Increase In Calcium Concentration Research Articles

RANKL, but Not R-Spondins, Is Involved in Vascular Smooth Muscle Cell Calcification through LGR4 Interaction

Background: Vascular calcification has a global health impact that is closely linked to bone loss. The Receptor Activator of Nuclear Factor Kappa B (RANK)/RANK ligand (RANKL)/osteoprotegerin (OPG) system, fundamental for bone metabolism, also plays an important role in vascular calcification. The Leucine-rich repeat-containing G-protein-coupled receptor 4 (LGR4), a novel receptor for RANKL, regulates bone remodeling, and it appears to be involved in vascular calcification. Besides RANKL, LGR4 interacts with R-spondins (RSPOs), which are known for their roles in bone but are less understood in vascular calcification. Methods: Studies were conducted in rats with chronic renal failure fed normal or high phosphorus diets for 18 weeks, with and without control of circulating parathormone (PTH) levels, resulting in different degrees of aortic calcification. Additionally, vascular smooth muscle cells (VSMCs) were cultured under non-calcifying (1 mM phosphate) and calcifying (3 mM phosphate) media with different concentrations of PTH. To explore the role of RANKL in VSMC calcification, increasing concentrations of soluble RANKL were added to non-calcifying and calcifying media. The effects mediated by RANKL binding to its receptor LGR4 were investigated by silencing the LGR4 receptor in VSMCs. Furthermore, the gene expression of the RANK/RANKL/OPG system and the ligands of LGR4 was assessed in human epigastric arteries obtained from kidney transplant recipients with calcification scores (Kauppila Index). Results: Increased aortic calcium in rats coincided with elevated systolic blood pressure, upregulated Lgr4 and Rankl gene expression, downregulated Opg gene expression, and higher serum RANKL/OPG ratio without changes in Rspos gene expression. Elevated phosphate in vitro increased calcium content and expression of Rankl and Lgr4 while reducing Opg. Elevated PTH in the presence of high phosphate exacerbated the increase in calcium content. No changes in Rspos were observed under the conditions employed. The addition of soluble RANKL to VSMCs induced genotypic differentiation and calcification, partly prevented by LGR4 silencing. In the epigastric arteries of individuals presenting vascular calcification, the gene expression of RANKL was higher. Conclusions: while RSPOs show minimal impact on VSMC calcification, RANKL, interacting with LGR4, drives osteogenic differentiation in VSMCs, unveiling a novel mechanism beyond RANKL-RANK binding.

The Potential of QP3VH-Chitosan Peptide as Biomimetic Remineralization in Early Dental Caries Treatment: An In Vitro Study.

The development of remineralization biomimetics using organic peptide molecules is expected to resemble the hydroxyapatite (HA) mineralization process in tooth enamel. The development of an amelogenin derivative peptide combined with antimicrobial peptide was designed, resulting in QP3VH. This combination then was mixed with chitosan as a carrier. This study aimed to evaluate the biomimetic efficacy of QP3VH as a remineralizing agent combined with chitosan. Fifty deciduous mandibular incisor enamel samples were used in this study. The artificial enamel lesions were created on a buccal surface and were randomly assigned to five groups of 10 each according to the remineralizing agent used: QP3VH, NaF, QP3VH + NaF, QP3VH + CS (QP3VH + chitosan), and saline distilled water (SDW). Each group was performed pH cycling for seven days. Enamel surface morphology and evaluation of mineral content Ca/P (calcium and phosphate) using scanning electron microscopy and energy dispersive X-ray analysis. The assessment was carried out, after demineralization, and after application with remineralization agents. Data were analyzed using a one-way analysis of variance followed by least significance difference post-hoc test. The paired t-test was utilized to compare the demineralization and remineralization results. The significance level used was 95%. The remineralized group exhibited a significant increase in calcium and phosphate content on the enamel surface (p <0.05), and QP3VH + CS produced the maximum Ca/P mass percent after remineralization. Combining QP3VH with chitosan produces greatest remineralization than QP3VH, QP3VH + NaF, Naf, and SDW; therefore, QP3VH peptide has potential as a remineralizing agent, in the future.

Hydric environment and chemical composition shape non-avian reptile eggshell absorption.

