Cell-cell contacts via N-cadherin induce a regulatory renin secretory phenotype in As4.1 cells.

The lack of a clonal renin-secreting cell line has greatly hindered the investigation of the regulatory mechanisms of renin secretion at the cellular, biochemical, and molecular levels. In the present study, we investigated whether it was possible to induce phenotypic switching of the renin-expressing clonal cell line As4.1 from constitutive inactive renin secretion to regulated active renin secretion. When grown to postconfluence for at least two days in media containing fetal bovine serum or insulin-like growth factor-1, the formation of cell-cell contacts via N-cadherin triggered downstream cellular signaling cascades and activated smooth muscle-specific genes, culminating in phenotypic switching to a regulated active renin secretion phenotype, including responding to the key stimuli of active renin secretion. With the use of phenotype-switched As4.1 cells, we provide the first evidence that active renin secretion via exocytosis is regulated by phosphorylation/dephosphorylation of the 20 kDa myosin light chain. The molecular mechanism of phenotypic switching in As4.1 cells described here could serve as a working model for full phenotypic modulation of other secretory cell lines with incomplete phenotypes.

Teaching cardiac excitation-contraction coupling using a mathematical computer simulation model of human ventricular myocytes.

To understand the excitation-contraction (E-C) coupling of cardiomyocytes, including the electrophysiological mechanism of their characteristically long action potential duration, is one of the major learning goals in medical physiology. However, the integrative interpretation of the responses occurring during the contraction-relaxation cycle is challenging due to the dynamic interaction of underlying factors. Starting in 2017, we adopted the mathematical computer simulation model of human ventricular myocyte (Cardiac E-C_Sim), hypothesizing that this educational technology may facilitate students' learning of cardiac physiology. Here, we describe the overall process for the educational application of Cardiac E-C_Sim in the human physiology practicum of Seoul National University College of Medicine. We also report the results from questionnaires covering detailed assessment of the practicum class. The analysis of results and feedback opinions enabled us to understand how the students had approached the problem-solving process. As a whole, the students could better accomplish the learning goals using Cardiac E-C_Sim, followed by constructive discussions on the complex and dynamic mechanisms of cardiac E-C coupling. We suggest that the combined approach of lecture-based teaching and computer simulations guided by a manual containing clinical context would be broadly applicable in physiology education.

Toward a grey box approach for cardiovascular physiome.

The physiomic approach is now widely used in the diagnosis of cardiovascular diseases. There are two possible methods for cardiovascular physiome: the traditional mathematical model and the machine learning (ML) algorithm. ML is used in almost every area of society for various tasks formerly performed by humans. Specifically, various ML techniques in cardiovascular medicine are being developed and improved at unprecedented speed. The benefits of using ML for various tasks is that the inner working mechanism of the system does not need to be known, which can prove convenient in situations where determining the inner workings of the system can be difficult. The computation speed is also often higher than that of the traditional mathematical models. The limitations with ML are that it inherently leads to an approximation, and special care must be taken in cases where a high accuracy is required. Traditional mathematical models are, however, constructed based on underlying laws either proven or assumed. The results from the mathematical models are accurate as long as the model is. Combining the advantages of both the mathematical models and ML would increase both the accuracy and efficiency of the simulation for many problems. In this review, examples of cardiovascular physiome where approaches of mathematical modeling and ML can be combined are introduced.

Neuronal nitric oxide synthase modulation of intracellular Ca2+ handling overrides fatty acid potentiation of cardiac inotropy in hypertensive rats.

