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This study evaluated the efficacy of intravenous (IV) calcium pretreatment for preventing diltiazem-induced hypotension and assessed its safety in adult patients with atrial fibrillation (AF)/atrial flutter (AFL) with rapid ventricular response (RVR). This randomized, double-blind, placebo-controlled trial included 217 adults with AF/AFL and a ventricular rate > 120 beats per minute, who were randomized into three groups: those who received an IV NaCl 0.9 % placebo pretreatment prior to IV diltiazem (PD; 73 patients) and those who received 90 mg (C90D; 71 patients) and 180 mg (C180D; 73 patients) IV calcium chloride pretreatment before IV diltiazem. We compared participants' systolic blood pressure (SBP) and heart rate (HR) at baseline and at 5, 10, and 15 min post-treatment, as well as the incidence of adverse events (e.g., hypotension, urticaria, nausea) among the groups. The PD and C90D pretreatment groups had significantly lower HR measurements at 10 and 15 min compared to the C180D group. In addition, at 5 min, the mean SBP in the PD group was significantly lower compared to the C90D and C180D groups. At 10 min, the mean SBP was significantly higher in the C180D group than in the other groups. Furthermore, at 15 min, the mean SBP was significantly higher in both the C90D and C180D groups than in the PD group. There were no significant differences between the calcium pretreatment and placebo groups in terms of the need for additional diltiazem doses or the incidence of adverse events. IV calcium pretreatment effectively prevents diltiazem-induced hypotension in patients with AF/AFL with RVR without compromising the efficacy of diltiazem in achieving and maintaining ventricular rate control. Trial registry: National Library of Medicine Clinical Trial Registry; No.: NCT06494007; URL: https://clinicaltrials.gov/study/NCT06494007

The alizarin red S assay is considered the gold standard for quantification of osteoblast mineralization and is thus widely used among scientists. However, there are several restrictions to this method, e.g., moderate sensitivity makes it difficult to uncover slight but significant effects of potentially clinically relevant substances. Therefore, an adaptation of the staining method is appropriate and might be obtained by increasing the mineralization ability of osteoblasts. In this study, cell culture experiments with human (SaOs-2) and murine (MC3T3-E1) osteoblasts were performed under the addition of increasing concentrations of calcium chloride (1, 2.5, 5, and 10 mM) or calcitonin (1, 2.5, 5, and 10 nM). After three or four weeks, the mineralization matrix was stained with alizarin red S and the concentration was quantified photometrically. Only calcium chloride was able to significantly increase mineralization, and therefore enhanced the sensitivity of the alizarin red S staining in a dose-dependent manner in both osteoblastic cell lines as well as independent of the cell culture well surface area. This cost- and time-efficient optimization enables a more sensitive analysis of potentially clinically relevant substances in future bone research.

Pancreatic cancer typically spreads rapidly and has poor survival rates. Here, we report that the calcium-activated chloride channel TMEM16A is a biomarker for pancreatic cancer with a poor prognosis. TMEM16A is up-regulated in 75% of cases of pancreatic cancer and high levels of TMEM16A expression are correlated with low patient survival probability. TMEM16A up-regulation is associated with the ligand-dependent EGFR signaling pathway. In vitro, TMEM16A is required for EGF-induced store-operated calcium entry essential for pancreatic cancer cell migration. TMEM16A also has a profound impact on phosphoproteome remodeling upon EGF stimulation. Moreover, molecular actors identified in this TMEM16A-dependent EGFR-induced calcium signaling pathway form a gene set that makes it possible not only to distinguish neuro-endocrine tumors from other forms of pancreatic cancer, but also to subdivide the latter into three clusters with distinct genetic profiles that could reflect their molecular underpinning.

Abstract Calcite precipitation by non-growing cells occurs more rapidly in the laboratory than with proliferative cells. Sporosarcina pasteurii was grown in urea-containing nutrient broth, and then harvested, washed, and resuspended in a solution. This bacterial suspension was mixed with an equal volume of a reactant solution containing calcium chloride and urea at different concentrations. The precipitate formed was collected, dried, and analyzed using X-ray diffraction (XRD), energy-dispersive X-ray spectroscopy, field emission scanning electron microscopy, and particle size analysis (PSA). This study investigated the effects of urea and calcium chloride concentrations on the type and size of the crystals formed. The most effective concentrations of urea and calcium chloride were both 0.3 M. Semi-quantitative XRD analysis revealed that 97% of the precipitate was calcite, with only 3% being vaterite at these concentrations. PSA results indicated that, in a 0.3 M solution of urea and calcium chloride, most particles ranged in size from 0 to 60 nm. The results show that higher reactant concentrations lead to more precipitate in the slurry, but with a lower percentage of calcite; additionally, the size of the calcite crystals also increases. This study not only reaffirms the ability of non-growing cells to precipitate calcite but also investigates the precise relationship between reactant concentrations, crystal type, and particle size. These insights provide a deeper understanding of bio-slurry and its potential applications in fields like concrete and soil enhancement. This study investigates how reactant concentrations affect calcite precipitation in non-growing cells of  S. pasteurii and their capacity to produce calcite in a solution containing these reactants.