The amniotic egg fulfils a critical role in reproduction by serving as an interface between the external environment and the embryo. Because non-avian reptiles are rarely incubated, they must be heated by, and absorb water from, the oviposition site for the developing embryo. The mechanisms by which they absorb sufficient, but not excess, water and how these mechanisms vary with local habitat is largely unknown, despite its significance to their evolution. Here, we first performed histology, Fourier transform infra-red spectroscopy and dynamic vapor sorption experiments to elucidate the mechanisms of eggshell absorption for 56 reptile species. Then, we used phylogenetic comparative analyse to test the hypothesis that absorptive capacity of reptile eggshells increases with aridity of the environment. We found that water absorption increases in the presence of a superficial mucopolysaccharide layer and decreases with increased calcium content. We found that eggs from arid environments have highly absorbent eggshells, but only in species with weakly calcified shells. Our results suggest that reptile eggshells have over evolutionary time tuned absorptive capacity to environmental moisture level. Since these eggs often must sustain conflicting constraints, they may serve as inspirations for new biomimetic materials, such as water filtering membranes or humidity sensors.

Molecular insight into the structural and mechanical properties of Ca-based geopolymers

Ca-based geopolymers are a principal product of introducing calcium into geopolymers. Analyzing Ca-based geopolymers structural and mechanical properties aids in optimizing mix design of geopolymers. In this paper, partially reacted low-polymerization and fully reacted high-polymerization models were constructed to examine the structural and mechanical properties of Ca-based geopolymers with varying calcium contents via molecular dynamics simulations. Results show that Ca-based geopolymers contain pentacoordinate aluminum and tricluster oxygen which is more brittle and makes the structure loose. In low-polymerization models, an increase in calcium content accelerates the polymerization rate by overcoming the low-temperature reaction barrier. However, calcium content hinders the formation of pentacoordinate aluminum, decreasing the final degree of polymerization, resulting in weakened mechanical properties. In high-polymerization models, increased calcium decreases tricluster oxygen content, resulting in improved mechanical properties. While the hydrolysis reaction reduces mechanical properties, the weakening effect of water is greater than the enhancing effect of calcium.

The Effect of Calcium Ions on the Electrophysiological Properties of Single ANO6 Channels.

Proteins belonging to the anoctamin (ANO) family form calcium-activated chloride channels (CaCCs). The most unusual member of this family, ANO6 (TMEM16F), simultaneously exhibits the functions of calcium-dependent scramblase and the ion channel. ANO6 affects the plasma membrane dynamics and phosphatidylserine transport; it is also involved in programmed cell death. The properties of ANO6 channels remain the subject of debate. In this study, we investigated the effect of variations in the intracellular and extracellular concentrations of calcium ions on the electrophysiological properties of endogenous ANO6 channels by recording single ANO6 channels. It has been demonstrated that (1) a high calcium concentration in an extracellular solution increases the activity of endogenous ANO6 channels, (2) the permeability of endogenous ANO6 channels for chloride ions is independent of the extracellular concentration of calcium ions, (3) that an increase in the intracellular calcium concentration leads to the activation of endogenous ANO6 channels with double amplitude, and (4) that the kinetics of the channel depend on the plasma membrane potential rather than the intracellular concentration of calcium ions. Our findings give grounds for proposing new mechanisms for the regulation of the ANO6 channel activity by calcium ions both at the inner and outer sides of the membrane.

Modulation by Astrocytes of AMPA Currents in the NTS Neurons of Mice Is Reduced by Short Term-Sustained Hypoxia

