Potential Channels Research Articles

Investigating Contributions of Canonical Transient Receptor Potential Channel 3 to Hippocampal Hyperexcitability and Seizure-Induced Neuronal Cell Death.

Canonical transient receptor potential channel 3 (TRPC3) is the most abundant TRPC channel in the brain and is highly expressed in all subfields of the hippocampus. Previous studies have suggested that TRPC3 channels may be involved in the hyperexcitability of hippocampal pyramidal neurons and seizures. Genetic ablation of TRPC3 channel expression reduced the intensity of pilocarpine-induced status epilepticus (SE). However, the underlying cellular mechanisms remain unexplored and the contribution of TRPC3 channels to SE-induced neurodegeneration is not determined. In this study, we investigated the contribution of TRPC3 channels to the electrophysiological properties of hippocampal pyramidal neurons and hippocampal synaptic plasticity, and the contribution of TRPC3 channels to seizure-induced neuronal cell death. We found that genetic ablation of TRPC3 expression did not alter basic electrophysiological properties of hippocampal pyramidal neurons and had a complex impact on epileptiform bursting in CA3. However, TRPC3 channels contribute significantly to long-term potentiation in CA1 and SE-induced neurodegeneration. Our results provided further support for therapeutic potential of TRPC3 inhibitors and raised new questions that need to be answered by future studies.

How does digital transformation affect the total factor productivity of China's A-share listed enterprises in the mineral resource-based sector?

In light of the escalating constraints on resources and the complexities of sustainable development, the conventional extensive production approaches employed by mineral resource-based enterprises are ill-suited to fulfill the demands for efficient and eco-friendly operations. Such circumstances call for the adoption of novel development paradigms that promote sustainable practices. Particularly, the digital transformation has become a pivotal pathway towards the realization of high-quality development within the context of the ongoing technological revolution and the shift in economic frameworks. Based on the comprehensive dataset of the annual reports and input-output data of Chinese A-share listed enterprises between 2009 and 2021, this study seeks to examine the net impact and potential channels through which digital transformation affects the total factor productivity of these specific enterprises. The empirical results confirm the promoting effect of digital transformation on enterprises' total factor productivity. This highlight persists valid after a series of thorough robustness checks. Notably, this beneficial role can be achieved by the enhancement of corporate capabilities, the mitigation of financial constraints, and the improvement of economic advantages. And the heterogeneity in this promoting effect is evident across enterprises with different characteristics. The positive moderating influence of government competitions in this context is a factor that warrants consideration. Thereby, the research highlights not only furnish a rigorous scientific foundation and direction guide for the strategic formulation of mineral resource-based enterprises, but also proffer critical insights for policymakers to accelerate the modernization endeavor of resource-based cities as well as entire regional domains.

TRPV4 affects visual signals in photoreceptors and rod bipolar cells.

Mechanical sensitive channels expressed in mammalian retinas are effectors of elevated pressure stresses, but it is unclear how their activation affects visual function in pressure-related retinal disorders. This study investigated the role of the transient potential channel vanilloid TRPV4 in photoreceptors and rod bipolar cells (RBCs) with immunohistochemistry, confocal microscopy, electroretinography (ERG), and patch-clamp techniques. TRPV4 immunoreactivity (IR) was found in the outer segments of photoreceptors, dendrites and somas of PKCα-positive RBCs and other BCs, plexiform layers, and retinal ganglion cells (RGCs) in wild-type mice. TRPV4-IR was largely diminished in the retinas of homozygous TRPV4 transgenic mice. Genetically suppressing TRPV4 expression moderately but significantly enhanced the amplitude of ERG a- and b-waves evoked by scotopic and mesopic lights (0.55 to 200 Rh*rod-1 s-1) and photopic lights (105-106 Rh*rod-1 s-1) compared to wild-type mice in fully dark-adapted conditions. The implicit time evoked by dim lights (0.55 to 200 Rh*rod-1 s-1) was significantly decreased for b-waves and elongated for a-waves in the transgenic mice. ERG b-wave evoked by dim lights is primarily mediated by RBCs, and under voltage-clamp conditions, the latency of the light-evoked cation current in RBCs of the transgenic mice was significantly shorter compared to wild-type mice. About 10% of the transgenic mice had one eye undeveloped, and the percentage was significantly higher than in wild-type mice. The data indicates that TRPV4 involves ocular development and is expressed and active in outer retinal neurons, and interventions of TRPV4 can variably affect visual signals in rods, cones, RBCs, and cone ON BCs.