Cardiac neuronal nitric oxide synthase (nNOS) is an important molecule that regulates intracellular Ca2+ homeostasis and contractility of healthy and diseased hearts. Here, we examined the effects of nNOS on fatty acid (FA) regulation of left ventricular (LV) myocyte contraction in sham and angiotensin II (Ang II)-induced hypertensive (HTN) rats. Our results showed that palmitic acid (PA, 100 µM) increased the amplitudes of sarcomere shortening and intracellular ATP in sham but not in HTN despite oxygen consumption rate (OCR) was increased by PA in both groups. Carnitine palmitoyltransferase-1 inhibitor, etomoxir (ETO), reduced OCR and ATP with PA in sham and HTN but prevented PA potentiation of sarcomere shortening only in sham. PA increased nNOS-derived NO only in HTN. Inhibition of nNOS with S-methyl-L-thiocitrulline (SMTC) prevented PA-induced OCR and restored PA potentiation of myocyte contraction in HTN. Mechanistically, PA increased intracellular Ca2+ transient ([Ca2+]i) without changing Ca2+ influx via L-type Ca2+ channel (I-LTCC) and reduced myofilament Ca2+ sensitivity in sham. nNOS inhibition increased [Ca2+]i, I-LTCC and reduced myofilament Ca2+ sensitivity prior to PA supplementation; as such, normalized PA increment of [Ca2+]i. In HTN, PA reduced I-LTCC without affecting [Ca2+]i or myofilament Ca2+ sensitivity. However, PA increased I-LTCC, [Ca2+]i and reduced myofilament Ca2+ sensitivity following nNOS inhibition. Myocardial FA oxidation (18F-fluoro-6-thia-heptadecanoic acid, 18F-FTHA) was comparable between groups, but nNOS inhibition increased it only in HTN. Collectively, PA increases myocyte contraction through stimulating [Ca2+]i and mitochondrial activity in healthy hearts. PA-dependent cardiac inotropy was limited by nNOS in HTN, predominantly due to its modulatory effect on [Ca2+]i handling.

Body composition and personality traits in so-Yang type males.

BACKGROUND: The purpose of the present study was to examine the body composition of So-Yang type males according to Sasang constitutional medicine, which is popular in Korea. Different Sasang constitutional types are associated with different muscle distributions, body shapes, and disease susceptibilities. We used the Sasang Personality Questionnaire (SPQ) as a measure of the temperament of each Sasang type. METHODS: In total, 953 subjects aged over 20 years were recruited in Korea. We collected anthropometric parameters and bioimpedence information from the subjects and administered the SPQ. A logistic regression was conducted to calculate propensity scores. RESULTS: The percentage of skeletal muscle mass in So-Yang (SY) and non-So-Yang (non-SY) males was 45.8 ± 2.7 and 44.2 ± 3.3, respectively, before matching and 45.8 ± 2.6 and 44.9 ± 3.0, respectively, after propensity score matching. The extracellular water (ECW)/intracellular water (ICW) and extracellular water (ECW)/total body water (TBW) ratios and SPQ scores were significantly different between the SY and non-SY types. CONCLUSIONS: This study suggested that the SY type may be significantly and independently associated with body composition and could be associated with personality.

Cardiac inotropy, lusitropy, and Ca2+ handling with major metabolic substrates in rat heart.

Fatty acid (FA)-dependent oxidation is the predominant process for energy supply in normal heart. Impaired FA metabolism and metabolic insufficiency underlie the failing of the myocardium. So far, FA metabolism in normal cardiac physiology and heart failure remains undetermined. Here, we evaluate the mechanisms of FA and major metabolic substrates (termed NF) on the contraction, relaxation, and Ca2+ handling in rat left ventricular (LV) myocytes. Our results showed that NF significantly increased myocyte contraction and facilitated relaxation. Moreover, NF increased the amplitudes of diastolic and systolic Ca2+ transients ([Ca2+]i), abbreviated time constant of [Ca2+]i decay (tau), and prolonged the peak duration of [Ca2+]i. Whole-cell patch-clamp experiments revealed that NF increased Ca2+ influx via L-type Ca2+ channels (LTCC, ICa-integral) and prolonged the action potential duration (APD). Further analysis revealed that NF shifted the relaxation phase of sarcomere lengthening vs. [Ca2+]i trajectory to the right and increased [Ca2+]i for 50 % of sarcomere relengthening (EC50), suggesting myofilament Ca2+ desensitization. Butanedione monoxime (BDM), a myosin ATPase inhibitor that reduces myofilament Ca2+ sensitivity, abolished the NF-induced enhancement of [Ca2+]i amplitude and the tau of [Ca2+]i decay, indicating the association of myofilament Ca2+ desensitization with the changes in [Ca2+]i profile in NF. NF reduced intracellular pH ([pHi]). Increasing [pH]i buffer capacity with HCO3/CO2 attenuated Δ [pH]i and reversed myofilament Ca2+ desensitization and Ca2+ handling in NF. Collectively, greater Ca2+ influx through LTCCs and myofilament Ca2+ desensitization, via reducing [pH]i, are likely responsible for the positive inotropic and lusitropic effects of NF. Computer simulation recapitulated the effects of NF.

