Free Calcium Research Articles

The influence of metal ions on aqueous dispersions of graphite oxide

ABSTRACT The residual metallic impurities in the graphite oxide (GO) dominate their properties and have a negative influence on the potential applications. The swelling of the GO suspensions is mainly affected by electrostatic forces and special interactions between water molecules, metal ions and hydrophilic groups. Bivalent and trivalent cations give rise to ionic crosslinking of the charged network within the particles. The CaCl2 solution has the best purification effect. A new purification method is proposed in the purification process, the origin metal ions in GO suspensions are replaced by homogeneous metal ions first, and GO suspensions are desalted by electrodialysis device. Free ionic calcium is removed by electrodialysis, and only the calcium that interact with GO particles is remained. The content of fixed calcium is 0.0049 g/g (nearly clear) after washing with 1.5 mmol/L CaCl2 solution and electrodialyzing for 20 min. The new method improves the purification efficiency and save energy.

Self-Assembly of Microstructured Protein Coatings with Programmable Functionality for Fluorescent Biosensors.

Proteins, as genetically programmable functional macromolecules, hold immense potential as biocompatible self-assembling building blocks, owing to their versatility in building coating materials and programming their functionality genetically. In this study, we demonstrate a modular self-assembly of protein coatings that are genetically programmable for a biosensor application. We designed and produced recombinant fusion protein building blocks to form microstructured coatings on diverse substrates, such as glass or polymers, through thermally triggered liquid-liquid phase separation and an orthogonal high-affinity coiled-coil interaction. We incorporated fluorescence proteins into coatings and controlled the protein density to enable fluorescence imaging and quantification in a low-resource setting. Then, we created a coating for a calcium biosensor using a genetically engineered calcium indicator protein. This protein coating served as the foundation for our smartphone-based fluorescent biosensor, which successfully measured free calcium concentrations in the millimolar range at which extracellular calcium homeostasis is maintained. Using this fluorescent biosensor, we were able to detect abnormal physiological conditions, such as mild or moderate hypercalcemia. We envision that this modular and genetically programmable functional protein coating platform could be extended to the development of highly accessible, low-cost fluorescent biosensors for a variety of targets.

Acidic Stress Induces Cytosolic Free Calcium Oscillation, and an Appropriate Low pH Helps Maintain the Circadian Clock in Arabidopsis.

Acidic stress is a formidable environmental factor that exerts adverse effects on plant growth and development, ultimately leading to a potential reduction in agricultural productivity. A low pH triggers Ca2+ influx across the plasma membrane (PM), eliciting distinct responses under various acidic pH levels. However, the underlying mechanisms by which Arabidopsis plant cells generate stimulus-specific Ca2+ signals in response to acidic stress remain largely unexplored. The experimentally induced stimulus may elicit spikes in cytosolic free Ca2+ concentration ([Ca2+]i) spikes or complex [Ca2+]i oscillations that persist for 20 min over a long-term of 24 h or even several days within the plant cytosol and chloroplast. This study investigated the increase in [Ca2+]i under a gradient of low pH stress ranging from pH 3.0 to 6.0. Notably, the peak of [Ca2+]i elevation was lower at pH 4.0 than at pH 3.0 during the initial 8 h, while other pH levels did not significantly increase [Ca2+]i compared to low acidic stress conditions. Lanthanum chloride (LaCl3) can effectively suppress the influx of [Ca2+]i from the apoplastic to the cytoplasm in plants under acid stress, with no discernible difference in intracellular calcium levels observed in Arabidopsis. Following 8 h of acid treatment in the darkness, the intracellular baseline Ca2+ levels in Arabidopsis were significantly elevated when exposed to low pH stress. A moderately low pH, specifically 4.0, may function as a spatial-temporal input into the circadian clock system. These findings suggest that acid stimulation can exert a continuous influence on intracellular calcium levels, as well as plant growth and development.