Gallic acid pretreatment mitigates parathyroid ischemia–reperfusion injury through signaling pathway modulation

Thyroid surgery often results in ischemia–reperfusion injury (IRI) to the parathyroid glands, yet the mechanisms underlying this and how to ameliorate IRI remain incompletely explored. Our study identifies a polyphenolic herbal extract—gallic acid (GA)—with antioxidative properties against IRI. Through flow cytometry and CCK8 assays, we investigate the protective effects of GA pretreatment on a parathyroid IRI model and decode its potential mechanisms via RNA-seq and bioinformatics analysis. Results reveal increased apoptosis, pronounced G1 phase arrest, and significantly reduced cell proliferation in the hypoxia/reoxygenation group compared to the hypoxia group, which GA pretreatment mitigates. RNA-seq and bioinformatics analysis indicate GA’s modulation of various signaling pathways, including IL-17, AMPK, MAPK, transient receptor potential channels, cAMP, and Rap1. In summary, GA pretreatment demonstrates potential in protecting parathyroid cells from IRI by influencing various genes and signaling pathways. These findings offer a promising therapeutic strategy for hypoparathyroidism treatment.

Does digital transformation matter for trade credit provision? Evidence from China

This study demonstrated that digital transformation can promote the supply of trade credit in China. The results are robust to different measures of trade credit provision and endogeneity. The impact of digital transformation on trade credit provision is more pronounced among firms in regions with lower marketization and worse information environment. This study investigates this potential channel and finds that digital transformation increases trade credit provision by improving firms' financing availability. Overall, the results are consistent with the financial theory of trade credit.

Signal Transduction of Transient Receptor Potential TRPM8 Channels: Role of PIP5K, Gq-Proteins, and c-Jun.

Transient receptor potential melastatin-8 (TRPM8) is a cation channel that is activated by cold and "cooling agents" such as menthol and icilin, which induce a cold sensation. The stimulation of TRPM8 activates an intracellular signaling cascade that ultimately leads to a change in the gene expression pattern of the cells. Here, we investigate the TRPM8-induced signaling pathway that links TRPM8 channel activation to gene transcription. Using a pharmacological approach, we show that the inhibition of phosphatidylinositol 4-phosphate 5 kinase α (PIP5K), an enzyme essential for the biosynthesis of phosphatidylinositol 4,5-bisphosphate, attenuates TRPM8-induced gene transcription. Analyzing the link between TRPM8 and Gq proteins, we show that the pharmacological inhibition of the βγ subunits impairs TRPM8 signaling. In addition, genetic studies show that TRPM8 requires an activated Gα subunit for signaling. In the nucleus, the TRPM8-induced signaling cascade triggers the activation of the transcription factor AP-1, a complex consisting of a dimer of basic region leucine zipper (bZIP) transcription factors. Here, we identify the bZIP protein c-Jun as an essential component of AP-1 within the TRPM8-induced signaling cascade. In summary, with PIP5K, Gq subunits, and c-Jun, we identified key molecules in TRPM8-induced signaling from the plasma membrane to the nucleus.

Dynamic molecular portraits of ion-conducting pores characterize functional states of TRPV channels

Structural biology is solving an ever-increasing number of snapshots of ion channel conformational ensembles. Deciphering ion channel mechanisms, however, requires understanding the ensemble dynamics beyond the static structures. Here, we present a molecular modeling-based approach characterizing the ion channel structural intermediates, or their “dynamic molecular portraits”, by assessing water and ion conductivity along with the detailed evaluation of pore hydrophobicity and residue packing. We illustrate the power of this approach by analyzing structures of few vanilloid-subfamily transient receptor potential (TRPV) channels. Based on the pore architecture, there are three major states that are common for TRPVs, which we call α-closed, π-closed, and π-open. We show that the pore hydrophobicity and residue packing for the open state is most favorable for the pore conductance. On the contrary, the α-closed state is the most hydrophobic and always non-conducting. Our approach can also be used for structural and functional classification of ion channels.

