Rate Of Ca2 Research Articles

Effects of exogenous calcium and calcium inhibitor on physiological characteristics of winter turnip rape (Brassica rapa) under low temperature stress

Low temperature is one of the environmental factors that restrict the growth and geographical distribution of Brassica. To investigate the effects of exogenous calcium and calcium inhibitor on the ability of winter turnip rapeseed (Brassica rapa L.) to withstand low temperatures (4℃), we used a strong cold-resistant variety Longyou 7 (L7) and a weak cold-resistant variety Longyou 99 (L99) as the materials. The seedlings were treated with CaCl2 (20 mmol·L-1) and calcium inhibitor LaCl3 (10 mmol·L-1) at 0 h (CK), 6 h, 12 h, 24 h and 48 h after 4℃ treatments. Physiological characteristics, Ca2+ flux and Ca2+ concentration in roots after treatments were analyzed. Results illustrated that under 4℃ treatment, activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) increased by both CK and exogenous CaCl2 treatments. Contents of soluble protein (SP) and proline (Pro) increased, while contents of malondialdehyde (MDA) decreased, resulting in reduced membrane lipid peroxidation. But enzyme activity decreased and MDA content increased following treatment with exogenous LaCl3. The rate of Ca2+ flow showed a higher uptake in L7 roots compared with L99. L99 showed Ca2+ efflux with a rate of 30.21 pmol‧cm-2‧s-1, whereas L7 showed short efflux then returned to influx. Calcium ion content in roots decreased in both cultivars after CaCl2 treatment. Results of RNA-seq revealed that genes were differentially expressed in response to low temperatures, hormones, photosystem II, chloroplasts, DNA replication, ribosomal RNA processing, and translation. This study found significant expression genes related to cellular signal transduction (MAPK signaling pathway) and material metabolism (nitrogen metabolism, glycerol ester metabolism).It was also analyzed by WGCNA that two modules had the strongest correlation with physiological indicators. Eight candidate genes were identified among MAPK signaling pathway and the two modules.

Restoring Atrial T-Tubules Augments Systolic Ca Upon Recovery From Heart Failure.

Transverse (t)-tubules drive the rapid and synchronous Ca2+ rise in cardiac myocytes. The virtual complete atrial t-tubule loss in heart failure (HF) decreases Ca2+ release. It is unknown if or how atrial t-tubules can be restored and how this affects systolic Ca2+. HF was induced in sheep by rapid ventricular pacing and recovered following termination of rapid pacing. Serial block-face scanning electron microscopy and confocal imaging were used to study t-tubule ultrastructure. Function was assessed using patch clamp, Ca2+, and confocal imaging. Candidate proteins involved in atrial t-tubule recovery were identified by western blot and expressed in rat neonatal ventricular myocytes to determine if they altered t-tubule structure. Atrial t-tubules were lost in HF but reappeared following recovery from HF. Recovered t-tubules were disordered, adopting distinct morphologies with increased t-tubule length and branching. T-tubule disorder was associated with mitochondrial disorder. Recovered t-tubules were functional, triggering Ca2+ release in the cell interior. Systolic Ca2+, ICa-L, sarcoplasmic reticulum Ca2+ content, and sarcoendoplasmic reticulum Ca2+ ATPase function were restored following recovery from HF. Confocal microscopy showed fragmentation of ryanodine receptor staining and movement away from the z-line in HF, which was reversed following recovery from HF. Acute detubulation, to remove recovered t-tubules, confirmed their key role in restoration of the systolic Ca2+ transient, the rate of Ca2+ removal, and the peak L-type Ca2+ current. The abundance of telethonin and myotubularin decreased during HF and increased during recovery. Transfection with these proteins altered the density and structure of tubules in neonatal myocytes. Myotubularin had a greater effect, increasing tubule length and branching, replicating that seen in the recovery atria. We show that recovery from HF restores atrial t-tubules, and this promotes recovery of ICa-L, sarcoplasmic reticulum Ca2+ content, and systolic Ca2+. We demonstrate an important role for myotubularin in t-tubule restoration. Our findings reveal a new and viable therapeutic strategy.