Hypoxia due to reduction in the partial pressure of oxygen in the arterial blood (PaO2) activates the afferents of peripheral chemoreflex to produce cardiovascular and respiratory adjustments to bring PaO2 back to the normal range. Herein we evaluated the possible changes of astrocytic modulation on glutamatergic transmission in NTS neurons of mice submitted to short term-sustained hypoxia (SH). To reach this goal, we used Male C57Bl/6J mice (18-20g), which were submitted to SH (FiO2 0.1) or normoxia for 24 hours. Brainstem slices and whole-cell patch clamp technique were used to evaluate the synaptic transmission in NTS neurons. The intracellular calcium concentration in NTS astrocytes was evaluated by multiphoton microscopy and transgenic Aldh-CRE-GCAMP6f mice (Institutional Ethical Committee Approvals: #016/2018 and #1109/2022R1). SH increased the amplitude of TS-evoked AMPA currents [control: -72.8 ± 13.3 pA ( n = 12) vs SH: -120 ± 14.4 pA ( n = 21)] in NTS neurons with no changes on the rise- or decay-time. SH also increased the amplitude [control: 35.88 ± 7.9 pA ( n = 12) vs SH: 72.74 ± 15.47 pA ( n = 10)] and decay time [control: 61.8 ± 15.2 ms ( n = 9) vs SH: 286 ± 86.7 ms ( n = 10)] of TS evoked-NMDA currents in neurons, while the amplitude of extrasynaptic NMDA currents [control: 28.7 ± 7.4 pA ( n = 10) vs SH: 24.7 ± 3.2 pA ( n = 7)] was not affected. Perfusion with fluoracetate (FAC), a glial metabolism inhibitor, decreased the amplitude of TS-evoked AMPA currents in control group [aCSF: -54.29 ± 16.1 pA vs aCSF + FAC: -24.43 ± 6.82 pA ( n =9)] without affecting AMPA currents in SH neurons [aCSF: -124.5 ± 35.27 pA vs aCSF + FAC: -130.5 ± 50.16 pA ( n =8)]. The passive properties of astrocytes did not change after SH [RMP: -83.73 ± 11.85 mV ( n =10) vs −83.63 ± 11.86 mV ( n =10); Rinput: 123 ± 127.5 MΩ ( n =9) vs 208 ± 212.7 MΩ ( n =10)]. The increase in intracellular calcium concentration induced by AMPA in the astrocytes of control and SH mice was similar [Control: 112.8 ± 15.41 ΔF/F0 (n=68 ROIs) vs SH: 56.38 ± 18.31 ΔF/F0 (n=13 ROIs)]. The data shows that SH increased the evoked glutamatergic currents in NTS neurons of mice, probably due to a reduction in the astrocytic modulation of excitatory transmission by mechanisms apparently independent of astrocytic intracellular calcium. FAPESP (2018/15957-2, 2018/02336-0 and 2022/05237-8), CNPq (309338/2020-4) and CAPES-PROEX (23038.006922/2021-31). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Molecular recording of calcium signals via calcium-dependent proximity labeling.

Calcium ions serve as key intracellular signals. Local, transient increases in calcium concentrations can activate calcium sensor proteins that in turn trigger downstream effectors. In neurons, calcium transients play a central role in regulating neurotransmitter release and synaptic plasticity. However, it is challenging to capture the molecular events associated with these localized and ephemeral calcium signals. Here we present an engineered biotin ligase that generates permanent molecular traces in a calcium-dependent manner. The enzyme, calcium-dependent BioID (Cal-ID), biotinylates nearby proteins within minutes in response to elevated local calcium levels. The biotinylated proteins can be identified via mass spectrometry and visualized using microscopy. In neurons, Cal-ID labeling is triggered by neuronal activity, leading to prominent protein biotinylation that enables transcription-independent activity labeling in the brain. In summary, Cal-ID produces a biochemical record of calcium signals and neuronal activity with high spatial resolution and molecular specificity.

Mechanisms driving fruit cracking in ‘Sinhwa’ pears (Pyrus pyrifolia Nakai) and effect of foliar fertilizer application on fruit quality