Bound states in bent soft waveguides

The aim of this paper is to show that a two-dimensional Schrödinger operator with the potential in the form of a ‘ditch’ of a fixed profile can have a geometrically induced discrete spectrum. This occurs if such a potential channel has a single or multiple bends, being straight outside a compact. Moreover, under stronger geometric restrictions, the claim remains true in the presence of a potential bias at one of the channel ‘banks.’

Can a not-for-profit minority institutional investor affect corporate tax avoidance? Evidence from China

ABSTRACT We investigate the impact of the China Securities Investor Service Center (CSISC), a not-for-profit minority institutional investor, on corporate tax avoidance. Employing a difference-in-differences (DID) model based on a sample of Chinese listed companies during 2014–2017, we find that CSISC shareholding reduces corporate tax avoidance. We check this finding’s robustness using alternative measures of tax avoidance, alternative sampling, placebo tests, and parallel trend analysis, and our results still hold. In addition, we find the CSISC’s negative impact on corporate tax avoidance is more pronounced in state-owned enterprises (SOEs), firms with worse internal and external corporate governance, and regions with stricter tax enforcement. Further analyses reveal that the improved information environment is a potential channel through which the CSISC inhibits corporate tax avoidance. This paper enriches the literature on the impact of minority shareholder protection mechanism and the determinants of corporate tax avoidance.

2-Aminoethoxydiphenyl borate prolongs atrial and shortens ventricular action potential duration through modulation of ion channels in the sarcolemma

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): PPP allowance, Health holland ZonmW Background 2-Aminoethoxydiphenyl Borate (2-APB) has been shown to prevent atrial fibrillation in animal models, supposedly through its inhibitory action on the intracellular IP3 receptor and subsequent calcium regulation. However, 2-APB may modulate other ion channels or transporters and the cellular mechanisms associated with the prevention of atrial fibrillation are not well established. We aimed at unravelling the cellular effects of 2-APB on atrial and ventricular action potential and currents. Methods Rabbit (New Zealand White) left atrial and ventricular cardiomyocytes were isolated from Langendorff-perfused hearts by enzymatic dissociation. Adult human atrial cardiomyocytes were obtained by enzymatic dissociation from left atrial appendages (LAA) routinely removed from AF patients undergoing minimally invasive surgical pulmonary vein isolation. Action potentials (APs) and K+ currents were recorded at 36.5 °C with the amphotericin-B-perforated or whole cell patch clamp technique. RNA was isolated from left atrial and ventricular tissue and sequenced on an Illumina HiSeq4000 platform. Results In rabbit atrial cardiomyocytes, extracellular application of 5 uM 2-APB had no effect on action potential duration (APD), but extracellular application of 50 uM 2-APB prolonged the APD by 40% and inhibited an outward K+-current. Inversely, extracellular application of 5 uM 2-APB shortened the APD of rabbit left ventricular cardiomyocytes by 20% while extracellular application 50 uM 2-APB did not prolong the APD. Additionally, 2-APB did not modulate K+ currents in ventricular rabbit cardiomyocytes. Intracellular application (in the patch pipet) of 50 uM 2-APB did not have any effects on atrial APD, suggesting that IP3 receptor inhibition is not involved. However, subsequent application of 50 uM 2-APB in the bath prolonged the APD by 40% indicating that 2-APB prolongs atrial APD by inhibiting one or more K+ channel types in the sarcolemmal membrane from the extracellular site. RNA sequencing analysis revealed potential targets channel including TRP and Kv channels differentially expressed in atria versus ventricle. In human atrial cardiomyocytes from AF patients, application of 50 uM 2-APB prolonged APD by 25%. Conclusions 2-APB prolongs rabbit and human atrial APD and shortens rabbit ventricular APD in a concentration dependent manner, most likely due to modulation of sarcolemmal K+ channels, differentially expressed in the two cell types. Pharmacological studies with 2-APB may reveal novel cellular mechanisms identifying potential new targets for the treatment of atrial fibrillation.

Financial instability in Europe: Does geopolitical risk from proximate countries and trading partners matter?

Using a spatial econometric approach, we measure 28 European countries' foreign geopolitical risks (GPR) from their proximate countries, export destinations, and import origins from January 1985 to January 2024 at a monthly frequency. Based on difference GMM estimation on dynamic panel data model, we find that the foreign GPRs increase Europe's financial instability through potential channels of surging energy inflation and capital flight to the U.S. Compared with high-income countries with a bank-based financial system, low-income countries with a market-based financial system are more fragile to the foreign GPRs. These findings can help European financial authorities improve prudential regulation policies.

