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This study aimed to evaluate the effects of chicken eggshell powder rich in calcium and the inorganic form of calcium carbonate on osteoporotic bone structure using an animal model of ovariectomized rats. Animals were divided into four groups: sham-operated rats (SHAM group); ovariectomized rats untreated (OVX group); OVX rats treated with eggshell powder (Biomin H® OVX + ECa group; total Ca content 1.5%); and OVX rats receiving inorganic calcium carbonate (calcium carbonate precipitated, total Ca content 1.5%; OVX + ICa group) during 8 weeks. Ovariectomy increased total body weight, plasma alkaline phosphatase activity, cortical bone thickness, periosteal bone apposition, and considerably worsened the trabecular bone microarchitecture. Calcium supplementation in both OVX + ECa and OVX + ICa groups elevated blood supply in the cortical bone when compared to OVX rats. Treatment with eggshell powder, unlike the OVX + ICa group, significantly lowered bone resorption, increased plasma calcium level, relative volume of trabecular bone, and secondary osteon population density, which together contributes to the enhanced bone strength. In addition, sizes of Haversian canals and secondary osteons in OVX + ECa group reached the values of SHAM group. Our results suggest that chicken eggshell powder more effectively ameliorates bone loss in ovariectomized rats than precipitated calcium carbonate.

Introduction: Imperfect hemostasis after arteriovenous fistula (AVF) and arteriovenous graft (AVG) cannulation can cause a hematoma or pseudoaneurysm and leads to poor satisfaction. We hypothesized that a hydrogel-coated needle would effectively and rapidly stop bleeding after vascular cannulation in a rat AVF and AVG model., Method: A hydrogel comprised of sodium alginate (SA), hyaluronic acid (HA), and calcium carbonate was coated onto the surface of suture needles using a rotating system. The needles were observed using scanning electron microscopy (SEM) and immunofluorescence. Rat AVF with or without renal failure and AVG were punctured using bare and hydrogel-coated needles. The tissues were examined by histology., Result: The hydrogel was successfully coated onto the surface of 30 G needles and confirmed by SEM. Hydrogel-coated needles rapidly stopped bleeding after AVF and AVG cannulation in rat., Conclusion: In this preliminary animal research, hydrogel-coated needles can stop AVF and AVG puncture-site bleeding; but additional clinical studies are needed to justify whether it is still effective in clinical., (Copyright © 2021 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

The nutritional values of limestone, oyster shell and extruded eggshells were evaluated using different methods. In total, 120 ready-to-lay pullets, 18-week-old ISA Brown were distributed into six dietary groups, namely D 1 (4% limestone), D 2 (8% limestone), D 3 (4% oyster shell), D 4 (8% oyster shell), D 5 (4% eggshell) and D 6 (8% eggshell), to assess the effect of calcium sources on egg production, egg quality, dry matter and cholesterol content of the egg. Kitchen-extruded eggshell contained 98.52, 4.24, 29.75 and 14.82% DM, CP, Ca and P, whereas hatchery-extruded eggshell contained 99.20, 13.80, 25.53 and 13.87% DM, CP, Ca and P, respectively. Limestone and Oyster shells contained 99.60 and 99.51% DM, and 37.12 and 35.20% Ca, respectively. Body weight, egg, hen day and egg mass production, and FCR did not differ among diets (p > .05). Egg production tended to increase with the increase of hen-housed egg production (p < .001) in D 6 , followed by D 2 , D 5 , D 3 , D 4 and D 1 , respectively. The lowest production cost and the highest net profit were observed in D 6 , followed by D 2 , D 4 , D 5 , D 1 and D 3 , respectively. Diet with 8% Ca sources performed better than the diets with 4% Ca sources in terms of egg quality and dry matter content, where D 6 was comparable to D 2 or D 4 . The weight of egg, albumen, yolk, eggshell, dry yolk and yolk-albumen ratio increased, while dry albumen and eggshell weight, eggshell strength and thickness, Haugh unit, yolk index and egg-specific gravity decreased with the increase in bird's age. The cholesterol content of yolk was statistically similar among diets. Therefore, no adverse effect of calcium sources on the production of laying hen was observed. Of these, extruded eggshell especially the 8% extruded eggshell may be beneficial to use in the diet of laying hen for producing a quality, safe and profitable egg., (© 2021 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Malaria is still a global health problem. Plasmodium is a single-cell protozoan parasite that causes malaria and is transmitted to humans through the female Anopheles mosquito. The previous study showed that Sonchus arvensis L. callus has antiplasmodial activity. Several treatments are needed for callus quality improvement for antimalarial compound production. This study aimed to examine the effect of dolomite [CaMg(CO3)2] on growth (morpho-anatomical structure and biomass), secondary metabolite production, and in vitro antiplasmodial activity of S. arvensis L. callus. In this study, leaf explants were grown in Murashige and Skoog medium with a combination of 2,4-dichlorophenoxyacetic acid (2,4-D, one mg/L) and 6-benzyl amino purine (BAP, 0.5 mg/L) with dolomite (50, 75, 100, 150, and 200 mg/L). The 21 days callus ethanolic and methanolic extract were analyzed by gas chromatography-mass spectrometry (GC-MS) and thin-layer chromatography (TLC). The antiplasmodial test was performed on a blood culture infected with Plasmodium falciparum strain 3D7 using the Rieckmann method. The results showed that dolomite significantly affected callus growth, metabolite profile, and in vitro antiplasmodial activity. Dolomite (150 mg/L) showed the highest biomass (0.590 ± 0.136 g fresh weight and 0.074 ± 0.008 g dry weight). GC-MS analysis detected four compounds from callus ethanolic extract. Pelargonic acid, decanoic acid, and hexadecanoic acid were major compounds. One new terpenoid compound is based on TLC analysis. S. arvensis L. callus has antiplasmodial activity with the IC50 value of 5.037 μg/mL. It was three times lower than leaf methanolic extract and five times lower than leaf ethanolic extract., Competing Interests: The authors have declared that no competing interests exist.