Fruit cracking poses a significant challenge to ‘Sinhwa’ pear cultivation and has potential economic repercussions for growers due to its adverse effects on yield and fruit quality. This study delves into the phenomenon of fruit cracking in ‘Sinhwa’ pears by conducting a comprehensive analysis of its causes and exploring the efficacy of foliar fertilization as a mitigation strategy. To reveal the conditions contributing to fruit cracking in ‘Sinhwa’ pears, moisture change regimes were systematically applied to 5-year-old ‘Sinhwa' trees from 2018 to 2020 during post-flowering and harvest periods. Alternating wet and dry conditions resulted in the highest fruit cracking rates, thus highlighting the importance of consistent soil moisture levels for mitigating cracking. Furthermore, the timing of fruit cracking and its histological characteristics in ‘Sinhwa’ pears were examined to understand the underlying causes. Microcracks appeared approximately 60 days after full bloom and preceded visible cracking, although notable changes in epidermis thickness and stone cell layer distribution were observed. Analysis of the nutrient content revealed differences in calcium and potassium levels between intact and cracked fruits, thus providing insights into potential nutritional influences on fruit quality. Potential mitigation strategies included foliar application of 0.3 % calcium chloride and 0.5 % potassium nitrate, and they demonstrated promising results in reducing fruit cracking rates compared to the control. Fruit characteristics, including mass and hardness, varied among the treatment groups, with treated trees showing notable increases in calcium content, which were correlated with decreases in fruit cracking incidence. To understand the molecular mechanisms underlying the effectiveness of foliar fertilization, gene expression levels associated with fruit cracking were analyzed by real-time PCR. Higher expression of heat-related expansion genes (Exp1 and Exp2) was observed in the treated fruits, particularly those in the calcium chloride and potassium nitrate treatments, suggesting the role of these elements in mitigating fruit cracking. This study provides valuable insights into the causes of fruit cracking in ‘Sinhwa’ pears and offers practical mitigation strategies, such as foliar fertilization. Moreover, the findings underscore the importance of maintaining consistent soil moisture levels and optimizing nutrient management practices to enhance fruit quality and minimize economic losses in ‘Sinwha’ pear cultivation.

Elevated intracellular Ca2+ functions downstream of mitodysfunction to induce Wallerian-like degeneration and necroptosis in organophosphorus-induced delayed neuropathy

Neurotoxic organophosphorus compounds can induce a type of delayed neuropathy in humans and sensitive animals, known as organophosphorus-induced delayed neuropathy (OPIDN). OPIDN is characterized by axonal degeneration akin to Wallerian-like degeneration, which is thought to be caused by increased intra-axonal Ca2+ concentrations. This study was designed to investigate that deregulated cytosolic Ca2+ may function downstream of mitodysfunction in activating Wallerian-like degeneration and necroptosis in OPIDN. Adult hens were administrated a single dosage of 750 mg/kg tri-ortho-cresyl phosphate (TOCP), and then sacrificed at 1 day, 5 day, 10 day and 21 day post-exposure, respectively. Sciatic nerves and spinal cords were examined for pathological changes and proteins expression related to Wallerian-like degeneration and necroptosis. In vitro experiments using differentiated neuro-2a (N2a) cells were conducted to investigate the relationship among mitochondrial dysfunction, Ca2+ influx, axonal degeneration, and necroptosis. The cells were co-administered with the Ca2+-chelator BAPTA-AM, the TRPA1 channel inhibitor HC030031, the RIPK1 inhibitor Necrostatin-1, and the mitochondrial-targeted antioxidant MitoQ along with TOCP. Results demonstrated an increase in cytosolic calcium concentration and key proteins associated with Wallerian degeneration and necroptosis in both in vivo and in vitro models after TOCP exposure. Moreover, co-administration with BATPA-AM or HC030031 significantly attenuated the loss of NMNAT2 and STMN2 in N2a cells, as well as the upregulation of SARM1, RIPK1 and p-MLKL. In contrast, Necrostatin-1 treatment only inhibited the TOCP-induced elevation of p-MLKL. Notably, pharmacological protection of mitochondrial function with MitoQ effectively alleviated the increase in intracellular Ca2+ following TOCP and mitigated axonal degeneration and necroptosis in N2a cells, supporting mitochondrial dysfunction as an upstream event of the intracellular Ca2+ imbalance and neuronal damage in OPIDN. These findings suggest that mitochondrial dysfunction post-TOCP intoxication leads to an elevated intracellular Ca2+ concentration, which plays a pivotal role in the initiation and development of OPIDN through inducing SARM1-mediated axonal degeneration and activating the necroptotic signaling pathway.