Host-Cell-Dependent Roles of E-Cadherin in Serratia Invasion.

Bacteria use cell surface proteins to mediate host-pathogen interactions. Proteins responsible for cell adhesion, including E-cadherin, serve as receptors for entry into the host cell. We have previously shown that an increase in eukaryotic cell sensitivity to Serratia grimesii correlates with an increase in E-cadherin expression. On the other hand, Serratia proteamaculans invasion involves the EGFR, which can interact with E-cadherin on the surface of host cells. Therefore, we investigated the role of E-cadherin in Serratia invasion into M-HeLa and Caco-2 cells. Bacterial infection increased E-cadherin expression in both cell lines. Moreover, E-cadherin was detected in the Caco-2 cells in a full-length form and in the M-HeLa cells in only a truncated form in response to incubation with bacteria. Transfection with siRNA targeting E-cadherin inhibited S. proteamaculans invasion only into the Caco-2 cells. Thus, only full-length E-cadherin is involved in S. proteamaculans invasion. On the other hand, transfection with siRNA targeting E-cadherin inhibited S. grimesii invasion into both cell lines. Thus, not only may full-length E-cadherin but also truncated E-cadherin be involved in S. grimesii invasion. Truncated E-cadherin can be formed as a result of cleavage by bacterial proteases or the Ca2+-activated cellular protease ADAM10. The rate of Ca2+ accumulation in the host cells depends on the number of bacteria per cell upon infection. During incubation, Ca2+ accumulates only when more than 500 S. grimesii bacteria are infected per eukaryotic cell, and only under these conditions does the ADAM10 inhibitor reduce the sensitivity of the cells to bacteria. An EGFR inhibitor has the same quantitative effect on S. grimesii invasion. Apparently, as a result of infection with S. grimesii, Ca2+ accumulates in the host cells and may activate the ADAM10 sheddase, which can promote invasion by cleaving E-cadherin and, as a result, triggering EGFR signaling. Thus, the invasion of S. proteamaculans can only be promoted by full-length E-cadherin, and S. grimesii invasion can be promoted by both full-length and truncated E-cadherin.

Positive Allosteric Modulators of SERCA Pump Restore Dendritic Spines and Rescue Long-Term Potentiation Defects in Alzheimer's Disease Mouse Model.

Alzheimer's disease (AD) is a neurodegenerative disorder that affects memory formation and storage processes. Dysregulated neuronal calcium (Ca2+) has been identified as one of the key pathogenic events in AD, and it has been suggested that pharmacological agents that stabilize Ca2+ neuronal signaling can act as disease-modifying agents in AD. In previous studies, we demonstrated that positive allosteric regulators (PAMs) of the sarco/endoplasmic reticulum Ca2+ ATPase (SERCA) pump might act as such Ca2+-stabilizing agents and exhibit neuroprotective properties. In the present study, we evaluated effects of a set of novel SERCA PAM agents on the rate of Ca2+ extraction from the cytoplasm of the HEK293T cell line, on morphometric parameters of dendritic spines of primary hippocampal neurons in normal conditions and in conditions of amyloid toxicity, and on long-term potentiation in slices derived from 5xFAD transgenic mice modeling AD. Several SERCA PAM compounds demonstrated neuroprotective properties, and the compound NDC-9009 showed the best results. The findings in this study support the hypothesis that the SERCA pump is a potential therapeutic target for AD treatment and that NDC-9009 is a promising lead molecule to be used in the development of disease-modifying agents for AD.

