Calcium Cycling Research Articles

The IgCAM CAR Regulates Gap Junction-Mediated Coupling on Embryonic Cardiomyocytes and Affects Their Beating Frequency.

The IgCAM coxsackie-adenovirus receptor (CAR) is essential for embryonic heart development and electrical conduction in the mature heart. However, it is not well-understood how CAR exerts these effects at the cellular level. To address this question, we analyzed the spontaneous beating of cultured embryonic hearts and cardiomyocytes from wild type and CAR knockout (KO) embryos. Surprisingly, in the absence of the CAR, cultured cardiomyocytes showed increased frequencies of beating and calcium cycling. Increased beatings of heart organ cultures were also induced by the application of reagents that bind to the extracellular region of the CAR, such as the adenovirus fiber knob. However, the calcium cycling machinery, including calcium extrusion via SERCA2 and NCX, was not disrupted in CAR KO cells. In contrast, CAR KO cardiomyocytes displayed size increases but decreased in the total numbers of membrane-localized Cx43 clusters. This was accompanied by improved cell-cell coupling between CAR KO cells, as demonstrated by increased intercellular dye diffusion. Our data indicate that the CAR may modulate the localization and oligomerization of Cx43 at the plasma membrane, which could in turn influence electrical propagation between cardiomyocytes via gap junctions.

Quantitative interactome proteomics reveals Plin5 as a novel partner of sarcoplasmic/endoplasmic reticulum Ca2+ATPase 2a (SERCA2a) in the regulation of calcium cycling

Quantitative interactome proteomics reveals Plin5 as a novel partner of sarcoplasmic/endoplasmic reticulum Ca2+ATPase 2a (SERCA2a) in the regulation of calcium cycling

A novel (CaO/CeO2)@CeO2 composite adsorbent based on microinjection titration-calcination strategy for CO2 adsorption

A simultaneous titration-calcination strategy was proposed to synthesize core–shell type (CaCO3/Ce(OH)CO3)@Ce(OH)CO3 precursors by titrating Ca2+ and Ce3+ into aqueous CO2 storage material (CO2SM) solution simultaneously without adding any templating agents or surfactants. After calcination at 900 °C, the novel core–shell type (CaO/CeO2)@CeO2 composite adsorbent with the strong stability of adsorption cycle was synthesized. The effects of Ca/Ce doping ratio, CO2SM dosage, drop rate and temperature conditions on the CO2 adsorption performance on the composite adsorbent during the calcium cycle were investigated. The synthetic adsorbent with a Ca/Ce doping ratio of 24: 1 had the strongest CO2 capture performance during the calcium cycle, and it mainly consisted of CaO and CeO2. The CO2 adsorption amount still maintained at about 430 mg/g after 15 cycles under the adsorption–desorption conditions at 750 °C and 900 °C. (CaO/CeO2)@CeO2 composite adsorbent is a core–shell structure with CeO2 as the shell and CaO/CeO2 as the liner, in which CeO2 is uniformly dispersed in CaO pore channels, which gives the adsorbent a stable pore structure and enhances the anti-sintering performance of Ca-based adsorbent. Due to the special property of CeO2, it can also promote the ion migration during the carbonation process, thus maintaining the adsorption activity. Density functional theory (DFT) calculations successfully explain why CeO2 can be used as an inert dopant to promote the adsorption of CO2. After that, the synthesis mechanism of core–shell adsorbent and the reaction mechanism of CO2 adsorption are proposed in conjunction with the series characterization. This work provides a new strategy for the efficient synthesis of Ca-based adsorbents with special morphology and a new idea for CO2 capture.

Modulation of Mitochondrial Bioenergetics by Polydopamine Nanoparticles in Human iPSC-Derived Cardiomyocytes.