Protein disulfide isomerase 1 (PDIA1) regulates platelet-derived extracellular vesicle release

BackgroundProtein disulfide isomerase 1 (PDIA1) and 3 (PDIA3) regulate platelet activation and thrombus formation. However, their role in the formation of platelet-derived extracellular vesicles (pEVs) remains unknown. AimTo characterise the effects of PDIA1 and PDIA3 inhibition on pEV formation in washed murine platelets in response to platelet glycoprotein VI (GPVI) receptor or intracellular calcium signal activation. MethodsWashed platelets were isolated from C57BL/6 mice and activated using convulxin or the calcium ionophore A23187. Then, the resulting pEVs were analysed using nano flow cytometry (FC), platelet aggregation was measured by FC and a 96-well plate-based assay, and intracellular free calcium concentration ([Ca2+]i) was measured by indicator fluorescence. Platelet PDIs were blocked by a classic selective PDIA1 inhibitor (bepristat 2a) and sulphonamides of aziridine-2-carboxylic acid derivatives, novel PDI inhibitors relatively selective for PDIA1 or PDIA3 (C-3389 and C-3399, respectively). Clinically relevant antiplatelet drugs were used for comparison. ResultsConvulxin and A23187 concentration-dependently induced pEV formation. However, unlike convulxin, platelet activation by A23187 did not stimulate their aggregation. Bepristat 2a, C-3389 and C-3399 inhibited convulxin-induced pEV release accompanied by the reduction of [Ca2+]i. In contrast, only bepristat 2a inhibited A23187-induced pEV release, but without effect on [Ca2+]i. Cangrelor and tirofiban, but not acetylsalicylic acid (ASA), inhibited convulxin-induced pEV release, but neither of them inhibited A23187-induced pEV release. ConclusionThe inhibition of PDIA1 represents a novel approach to inhibit pEV formation by a mechanism independent of platelet aggregation and calcium signaling.

Exploring fungal potential for microbial-induced calcite precipitation (MICP) in bio-cement production

IntroductionMicrobial-induced calcite precipitation (MICP) involves various microorganisms, such as bacteria, fungi, and algae. This study focuses on producing bio-cement using fungal species and selecting potential candidates isolated from alkaline soil of different regions of Punjab, namely, Majha, Malwa, and Doaba.MethodsThe selection of fungi isolates capable of bio-cement production involves several tests, including a urease assay and calcium precipitation. Isolates having high urease enzyme production and the ability to perform calcite precipitation are selected for instrumental analyses such as X-ray diffraction (XRD) and scanning electron microscopy (SEM). The isolates selected for further analysis are S1 (3) with 8.879 ± 2.94 µg/ml, S1 (18) with 8.421 ± 0.13 µg/ml, and S4 (1) with 10.057 ± 0.45 µg/ml urease activity and least free calcium ions that are 2.337 ± 0.5 µg/ml, 3.339 ± 0.5 µg/ml, and 4.074 ± 0.1 µg/ml respectively.Results and discussionCalcite precipitation is confirmed through XRD and field emission scanning electron microscopy (FESEM). XRD images showing calcite precipitation with sharp crystalline peaks for S1 (3), S1 (18), and S4 (1) are shown. The calcite precipitation is evident in the micrographs of FESEM. These combined results confirm the potential of urease-positive fungi to facilitate calcite production, which could lead to bio-cement development in future research.

Mitochondrial-targeted plastoquinone therapy ameliorates early onset muscle weakness that precedes ovarian cancer cachexia in mice.

Cancer cachexia, and the related loss of muscle and strength, worsens quality of life and lowers overall survival. Recently, a novel 'pre-atrophy' muscle weakness was identified during early-stage cancer. While mitochondrial stress responses are associated with early-stage pre-atrophy weakness, a causal relationship has not been established. Using a robust mouse model of metastatic epithelial ovarian cancer (EOC)-induced cachexia, we found the well-established mitochondrial-targeted plastoquinone SkQ1 partially prevents pre-atrophy weakness in the diaphragm. Furthermore, SkQ1 improved force production during atrophy without preventing atrophy itself in the tibialis anterior and diaphragm. EOC reduced flexor digitorum brevis (FDB) force production and myoplasmic free calcium ([Ca 2+ ] i ) during contraction in single muscle fibers, both of which were prevented by SkQ1. Remarkably, changes in mitochondrial reactive oxygen species and pyruvate metabolism were heterogeneous across time and between muscle types which highlights a considerable complexity in the relationships between mitochondria and muscle remodeling throughout EOC. These discoveries identify that muscle weakness can occur independent of atrophy throughout EOC in a manner that is linked to improved calcium handling. The findings also demonstrate that mitochondrial-targeted therapies exert a robust effect in preserving muscle force during the early pre-atrophy period and in late-stage EOC once cachexia has become severe.