Evaluation of the thiazide challenge test to differentiate primary from hypercalciuria-related hyperparathyroidism.

treatment of primary hyperparathyroidism (PHPT) and secondary hyperparathyroidism due to idiopathic hypercalciuria (SHPT-IH) is markedly different. Robust diagnostic tools to differentiate between both entities are however lacking. evaluate the thiazide challenge test (TCT) in clinical practice, its aid in clinical decision making, evaluate the accuracy (sensitivity, specificity) and potentially useful parameters of the TCT. monocentric observational retrospective cohort study from January 2017 to November 2023. outpatient, Ghent University Hospital (Belgium). 25 adult patients with hypercalciuria, elevated parathyroid hormone (PTH), and high-normal or elevated serum calcium that underwent a TCT. TCT. serum, urinary biochemical parameters before and after testing, clinical and imaging outcomes, treatment, and follow-up. patients with a TCT-based working diagnosis of PHPT show greater increases in albumin-adjusted calcium and total serum calcium concentration than patients with SHPT-IH (+0,11 ± 0,10 vs. + 0,0071 ± 0,10mmol/l; p = 0,025 and +0,14 ± 0,12 vs. + 0,012 ± 0,15mmol/l; p = 0,024 respectively). The TCT-based working diagnosis of PHPT has a sensitivity of 81,8%, a specificity of 77,8% and a likelihood ratio of 3,68 of estimating a correct final diagnosis.Urinary calcium excretion, PTH, calcium-phosphorous ratio, PTH-inhibition rate, and parathyroid function index do not differ significantly in patients with PHPT compared to those with SHPT-IH. the TCT aids in discriminating patients with PHPT from those with SHPT-IH based on a rise in serum calcium. Other parameters are not different between both groups. Larger prospective trials are necessary to further define the diagnostic potential of the TCT, its most appropriate biochemical outcome variables, and decision cut-offs.

Essential Nutrients and White Matter Hyperintensities: A Two-Sample Mendelian Randomization Study.

Stroke and dementia have been linked to the appearance of white matter hyperintensities (WMHs). Meanwhile, diffusion tensor imaging (DTI) might capture the microstructural change in white matter early. Specific dietary interventions may help to reduce the risk of WMHs. However, research on the relationship between specific nutrients and white matter changes is still lacking. We aimed to investigate the causal effects of essential nutrients (amino acids, fatty acids, mineral elements, and vitamins) on WMHs and DTI measures, including fraction anisotropy (FA) and mean diffusivity (MD), by a Mendelian randomization analysis. We selected single nucleotide polymorphisms (SNPs) associated with each nutrient as instrumental variables to assess the causal effects of nutrient-related exposures on WMHs, FA, and MD. The outcome was from a recently published large-scale European Genome Wide Association Studies pooled dataset, including WMHs (N = 18,381), FA (N = 17,663), and MD (N = 17,467) data. We used the inverse variance weighting (IVW) method as the primary method, and sensitivity analyses were conducted using the simple median, weighted median, and MR-Egger methods. Genetically predicted serum calcium level was positively associated with WMHs risk, with an 8.1% increase in WMHs risk per standard deviation unit increase in calcium concentration (OR = 1.081, 95% CI = 1.006-1.161, p = 0.035). The plasma linoleic acid level was negatively associated with FA (OR = 0.776, 95% CI = 0.616-0.978, p = 0.032). Our study demonstrated that genetically predicted calcium was a potential risk factor for WMHs, and linoleic acid may be negatively associated with FA, providing evidence for interventions from the perspective of gene-environment interactions.

Comprehensive effect of increased calcium content in coal on the selenium emission from coal-fired power plants: Combined laboratory and field experiments

Coal combustion is the major contributor to global toxic selenium (Se) emissions. Inorganic elements in coals significantly affect Se partitioning during combustion. This work confirmed that the calcium (Ca) in ash had a stronger relationship with Se retention at 1300 °C than other major elements. Ca oxide chemically reacted with gaseous Se, and its sintering densification slightly affected Se adsorption capacities (44.45 −1840.71→35.17 −1540.15 mg/kg) at 300 − 1300 °C. Therefore, Ca in coals was identified as having potential for hindering gaseous Se emissions, and coals with increased Ca contents (2.74→5.19 wt%) were used in a 350 MW unit. The decreased Se mass distribution (3.54%→2.63%) in flue gas at air preheater inlet (320 −362 °C) confirmed the effectiveness of increased Ca content on gaseous Se emission reduction. More gaseous Se further condensed and was chemically adsorbed by fly ash when passed through an electrostatic precipitator, resulting in a significant increase in the Se content of fly ash. Additionally, the corresponding Se leaching ratio decreased from 4.88 − 35.74% to 1.87 − 26.31%, indicating enhanced stability of Se enriched in fly ash. This research confirmed the feasibility and environmental safety of sequestration of gaseous Se from flue gas to fly ash by increasing the Ca content in coals.