Myocardial infarction (MI) leads to the formation of an akinetic scar on the heart muscle causing impairment in cardiac contractility and conductance, leading to cardiac remodeling and heart failure (HF). The current pharmacological approaches for attenuating MI are limited and often come with long-term adverse effects. Therefore, there is an urgent need to develop novel multimodal therapeutics capable of modulating cardiac activity without causing any major adverse effects. In the current study, we have demonstrated the applicability of polydopamine nanoparticles (PDA-NPs) as a bioactive agent that can enhance the contractility and beat propagation of human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Treatment of hiPSC-CMs with PDA-NPs demonstrated accumulation of the latter into mitochondria and significantly enhanced time-dependent adenosine triphosphate (ATP) production in these cells, indicating improved mitochondrial bioenergetics. Furthermore, the effect of PDA-NPs on hiPSC-CM activity was evaluated by measuring calcium transients. Treatment with PDA-NPs increased the calcium cycling in hiPSC-CMs in a temporal manner. Our results demonstrated a significant reduction in peak amplitude, transient duration, time to peak, and transient decay time in the PDA-NPs-treated hiPSC-CMs as compared to untreated hiPSC-CMs. Additionally, treatment of isolated perfused rat heart ex vivo with PDA-NPs demonstrated cardiotonic effects on the heart and significantly improved the hemodynamic function, suggesting its potential for enhancing whole heart contractility. Lastly, the gene expression analysis data revealed that PDA-NPs significantly upregulated cardiac-specific genes (ACADM, MYL2, MYC, HCN1, MYL7, GJA5, and PDHA1) demonstrating the ability to modulate genetic expression of cardiomyocytes. Taken together, these findings suggest PDA-NPs capability as a versatile nanomaterial with potential uses in next-generation cardiovascular applications.

Modeling Calcium Cycling in the Heart: Progress, Pitfalls, and Challenges.

Intracellular calcium (Ca) cycling in the heart plays key roles in excitation-contraction coupling and arrhythmogenesis. In cardiac myocytes, the Ca release channels, i.e., the ryanodine receptors (RyRs), are clustered in the sarcoplasmic reticulum membrane, forming Ca release units (CRUs). The RyRs in a CRU act collectively to give rise to discrete Ca release events, called Ca sparks. A cell contains hundreds to thousands of CRUs, diffusively coupled via Ca to form a CRU network. A rich spectrum of spatiotemporal Ca dynamics is observed in cardiac myocytes, including Ca sparks, spark clusters, mini-waves, persistent whole-cell waves, and oscillations. Models of different temporal and spatial scales have been developed to investigate these dynamics. Due to the complexities of the CRU network and the spatiotemporal Ca dynamics, it is challenging to model the Ca cycling dynamics in the cardiac system, particularly at the tissue sales. In this article, we review the progress of modeling of Ca cycling in cardiac systems from single RyRs to the tissue scale, the pros and cons of the current models and different modeling approaches, and the challenges to be tackled in the future.

Characteristics of Calcium Isotopes at Different Water Depths and Their Palaeoenvironmental Significance for Carbonate Rocks of the Permian-Triassic Boundary in Chibi, Southern China

Calcium isotopes of carbonate rocks can trace calcium cycles and record changes in the marine environment. As published calcium isotope profiles of carbonate rocks at the Permian-Triassic boundary are rare, comparative studies on deep-water profiles were lacking for the major extinction event that occurred during this time. We present sections of different water depths in the Chibi area of southern China that we have selected for a comparative study. We analyzed carbon isotopes, calcium isotopes, as well as major and trace elements of carbonates from two sections (Chibi North and Chibi West) to obtain information on the volcanic activity, ocean acidification, as well as sea level rise and fall in the Chibi area during the mass extinction period. All carbon and calcium isotopes of carbonates from both sections are all negative after the mass extinction boundary. Carbonates from the Chibi North section have higher δ44/40Ca values and lower Sr/Ca ratios than the rocks from the Chibi West section. We propose that the negative bias of the calcium isotopes in the two sections result from diagenesis. Diagenesis transforms primary aragonite into calcite, showing the characteristics of high δ44/40Ca value and low Sr/Ca. By comparing our data with three published profiles of shallow-water carbonate rock, we recognize that calcium isotopes record gradients at different water depths. In the slope environment, the enhancement of pore fluid action near the coast caused an increase of the fluid buffer alteration, and we propose that a regression event occurred in the Chibi region during the Late Permian.

Authigenic Formation of Clay Minerals in the Abyssal North Pacific

AbstractPresent estimates of the biogeochemical cycles of calcium, strontium, and potassium in the ocean reveal large imbalances between known input and output fluxes. Using pore fluid, incubation, and solid sediment data from North Pacific multi‐corer cores we show that, contrary to the common paradigm, the top centimeters of abyssal sediments can be an active site of authigenic precipitation of clay minerals. In this region, clay authigenesis is the dominant sink for potassium and strontium and consumes nearly all calcium released from benthic dissolution of calcium carbonates. These observations support the idea that clay authigenesis occurring over broad regions of the world ocean may be a major buffer for ocean chemistry on the time scale of the ocean overturning circulation, and key to the long‐term stability of Earth's climate.