Nutrient Consumption Patterns of Saccharomyces cerevisiae and Their Application in Fruit Wine Fermentation

This study aimed to evaluate the nutritional requirements of Saccharomyces cerevisiae to improve low ethanol production in some fruit wines. The growth kinetics, ethanol production and nutrient requirements of S. cerevisiae were analyzed in chemically defined media. The results revealed that Ca2⁺, Fe2⁺, Co2⁺, Mo2⁺, Cu2⁺ and BO₃3− were predominantly utilized during the late lag phase, whereas free amino acids, nicotinic acid, calcium pantothenate, Na⁺ and Mg2⁺ were mainly consumed during the logarithmic phase. Compared with the control medium, supplementation with threonine, inositol, calcium pantothenate, thiamine hydrochloride, riboflavin, biotin, MgSO4 or KH2PO4 significantly increased the ethanol content by 1.10-fold (p < 0.05). Furthermore, adding key nutrients to noni-, guava- and mango juice significantly shortened the fermentation time and increased the final alcohol content of the fruit wines (p < 0.05). This study provides scientific insights and effective methods for shortening fermentation time and increasing alcohol content with S. cerevisiae in some fruit wines.

Inhibitory effects of chlorhexidine-loaded calcium carbonate nanoparticles against dental implant infections

This study aimed to design sustained released biodegradable calcium carbonate nanoparticles loaded with chlorhexidine (CHX-loaded NPs) and to investigate the early osteogenic differentiation and antimicrobial effects on the important bacteria involved in infections of dental implants. The microemulsion method was used to prepare the calcium carbonate nanoparticles loaded with chlorhexidine. The prepared nanoparticles were characterized using conventional methods. The release pattern determination and the biodegradation test were performed for the prepared nanoparticles. For the early osteogenic differentiation test of the prepared nanoparticles, alkaline phosphatase (ALP) activity was detected in human dental pulp stem cells (HDPSCs). The antimicrobial effects of the nanoparticles were evaluated against Escherichia coli, Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, and Pseudomonas aeruginosa. The sizes of free calcium carbonate nanoparticles and CHX-loaded NPs were 105 ± 1.63 and 118 ± 1.47 nm and their zeta potentials were − 27 and − 36, respectively. A 50% degradation of nanoparticles was achieved after 100 days. These nanoparticles showed a two-stage sustained release pattern in vitro. Microscopic images revealed that the morphology of free calcium carbonate nanoparticles primarily took on a spherical calcite form, while CHX-loaded NPs predominantly exhibited a cauliflower-like vaterite polymorph. The nanoparticles increased the activity of ALP in cells in two weeks significantly (p < 0.05). Antimicrobial and antibiofilm results showed an efficient effect of the prepared nanoparticle against the studied bacteria. Calcium carbonate nanoparticles are an efficient multifunctional vector for chlorhexidine and can be used as a bioactive antibacterial agent against various oral microorganisms to prevent implant infections.

Canonical Transient Receptor Potential (TRPC) Channels in Cardiovascular Pathology and their Modulators.

Ion channels play a crucial role in various aspects of cardiac function, such as regulating rhythm and contractility. As a result, they serve as key targets for therapeutic interventions in cardiovascular diseases. Cell function is substantially influenced by the concentration of free cytosolic calcium (Ca2+) and the voltage across the plasma membrane. These characteristics are known to be regulated by Ca2+-permeable non-selective cationic channels, although our knowledge of these channels is still inadequate. The transient receptor potential (TRP) superfamily comprises of many non-selective cation channels with diverse Ca2+ permeability. Canonical or classical TRP (TRPC) channels are a subgroup of the TRP superfamily that are expressed ubiquitously in mammalian cells. TRPC channels are multidimensional signalling protein complexes that play essential roles in a variety of physiological and pathological processes in humans, including cancer, neurological disorders, cardiovascular diseases, and others. The objective of this article is to focus on the role that TRPC channels play in the cardiovascular system. The role of TRPC channels will be deeply discussed in cardiovascular pathology. Together, a critical element in developing novel treatments that target TRPC channels is comprehending the molecular mechanisms and regulatory pathways of TRPC channels in related cardiovascular diseases and conditions.

Calcium Imaging in Brain Tissue Slices.

Calcium imaging is a method that was first developed in the mid-1970s yet kept developing until current days to allow accurate measurement of free calcium ions in tissues. This widely used method has provided significant advances to our understanding of cellular signal transduction, including the discovery of subcellular compartmentalization of neurons and astrocytes, the identification of multiple signaling pathways, and mapping the functional connectivity between astrocytes and neuronal networks. Here we describe a method for the loading and imaging of cell-permeable AM ester calcium-sensitive dyes for the in vitro measurement of free intracellular Ca2+ ions in acute brain slices.