The role of the calcium-sensing receptor in the regulation of parathyroid hormone secretion in physiology and in calcitropic diseases

Parathyroid hormone (PTH) plays a key role in the regulation of calcium-phosphate metabolism. The secretion of PTH is regulated by calcium-sensing receptor (CaSR), which primarily expressed in the parathyroid glands and the renal tubules of the kidney. Increase of calcium concentration in extracellular matrix of cells is causing activation of the CaSR. Activated CaSR inhibits secretion of PTH and increases urinary calcium excretion. All CaSR effects leads to prevent development of hypercalcemia complications. Downregulation of the CASR expression and/or altered CaSR functioning leads to dysregulation of PTH synthesis. It may be the underlying cause of the development of primary and secondary hyperparathyroidism, as well as a number of hereditary diseases associated with loss- and gain-of-function mutations of the CaSR. In this paper we discusses the function of the CaSR in physiology and also the potential mechanisms that can impaired CaSR-induced signaling in various calcitropic diseases.

Label-Free Single Microparticles and Cell Aggregates Sorting in Continuous Cell-Based Manufacturing.

There is a paradigm shift in biomanufacturing toward continuous bioprocessing but cell-based manufacturing using adherent and suspension cultures, including microcarriers, hydrogel microparticles, and 3D cell aggregates, remains challenging due to the lack of efficient in-line bioprocess monitoring and cell harvesting tools. Herein, a novel label-free microfluidic platform for high throughput (≈50 particles/sec) impedance bioanalysis of biomass, cell viability, and stem cell differentiation at single particle resolution is reported. The device is integrated with a real-time piezo-actuated particle sorter based on user-defined multi-frequency impedance signatures. Biomass profiling of Cytodex-3 microcarriers seeded with adipose-derived mesenchymal stem cells (ADSCs) is first performed to sort well-seeded or confluent microcarriers for downstream culture or harvesting, respectively. Next, impedance-based isolation of microcarriers with osteogenic differentiated ADSCs is demonstrated, which is validated with a twofold increase of calcium content in sorted ADSCs. Impedance profiling of heterogenous ADSCs-encapsulated hydrogel (alginate) microparticles and 3D ADSC aggregate mixtures is also performed to sort particles with high biomass and cell viability to improve cell quality. Overall, the scalable microfluidic platform technology enables in-line sample processing from bioreactors directly and automated analysis of cell quality attributes to maximize cell yield and improve the control of cell quality in continuous cell-based manufacturing.

FLIM-FRET-based analysis of S100A11/annexin interactions in living cells.

Proteins achieve their biological functions in cells by cooperation in protein complexes. In this study, we employed fluorescence lifetime imaging microscopy (FLIM)-based Förster resonance energy transfer (FRET) measurements to investigate protein complexes comprising S100A11 and different members of the annexin (ANX) family, such as ANXA1, ANXA2, ANXA4, ANXA5, and AnxA6, in living cells. Using an S100A11 mutant without the capacity for Ca2+ binding, we found that Ca2+ binding of S100A11 is important for distinct S100A11/ANXA2 complex formation; however, ANXA1-containing complexes were unaffected by this mutant. An increase in the intracellular calcium concentration induced calcium ionophores, which strengthened the ANXA2/S100A11 interaction. Furthermore, we were able to show that S100A11 also interacts with ANXA4 in living cells. The FLIM-FRET approach used here can serve as a tool to analyze interactions between S100A11 and distinct annexins under physiological conditions in living cells.