An iterated map model with CaMKII feedback in modeling the force frequency relationship of a cardiac cell

Experiment data on isolated rat hearts shows that the transient behaviors after switching pacing intervals are very complicated with increasing, decreasing, and rebound of the contraction force. The strength of contraction in the heart muscle is strongly related to intracellular free Ca2+ mediated by an action potential. This behavior can be explained by calcium cycling inside the excitable cardiac myocytes coupled with their action potential. The previous and recently proposed models can only explain a short period of time after changing the pacing frequency. Our aim is to develop a simple feedback model based on the role of the enzyme CaMKII to describe the whole dynamic picture captured from experiments.

Mechanisms of spontaneous Ca2+ release-mediated arrhythmia in a novel 3D human atrial myocyte model: I. Transverse-axial tubule variation.

Intracellular calcium (Ca2+ ) cycling is tightly regulated in the healthy heart ensuring effective contraction. This is achieved by transverse (t)-tubule membrane invaginations that facilitate close coupling of key Ca2+ -handling proteins such as the L-type Ca2+ channel and Na+ -Ca2+ exchanger (NCX) on the cell surface with ryanodine receptors (RyRs) on the intracellular Ca2+ store. Although less abundant and regular than in the ventricle, t-tubules also exist in atrial myocytes as a network of transverse invaginations with axial extensions known as the transverse-axial tubule system (TATS). In heart failure and atrial fibrillation, there is TATS remodelling that is associated with aberrant Ca2+ -handling and Ca2+ -induced arrhythmic activity; however, the mechanism underlying this is not fully understood. To address this, we developed a novel 3D human atrial myocyte model that couples electrophysiology and Ca2+ -handling with variable TATS organization and density. We extensively parameterized and validated our model against experimental data to build a robust tool examining TATS regulation of subcellular Ca2+ release. We found that varying TATS density and thus the localization of key Ca2+ -handling proteins has profound effects on Ca2+ handling. Following TATS loss, there is reduced NCX that results in increased cleft Ca2+ concentration through decreased Ca2+ extrusion. This elevated Ca2+ increases RyR open probability causing spontaneous Ca2+ releases and the promotion of arrhythmogenic waves (especially in the cell interior) leading to voltage instabilities through delayed afterdepolarizations. In summary, the present study demonstrates a mechanistic link between TATS remodelling and Ca2+ -driven proarrhythmic behaviour that probably reflects the arrhythmogenic state observed in disease. KEY POINTS: Transverse-axial tubule systems (TATS) modulate Ca2+ handling and excitation-contraction coupling in atrial myocytes, with TATS remodelling in heart failure and atrial fibrillation being associated with altered Ca2+ cycling and subsequent arrhythmogenesis. To investigate the poorly understood mechanisms linking TATS variation and spontaneous Ca2+ release, we built, parameterized and validated a 3D human atrial myocyte model coupling electrophysiology and spatially-detailed subcellular Ca2+ handling governed by the TATS. Simulated TATS loss causes diastolic Ca2+ and voltage instabilities through reduced Na+ -Ca2+ exchanger-mediated Ca2+ removal, cleft Ca2+ accumulation and increased ryanodine receptor open probability, resulting in spontaneous Ca2+ release and promotion of arrhythmogenic waves and delayed afterdepolarizations. At fast electrical rates typical of atrial tachycardia/fibrillation, spontaneous Ca2+ releases are larger and more frequent in the cell interior than at the periphery. Our work provides mechanistic insight into how atrial TATS remodelling can lead to Ca2+ -driven instabilities that may ultimately contribute to the arrhythmogenic state in disease.

Carbon-calcium composite conversion of calcium carbide-acetylene system: On the imperative roles of carbon capture and solid waste recycling