A novel integrated soft sensing model for online cement clinker quality monitoring based on fuzzy fine-grained classification

The content of free calcium oxide (f-CaO) in cement clinker is an important index for cement quality. For the problems of multiple working conditions and unbalanced distribution of sample labels in cement clinker production, a soft sensing method of cement clinker f-CaO content based on fuzzy fine-grained classification (FF) is proposed. First, a divide-and-conquer strategy is used to divide the samples into high, medium, and low subsets according to cement clinker f-CaO and extract the fine-grained features under diverse types of multiple production conditions. Second, fuzzy classification based on the membership function is used in the FF model to solve the uncertainty of the sample categories. To ensure the rationality of the classification, the fuzzy membership rule is combined with a convolutional neural network to implement the fuzzy classification method. Finally, different feature extraction methods are proposed to be selected according to the data size of various categories of samples. After experimental validation, the evaluation metrics of RMSE decreased by 4.5 % and R2increased by 17.8 % for the direct classification model compared to the single model. The RMSE of the fuzzy classification model over the direct classification model was again reduced by 2.34 % and R2was again improved by 7.24 %, showing the effectiveness of the proposed soft measurement model.

Computational Fuzzy Modelling Approach to Analyze Neuronal Calcium Dynamics With Intracellular Fluxes.

Mathematical neuroscience investigates how calcium distribution in nerve cells affects the neurological system. The interaction of numerous systems is necessary for the operation of several cellular processes in neuron cells, such as calcium, buffer, ER etc. The dynamics of interacting parameters give useful information on neural cell function. This work uses a mathematical model to analyze the dynamic interactions of buffer and ER inside neurons, considering their spatial properties. While buffers bind to calcium ions and lower their concentration, the endoplasmic reticulum (ER) serves as a reservoir, holding a significant number of free calcium ions. The uncertainty of initial values of calcium concentration poses challenges for researchers to develop calcium signaling models. In this article, we examined the exact solution and approximate solution of the mathematical model that was analyzed using the fuzzy undetermined coefficient approach. MATLAB is being used to perform the simulation. Endoplasmic reticulum and buffer have been found to have a substantial impact on calcium signaling. Fuzzy differential equation Provides a useful tool for evaluating complicated processes with imprecise values when ordinary differential equations perform not precisely. They allow for the examination of dynamic processes under fuzzy settings, which contributes to advances research.

Analysis of changes in serum calcium and magnesium levels during plateletpheresis on different apheresis machines in healthy donors: A cross-sectional study

Abstract: BACKGROUND: The most common observed complication of plateletpheresis procedure is hypocalcemia due to citrate’s chelation of free calcium. Magnesium is also affected by citrate infusion. We have evaluated the impact of plateletpheresis on the total serum calcium (Ca++) and magnesium (Mg++) levels of the donor. MATERIALS AND METHODS: One hundred and thirty-seven healthy plateletpheresis donors were included in the study. Samples were collected from the donor at 0 min (baseline), 30 min, and 60 min, at the end of the procedure and 30 min after completion of the procedure. The pattern of changes in serum Ca++ and serum Mg++ levels at different time points for different apheresis machines (Fenwal Amicus, Fresenius COM.TEC, Spectra Optia, and Hemonetics MCS+) was studied. RESULTS: A significant decline in serum Ca++ and serum Mg++ levels from baseline to the end of the procedure was observed. This decline was observed maximum for Hemonetics MCS+. The baseline values were regained 30 min after completion of the procedure. Difference from baseline values 30 min after the procedure was lowest in amicus in the case of serum Ca++ and COM.TEC in case of serum Mg++. Twenty donors (14.60%) experienced adverse donor reactions. CONCLUSION: Serum calcium and serum magnesium levels decreased transiently during the procedure and regained normal values after 30 min of the completion of the plateletpheresis procedure on all four apheresis machines (whether based on continuous or intermittent centrifugation). Hence, it is concluded from our study that plateletpheresis is a well-tolerated and safe procedure.

Quantification of cytosolic 'free' calcium in isolated coral cells with confocal microscopy.