Three-dimensional bioprinting allows for the fabrication of structures mimicking tissue architecture. This study aimed to develop a gelatin-based bioink for a bioprinted simplified skin model. The bioink printability and chemical-physical properties were evaluated by varying the concentrations of gelatin (10, 15, and 20%) in a semi-crosslinked alginate-based bioink and calcium chloride (100, 150, and 200 mM) in post-printing crosslinking. For increasing the gelatin concentration, the gelatin-based formulations have a shear thinning behavior with increasing viscosity, and the filament bending angle increases, the spreading ratio value approaches 1, and the shape fidelity and the printing resolution improve. However, the formulation containing 20% of gelatin was not homogeneous, resulting also in poor printability properties. The morphology of the pores, degradation, and swelling depend on gelatin and CaCl2 concentrations, but not in a significant way. The samples containing 15% of gelatin and crosslinked with 150 mM CaCl2 have been selected for the bioprinting of a bilayer skin model containing human fibroblasts and keratinocytes. The model showed a homogeneous distribution of viable and proliferating cells over up to 14 days of in vitro culture. The gelatin-based bioink allowed for the 3D bioprinting of a simplified skin model, with potential applications in the bioactivity of pro-reparative molecules and drug evaluation.

Abstract The efficacy of β-agonists in asthma is severely limited by β-adrenoceptor desensitization, which results in poorly managed symptoms and refractory bronchoconstriction. Thus, there is a need to identify novel therapeutic pathways and to clarify the relationship between novel therapeutics and functional β-adrenoceptor responsiveness. We have previously demonstrated that acute antagonism of the calcium-activated chloride channel, transmembrane member 16A (TMEM16A), relaxes airway smooth muscle (ASM). We sought to determine the efficacy and role of TMEM16A antagonism in the context of desensitization β-adrenoceptor responsiveness. For these studies, we exposed murine tracheal rings on wire myography and precision-cut lung slices to contractile mediators in the presence or absence of TMEM16A antagonists and β-agonists with or without prior β-adrenoceptor desensitization. Contractile studies were also performed with human tracheal and bronchial ASM. Finally, the ability of TMEM16A antagonism to prevent desensitization of β2-adrenoceptor–induced cAMP synthesis was measured in human ASM cells. From these studies, we demonstrate that acute TMEM16A antagonism is effective in relaxing β-agonist–desensitized ASM in central and peripheral murine ASM and human ASM. Furthermore, we demonstrate that chronic pretreatment with TMEM16A antagonists prevents functional desensitization of β-agonist responsiveness in mouse and human upper airways and prevents desensitization of β-agonist–mediated cAMP production in human ASM cells. Taken together, the present study demonstrates a favorable therapeutic profile of TMEM16A antagonism for ASM relaxation despite functional desensitization of β-agonist responsiveness, which may be a novel therapeutic approach in the face of β-adrenoceptor tachyphylaxis.

The expression levels of anoctamin 1 (ANO1, TMEM16A), a calcium-activated chloride channel (CaCC), are significantly increased in several tumors, and inhibition of ANO1 is known to reduce cell proliferation and migration. Here, we performed cell-based screening of a collection of natural products and drug-like compounds to identify inhibitors of ANO1. As a result of the screening, idebenone, miconazole and plumbagin were identified as novel ANO1 inhibitors. Electrophysiological studies showed that idebenone, a synthetic analog of coenzyme Q10, completely blocked ANO1 activity in FRT cells expressing ANO1 without any effect on intracellular calcium signaling and CFTR, a cAMP-regulated chloride channel. The CaCC activities in PC-3 and CFPAC-1 cells expressing abundant endogenous ANO1 were strongly blocked by idebenone. Idebenone inhibited cell proliferation and induced apoptosis in PC-3 and CFPAC-1 cells, but not in A549 cells, which do not express ANO1. These data suggest that idebenone, a novel ANO1 inhibitor, has potential for use in cancer therapy.