CALCIUM-ACCUMULATING ABILITY OF RAT LIVER MITOCHONDRIA DURING HYPOTHERMIA OF VARIOUS DURATIONS

Calcium is one of the most important intracellular messengers that regulate physiological and biochemical processes in the cell. Mitochondria are able to deposit calcium ions and are involved in the regulation of the calcium signal. Hypothermic conditions in homoiothermic animals can lead to disruption of this important function of mitochondria and pathological consequences. The aim of this study is to study the effects of moderate (30°C) hypothermia of varying duration on the calcium-accumulating capacity of rat liver mitochondria. The experiments were carried out on male rats Wistar. Hypothermia was induced by external cooling of the animals in Plexiglas chambers with a jacket through which cold water circulated. Mitochondria were isolated from the liver of decapitated rats by differential centrifugation. The calcium-accumulating capacity of mitochondria was assessed by the kinetics of calcium-induced swelling of mitochondria and their calcium capacity. A study of the kinetics of calcium-induced swelling of mitochondria showed that during short-term moderate (30°C) hypothermia, the rate of swelling of mitochondria decreases, prolongation of hypothermia to 1 h contributes to a further decrease in the swelling rate, and to 3 h, its normalization. A positive correlation was found between the rate of calcium-induced swelling and the calcium capacity of mitochondria (r = 0.79). Thus, with prolonged exposure to the cold factor in rats, a number of compensatory-adaptive reactions are activated. The decrease in the rate of Ca2+ – induced swelling and calcium capacity of mitochondria at the initial stages of hypothermia may be associated with the formation of mitochondrial pores and is reversible.

Brain vitamin D3-auto/paracrine system in relation to structural, neurophysiological, and behavioral disturbances associated with glucocorticoid-induced neurotoxicity.

Vitamin D3 (VD3) is a potent para/autocrine regulator and neurosteroid that can strongly influence nerve cell function and counteract the negative effects of glucocorticoid (GC) therapy. The aim of the study was to reveal the relationship between VD3 status and behavioral, structural-functional and molecular changes associated with GC-induced neurotoxicity. Female Wistar rats received synthetic GC prednisolone (5 mg/kg b.w.) with or without VD3 (1000 IU/kg b.w.) for 30 days. Behavioral, histological, physiological, biochemical, molecular biological (RT-PCR, Western blotting) methods, and ELISA were used. There was no difference in open field test (OFT), while forced swim test (FST) showed an increase in immobility time and a decrease in active behavior in prednisolone-treated rats, indicative of depressive changes. GC increased the perikaryon area, enlarged the size of the nuclei, and caused a slight reduction of cell density in CA1-CA3 hippocampal sections. We established a GC-induced decrease in the long-term potentiation (LTP) in CA1-CA3 hippocampal synapses, the amplitude of high K+-stimulated exocytosis, and the rate of Ca2+-dependent fusion of synaptic vesicles with synaptic plasma membranes. These changes were accompanied by an increase in nitration and poly(ADP)-ribosylation of cerebral proteins, suggesting the development of oxidative-nitrosative stress. Prednisolone upregulated the expression and phosphorylation of NF-κB p65 subunit at Ser311, whereas downregulating IκB. GC loading depleted the circulating pool of 25OHD3 in serum and CSF, elevated VDR mRNA and protein levels but had an inhibitory effect on CYP24A1 and VDBP expression. Vitamin D3 supplementation had an antidepressant-like effect, decreasing the immobility time and stimulating active behavior. VD3 caused a decrease in the size of the perikaryon and nucleus in CA1 hippocampal area. We found a recovery in depolarization-induced fusion of synaptic vesicles and long-term synaptic plasticity after VD3 treatment. VD3 diminished the intensity of oxidative-nitrosative stress, and suppressed the NF-κB activation. Its ameliorative effect on GC-induced neuroanatomical and behavioral abnormalities was accompanied by the 25OHD3 repletion and partial restoration of the VD3-auto/paracrine system. GC-induced neurotoxicity and behavioral disturbances are associated with increased oxidative-nitrosative stress and impairments of VD3 metabolism. Thus, VD3 can be effective in preventing structural and functional abnormalities in the brain and behavior changes caused by long-term GC administration.