Coupled Ca2+/H+ transport by cytoplasmic buffers regulates local Ca2+ and H+ ion signaling.

Ca(2+) signaling regulates cell function. This is subject to modulation by H(+) ions that are universal end-products of metabolism. Due to slow diffusion and common buffers, changes in cytoplasmic [Ca(2+)] ([Ca(2+)]i) or [H(+)] ([H(+)]i) can become compartmentalized, leading potentially to complex spatial Ca(2+)/H(+) coupling. This was studied by fluorescence imaging of cardiac myocytes. An increase in [H(+)]i, produced by superfusion of acetate (salt of membrane-permeant weak acid), evoked a [Ca(2+)]i rise, independent of sarcolemmal Ca(2+) influx or release from mitochondria, sarcoplasmic reticulum, or acidic stores. Photolytic H(+) uncaging from 2-nitrobenzaldehyde also raised [Ca(2+)]i, and the yield was reduced following inhibition of glycolysis or mitochondrial respiration. H(+) uncaging into buffer mixtures in vitro demonstrated that Ca(2+) unloading from proteins, histidyl dipeptides (HDPs; e.g., carnosine), and ATP can underlie the H(+)-evoked [Ca(2+)]i rise. Raising [H(+)]i tonically at one end of a myocyte evoked a local [Ca(2+)]i rise in the acidic microdomain, which did not dissipate. The result is consistent with uphill Ca(2+) transport into the acidic zone via Ca(2+)/H(+) exchange on diffusible HDPs and ATP molecules, energized by the [H(+)]i gradient. Ca(2+) recruitment to a localized acid microdomain was greatly reduced during intracellular Mg(2+) overload or by ATP depletion, maneuvers that reduce the Ca(2+)-carrying capacity of HDPs. Cytoplasmic HDPs and ATP underlie spatial Ca(2+)/H(+) coupling in the cardiac myocyte by providing ion exchange and transport on common buffer sites. Given the abundance of cellular HDPs and ATP, spatial Ca(2+)/H(+) coupling is likely to be of general importance in cell signaling.

Effects of streptozotocin and unilateral nephrectomy on L-type Ca²âº channels and membrane capacitance in arteriolar smooth muscle cells.

Diabetes mellitus and hypertension are common diseases frequently coexisting. Although augmentation of L-type Ca(2+) channel (ICaL) activity has been reported in vascular smooth muscle cells (VSMCs) of a spontaneously hypertensive rat model, no study on ICaL has been conducted for coexisting hypertension and diabetes. Sprague Dawley rats were assigned to four groups: a sham-operated control group (CG), a unilateral nephrectomy group (UNG), a streptozotocin (STZ)-induced type 1 diabetic group (SDG) and a coexisting hypertension and diabetes group (DHG), which underwent nephrectomy and received STZ injection. Blood pressure (BP) was significantly lower in the CG than in the other three groups. The membrane capacitance of VSMCs was nearly doubled in the SDG and DHG but not in the UNG. The ICaL was increased approximately 2-fold in both the UNG and SDG and approximately 4-fold in the DHG. The current density of ICaL was increased approximately 2-fold in the UNG and DHG, while no significant increase was seen in the SDG. The rate of Ca(2+) removal was inhibited significantly, by ~33 %, in the DHG. In conclusion, the effects of hypertension and diabetes on ICaL were apparently additive, and the vascular consequences of combined diabetes and hypertension may be caused by an elevated ICaL and slowed Ca(2+) removal.

A Novel Nicotinamide Adenine Dinucleotide Correction Method for Mitochondrial Ca(2+) Measurement with FURA-2-FF in Single Permeabilized Ventricular Myocytes of Rat.