Interorgan communication in neurogenic heterotopic ossification: the role of brain-derived extracellular vesicles

Brain-derived extracellular vesicles participate in interorgan communication after traumatic brain injury by transporting pathogens to initiate secondary injury. Inflammasome-related proteins encapsulated in brain-derived extracellular vesicles can cross the blood‒brain barrier to reach distal tissues. These proteins initiate inflammatory dysfunction, such as neurogenic heterotopic ossification. This recurrent condition is highly debilitating to patients because of its relatively unknown pathogenesis and the lack of effective prophylactic intervention strategies. Accordingly, a rat model of neurogenic heterotopic ossification induced by combined traumatic brain injury and achillotenotomy was developed to address these two issues. Histological examination of the injured tendon revealed the coexistence of ectopic calcification and fibroblast pyroptosis. The relationships among brain-derived extracellular vesicles, fibroblast pyroptosis and ectopic calcification were further investigated in vitro and in vivo. Intravenous injection of the pyroptosis inhibitor Ac-YVAD-cmk reversed the development of neurogenic heterotopic ossification in vivo. The present work highlights the role of brain-derived extracellular vesicles in the pathogenesis of neurogenic heterotopic ossification and offers a potential strategy for preventing neurogenic heterotopic ossification after traumatic brain injury.Brain-derived extracellular vesicles (BEVs) are released after traumatic brain injury. These BEVs contain pathogens and participate in interorgan communication to initiate secondary injury in distal tissues. After achillotenotomy, the phagocytosis of BEVs by fibroblasts induces pyroptosis, which is a highly inflammatory form of lytic programmed cell death, in the injured tendon. Fibroblast pyroptosis leads to an increase in calcium and phosphorus concentrations and creates a microenvironment that promotes osteogenesis. Intravenous injection of the pyroptosis inhibitor Ac-YVAD-cmk suppressed fibroblast pyroptosis and effectively prevented the onset of heterotopic ossification after neuronal injury. The use of a pyroptosis inhibitor represents a potential strategy for the treatment of neurogenic heterotopic ossification.

The effectiveness of Bio-plus2B®, Techno Mos® and their mixture on the rate of egg production, egg characteristics, retention of nutrients and blood metabolites through the early period of production

BackgroundThe trend of using probiotic, prebiotic and their mixture as alternative feed additives which works as growth promoters in poultry diets to increase the productive performance and the immunity of the flock still have an importance consideration. So that the aim of this study is to estimate the impact of Bio-plus2B® (probiotic), Techno Mos® (prebiotic) or their mixture (synbiotic) on egg production, egg and shell quality, some blood metabolites and retention of nutrients between 28 and 40 weeks of age. The hens (ISA brown laying) were allocated randomly in 40 cages; 10 cages/treatment with two hens each. The treatments were the control (T1), T2 (Probiotic: 1 g Bio-plus2B® (Bacillus licheniformis plus Bacillus subtilis)/kg feed), T3 (Prebiotic: 1 g Techno Mos® (Mannanoligosaccarides (MOS) and 1,3 B-glucan) /kg feed) and T4 (Synbiotic: 1 g Bio-plus2B® plus 1 g Techno Mos®/ kg feed).ResultsHen-day egg production% and mass were significantly increased (P < 0.05) with T2 and T4 treatments. The experimental treatments recorded an increase in albumen index, Haugh unit (P < 0.01), shell thickness (P < 0.05), the retention of crude protein (CP), crude fiber (CF) and ether extract (EE) (P < 0.05), plasma globulin, albumin and total protein (P < 0.001) versus to the untreated group, while egg weight was not affected. Synbiotic treatment showed a significant (P < 0.001) increase in shell calcium content. T3 and T4 treatments were significantly decreased plasma cholesterol (P < 0.001) and low density lipoprotein (LDL) (P < 0.05). Alanine transaminase (ALT) was significantly (P < 0.001) decreased and estradiol hormone was increased (P < 0.001) in the experimental groups versus to the control.ConclusionsIt concluded that adding probiotic and/or prebiotic in the early age laying hens diets had beneficial effects for productivity with improving the egg shell thickness.

Methadone Requires the Co-Activation of μ-Opioid and Toll-Like-4 Receptors to Produce Extracellular DNA Traps in Bone-Marrow-Derived Mast Cells.