Despite its prominent role as an intracellular messenger in all organisms, cytosolic free calcium ([Ca2+]i) has never been quantified in corals or cnidarians in general. Ratiometric calcium dyes and cell imaging have been key methods in successful research on [Ca2+]i in model systems, and could be applied to corals. Here, we developed a procedure to quantify [Ca2+]i in isolated cells from the model coral species Stylophora pistillata using Indo-1 and confocal microscopy. We quantified [Ca2+]i in coral cells with and without intracellular dinoflagellate symbionts, and verified our procedure on cultured mammalian cells. We then used our procedure to measure changes in [Ca2+]i in coral cells exposed to a classic inhibitor of [Ca2+]i regulation, thapsigargin, and also used it to record elevations in [Ca2+]i in coral cells undergoing apoptosis. Our procedure paves the way for future studies into intracellular calcium in corals and other cnidarians.

Myogenic response of the cerebral vessel: a mathematical model

The myogenic response is most typical for small arterial vessels and consists in constriction of the vessel with an increase in transmural pressure, the pressure difference between the inner and outer surfaces of the vascular wall. There are many models describing this response. In models of last decades, in addition to macrostretchings and macrostrains, their microanalogues are introduced into consideration, that significantly complicates the description. In the presented work a simple model of the wall of the small cerebral artery is described, which includes the main parameters that control the myogenic response and corresponds to physiological processes underlining this response. It is believed that active stresses depend on stretching and the concentration of free calcium ions in the cytoplasm of the smooth muscle cell. In turn, the calcium content depends on the membrane potential, determined by the pressure. An axisymmetric stationary problem is considered; the wall material in the non-activated state is assumed to be pseudoelastic. Calculations showed a qualitative difference in the dependence of active stress on stretching and its distribution in the radial direction before and after the development of the vascular response. The calculated diameter values before and after exposure to substances affecting the polarization of the smooth muscle cell membrane are within the range of experimental values. These results indicate that the introduction of the dependence of calcium concentration on membrane potential expands the area of applicability of simple models of this kind for evaluating the value of vascular diameter.

Therapeutic potential of monoterpene molecules acts against 7KCh-mediated oxidative stress and neuroinflammatory amyloidogenic signalling pathways

Alzheimer’s disease (AD) is a degenerative disorder characterised by amyloid-beta aggregates activated by the accumulation of lipid molecules and their derivatives, especially 7-ketocholesterol (7KCh), an oxidised lipid that plays a great part in the progression of AD. The current therapeutics need bio-potential molecules and their biomedical application preventing 7KCh-induced cytotoxicity. In this study, bornyl acetate (BA) and menthol (ME), the natural monoterpenes were investigated for their neuroprotective effects against 7KCh-induced SH-SY5Y cells and their effects were compared to the standard drug galantamine (GA). 7KCh-induced changes like lipid accumulation, amyloid generation, free radical generation, acetylcholinesterase levels, calcium accumulation and mitochondrial membrane integrity were analysed in SH-SY5Y cells with or without BA and ME treatment. Furthermore, various mediators involved in the amyloidogenic, inflammatory and apoptotic pathways were studied. In our results, the cells induced with 7KCh upon co-treatment with BA and ME significantly reduced lipid accumulation and amyloid generation through toll-like receptor (TLR) 4 suppression and enhanced ATP binding cassette (ABCA) 1-mediated clearance. Co-treatment with BA and ME concurrently regulated oxidative stress, acetylcholinesterase activity, mitochondrial membrane potential and intracellular calcification altered by 7KCh-induced SH-SY5Y cells. Moreover, 7KCh-induced cells showed elevated mRNA levels of misfolded protein markers and apoptotic mediators which were significantly downregulated by BA and ME co-treatment. In addition, the protein expression of amyloidogenic, proinflammatory as well as pro-apoptotic markers was decreased by BA and ME co-treatment in 7KCh-induced cells. Overall, BA and ME mediated inhibition of amyloidogenic activation and cell survival against 7KCh-induced inflammation, thereby preventing the onset and progression of AD in comparison to GA.