Fura-2 analogs are ratiometric fluoroprobes that are widely used for the quantitative measurement of [Ca(2+)]. However, the dye usage is intrinsically limited, as the dyes require ultraviolet (UV) excitation, which can also generate great interference, mainly from nicotinamide adenine dinucleotide (NADH) autofluorescence. Specifically, this limitation causes serious problems for the quantitative measurement of mitochondrial [Ca(2+)], as no available ratiometric dyes are excited in the visible range. Thus, NADH interference cannot be avoided during quantitative measurement of [Ca(2+)] because the majority of NADH is located in the mitochondria. The emission intensity ratio of two different excitation wavelengths must be constant when the fluorescent dye concentration is the same. In accordance with this principle, we developed a novel online method that corrected NADH and Fura-2-FF interference. We simultaneously measured multiple parameters, including NADH, [Ca(2+)], and pH/mitochondrial membrane potential; Fura-2-FF for mitochondrial [Ca(2+)] and TMRE for Ψm or carboxy-SNARF-1 for pH were used. With this novel method, we found that the resting mitochondrial [Ca(2+)] concentration was 1.03 µM. This 1 µM cytosolic Ca(2+) could theoretically increase to more than 100 mM in mitochondria. However, the mitochondrial [Ca(2+)] increase was limited to ~30 µM in the presence of 1 µM cytosolic Ca(2+). Our method solved the problem of NADH signal contamination during the use of Fura-2 analogs, and therefore the method may be useful when NADH interference is expected.

Pumping Ca2+ up H+ gradients: a Ca2(+)-H+ exchanger without a membrane.

Cellular processes are exquisitely sensitive to H+ and Ca2+ ions because of powerful ionic interactions with proteins. By regulating the spatial and temporal distribution of intracellular [Ca2+] and [H+], cells such as cardiac myocytes can exercise control over their biological function. A well-established paradigm in cellular physiology is that ion concentrations are regulated by specialized, membrane-embedded transporter proteins. Many of these couple the movement of two or more ionic species per transport cycle, thereby linking ion concentrations among neighbouring compartments. Here, we compare and contrast canonical membrane transport with a novel type of Ca(2+)-H+ coupling within cytoplasm, which produces uphill Ca2+ transport energized by spatial H+ ion gradients, and can result in the cytoplasmic compartmentalization of Ca2+ without requiring a partitioning membrane. The mechanism, demonstrated in mammalian myocytes, relies on diffusible cytoplasmic buffers, such as carnosine, homocarnosine and ATP, to which Ca2+ and H+ ions bind in an apparently competitive manner. These buffer molecules can actively recruit Ca2+ to acidic microdomains, in exchange for the movement of H+ ions. The resulting Ca2+ microdomains thus have the potential to regulate function locally. Spatial cytoplasmic Ca(2+)-H+ exchange (cCHX) acts like a 'pump' without a membrane and may be operational in many cell types.

Potassium currents in isolated deiters' cells of Guinea pig.

Deiters' cells are the supporting cells in organ of Corti and are suggested to play an important role in biochemical and mechanical modulation of outer hair cells. We successfully isolated functionally different K(+) currents from Deiters' cells of guinea pig using whole cell patch clamp technique. With high K(+) pipette solution, depolarizing step pulses activated strongly outward rectifying currents which were dose-dependently blocked by clofilium, a class III anti-arrhythmic K(+) channel blocker. The remaining outward current was transient in time course whereas the clofilium-sensitive outward current showed slow inactivation and delayed rectification. Addition of 5 mM tetraethylammonium (TEA) further blocked the remaining current leaving a very fast inactivating transient outward current. Therefore, at least three different types of K(+) current were identified in Deiters' cells, such as fast activating and fast inactivating current, fast activating slow inactivating current, and very fast inactivating transient outward current. Physiological role of them needs to be established.

Improving the hERG model fitting using a deep learning-based method.

The hERG channel is one of the essential ion channels composing the cardiac action potential and the toxicity assay for new drug. Recently, the comprehensive in vitro proarrhythmia assay (CiPA) was adopted for cardiac toxicity evaluation. One of the hurdles for this protocol is identifying the kinetic effect of the new drug on the hERG channel. This procedure included the model-based parameter identification from the experiments. There are many mathematical methods to infer the parameters; however, there are two main difficulties in fitting parameters. The first is that, depending on the data and model, parametric inference can be highly time-consuming. The second is that the fitting can fail due to local minima problems. The simplest and most effective way to solve these issues is to provide an appropriate initial value. In this study, we propose a deep learning-based method for improving model fitting by providing appropriate initial values, even the right answer. We generated the dataset by changing the model parameters and trained our deep learning-based model. To improve the accuracy, we used the spectrogram with time, frequency, and amplitude. We obtained the experimental dataset from https://github.com/CardiacModelling/hERGRapidCharacterisation. Then, we trained the deep-learning model using the data generated with the hERG model and tested the validity of the deep-learning model with the experimental data. We successfully identified the initial value, significantly improved the fitting speed, and avoided fitting failure. This method is useful when the model is fixed and reflects the real data, and it can be applied to any in silico model for various purposes, such as new drug development, toxicity identification, environmental effect, etc. This method will significantly reduce the time and effort to analyze the data.

