenyl]propane (BAPDP) with various commercial dianhydrides were prepared for comparison and structure-property relationships study. The structures of the polyimides were characterized by Fourier transform infrared (FT-IR) spectrometer, wide-angle X-ray diffractograms (XRD) and elemental analysis. Film properties including solubility, optical transparency, water uptake, thermal and mechanical properties were also evaluated. The introduction of pyridine and kinky structure into the backbones that polyimides presented good optical properties with 91-97% transparent at 500 nm and a low cut-off wavelength at 353-398 nm. Moreover, phenyl pendant groups of the polyimides showed high glass transition temperatures (Tg ) in the range of 257-281 °C. These results suggest that the incorporating pyridine, kinky and bulky substituents to polymer backbone can improve the optical transparency effectively without sacrificing the thermal properties.

Evaluation of electrocardiograms recorded in cynomolgus monkeys with short- and long-term intracardiac lead implantations.

INTRODUCTION: minimally invasive placement of intracardiac (IC) ECG leads in monkeys has greatly improved signal quality and the ability to interpret these ECGs. However, information on characteristics of the ECGs recorded using the IC lead is not available in the literature. There are concerns about the potential impact of IC lead placement on the ECG waveform and cardiac function as a result of potential irritation or trauma resulting from the placement and/or long term residence of the IC lead. The purposes of this study were to characterize IC ECG morphology, to obtain information on the recovery processes after IC ECG lead implantation, and to evaluate the IC ECG model application to safety pharmacology studies. METHODS: the telemetry transmitter, arterial blood pressure catheter and IC ECG lead were implanted in 40 cynomolgus monkeys, two of which were also implanted with subcutaneous (SC) ECG leads. The data of IC ECG, heart rate (HR) and mean arterial blood pressure (MABP) were collected telemetrically for a period of 1-12 months after implantation, and measured using computer softwares. RESULTS: the IC ECG waveforms varied greatly from those of SC ECG. There was no clearly identifiable S-T segment, and T waves were biphasic in the majority of IC ECGs. The morphology of IC ECG was diversified among animals, progressively changed in the first 2 weeks post-surgery and stabilized approximately 3 weeks post-surgery. MABP and HR were elevated after implant surgery, but recovered to the levels comparable to those of SC in approximately 1 and 4 weeks, respectively. The IC ECG values obtained during week 8 to 10 (HR=134+/-25 bpm, PR interval=87+/-13 ms, QRS interval=40+/-7 ms, and QT interval=246+/-30 ms, QTcF=318+/-28 ms) were comparable to those from SC ECG. DISCUSSION: the IC ECG provides a clear ECG signal with values comparable to, and waveforms different from, SC recordings. The complicated surgical procedure with long substantial recovery time, high incidence of IC lead malfunction, and high costs for IC leads may limit application of the IC ECG model in safety pharmacology studies.

Heterogeneous gap junction remodeling in reentrant circuits in the epicardial border zone of the healing canine infarct.

BACKGROUND: The epicardial border zone (EBZ) of surviving myocytes in the healing, 4- to 5-day-old canine infarct is an arrhythmogenic substrate characterized by both structural and functional remodeling of Cx43. Unknown is whether the remodeling of gap junction conductance is heterogeneous in the EBZ like that of sarcolemmal ion channel remodeling and how remodeling of the gap junction influences conduction and anisotropy. METHODS AND RESULTS: Ventricular tachycardia was initiated by programmed stimulation in healing canine infarcted hearts. Reentrant circuits were mapped and the central common pathway (CCP) and outer pathway (OP) regions localized. Epimyocardium removed from the CCP was disaggregated to generate myocyte pairs for conductance measurements. Cx43 distribution was determined by immunofluorescent confocal microscopy. While transverse coupling (gap junction conductance) was markedly decreased in OP cells, CCP cells with lateralized Cx43 gap junctions showed normal conductance. Longitudinal coupling in both OP and CCP was no different than normal. Consistent with conductance measurements, the anisotropic ratio in the CCP was similar to that of normal tissue. In the OP it was increased. Despite normal longitudinal and transverse conductance and anisotropic ratio, longitudinal and transverse conduction velocities were decreased in the CCP with respect to normal epicardium, possibly as a result of the remodeling of sarcolemmal ion channels in this region. CONCLUSIONS: Gap junction conductance and distribution is heterogeneous in different regions of reentrant circuits. Lateralization of Cx43 gap junctions in CCP of reentrant circuits is associated with normal transverse conductance between cell pairs. In contrast, absence of lateralization in OP is associated with reduced transverse conductance. Despite normal anisotropic ratio, conduction velocity in CCP region remains slower than normal. This suggests that the effects of Cx43 remodeling in the infarcted heart should be interpreted in conjunction with other types of remodeling occurring in the EBZ (i.e. sarcolemmal ion channels).

Dimethyl sulfoxide effects on hERG channels expressed in HEK293 cells.