Temperature effects on calcium partition kinetics in pasteurised skim milk during storage

Calcium (Ca) dynamics between casein micelles (CM) and serum phase (SP) in pasteurised skim milk were characterised by temperature dependence (4–35 °C) of Ca partition between phases and rate of formation of ionic calcium (Ca2+) in SP. Increase in Ca2+ during storage was characterised by two kinetic models: (1) 4–20 °C, with two parallel first order reactions, a fast dominating for first 3 h and a slower between 3 and 8 h, negative energies of activation (−22 kJ mol−1 and −48 kJ mol−1, respectively) explained by endothermic binding of Ca2+ to CM for the slow reaction and by endothermic Ca2+ binding to ligands in SP; (2) 4–35 °C, decrease in rate of Ca2+ formation with increasing temperature assigned to reversible endothermic association of serum Ca2+ to phosphate and β-casein. β-Casein release from CM decreased like Ca2+, suggesting interaction between β-casein and Ca2+ during transfer from CM to SP.

Plasma Membrane Ca2+ ATPase Activity Enables Sustained Store-operated Ca2+ Entry in the Absence of a Bulk Cytosolic Ca2+ Rise.

In many cell types, the rise in cytosolic Ca2+ due to opening of Ca2+ release-activated Ca2+ (CRAC) channels drives a plethora of responses, including secretion, motility, energy production, and gene expression. The amplitude and time course of the cytosolic Ca2+ rise is shaped by the rates of Ca2+ entry into and removal from the cytosol. However, an extended bulk Ca2+ rise is toxic to cells. Here, we show that the plasma membrane Ca2+ ATPase (PMCA) pump plays a major role in preventing a prolonged cytosolic Ca2+ signal following CRAC channel activation. Ca2+ entry through CRAC channels leads to a sustained sub-plasmalemmal Ca2+ rise but bulk Ca2+ is kept low by the activity of PMCA4b. Despite the low cytosolic Ca2+, membrane permeability to Ca2+ is still elevated and Ca2+ continues to enter through CRAC channels. Ca2+-dependent NFAT activation, driven by Ca2+ nanodomains near the open channels, is maintained despite the return of bulk Ca2+ to near pre-stimulation levels. Our data reveal a central role for PMCA4b in determining the pattern of a functional Ca2+ signal and in sharpening local Ca2+ gradients near CRAC channels, whilst protecting cells from a toxic Ca2+ overload.

Development of nanocomposite based on hydroxyapatite/hematite/graphene oxide for medical applications

Developing bioceramics-based nanomaterials for scaffolding is still a challenge. Three biocompatible materials are combined in one nanocomposite to fabricate antibacterial nanocomposite with higher mechanical properties. The nanocomposite contains hydroxyapatite (HAP), hematite (Fe2O3), and graphene oxide. The nanocomposite is examined chemically, structurally, and in vitro. The structural investigations showed a second minor phase with Fe2O3 which is FeO(OH). TEM investigated the shape morphology. There are two types of nanorods are detected which are small ones that belong to HAP, and long nanorods that belong to Fe2O3. Further, the nanosheets of GO are determined in addition to nano ellipsoids of FeOOH. The surface roughness is enhanced and Rq raised from 4.6 to 5.3 from pure HAP to HAP/Fe2O3/GO. The released ions rate of Fe3+ was constant for the first and second 12 h with a value of 2 ppm/h. The rate of Ca2+ decreased from 8 to 2.25 ppm/h for the first and second 12 h. Moreover, mechanical properties are enhanced and microhardness increased from 2.5 ± 0.1 to 3.1 ± 0.2 GPa. In addition, the shear modulus changed from 16.503 ± 1 to 20.45 ± 0.1 GPa. The antibacterial properties enhanced against Escherichia coli and Staphylococcus aureus and reached 16.5 ± 0.5 and 17.2 ± 0.7 mm without showing cytotoxicity toward the human osteoblast cells.