Microbiome profile and nutritional benefits of traditional overnight soaked cooked rice

This study explores the microbial composition, nutrient content, and health benefits of "Panta Bhat," a traditional dish made from cooked rice soaked overnight, commonly consumed in Southeast Asia, particularly in Bangladesh and India. The research aims to evaluate its potential as an affordable food to enhance nutrition and combat malnutrition in population where rice is a staple. The findings reveal that soaking cooked rice overnight initiates mild fermentation, leading to the consistent growth of beneficial bacteria from the Bacillota phylum. 16S rRNA sequencing identified two Gram-positive bacteria, Leuconostoc lactis and Weissella confusa, known for their probiotic and lactic acid-producing capabilities. Furthermore, mineral analysis via ICP-MS showed significant increases in free iron, zinc, manganese, boron, calcium, magnesium, and potassium in soaked rice compared to unsoaked rice. Notably, there was a significant reduction in glycaemic response after consuming soaked rice (P < 0.01). This study suggests that overnight-soaked cooked rice is a promising source of probiotics and enhanced micronutrients, making it a valuable and cost-effective, low-glycaemic food option.

The effect of calcium stearate on the microbiological corrosion of cement stone concrete

To prevent biofouling of cement stone and its damage by fungal microorganisms, it is proposed to introduce 0,5 wt. % calcium stearate into the cement mixture. To ensure volumetric hydrophobization of cement stone, the additive is crushed to nanoparticles. The cement stone was cured in the air for 28 days. To study fungal corrosion, the surface of the cement stone was treated with a suspension of pores of Aspergillus niger fungi. The hydrophobic surface of the cement stone was not biofouled by fungal microorganisms Aspergillus niger during 6 months of the samples being in a humid environment, and black mold foci developed on the surface of ordinary cement stone during this period of time. The action of fungi and their waste products caused a decrease in the amount of calcium in the cement stone by 9 %, and had no effect on the hydrophobized cement stone. Due to the immunity of cement stone with a hydrophobizer to the action of microorganisms and water, free calcium hydroxide is not removed from the structure, but some amount is washed out of the surface layer and the pore liquid. A significant slowdown in mass transfer in cement stone under the action of liquids is provided by the hydrophobicity of the surface of cement stone and the walls of pores and capillaries, imparted by calcium stearate, as well as partial colmatation of the pore structure by means of the introduced additive.

Properties and hydration mechanism of slow-setting and expansive cement based on CFB ash and slag

Aiming at the problems of long construction time and easy shrinkage cracking of pavement base, the slow-setting and expansive cement (SEC) was prepared using circulating fluidized bed ash and slag (CFBA and CFBS), S95 ground granulated blast furnace slag (GGBFS), and silica fume (SF) as supplementary cementitious material in this paper. By adjusting the content of supplementary cementitious material, the ratio of ash and slag, the performance of SEC was evaluated by the indexes of water consumption of standard consistency, setting time, flexural and compressive strength. The hydration products of mortar samples and the influence mechanism were studied by means of XRD, TG-DTG, FTIR, and SEM-EDS. The results showed that when the SF content was 6 %, the compressive strength of the mortar specimens reached its maximum (35.19 MPa) at 28 days, which increased by 23.04 % compared to the control group. Meanwhile, the free calcium oxide and gypsum in CFBA reacted under alkaline conditions to form ettringite (AFt) and C-S-H gel, which caused the volume expansion of cement and led to a decrease in shrinkage within the cement system. SEC has higher mechanical properties and much smaller shrinkage cracks than ordinary cement. Therefore, replacing part of the cement by solid waste such as CFBS to prepare SEC to compensate for the shrinkage of the pavement base is an effective way to reduce pavement cracking.

Deciphering simplified regional anticoagulation with citrate in intermittent hemodialysis: a clinical and computational study

Regional citrate anticoagulation use in intermittent hemodialysis is limited by the increased risk of metabolic complications due to faster solute exchanges than with continuous renal replacement therapies. Several simplifications have been proposed. The objective of this study was to validate a mathematical model of hemodialysis anticoagulated with citrate that was then used to evaluate different prescription scenarios on anticoagulant effectiveness (free calcium concentration in dialysis filter) and calcium balance. A study was conducted in hemodialyzed patients with a citrate infusion into the arterial line and a 1.25 mmol/L calcium dialysate. Calcium and citrate concentrations were measured upstream and downstream of the citrate infusion site and in the venous line. The values measured in the venous lines were compared with those predicted by the model using Bland and Altman diagrams. The model was then used with 22 patients to make simulations. The model can predict the concentration of free calcium, bound to citrate or albumin, accurately. Irrespective of the prescription scenario a decrease in free calcium below 0.4 mmol/L was obtained only in a fraction of the dialysis filter. A zero or slightly negative calcium balance was observed, and should be taken into account in case of prolonged use.