Interaction between TRP channels and anoctamins

Anoctamin 1 (ANO1) binds to transient receptor potential (TRP) channels (protein-protein interaction) and then is activated by TRP channels (functional interaction). TRP channels are non-selective cation channels that are expressed throughout the body and play roles in multiple physiological functions. Studies on TRP channels increased after the identification of TRP vanilloid 1 (TRPV1) in 1997. Calcium-activated chloride channel anoctamin 1 (ANO1, also called TMEM16A and DOG1) was identified in 2008. ANO1 plays a major role in TRP channel-mediated functions, as first shown in 2014 with the demonstration of a protein-protein interaction between TRPV4 and ANO1. In cells that co-express TRP channels and ANO1, calcium entering cells through activated TRP channels causes ANO1 activation. Therefore, in many tissues, the physiological functions related to TRP channels are modulated through chloride flux associated with ANO1 activation. In this review, we summarize the latest understanding of TRP-ANO1 interactions, particularly interaction of ANO1 with TRPV4, TRP canonical 6 (TRPC6), TRPV3, TRPV1, and TRPC2 in the salivary glands, blood vessels, skin keratinocytes, primary sensory neurons, and vomeronasal organs, respectively.

Resonance of Low-frequency Electromagnetic and Ion-sound Modes in the Solar Wind

Parker Solar Probe measurements have recently shown that coherent fast magnetosonic and Alfvén ion-cyclotron waves are abundant in the solar wind and can be accompanied by higher-frequency electrostatic fluctuations. In this Letter we reveal the nonlinear process capable of channelling the energy of low-frequency electromagnetic to higher-frequency electrostatic fluctuations observed on board Parker Solar Probe. We present Hall-MHD simulations demonstrating that low-frequency electromagnetic fluctuations can resonate with the ion-sound mode, which results in steepening of plasma density fluctuations, electrostatic spikes, and harmonics in the electric field spectrum. The resonance can occur around the wavenumber determined by the ratio between local sound and Alfvén speeds, but only in the case of oblique propagation to the background magnetic field. The resonance wavenumber, its width, and steepening timescale are estimated, and all indicate that the revealed two-wave resonance can frequently occur in the solar wind. This process can be a potential channel of energy transfer from cyclotron resonant ions producing the electromagnetic fluctuations to Landau resonant ions and electrons absorbing the energy of the higher-frequency electrostatic fluctuations.

A novel gain-of-function mutation in transient receptor potential C6 that causes podocytes injury.

Podocyte injury plays a vital role in focal segmental glomerulosclerosis (FSGS), and apoptosis is one of its mechanisms. The transient receptor potential channel 6 (TRPC6) is highly expressed in podocytes and mutations mediate podocyte injury. We found TRPC6 gene mutation (N110S) was a new mutation and pathogenic in the preliminary clinical work. The purpose of this study was to investigate the potential mechanism of mutation in TRPC6 (TRPC6-N110S) in the knock-in gene mouse model and in immortalized mouse podocytes (MPC5). Transmission electron microscopy was used to evaluate renal injury morphology. We measured 24-hour urinary albumin-to-creatinine ratios and major biochemical parameters such as serum albumin, urea nitrogen, and total cholesterol. The results of CCK-8 assay and apoptosis experiments showed that the TRPC6-N110S overexpression group had slower proliferative activity and increased apoptosis than the control group. FluO-3 assay revealed increased calcium influx in the TRPC6-N110S overexpression group. Podocin level was decreased in TRPC6-N110S group, while TRPC6 and desmin levels were increased in TRPC6-N110S group. The 24 h uACR at 6 weeks was significantly higher in the pure-zygotes group than in the WT and heterozygotes groups, and this difference was found at 8 and 10 weeks.TRPC6 levels showed no significant difference between homozygote and WT mice. Compared to homozygote group, expression of podocin and nephrin were increased in WT, but levels of desmin was decreased in WT. Our results suggest that this new mutation causes podocyte injury probably by enhancing calcium influx and podocyte apoptosis, accompanied by increased proteinuria and decreased expression of nephrin and podocin.