Trehalose and alginate oligosaccharides enhance the stability of myofibrillar proteins in shrimp ( Litopenaeus vannamei ) muscle during frozen storage

Saccharin with antifreeze effects can reduce protein loss in some aquatic products and effectively preserve taste and quality characteristics. However, the mechanisms responsible for stabilizing these types of food products are still unknown. Here, we use the trehalose fragment index to study the effects of trehalose and alginate oligosaccharides on frozen shrimp meat over 120 days. The stability of muscle protein in Litopenaeus vannamei was evaluated using the myofibrillar fragmentation index (MFI), differential scanning calorimetry (DSC), and SDS-PAGE. We found that the role of saccharin is markedly retarded by the decrease in the rate of Ca2+-ATPase activity, and that it effectively inhibited increases in the total sulfhydryl, active sulfhydryl, and carbonyl content, as well as changes in the water migration of myofibrillar protein. The SDS-PAGE results further confirmed that trehalose and alginate oligosaccharide treatments can effectively inhibit degradation of myofibrillar proteins. Our study aims to reduce the quality loss of aquatic products in cold chain logistics and provide a theoretical basis for controlling associated factors. Novelty impact statement The stability of muscle protein in Litopenaeus vannamei was evaluated using the myofibrillar fragmentation index (MFI), differential scanning calorimetry (DSC), and SDS-PAGE. We found that the role of saccharin is markedly retarded by the decrease in the rate of Ca2+-ATPase activity, and that it effectively inhibited increases in the total sulfhydryl, active sulfhydryl, and carbonyl content, as well as changes in the water migration of myofibrillar protein. The SDS-PAGE results further confirmed that trehalose and alginate oligosaccharide treatments can effectively inhibit degradation of myofibrillar proteins.

Spontaneous and action potential-evoked Ca2+ release from endoplasmic reticulum in neocortical synaptic boutons

Although the endoplasmic reticulum (ER) is present throughout axons, and IP3 and ryanodine receptors are widely expressed in nerve terminals, whether Ca2+ release from presynaptic stores contributes to action potential (AP)-evoked Ca2+ transients remains controversial. We investigated the release of Ca2+ from ER stores in boutons en passant of neocortical layer 5 pyramidal neurons. A hallmark of these stores is that they spontaneously release Ca2+ at a low frequency. Using a high-affinity Ca2+ indicator, we documented and characterised such spontaneous Ca2+ transients (sCaTs), which occurred at a rate of ~0.2 per min and raised the intracellular Ca2+ concentration ([Ca2+]i) by ~2 µM in the absence of exogenous buffers. Caffeine increased the average frequency of sCaTs by 90%, without affecting their amplitude and decay kinetics. Therefore, presynaptic ryanodine receptors were likely involved. To determine if presynaptic ER stores contribute to intracellular Ca2+ accumulation during repetitive stimulation, we measured [Ca2+]i during 2 s long trains of APs evoked at 10–50 Hz. We found that for frequencies <20 Hz, [Ca2+]i reached a steady state within ~500 ms after stimulation onset. However, for higher frequencies, [Ca2+]i continued to increase with AP number, suggesting that the rate of Ca2+ entry exceeded the rate of clearance. Comparison between measured and predicted values indicates supralinear summation of Ca2+. Block of the sarco/endoplasmic reticulum Ca2+-ATPase reduced the supralinearity of summation, without reducing the amplitude of a single AP-evoked Ca2+ transient. Together, our results implicate presynaptic ER stores as a source of Ca2+ during repetitive stimulation.

Long-Term Tracking and Dynamically Quantifying of Reversible Changes of Extracellular Ca2+ in Multiple Brain Regions of Freely Moving Animals.

Understanding physiological and pathological processes in the brain requires tracking the reversible changes in chemical signals with long-term stability. We developed a new anti-biofouling microfiber array to real-time quantify extracellular Ca2+ concentrations together with neuron activity across many regions in the mammalian brain for 60 days, in which the signal degradation was < ca. 8 %. The microarray with high tempo-spatial resolution (ca. 10 μm, ca. 1.3 s) was implanted into 7 brain regions of free-moving mice to monitor reversible changes of extracellular Ca2+ upon ischemia-reperfusion processes. The changing sequence and rate of Ca2+ in 7 brain regions were different during the stroke. ROS scavenger could protect Ca2+ influx and neuronal activity after stroke, suggesting the significant influence of ROS on Ca2+ overload and neuron death. We demonstrated this microarray is a versatile tool for investigating brain dynamic during pathological processes and drug treatment.