Rose flowers ( Rosa hybrida L.) are highly perishable and have a limited vase life. This study evaluated the effects of preharvest foliar applications of γ-aminobutyric acid (GABA) and calcium chloride (CaCl 2 ), individually and combined, on antioxidant responses and vase life of cut Jumilia rose flowers. Treatments included foliar sprays of GABA at 0, 20, 40, and 60 mM and CaCl 2 at 0, 0.75%, and 1.5%, applied in a factorial design within a completely randomized setup before harvest. Results showed GABA and CaCl 2 interaction (especially, 60 mM GABA and 1.5% CaCl 2 ) significantly increased enzymatic antioxidants including superoxide dismutase, catalase, and peroxidase, as well as non-enzymatic antioxidants such as flavonoids, carotenoids, phenolics, and antioxidant activity in petals compared to control. SOD activity in roses, treated with CaCl 2 (1.5%) and GABA (60 mM), peaked at 7.86 units. mg −1 protein min −1 , showing a nearly 2.93-fold increase over the control (2.68 units. mg −1 protein min −1 ). A parallel trend was observed for CAT activity. These treatments also reduced petal malondialdehyde content and polyphenol oxidase activity. Protein content and vase life duration increased in all treatments. Plants treated with a combination of GABA (20 mM) and CaCl 2 (0.75%), GABA (60 mM) and CaCl 2 (1.5%), or GABA (40 mM) individually exhibited the longest vase life duration. The co-application of GABA and CaCl 2 improved the antioxidant activity and postharvest quality of cut roses by reducing PPO activity and MDA contents, increasing protein content and prolonging vase life. This treatment is a potential postharvest strategy to improve antioxidant capacity and delay senescence in cut roses.

Hydrated salt phase change materials (PCMs) can play an important role in the temperature regulation of buildings by storing and releasing latent heat. However, hydrated salt PCMs are affected by phase separation, supercooling, and leakage, which greatly limit their application. In this study, an innovative modified calcium chloride hexahydrate (CaCl2·6H2O) PCM was prepared with potassium chloride (KCl) as the temperature regulator and strontium hydroxide octahydrate (Sr(OH)2·8H2O) as the nucleating agent. Subsequently, 15 wt% expanded graphite (EG) was immersed into 85 wt% modified PCM to prepare form-stable composite PCM, which is suitable for space thermal regulation with a phase transition temperature of 26.25 °C, phase change enthalpy of 153.03 J/g and a thermal conductivity of 4.3893 W/(mK). After 500 heating–cooling cycles, the phase change temperature changed little, and enthalpy lost only 5.95%. The test chamber prepared with a composite PCM and polyvinyl chloride (PVC) board was studied to evaluate its temperature regulation performance. In the heating and cooling process, the indoor temperature is maintained at 24.5–27.5 °C for 180 to 652 min. Therefore, the prepared composite PCM can have a good application in the temperature management of buildings.Graphical abstract

The present study was performed to evaluate the effects of seed priming. This was done by soaking the seeds of two rapeseed cultivars, namely, ZY15 (tolerant to low temperature and drought) and HY49 (sensitive to low temperature and drought), for 12 h in varying solutions: distilled water, 138 mg/L salicylic acid (SA), 300 mg/L gibberellic acid (GA), 89.4 mg/L sodium nitroprusside (SNP), 3000 mg/L calcium chloride (CaCl 2 ), and 30 mg/L abscisic acid (ABA). Primed and non-primed seeds were left to germinate at 15°C and -0.15 MPa (T 15 W 15 ) and at 25°C and 0 MPa (T 25 W 0 ), respectively. The results showed that SA, GA, SNP, CaCl 2 , and ABA significantly improved the germination potential (GP), germination rate (GR), germination index (GI), stem fresh weight (SFW), stem dry weight (SDW), root length (RL), stem length (SL), and seed vigor index (SVI) under T 15 W 15 . For ZY15 seeds under T 25 W 0 , GA, SNP, CaCl 2 , and ABA priming reduced the average germination time (96% after 5 days) compared to that of the control (88% after 5 days). For ZY15 seeds under T 15 W 15 , SA, SNP, CaCl 2 , and ABA priming, with respect to the control and water-treated groups, shortened the average germination time (92% after 5 days) compared to that of the control (80% after 5 days). For HY49 seeds under T 25 W 0 , GA, SNP, CaCl 2 , and ABA priming reduced the average germination time (92% after 5 days) compared to that of the control (85% after 5 days). Similarly, for HY49 seeds under T 15 W 15 , GA priming shortened the average germination time (89% after 5 days) compared to that of the control (83% after 5 days). These priming agents increased the net photosynthesis, stomatal conductivity, and transpiration rate of rape seedlings under conditions of low temperature and drought stress, while also decreasing intercellular carbon dioxide (CO 2 ) concentrations. Additionally, SA, GA, SNP, CaCl 2 , and ABA increased superoxide dismutase concentrations (SOD) and ascorbic peroxidase (APX) activities of rape seedlings under stress conditions, while decreasing catalase (CAT) and peroxidase (POD) activities in ZY15 seedlings. In HY49, which is sensitive to low temperature and drought, all priming solutions, except for SNP, led to an increase in SOD activity levels and a decrease in CAT activity levels. Overall, SA, GA, SNP, and CaCl 2 increased the concentrations of indoleacetic acid (IAA), GA, ABA, and cytokinin (CTK) in seedlings under stress conditions. Moreover, compared to SA, CaCl 2 , and ABA, GA (300 mg/L) and SNP (300 mol/L) showed improved priming effects for ZY15 and HY49 under stress conditions.