Methadone is an effective and long-lasting analgesic drug that is also used in medication-assisted treatment for people with opioid use disorders. Although there is evidence that methadone activates μ-opioid and Toll-like-4 receptors (TLR-4s), its effects on distinct immune cells, including mast cells (MCs), are not well characterized. MCs express μ-opioid and Toll-like receptors (TLRs) and constitute an important cell lineage involved in allergy and effective innate immunity responses. In the present study, murine bone-marrow-derived mast cells (BMMCs) were treated with methadone to evaluate cell viability by flow cytometry, cell morphology with immunofluorescence and scanning electron microscopy, reactive oxygen species (ROS) production, and intracellular calcium concentration ([Ca2+]i) increase. We found that exposure of BMMCs to 0.5 mM or 1 mM methadone rapidly induced cell death by forming extracellular DNA traps (ETosis). Methadone-induced cell death depended on ROS formation and [Ca2+]i. Using pharmacological approaches and TLR4-defective BMMC cultures, we found that µ-opioid receptors were necessary for both methadone-induced ROS production and intracellular calcium increase. Remarkably, TLR4 receptors were also involved in methadone-induced ROS production as it did not occur in BMMCs obtained from TLR4-deficient mice. Finally, confocal microscopy images showed a significant co-localization of μ-opioid and TLR4 receptors that increased after methadone treatment. Our results suggest that methadone produces MCETosis by a mechanism requiring a novel crosstalk pathway between μ-opioid and TLR4 receptors.

Feasibility of increasing calcium content of drinking tap water following quality regulations to improve calcium intake at population level

Background Calcium intake is below recommendations in several parts of the world. Improving calcium intake has benefits not only for bone health but also helps to prevent pregnancy hypertension disorders. Calcium concentration of tap water is usually low The aim of the present study was to determine the maximum amount of calcium that can be added to tap water while complying with drinking water local regulations. Methods Tap water samples were collected from the Province of Buenos Aires (Argentina). Physicochemical properties and saturation index were measured. Different incremental concentrations of calcium chloride were added to the experimental aliquots. Results Baseline water had a mean calcium concentration of 22.00 ± 2.54 mg/L, water hardness of 89.9 ± 6.4 mg/L CaCO3, and a saturation index of -1.50 ± 0.11. After the addition of 0.4554 ± 0.0071 g of salt, water hard-ness reached 355.0 ± 7.1 mg/L CaCO3, a calcium concentration of 140.50 ± 2.12 mg/L, and a saturation index -0.53 ± 0.02. Conclusions This study shows that at laboratory level it is feasible to increase calcium concentration of drinking water by adding calcium chloride while complying with national standards. Calcium concentration of drinking tap water could be evaluated and minimum calcium concentration of tap water regulated so as to improve calcium intake in populations with low calcium intake.

Superwettable calcium ion exchanged carboxymethyl cellulose powder with self-gelation, tissue adhesion and bioabsorption for effective hemorrhage control

Water-soluble polymers can be used to develop self-gelling hemostatic powders, and powder wettability potentially plays the key role in determining their hemostatic efficacy. Herein, a superwettable calcium ion exchanged carboxymethyl cellulose (Ca-ex-CMC) powder is reported for effective hemorrhage control. Despite being water-soluble, Ca-ex-CMC powder can form a physically crosslinked hydrogel upon contact with water (self-gelation), and the rate of water permeating the hydrogel to achieve dissolution is extremely slow. With increased calcium content, Ca-ex-CMC powder shows not only improved procoagulant activity, but enhanced wettability which can facilitate the fusion of the powders with water, resulting in promoted water uptake and in-situ formation of strong hydrogel barriers with enhanced tissue adhesion. Applying Ca-ex-CMC powder to non-compressible (liver and tail) and massive (artery) hemorrhage wounds in rat and rabbit models can stanch bleeding instantly, because of complete sealing of the bleeding sites by enhanced in-situ self-gelation. Moreover, owing to water-solubility, Ca-ex-CMC powder shows outstanding bioabsorbability and histocompatibility after implantation into rabbit liver and gluteus. These findings not only support Ca-ex-CMC powder as a promising hemostatic agent for extensive application but identify the critical role of wettability in promoting hemostasis for self-gelling powders.