INTRODUCTION: Dimethyl sulfoxide (DMSO) is widely used as a solvent to facilitate formulation of test substances in cell perfusion solutions. However, DMSO concentration in bath (extracellular) solution is usually limited to 0.1-0.3% to avoid DMSO-induced changes in cell morphology and membrane properties due to elevation of osmolality. The purpose of this study was to examine whether DMSO-induced hyperosmotic effects on hERG expressing cells could be compensated by adding an equivalent amount of DMSO in pipette (intracellular) solution, to investigate DMSO effects on hERG channels, and to determine the impact of DMSO on the potency of hERG channel blockers. METHOD: Whole-cell patch clamp method was used to record hERG currents in HEK293 cells. DMSO at concentrations of 0.1% to 2% was applied to bath and pipette solutions. Various voltage protocols were used to examine DMSO effects on hERG channel properties and to evaluate DMSO impacts on the potency of terfenadine and E-4031. RESULTS: When DMSO was added simultaneously in bath and pipette solutions, normal cell morphology and the proper current recording conditions could be maintained with application of up to 2% DMSO. DMSO slightly shifted the current-voltage relationship, activation curve, and inactivation curve of the hERG channel to more positive voltages. DMSO had little effect on the concentration-response relationship of hERG channel blockers we assessed. The IC50 for terfenadine and E-4031 were not significantly changed in the presence of 0.3, 0.5, 1 and 2% DMSO. DISCUSSION: Our results demonstrate that changes in cell morphology induced by extracellular DMSO can be prevented by application of DMSO in pipette solution. By utilizing this approach, we successfully performed hERG current recordings using bath solution containing up to 2% DMSO. DMSO-induced shifts of the voltage-dependence of hERG channel gating had little impact on the potency of hERG channel blockers.

Estimation of potency of HERG channel blockers: impact of voltage protocol and temperature.

INTRODUCTION: The HERG channel is widely used for the assessment of proarrhythmic risk for new drugs. HERG channel blockers obstruct channel functions through various mechanisms, which usually show time dependence, voltage dependence, and state dependence. The voltage protocol and temperature may affect the estimation of drug potency, but limited information is available in this regard. The purpose of this study was to evaluate the influence of voltage protocol and temperature on predicting the potency of HERG channel blockers, and to determine electrophysiological approaches for new drugs screening studies. METHOD: Whole-cell patch-clamp electrophysiology was carried out by utilizing different voltage step protocols to examine the potency of compounds known to preferentially block the channel in the closed (ketoconazole and BeKm-1), open, and/or inactivated states (E-4031, astemizole, and terfenadine) in HEK293 cells transfected with HERG cDNA at room temperature and near-physiological temperature. RESULTS: Drug potency determined using different voltage protocols varied dependent on the mechanisms of drug actions. For most compounds, the IC(50) values obtained with a long pulse step protocol at room temperature were close to those determined with the voltage protocols designed to disclose their intrinsic potency. Relative to room temperature, the potency of E-4031, terfenadine, and ketoconazole was not changed at approximately 35 degrees C, but potency of astemizole was reduced. DISCUSSION: The long pulse step protocol with room temperature can be selected for HERG channel safety screening studies. Alternative voltage protocols or temperatures should be considered if HERG study results are not consistent with other cardiac safety assessments.

Cell coupling between ventricular myocyte pairs from connexin43-deficient murine hearts.

Mice with cardiac-restricted inactivation of the connexin43 gene (CKO mice) have moderate slowing of ventricular conduction and lethal arrhythmias. Mechanisms through which propagation is maintained in the absence of Cx43 are unknown. We evaluated gap junctional conductance in CKO ventricular pairs using dual patch clamp methods. Junctional coupling was reduced to 4+/-2 nS (side-to-side) and 11+/-2 nS (end-to-end), including 21% of cell-pairs with no detectable coupling, compared with 588+/-104 nS (side-to-side) and 558+/-92 nS (end-to-end) in control cell-pairs. Voltage dependence of control gap junctions was characteristic of Cx43. CKO conductance showed increased voltage dependence, suggesting low-level expression of other connexin isoforms. From theoretical models, this degree of CKO coupling is not expected to support levels of conduction persisting in vivo, suggesting the possibility that there are additional mechanisms for maintained propagation when gap junctional conductance is severely reduced.

Remodeling of gap junctional channel function in epicardial border zone of healing canine infarcts.

The epicardial border zone (EBZ) of canine infarcts has increased anisotropy because of transverse conduction slowing. It remains unknown whether changes in gap junctional conductance (Gj) accompany the increased anisotropy. Ventricular cell pairs were isolated from EBZ and normal hearts (NZ). Dual patch clamp was used to quantify Gj. At a transjunctional voltage (Vj) of +10 mV, side-to-side Gj of EBZ pairs (9.2+/-3.4 nS, n=16) was reduced compared with NZ side-to-side Gj (109.4+/-23.6 nS, n=14, P<0.001). Gj of end-to-end coupled cells was not reduced in EBZ. Steady-state Gj of both NZ and EBZ showed voltage dependence, described by a two-way Boltzmann function. Half-maximal activation voltage in EBZ was shifted to higher Vj in positive and negative directions. Immunoconfocal planimetry and quantification showed no change in connexin43 per unit cell volume or surface area in EBZ. Decreased side-to-side coupling occurs in EBZ myocytes, independent of reduced connexin43 expression, and is hypothesized to contribute to increased anisotropy and reentrant arrhythmias.