Analysis of factors affecting Ca(2+)-dependent inactivation dynamics of L-type Ca(2+) current of cardiac myocytes in pulmonary vein of rabbit.

L-type Ca(2+) channels (ICaLs) are inactivated by an increase in intracellular [Ca(2+)], known as Ca(2+)-dependent inactivation (CDI). CDI is also induced by Ca(2+) released from the sarcoplasmic reticulum (SR), known as release-dependent inhibition (RDI). As both CDI and RDI occur in the junctional subsarcolemmal nanospace (JSS), we investigated which factors are involved within the JSS using isolated cardiac myocytes from the main pulmonary vein of the rabbit. Using the whole-cell patch clamp technique, RDI was readily observed with the application of a pre-pulse followed by a test pulse, during which the ICaLs exhibited a decrease in peak current amplitude and a slower inactivation. A fast acting Ca(2+) chelator, 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA), abolished this effect. As the time interval between the pre-pulse and test pulse increased, the ICaLs exhibited greater recovery and the RDI was relieved. Inhibition of the ryanodine receptor (RyR) or the SR Ca(2+)-ATPase (SERCA) greatly attenuated RDI and facilitated ICaL recovery. Removal of extracellular Na(+),which inhibits the Na(+)-Ca(2+) exchange (Incx), greatly enhanced RDI and slowed ICaL recovery, suggesting that Incx critically controls the [Ca(2+)] in the JSS. We incorporated the Ca(2+)-binding kinetics of the ICaL into a previously published computational model. By assuming two Ca(2+)-binding sites in the ICaL, of which one is of low-affinity with fast kinetics and the other is of high-affinity with slower kinetics, the new model was able to successfully reproduce RDI and its regulation by Incx. The model suggests that Incx accelerates Ca(2+) removal from the JSS to downregulate CDI and attenuates SR Ca(2+) refilling. The model may be useful to elucidate complex mechanisms involved in excitation­contraction coupling in myocytes.

Variability, Mean, and Baseline Values of Metabolic Parameters in Predicting Risk of Type 2 Diabetes.

CONTEXT: The effect of baseline (B) and alteration of metabolic parameters (MPs), including plasma glucose (PG) testing, insulin resistance surrogates, and lipid profile and their mutual interactions on the development of type 2 diabetes mellitus (T2DM), has not been investigated systematically. OBJECTIVE: To access the association of the past variability (V), past mean (M), and B values of various MPs and their mutual interaction with the risk of T2DM. METHODS: A community-based, longitudinal analysis was conducted using the Korean Genome and Epidemiology Study comprising 3829 nondiabetic participants with completed MPs measurements during 3 biannually visits who were followed over the next 10 years. Outcomes included the incidence of T2DM during follow-up. RESULTS: Among predictors, PG concentrations measured during the oral glucose tolerance test were the most prominent T2DM determinants, in which the M of the average value of fasting PG (FPG), 1-hour, and 2-hour PGs had the strongest discriminative power (hazard ratios and 95% CI for an increment of SD: 3.00 (2.5-3.26), AUC: 0.82). The M values of MPs were superior to their B and V values in predicting T2DM, especially among postload PGs. Various mutual interactions between indices and among MPs were found. The most consistent interactants were the M values of high-density lipoprotein cholesterol and the M and V values of FPG. The findings were similar in normal glucose tolerance participants and were confirmed by sensitivity analyses. CONCLUSION: Postload PG, past alteration of measurements, and mutual interactions among indices of MPs are important risk factors for T2DM development.

Development of a Portable Respiratory Gas Analyzer for Measuring Indirect Resting Energy Expenditure (REE).