Directors’ and officers’ liability insurance and corporate digital transformation

ABSTRACT In the digital era, digital transformation has become imperative for enterprises to seek better development opportunities. Using a sample of Chinese A-share listed firms from 2007 to 2020, we document a positive relationship between directors’ and officers’ liability insurance (D&O insurance) and corporate digital transformation. This correlation is robust to a series of robustness tests. Further mechanism tests reveal that mitigating agency costs and enhancing risk-taking ability are two potential channels. Additional heterogeneity analyses show that the impact of D&O insurance on digital transformation is more pronounced for non-state-owned firms, firms perceiving more economic policy uncertainty, firms with less media coverage and firms in highly competitive industries. These findings suggest that companies should pay attention to the positive governance effect of D&O insurance and it’s more advisable for companies with severe agency conflict and weak risk-taking ability to purchase D&O insurance to facilitate digital transformation.

Impact of phase-correction of late-gadolinium enhancement images on quantification of scar metrics and in-silico VT-modelling in patients with an ischemic cardiomyopathy

Abstract Background Late gadolinium enhancement (LGE) is used to perform tissue characterization and has become the cornerstone to guide treatment of ventricular tachyarrhythmia (VT). Traditionally, LGE acquisition is reconstructed in two distinct forms: non-phase corrected magnitude (MIR) and phase-corrected inversion recovery (PSIR) images. However, the influence of MIR vs. PSIR on the quantification of scar metrics and the repercussion on estimating myocardial arrhythmogenicity have not been comprehensively explored. Aim This study investigated the influence of PSIR and MIR LGE on the quantification of scar metrics in a randomly selected cohort of patients with ischemic cardiomyopathy who had undergone imaging prior to ICD implantation and had received device therapy during follow-up. In addition, this study assessed how these image variations affect functional estimation of arrhythmogenicity using computational modelling. Methods ADAS 3D LV was used to generate scar maps from 2D PSIR and MIR LGE images in 10 patients. The extent of variation in scar-characteristics was evaluated by quantifying the borderzone (BZ), scar core, and conduction corridors. Electrical simulations were performed on the patient-specific anatomical models using the Virtual Induction and Treatment of Arrhythmias (VITA) framework. VITA was used to assess the electrical vulnerability of potential reentrant channels and provided the number of inducible VTs and their corresponding round-trip-time (RTT), an equivalent of VT cycle-length. Results Scar quantification using PSIR LGE resulted in a significantly larger BZ (22.1±23.7g vs. 6.2±2.7g, p <.001) and scar core (43.5±21.5g vs. 10.5±4g, p =.047). Furthermore, both the absolute number (6.8±4 vs. 3.4±1.5, p = .031) and weight (13±13.1g vs. 1.4±0.8g, p =.018) of conduction corridors were also higher using PSIR. VTs were inducible in 8 PSIR-models and 7 MIR-models. Despite large differences in scar characteristics, VITA metrics were not significantly different between PSIR and MIR derived models. Although, the absolute number of inducible VTs were larger for PSIR (6.2±5.4 vs. 3.4±3.1), non-parametric testing using Wilcoxon signed-rank test did not demonstrate a significant difference (Z = -1.334, p =.139). A comparable, non-significant difference was observed for mean- (117±78 vs. 79±60, p =.257) and max-RTT (181±141 vs. 120±103, p =.294) between PSIR and MIR derived models. Conclusion PSIR resulted in significantly larger BZ, scar core area, and conduction corridors. While the absolute, VITA-derived estimation of myocardial arrhythmogenicity, including inducible VTs and RTT, were higher in the PSIR-based quantification, these differences did not reach statistical significance. These results highlight important differences between two clinically applied image reconstruction techniques for the assessment of arrhythmogenic substrate, emphasizing the need for additional investigation and refinement of scar quantification techniques.

TRPM2 channels are essential for regulation of cytokine production in lung interstitial macrophages.