Long‐Term Tracking and Dynamically Quantifying of Reversible Changes of Extracellular Ca2+ in Multiple Brain Regions of Freely Moving Animals

AbstractUnderstanding physiological and pathological processes in the brain requires tracking the reversible changes in chemical signals with long‐term stability. We developed a new anti‐biofouling microfiber array to real‐time quantify extracellular Ca2+ concentrations together with neuron activity across many regions in the mammalian brain for 60 days, in which the signal degradation was &lt; ca. 8 %. The microarray with high tempo‐spatial resolution (ca. 10 μm, ca. 1.3 s) was implanted into 7 brain regions of free‐moving mice to monitor reversible changes of extracellular Ca2+ upon ischemia‐reperfusion processes. The changing sequence and rate of Ca2+ in 7 brain regions were different during the stroke. ROS scavenger could protect Ca2+ influx and neuronal activity after stroke, suggesting the significant influence of ROS on Ca2+ overload and neuron death. We demonstrated this microarray is a versatile tool for investigating brain dynamic during pathological processes and drug treatment.

Calcium is reduced in presynaptic mitochondria of motor nerve terminals during neurotransmission in SMA mice.

Spinal muscular atrophy (SMA) is an autosomal recessive degenerative motor neuron disease characterized by symmetrical muscle weakness and atrophy of limb and trunk muscles being the most severe genetic disease in children. In SMA mouse models, motor nerve terminals display neurotransmitter release reduction, endocytosis decrease and mitochondria alterations. The relationship between these changes is, however, not well understood. In the present study, we investigated whether the endocytosis impairment could be related to the functional alteration of the presynaptic mitochondria during action potential (AP) firing. To this aim, we generated a Synaptophysin-pHluorin (SypHy) transgenic mouse, crossed it with Taiwanese SMA mice, and recorded exo- and endocytosis and mitochondria Ca2+ signaling in real-time at ex vivo motor nerve terminals of Taiwanese-SypHy mice. The experiments were performed at the beginning of the motor symptoms to get an integrated view of the nerve terminal’s functional state before degeneration. Our electrophysiological and live imaging results demonstrated that the mitochondria’s capacity to increase matrix-free Ca2+ in SMA mice was significantly limited during nerve AP firing, except when the rate of Ca2+ entry to the cytosol was considerably reduced. These results indicate that both the mitochondrial Ca2+ signaling alterations and the secretion machinery defects are significant players in the dysfunction of the presynaptic terminal in SMA.

D,l-Methadone causes leukemic cell apoptosis via an OPRM1-triggered increase in IP3R-mediated ER Ca2+ release and decrease in Ca2+ efflux, elevating [Ca2+

The search continues for improved therapy for acute lymphoblastic leukemia (aLL), the most common malignancy in children. Recently, d,l-methadone was put forth as sensitizer for aLL chemotherapy. However, the specific target of d,l-methadone in leukemic cells and the mechanism by which it induces leukemic cell apoptosis remain to be defined. Here, we demonstrate that d,l-methadone induces leukemic cell apoptosis through activation of the mu1 subtype of opioid receptors (OPRM1). d,l-Methadone evokes IP3R-mediated ER Ca2+ release that is inhibited by OPRM1 loss. In addition, the rate of Ca2+ extrusion following d,l-methadone treatment is reduced, but is accelerated by loss of OPRM1. These d,l-methadone effects cause a lethal rise in [Ca2+]i that is again inhibited by OPRM1 loss, which then prevents d,l-methadone-induced apoptosis that is associated with activation of calpain-1, truncation of Bid, cytochrome C release, and proteolysis of caspase-3/12. Chelating intracellular Ca2+ with BAPTA-AM reverses d,l-methadone-induced apoptosis, establishing a link between the rise in [Ca2+]i and d,l-methadone-induced apoptosis. Altogether, our findings point to OPRM1 as a specific target of d,l-methadone in leukemic cells, and that OPRM1 activation by d,l-methadone disrupts IP3R-mediated ER Ca2+ release and rate of Ca2+ efflux, causing a rise in [Ca2+]i that upregulates the calpain-1-Bid-cytochrome C-caspase-3/12 apoptotic pathway.