As an important national basic industry in China, the production of calcium carbide faced with huge pressure on energy conservation and emission reduction because of the CO2 emission and solid waste carbide slag. Therefore, it is necessary to realize the recycling of carbon and calcium in the production process of calcium carbide. A novel system of carbon-calcium compound conversion for calcium carbide-acetylene production was proposed in this work, which combines two-stage carbon capture and calcium carbide waste slag reuse processes to achieve CO2 enrichment and calcium cycle. Based on the simulation data, the proposed system was comprehensively evaluated by material conversion, exergy and exergoeconomic analyses. It was found that the improved process performed better with an effective C, H, Ca atomic conversion rate for carbide furnace of 85.41% and CO2 capture efficiency of 90.35%, compared with the referenced process of 64.51% and 0, respectively. The exergoeconomic analysis results suggested that more focus should be put on carbide furnace, acetylene reactor, re-carbonization furnace, gasifier and calciner since they are the top five of capital investments and exergy destruction. Besides, the carbide furnace, acetylene reactor, re-carbonization furnace and gasifier have relatively lower exergoeconomic factor (fk) values of 1.00%, 0.93%, 2.27% and 3.07%, respectively, indicating that exergy destruction costs of these components can be decreased with the improvement of system thermodynamic and equipment performance. Furthermore, the calcium looping process formed based on the improved oxy-thermal method (OTM) process, using the captured CO2 to mineralize carbide slag to form another calcification cycle for the production of calcium carbide-acetylene, has a higher exergy efficiency of 48.97% than the referenced process of 47.85%, and also achieves the lowest carbon emissions and the obvious reduction in CaO input. Results revealed that the proposed calcium looping system with high-efficiency, low-carbon and clean for calcium carbide-acetylene production, could be a promising process for carbon emission reduction in practical applications.

Three-dimensional remodelling of the cellular energy distribution system during postnatal heart development.

The heart meets the high energy demands of constant muscle contraction and calcium cycling primarily through the conversion of fatty acids into adenosine triphosphate (ATP) by a large volume of mitochondria. As such, the spatial relationships among lipid droplets (LDs), mitochondria, the sarcotubular system and the contractile apparatus are critical to the efficient distribution of energy within the cardiomyocyte. However, the connectivity among components of the cardiac cellular energy distribution system during postnatal development remains unclear. Here, we use volume electron microscopy to demonstrate that the sarcomere branches uniting the myofibrillar network occur more than twice as frequently during early postnatal development as in mature cardiomyocytes. Moreover, we show that the mitochondrial networks arranged in parallel to the contractile apparatus are composed of larger, more compact mitochondria with greater connectivity to adjacent mitochondria in mature as compared with early postnatal cardiomyocytes. Finally, we find that connectivity among mitochondria, LDs and the sarcotubular network is greater in developing than in mature muscles. These data suggest that physical connectivity among cellular structures may facilitate the communication needed to coordinate developmental processes within the cardiac muscle cell. This article is part of the theme issue 'The cardiomyocyte: new revelations on the interplay between architecture and function in growth, health, and disease'.

Diabetes disturbs functional adaptation of the remote myocardium after ischemia/reperfusion

Diabetes mellitus type 2 is associated with adverse clinical outcome after myocardial infarction. To better understand the underlying causes we here investigated sarcomere protein function and its calcium-dependent regulation in the non-ischemic remote myocardium (RM) of diabetic mice (db/db) after transient occlusion of the left anterior descending coronary artery. Before and 24 h after surgery db/db and non-diabetic db/+ underwent magnetic resonance imaging followed by histological and biochemical analyses of heart tissue. Intracellular calcium transients and sarcomere function were measured in isolated cardiomyocytes. Active and passive force generation was assessed in skinned fibers and papillary muscle preparations. Before ischemia and reperfusion (I/R), beat-to-beat calcium cycling was depressed in diabetic cardiomyocytes. Nevertheless, contractile function was preserved owing to increased myofilament calcium sensitivity and higher responsiveness of myocardial force production to β-adrenergic stimulation in db/db compared to db/+. In addition, protein kinase C activity was elevated in db/db hearts leading to strong phosphorylation of the titin PEVK region and increased titin-based tension of myofilaments. I/R impaired the function of whole hearts and RM sarcomeres in db/db to a larger extent than in non-diabetic db/+, and we identified several reasons. First, the amplitude and the kinetics of cardiomyocyte calcium transients were further reduced in the RM of db/db. Underlying causes involved altered expression of calcium regulatory proteins. Diabetes and I/R additively reduced phospholamban S16-phosphorylation by 80% (P < 000.1) leading to strong inhibition of the calcium ATPase SERCA2a. Second, titin stiffening was only observed in the RM of db/+, but not in the RM of db/db. Finally, db/db myofilament calcium sensitivity and force generation upon β-adrenergic stimulation were no longer enhanced over db/+ in the RM. The findings demonstrate that impaired cardiomyocyte calcium cycling of db/db hearts is compensated by increased myofilament calcium sensitivity and increased titin-based stiffness prior to I/R. In contrast, sarcomere function of the RM 24 h after I/R is poor because both these compensatory mechanisms fail and myocyte calcium handling is further depressed.