OBJECTIVE: A rapidly growing home healthcare market has resulted in the development of many portable or wearable products. Most of these products measure, estimate, or calculate physiologic signals or parameters, such as step counts, blood pressure, or electrocardiogram. One of the most important applications in home healthcare is monitoring one's metabolic state since the change of metabolic state could reveal minor or major changes in one's health condition. A simple and noninvasive way to measure metabolism is through breath monitoring. With breath monitoring by breath gas analysis, two important indicators like the respiratory quotient (RQ) and resting energy exposure (REE) can be calculated. Therefore, we developed a portable respiratory gas analyzer for breath monitoring to monitor metabolic state, and the performance of the developed device was tested in a clinical trial. Approach. The subjects consisted of 40 healthy men and women. Subjects begin to measure exhalation gas using Vmax 29 for 15 minutes. After that, subjects begin to measure exhalation gas via the developed respiratory gas analyzer. Finally, the recorded data on the volume of oxygen (VO2), volume of carbon dioxide (VCO2), RQ, and REE were used to validate correlations between Vmax 29 and the developed respiratory gas analyzer. RESULTS: The results showed that the root-mean-square errors (RMSE) values of VCO2, VO2, RQ, and REE are 0.0315, 0.0417, 0.504, and 0.127. Bland-Altman plots showed that most of the VCO2, VO2, RQ, and REE values are within 95% of the significance level. CONCLUSIONS: We have successfully developed and tested a portable respiratory gas analyzer for home healthcare. However, there are limitations of the clinical trial; the number of subjects is small in size, and the age and race of subjects are confined. The developed portable respiratory gas analyzer is a cost-efficient method for measuring metabolic state and a new application of home healthcare.

The mean of fasting, 1-h, and 2-h plasma glucose levels is superior to each separate index in predicting diabetes.

AIMS: The fasting, 1-h, and 2-h plasma glucose (PG) levels during oral glucose tolerance test represent different glucose metabolic functions. We examined whether averaging these PG indices (GLUM0.60.120) results in a better predictor of future type 2 diabetes (T2DM). METHODS: 7533 participants were followed up biannually for 12 years. Hazard ratios (HRs), area under the curve (AUC) of the receiver-operating characteristic, and the net reclassification index (NRI) for T2DM were calculated to compare the discriminative ability of GLUM0.60.120 versus other PG indices. RESULTS: The adjusted HRs and 95% confidence intervals for an increase in SD of GLUM0.60.120 was 2.50 (2.36-2.65) and 1.88 (1.73-2.04) in T2DM-free and normal glucose tolerance (NGT) participants, respectively. The AUC of GLUM0.60.120 was higher than that of fasting PG, 1-h, and 2-h PG values for T2DM-free (0.79 versus 0.67, 0.77, and 0.73) and NGT (0.73 versus 0.65, 0.72, and 0.61). The model using GLUM0.60.120 improved the classification of the models with fasting PG, 1-h, and 2-h PG values (NRI: 0.369, 0.272, and 0.282 for T2DM-free and 0.249, 0.131, and 0.351 for NGT participants with all p < 0.001). CONCLUSIONS: The mean of fasting, 1-h, and 2-h PG levels predicts future T2DM better than each index.

In silico models for evaluating proarrhythmic risk of drugs.

Safety evaluation of drugs requires examination of the risk of generating Torsade de Pointes (TdP) because it can lead to sudden cardiac death. Until recently, the QT interval in the electrocardiogram (ECG) has been used in the evaluation of TdP risk because the QT interval is known to be associated with the development of TdP. Although TdP risk evaluation based on QT interval has been successful in removing drugs with TdP risk from the market, some safe drugs may have also been affected due to the low specificity of QT interval-based evaluation. For more accurate evaluation of drug safety, the comprehensive in vitro proarrhythmia assay (CiPA) has been proposed by regulatory agencies, industry, and academia. Although the CiPA initiative includes in silico evaluation of cellular action potential as a component, attempts to utilize in silico simulation in drug safety evaluation are expanding, even to simulating human ECG using biophysical three-dimensional models of the heart and torso under the effects of drugs. Here, we review recent developments in the use of in silico models for the evaluation of the proarrhythmic risk of drugs. We review the single cell, one-dimensional, two-dimensional, and three-dimensional models and their applications reported in the literature and discuss the possibility of utilizing ECG simulation in drug safety evaluation.

Seasonal effects on resting energy expenditure are dependent on age and percent body fat.