Coupled reaction and solvent extraction process to utilize distiller waste using N235-isoamylol system

ABSTRACT Distiller waste produced from the Solvay Process, which mainly contains calcium chloride (CaCl2), is being discharged and has not yet been fully recycled. In this study, a coupled reaction and solvent extraction process was adopted to utilize distiller waste using the mixed organic solvent composed of N235 and isoamylol. High purity and good crystallinity calcite with an average of 40~50 μm were prepared in this process. The effect of operation conditions was firstly studied and the conversion rate of CaCl2 reached 83.40% in single-stage with the optimized conditions: reaction time of 3 h, N235/isoamylol volume ratio of 1, reaction temperature of 25°C, N235/CaCl2 molar ratio of 3, and the initial CaCl2 concentration of 1 mol·L−1. The enthalpy change of the overall reaction was −89.93 kJ·mol−1. Then, the phase equilibrium was measured and the theoretical extraction stage was predicted to be 3 with the target conversion rate of 90%. Three-stage counter-current extraction process showed that about 93.75% CaCl2 was mineralized into CaCO3 and the results had a good agreement with the McCabe-Thiele calculation. This indicated that multi-stage counter-current extraction can effectively improve the conversion rate of Ca2+ and related result was of great significance in treating distiller waste industrially.

Structural and Functional Features of Ca2+ Transport Systems in Liver Mitochondria of Rats with Experimental Hyperthyroidism.

We analyzed structural and functional features of the main mitochondrial Ca2+-transporting systems, mitochondrial Ca2+ uniporter complex (MCUC) and Ca2+-dependent cyclosporin A-sensitive mitochondrial permeability transition pore (MPT pore), in rats with hyperthyroid state. It was found that, the rate of Ca2+ accumulation by rat liver mitochondria in this pathology increases by 1.3 times, which can be associated with higher level of the channel-forming subunit of the uniporter MCU and lower content of dominant-negative subunit of this complex MCUb. At the same time, the level of the regulatory subunit MICU1 remained unchanged. It was shown that calcium retention capacity of liver mitochondria in rats with experimental hyperthyroidism decreased by 2 times in comparison with the control, which attested to reduced resistance of liver mitochondria of hyperthyroid rats to induction of the MPT pore. The observed changes are consistent with the data on increased amount of cyclophilin D, a mitochondrial matrix peptidyl-prolyl isomerase that is known to modulate the MPT pore opening and expression of the Ppif gene that encodes mitochondrial cyclophilin D in rats with experimental hyperthyroidism.

Effects of prolactin on ventricular myocyte shortening and calcium transport in the streptozotocin-induced diabetic rat