Altered skeletal muscle sarco-endoplasmic reticulum Ca2+-ATPase calcium transport efficiency after a thermogenic stimulus.

Exposure to predator threat induces a rapid and robust increase in skeletal muscle thermogenesis in rats. The central nervous system relays threat information to skeletal muscle through activation of the sympathetic nervous system, but muscle mechanisms mediating this thermogenesis remain unidentified. Given the relevance of sarcolipin-mediated futile calcium cycling through the sarco-endoplasmic reticulum Ca2+-ATPase (SERCA) pump to mammalian muscle nonshivering thermogenesis, we hypothesized that this plays a role in contextually induced muscle thermogenesis as well. This was assessed by measuring enzymatic activity of SERCA and sarcoplasmic reticulum Ca2+ transport, where the apparent coupling ratio (Ca2+ uptake rate divided by ATPase activity rate at a standard Ca2+ concentration) was predicted to decrease in association with muscle thermogenesis. Sprague-Dawley rats exposed to predator (ferret) odor (PO) showed a rapid decrease in the apparent coupling ratio in the soleus muscle, indicating SERCA uncoupling compared with control-odor-exposed rats. A rat model of high aerobic fitness and elevated muscle thermogenesis also demonstrated soleus muscle SERCA uncoupling relative to their obesity-prone, low-fitness counterparts. Both the high- and low-aerobic fitness rats showed soleus SERCA uncoupling with exposure to PO. Finally, no increase in sarcolipin expression in soleus muscle was detected with PO exposure. This dataset implicates muscle uncoupling of SERCA Ca2+ transport and ATP hydrolysis, likely through altered SERCA or sarcolipin function outside of translational regulation, as one contributor to the muscle thermogenesis provoked by exposure to predator threat. These data support the involvement of SERCA uncoupling in both muscle thermogenic induction and enhanced aerobic capacity.

Coupled dolomite and silica precipitation from continental weathering during deglaciation of the Marinoan Snowball Earth

Geological records show that multiple ice sheets have covered tropical areas toward the end of the Neoproterozoic. The Snowball Earth concept, one of the most prevalent hypotheses used to explain these records, contemplates a stable frozen Earth from the runaway ice-albedo effect. The Snowball Earth scenario eventually ended with intensive deglaciation due to extremely high pCO2 levels accumulated through millions of years. We used a geochemical model that integrated the carbon, silica, calcium, and magnesium cycles to understand changes in sedimentary mineral deposits and ocean chemistry during and after a Snowball Earth event. Intense chemical weathering in response to the extremely high levels of pCO2 at the beginning of deglaciation delivered an immense amount of calcium, magnesium, and dissolved silica into the ocean resulting in a spike of dolomite, as “cap dolostone”, and silica deposition. Previous models used CaCO3 to calculate carbonate precipitation due to an insufficient understanding of dolomite precipitation criteria and rates. By incorporating the magnesium cycle and coupling dolomite precipitation with dissolved silica levels, we provide an explanation for the occurrence and duration of the postglacial dolomite burial. This model also provides constraints on the relative intensity of silicate versus carbonate weathering based on the appearance of cap dolostone during deglaciation and the amplitude of pH change.

Statins Protect Against Early Stages of Doxorubicin-induced Cardiotoxicity Through the Regulation of Akt Signaling and SERCA2