Seasonal variation in resting energy expenditure (REE) is still under debate. This study investigated seasonal changes in REE and relevant factors among Korean adults. A total of 867 healthy volunteers (385 men and 482 women) aged 20-69 years were divided into four seasonal groups and subgroups based on age, body mass index (BMI), and percent body fat (PBF) quartiles. REE, body composition, glucose metabolism, thyroid hormones, and catecholamines were measured. The seasonal factor contributed to REE independent of anthropometric indices, with additional variation decreasing from 6% to 2% among younger and older persons, respectively. The adjusted REE in the winter was 5.4-13.9%, 7.8-14.3%, and 8.6-11.9% higher than that in the summer in the age, BMI, and PBF subgroups, respectively. T3 and log-transformed norepinephrine (NElog) were higher, whereas log-transformed epinephrine (EPIlog) was lower in the winter compared to the summer. The magnitude of the winter-summer difference in REE and T3 and of the summer-winter difference in EPIlog were reduced three-fold between the lowest and highest intervals of age and PBF, whereas the difference in NElog was constant across all age and PBF intervals. There was no obvious change in seasonal differences in REE or its relevant biomarkers across BMI intervals. In summary, season is an independent predictor of REE and its effect is attenuated by the increment of age and PBF but not BMI.

A new integrated method for analyzing heart mechanics using a cell-hemodynamics-autonomic nerve control coupled model of the cardiovascular system.

A model of the cardiovascular system coupling cell, hemodynamics, and autonomic nerve control function is proposed for analyzing heart mechanics. We developed a comprehensive cardiovascular model with multi-physics and multi-scale characteristics that simulates the physiological events from membrane excitation of a cardiac cell to contraction of the human heart and systemic blood circulation and ultimately to autonomic nerve control. A lumped parameter model is used to compute the systemic and pulmonary circulations interacting with the cardiac cell mechanism. For autonomic control of the cardiovascular system, we used the approach suggested by Heldt et al. [2002. Computational modeling of cardiovascular response to orthostatic stress. J. Appl. Physiol. 92, 1239-1254] (Heldt model), including baroreflex and cardiopulmonary reflexes. We assumed sympathetic and parasympathetic pathways for the nerve control system. The cardiac muscle response to these reflex control systems was implemented using the activation-level changes in the L-type calcium channel and sarcoplasmic/endoplasmic reticulum calcium ATPase function based on experimental observations. Using this model, we delineated the cellular mechanism of heart contractility mediated by nerve control function. To verify the integrated method, we simulated a 10% hemorrhage, which involves cardiac cell mechanics, circulatory hemodynamics, and nerve control function. The computed and experimental results were compared. Using this methodology, the state of cardiac contractility, influenced by diverse properties such as the afterload and nerve control systems, is easily assessed in an integrated manner.

Simulation of spontaneous action potentials of cardiomyocytes in pulmonary veins of rabbits.

Atrial fibrillation is the most prevalent arrhythmia, but the mechanisms by which it develops are not clear. Recently, over 90% of paroxysmal atrial fibrillation was found to be located inside the main pulmonary veins (PVs). We found that single cardiac myocytes isolated from the main PVs of rabbits generate spontaneous action potentials (SAP). We therefore assayed the electrical characteristics of these cardiomyocytes. Among the diverse ionic currents identified were INa, ICa,L, IK1, IKr, IKs, Ito, IKsus, Incx, Ipump, IKH and ICl,Ca. In contrast, IK1 was minimal, IKs could be detected only in the presence of 10 microM forskolin, and we were unable to detect If and ICa,T, the most important currents for pacemaking activity in sinoatrial node cells. To identify the main cause of SAP, we developed a model that can explain the electrical properties of these cardiomyocytes. After reconstructing the ionic currents based on experimental observations, we were able to use our model to successfully reconstruct the characteristics of the SAP of PV cardiomyocytes. The simulation showed that the major currents contributing to pacemaking depolarization were ICaL, IKr, a background current and Na+-K+ pump current. Deactivation kinetics of IKr was one of the major determinants of the rate of pacemaking depolarization. The steady state inactivation of Ito was shifted to the negative voltage and the activity of Ito was minimal in the range of the SAP. The major currents for the repolarization were IKr and Ipump. The amplitude of most currents in these cardiac myocytes was small and no currents did not exceed 30 pA during the SAP, indicating that slight activation of other inward or outward currents will have profound effects on the SAP. To our knowledge, this report is the first to show the simulation of SAP of PV cardiomyocytes. This model may help to study on the electrophysiological basis of paroxysmal atrial fibrillation originating from PVs.