Interstitial macrophages (IMs) are essential for organ homeostasis, inflammation, and autonomous immune response in lung tissues, which are achieved through polarization to a pro-inflammatory M1 and an M2 state for tissue repair. Their remote parenchymal localization and low counts, however, are limiting factors for their isolation and molecular characterization of their specific role during tissue inflammation. We isolated viable murine IMs in sufficient quantities by coculturing them with stromal cells and analyzed mRNA expression patterns of transient receptor potential (TRP) channels in naïve and M1 polarized IMs after application of lipopolysaccharide (LPS) and interferon γ. M-RNAs for the second member of the melastatin family of TRP channels, TRPM2, were upregulated in the M1 state and functional channels were identified by their characteristic currents induced by ADP-ribose, its specific activator. Most interestingly, cytokine production and secretion of interleukin-1α (IL-1α), IL-6 and tumor necrosis factor-α in M1 polarized but TRPM2-deficient IMs was significantly enhanced compared to WT cells. Activation of TRPM2 channels by ADP-ribose (ADPR) released from mitochondria by ROS-produced H2O2 significantly increases plasma membrane depolarization, which inhibits production of reactive oxygen species by NADPH oxidases and reduces cytokine production and secretion in a negative feedback loop. Therefore, TRPM2 channels are essential for the regulation of cytokine production in M1-polarized murine IMs. Specific activation of these channels may promote an anti-inflammatory phenotype and prevent a harmful cytokine storm often observed in COVID-19 patients.

Enterprise digital development and capacity utilization

PurposeTo enrich the research on the economic consequences of enterprise digital development from the perspective of capacity utilization.Design/methodology/approachUsing a sample of listed firms from 2010 to 2020, this paper exploits text analysis of annual reports to construct a proxy for enterprise digital development.FindingsResults show that enterprise digital development not only improves their own capacity utilization but also generates a positive spillover effect on the capacity utilization of peer firms and firms in the supply chain. Next, based on the incomplete information about market demand and potential competitors when making capacity-building decisions, the mechanism tests show that improving the accuracy of market forecasts and reducing investment surges are potential channels behind the baseline results. Cross-sectional tests show the baseline result is more pronounced when industries are highly homogeneous and when firms have access to less information.Originality/valueThis paper contributes to the research related to the economic consequences of digital development. With the development of the digital economy, the real effects of enterprise digital development have also triggered extensive interest and exploration. Existing studies mainly examine the impact on physical operations, such as specialization division of labor, innovation activities, business performance or total factor productivity (Huang, Yu, & Zhang, 2019; Yuan, Xiao, Geng, & Sheng, 2021; Wang, Kuang, & Shao, 2017; Li, Liu, & Shao, 2021; Zhao, Wang, & Li, 2021). These studies measure the economic benefits from the perspective of the supply (output) side but neglect the importance of the supply system to adapt to the actual market demand. In contrast, this paper focuses on capacity utilization, aimed at estimating the net economic effect of digital development by considering the supply-demand fit scenario. Thus, our findings enrich the relevant studies on the potential consequences of digital development.

Quantity or quality: Novel insights into the impact of digital finance on innovation

Despite prior literature shedding light on the role of digital finance in enhancing innovation quantity, research on whether and how it influences innovation quality remains limited. To address this gap, this study investigates the causal relationship between digital finance and the innovation performance of manufacturing enterprises, and identifies the inclusive effects of digital finance in China. We provide compelling evidence that digital finance generates significantly positive effects on innovation performance, encompassing both quantity and quality dimensions. Mechanism analyses demonstrate that financial constraints serve as a potential channel through which digital finance operates. We also observe the inclusive effects of digital finance on innovation performance. Specifically, the innovation improvements induced by digital finance are more pronounced for small and medium-sized enterprises (SMEs) and state-owned enterprises (SOEs).

Economic policy uncertainty and credit risk in microfinance: A cross‐country analysis

AbstractUsing a cross‐country data set of 670 microfinance institutions (MFIs) in nine countries from 1999 to 2018, this study examines the impact of economic policy uncertainty (EPU) on the credit risk of MFIs. The empirical results show that EPU significantly increases the credit risk of MFIs. Our findings are valid in a series of robustness checks. The moderating effects reveal that group lending can mitigate the impact of EPU on the credit risk of MFIs and that EPU has a weaker impact on nongovernmental organizations (NGOs) and cooperatives and credit unions (Coop/CUs). We explore potential channels through which credit risk is influenced by EPU from the perspectives of earnings volatility and cost. We find that EPU increases the credit risk of MFIs not only by reducing profitability and leverage levels and increasing earnings volatility but also by raising financing costs.