BackgroundDoxorubicin-induced cardiomyopathy (DICM) is one of the complications that can limit treatment for a significant number of cancer patients. In animal models, the administration of statins can prevent the development of DICM. Therefore, the use of statins with anthracyclines potentially could enable cancer patients to complete their chemotherapy without added cardiotoxicity. The precise mechanism mediating the cardioprotection is not well understood. The purpose of this study is to determine the molecular mechanism by which rosuvastatin confers cardioprotection in a mouse model of DICM. MethodsRosuvastatin was intraperitoneally administered into adult male mice at 100 μg/kg daily for 7 days, followed by a single intraperitoneal doxorubicin injection at 10 mg/kg. Animals continued to receive rosuvastatin daily for an additional 14 days. Cardiac function was assessed by echocardiography. Optical calcium mapping was performed on retrograde Langendorff perfused isolated hearts. Ventricular tissue samples were analyzed by immunofluorescence microscopy, Western blotting, and quantitative polymerase chain reaction. ResultsExposure to doxorubicin resulted in significantly reduced fractional shortening (27.4% ± 1.11% vs 40% ± 5.8% in controls; P < 0.001) and re-expression of the fetal gene program. However, we found no evidence of maladaptive cardiac hypertrophy or adverse ventricular remodeling in mice exposed to this dose of doxorubicin. In contrast, rosuvastatin-doxorubicin-treated mice maintained their cardiac function (39% ± 1.26%; P < 0.001). Mechanistically, the effect of rosuvastatin was associated with activation of Akt and phosphorylation of phospholamban with preserved sarcoplasmic/endoplasmic reticulum Ca2+ transporting 2 (SERCA2)-mediated Ca2+ reuptake. These effects occurred independently of perturbations in ryanodine receptor 2 function. ConclusionsRosuvastatin counteracts the cardiotoxic effects of doxorubicin by directly targeting sarcoplasmic calcium cycling.

Abstract P2097: Hypochloremia Alters Cardiac Physiology After Myocardial Ischemia-Reperfusion Injury

Ischemic heart disease (IHD) is one of the leading causes of death related due to heart failure (HF). It is mainly characterized by impairment of heart contraction-relaxation (rhythmicity) and cardiac output. While a myriad of cellular processes is associated with IHD, one of the major cellular aspects strongly associated is the electrolyte and ionic imbalance. Serum sodium levels have been a well-established adverse prognostic marker for patients with chronic heart failure. Recent evidences have suggested that the lower serum chloride (hypochloremia, &amp;lt90mM) is associated with increased mortality risk in patients with chronic HF independent of serum sodium levels. Serum chloride (Cl - ) is a major anion in physiology but its correlation to IHD has not been studied extensively. Hence, to better understand the role of hypochloremia in HF patients, we reviewed patients who received a left ventricular assist device (LVAD) placement at The Ohio State University Medical Center. Survival analysis demonstrated that patients with hypochloremia before LVAD placement had decreased survival at one year compared to patients with normal serum Cl - (p=0.0055). Moreover, our e x vivo studies with isolated rat hearts indicate hypochloremia aggravates myocardial infarction post ischemia-reperfusion (IR) injury. Hence, to elucidate Cl - mediated signaling mechanism we incorporated highly translational human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM). Our cellular studies demonstrate that physiological alteration of extracellular Cl - affects the function of beating hiPSC-CM. Hypochloremia significantly increases the beating rate by reducing the calcium cycling durations in hiPSC-CM. Studies are ongoing to determine the involvement of the chloride channel in maintaining chloride homeostasis. Taken together, our results for the first time establish the novel role and mechanism of chloride homeostasis in IR injury. These results will advance the need to search for effective therapeutic approaches to target novel Cl - channels in IHD.

Abstract P2105: Precision Medicine In Pediatric Calmodulinopathy

Calmodulinopathy is a life-threatening cardiac arrhythmia syndrome resulting from a single heterozygous mutation in any of the three calmodulin (CaM) genes. Most (68%) persons with a pathogenic CaM variant will experience a major arrhythmic event such as aborted cardiac arrest or sudden cardiac death by the age of five years. Calmodulinopathies result in heterogenous clinical features with one of the predominant clinical phenotypes being long QT syndrome (CaM-LQTS). Despite current treatments, 56% of calmodulinopathy patients experience breakthrough cardiac events, underscoring the need for improved therapeutics. A current barrier to effective treatment is our incomplete understanding of how calmodulin genetic variants contribute to pathological phenotypes. Human induced pluripotent stem cells (hiPSCs) can be differentiated into cardiomyocytes (hiPSC-CMs) while maintaining the patient’s genetic background. Application of hiPSC-CMs in the context of this disease allows us to better understand patient-specific heterogeneity at the cellular level. We have established a CaM-LQTS hiPSC-CM model using hiPSCs generated from five persons with a pathogenic CaM variant who present with CaM-LQTS. We hypothesize that we can model a CaM-LQTS phenotype using patient-specific hiPSC-CMs, identify clinically appropriate drugs that correct it, and determine underlying mechanisms that drive this phenotype. We have determined the cellular CaM-LQTS phenotype of each patient line using blinded electrophysiological, contractility, and calcium imaging studies and have identified FDA-approved drugs that alleviate these phenotypes. We are also performing experiments, with a focus on calcium cycling dysregulation, to identify underlying mechanisms that drive the cellular CaM-LQTS phenotype. In conclusion, these studies will provide a patient-specific methodology to identify necessary clinical treatments for CaM-LQTS and elucidate the drivers of this rare and severe disease.