The physiological role of prolactin (PRL) in the heart, and in particular the diabetic heart, are largely unknown. The effects of PRL on ventricular myocyte shortening and Ca2+ transport in the streptozotocin (STZ) – induced diabetic and in age-matched control rats were investigated. PRL receptor protein, myocyte shortening, intracellular [Ca2+], L-type Ca2+ current were measured by Western blot, cell imaging, fluorescence photometry and whole-cell patch-clamp techniques, respectively. Compared to normal Tyrode solution (NT), PRL (50 ng/ml) significantly (p < 0.05) increased the amplitude of shortening in myocytes from control (7.43 ± 0.38 vs. 9.68 ± 0.46 %) and diabetic (6.57 ± 0.24 vs. 8.91 ± 0.44 %) heart (n = 44–49 cells). Compared to NT, PRL (50 ng/ml) significantly increased the amplitude of Ca2+ transients in myocytes from control (0.084 ± 0.004 vs. 0.115 ± 0.007 Fura-2 ratio units) and diabetic (0.087 ± 0.007 vs. 0.112 ± 0.006 Fura-2 ratio units) heart (n = 36–50 cells). PRL did not significantly alter the amplitude of caffeine-evoked Ca2+ transients however, PRL significantly increased the fractional release of Ca2+ in myocytes from control (21 %) and diabetic (14 %) and heart. The rate of Ca2+ transient recovery following PRL treatment was significantly increased in myocytes from diabetic and control heart. Amplitude of L-type Ca2+ current was not significantly altered by diabetes or by PRL. PRL increased the amplitude of shortening and Ca2+ transients in myocytes from control and diabetic heart. Increased fractional release of sarcoplasmic reticulum Ca2+ may partly underlie the positive inotropic effects of PRL in ventricular myocytes from control and STZ-induced diabetic rat.

Transport of Ca2+ and Ca2+-Dependent Permeability Transition in the Liver and Heart Mitochondria of Rats with Different Tolerance to Acute Hypoxia.

The work examines the kinetic parameters of Ca2+ uptake via the mitochondrial calcium uniporter complex (MCUC) and the opening of the Ca2+-dependent permeability transition pore (MPT pore) in the liver and heart mitochondria of rats with high resistance (HR) and low resistance (LR) to acute hypoxia. We found that the rate of Ca2+ uptake by mitochondria of the liver and heart in HR rats is higher than that in LR rats, which is associated with a higher level of the channel-forming subunit MCU in liver mitochondria of HR rats and a lower content of the dominant-negative channel subunit MCUb in heart mitochondria of HR rats. It was shown that the liver mitochondria of HR rats are more resistant to the induction of the MPT pore than those of LR rats (the calcium retention capacity of liver mitochondria of HR rats was found to be 1.3 times greater than that of LR rats). These data correlate with the fact that the level of F0F1-ATP synthase, a possible structural element of the MPT pore, in the liver mitochondria of HR rats is lower than in LR rats. In heart mitochondria of rats of the two phenotypes, no statistically significant difference in the formation of the MPT pore was revealed. The paper discusses how changes in the expression of the MCUC subunits and the putative components of the MPT pore can affect Ca2+ homeostasis of mitochondria in animals with originally different tolerance to hypoxia and in hypoxia-induced tissue injury.

The Physicomechanical Deterioration Characteristics and Mesoscopic Damage Analysis of Sandstone under Acidic Environment

The physicomechanical deterioration characteristics of sandstone subjected to H2SO4, HCl, and H2O solutions of different pH values are studied by the method of long‐term accelerated immersion. The quantitative relationships between the damage variables based on CT (computer tomographic identification technology) numbers and the immersion time, the uniaxial compressive strength, the peak point strain, and the elastic modulus of rock samples are analyzed. The test results indicate that the pH value of immersion solutions, the dissolution rate of Ca2+ and Na+, and the quality change of rock samples show visible stage characteristics under acidic environment. With the soaking time extended, the pH value of solutions increases gradually, and the quality change of rock samples decreases gradually. The smaller the pH value of immersion solutions is, the higher the dissolution rate of Ca2+ and Na+ is. However, the cation dissolution rate under a weak acid environment with a high pH value has little difference with that under the distilled water (pH = 7). With the increase of the soaking time and the acidity, the compaction stage of rock samples becomes longer, the elastic stage becomes shorter, the deterioration degree of mechanical parameters becomes more extensive, and the destruction of sandstone samples shows ductility characteristics increasingly. The corrosion degree of corroded sandstone samples is quantitatively represented by microscopic damage variables based on CT numbers. The regression analysis results show that damage variables of acid‐corroded sandstone samples have a power function relationship with soaking time and an exponential function relationship with peak strength, peak point strain, and elastic modulus.