Contractility and Calcium Transient Maturation in the Human iPSC-Derived Cardiac Microfibers.

Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) are considered immature in the sarcomere organization, contractile machinery, calcium transient, and transcriptome profile, which prevent them from further applications in modeling and studying cardiac development and disease. To improve the maturity of hiPSC-CMs, here, we engineered the hiPSC-CMs into cardiac microfibers (iCMFs) by a stencil-based micropatterning method, which enables the hiPSC-CMs to be aligned in an end-to-end connection for prolonged culture on the hydrogel of physiological stiffness. A series of characterization approaches were performed to evaluate the maturation in iCMFs on both structural and functional levels, including immunohistochemistry, calcium transient, reverse-transcription quantitative PCR, cardiac contractility, and electrical pacing analysis. Our results demonstrate an improved cardiac maturation of hiPSC-CMs in iCMFs compared to micropatterned or random single hiPSC-CMs and hiPSC-CMs in a random cluster at the same cell number of iCMFs. We found an increased sarcomere length, better regularity and alignment of sarcomeres, enhanced contractility, matured calcium transient, and T-tubule formation and improved adherens junction and gap junction formation. The hiPSC-CMs in iCMFs showed a robust calcium cycling in response to the programmed and continuous electrical pacing from 0.5 to 7 Hz. Moreover, we generated the iCMFs with hiPSC-CMs with mutations in myosin-binding protein C (MYBPC3) to have a proof-of-concept of iCMFs in modeling cardiac hypertrophic phenotype. These findings suggest that the multipatterned iCMF connection of hiPSC-CMs boosts the cardiac maturation structurally and functionally, which will reveal the full potential of the application of hiPSC-CM models in disease modeling of cardiomyopathy and cardiac regenerative medicine.

Classification of various nutrient deficiencies in tomato plants through electrophysiological signal decomposition and sample space reduction

Plants leave testimonies of undergoing physical state by depicting distinct variations in their electrophysiological data. Adequate nutrition of plants signifies their role in the growth and a plentiful harvest. Plant signal data carries enough information to detect and analyse nutrient deficiency. Classification of nutrient deficiencies through signal decomposition and bilevel measurements has not been reported earlier. The proposed work explores tomato plants in four-time cycles (Early Morning, Morning, After Noon, Night) of macronutrients Calcium (Ca), Nitrogen (N) and micronutrients Manganese (Mn), Iron (Fe). Using the Empirical Mode Decomposition method (EMD), signals are decomposed into Intrinsic Mode Functions (IMF) in 10-levels. Further, Intrinsic mode functions are grouped into two clusters to extract descriptive data statistics and bi-level measurements. Then a novel sample selection method is proposed to achieve a better classification rate by reducing sample space. A binary classification model is built to train and test 15 features individually using discriminant analysis and naïve-Bayes classifier variants. The reported results achieved a classification rate up to 98% after 5-fold cross-validation. Attained findings endorse novel pathways for detection and classification of nutrient deficiencies in the early stages, consequently promoting prevention and treatment approaches earliest to the appearance of symptoms, also helping to enhance plant growth.

Characterizing arrhythmia using machine learning analysis of Ca2+ cycling inhuman cardiomyocytes.

SummaryAccurate modeling of the heart electrophysiology to predict arrhythmia susceptibility remains a challenge. Current electrophysiological analyses are hypothesis-driven models drawing conclusions from changes in a small subset of electrophysiological parameters because of the difficulty of handling and understanding large datasets. Thus, we develop a framework to train machine learning classifiers to distinguish between healthy and arrhythmic cardiomyocytes using their calcium cycling properties. By training machine learning classifiers on a generated dataset containing a total of 3,003 healthy derived cardiomyocytes and their various arrhythmic states, the multi-class models achieved >90% accuracy in predicting arrhythmia presence and type. We also demonstrate that a binary classifier trained to distinguish cardiotoxic arrhythmia from healthy electrophysiology could determine the key biological changes associated with that specific arrhythmia. Therefore, machine learning algorithms can be used to characterize underlying arrhythmic patterns in samples to improve in vitro preclinical models and complement current in vivo systems.