Calcium carbonate is used as bone-filling material due to its good biocompatibility, bioactivity, and bioabsorbability, but the prevalence of infectious complications associated with calcium carbonate has created a persisting challenge in the treatment of bone defect. Therefore, this greatly necessitate the need to endow calcium carbonate with antibacterial properties. In this study, calcium carbonate powders loaded with silver nanoparticles (Ag-CaCO 3 ) were prepared in attempt to serve as a novel antibacterial inorganic filler material. This objective was achieved using ultrasonic spray-pyrolysis (USSP) route to produce Ag-CaCO 3 with 1, 5 and 10 mol% silver. The size of silver nanoparticles on CaCO 3 microspheres could be regulated by adjusting silver concentration to facilitate effective release of Ag + ions. This was demonstrated in Ag-CaCO 3 (1), where the lowest silver content at 1 mol% achieved the highest Ag + ions release over 28 days. This in turn gave rise to effective antibacterial efficiency against Staphylococcus aureus and Escherichia coli. Furthermore, CaCO 3 (1) could also support osteoblast-like cells (MG-63) at a cell viability of 80%. Overall, this work extends the capabilities in employing USSP to produce inorganic filler materials with sustained antibacterial properties, bringing one step closer to the development of antibacterial products., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Biomineralization is the process by which living organisms generate organized mineral crystals. In human cells, this phenomenon culminates with the formation of hydroxyapatite, which is a naturally occurring mineral form of calcium apatite. The mechanism that explains the genesis within the cell and the propagation of the mineral in the extracellular matrix still remains largely unexplained, and its characterization is highly controversial, especially in humans. In fact, up to now, biomineralization core knowledge has been provided by investigations on the advanced phases of this process. In this study, we characterize the contents of calcium depositions in human bone mesenchymal stem cells exposed to an osteogenic cocktail for 4 and 10 days using synchrotron-based cryo-soft-X-ray tomography and cryo-XANES microscopy. The reported results suggest crystalline calcite as a precursor of hydroxyapatite depositions within the cells in the biomineralization process. In particular, both calcite and hydroxyapatite were detected within the cell during the early phase of osteogenic differentiation. This striking finding may redefine most of the biomineralization models published so far, taking into account that they have been formulated using murine samples while studies in human cell lines are still scarce.

The development of heterogeneous drug delivery systems leads to innovative strategies for targeted therapy of common pathologies, such as cancer, immunological and neurological disorders. Nowadays, it is possible to choose among a great variety of nanoparticles on the basis of the needs they have to satisfy. However, a candidate for the treatment of cardiovascular pathologies is still missing. In this context, a targeted therapy implies the conceptualization of nanoparticles that take active part in the treatment of vascular pathologies. The aim of this work was to provide a method to produce multi-layered calcium carbonate (CaCO 3 ) nanoparticles encapsulating a model protein, bovine serum albumin, with model antibodies on their surface. CaCO 3 nanoparticles were produced by the combination of complex coacervation and mineralization and were engineered using layer-by-layer technique with a polysaccharide, dextran sulfate, and a homo-poly-amino acid, poly-L-arginine. Morphology, biocompatibility, cellular uptake, influence on cell expression of the inflammatory marker matrix metalloproteinase-9, and hemocompatibility of the nanoparticles were studied. The presence of the dextran/poly-L-arginine layers did not negatively affect the nanoparticle overall characteristics and they did not trigger proinflammatory response in vitro. Taking together all the obtained results, we consider the proposed CaCO 3 nanoparticles as a promising tool in cardiovascular field., (Copyright © 2021. Published by Elsevier B.V.)

The acidic microenvironment of solid tumors induces the propagation of highly invasive and metastatic phenotypes. However, simulating these conditions in animal models present challenges that confound the effects of pH modulators on tumor progression. To recapitulate the tumor microenvironment and isolate the effect of pH on tumor viability, we developed a bifurcated microfluidic device that supports two different cell environments for direct comparison. RFP-expressing breast cancer cells (MDA-MB-231) were cultured in treatment and control chambers surrounded by fibrin, which received acid-neutralizing CaCO 3 nanoparticles (nanoCaCO 3 ) and cell culture media, respectively. Data analysis revealed that nanoCaCO 3 buffered the pH within the normal physiological range and inhibited tumor cell proliferation compared to the untreated control (p < 0.05). Co-incubation of cancer cells and fibroblasts, followed by nanoCaCO 3 treatment showed that the nanoparticles selectively inhibited the growth of the MDA-MB-231 cells and reduced cellular migration of these cells with no impact on the fibroblasts. Sustainable decrease in the intracellular pH of cancer cells treated with nanoCaCO 3 indicates that the extracellular pH induced cellular metabolic reprogramming. These results suggest that the nanoCaCO 3 can restrict the aggressiveness of tumor cells without affecting the growth and behavior of the surrounding stromal cells.

Background: Nanoparticles assembled from food-grade calcium carbonate have attracted attention because of their biocompatibility, digestibility, particle and surface features (such as size, surface area, and partial wettability), and stimuli-responsiveness offered by their acid-labile nature., Results: Herein, a type of edible oil-in-water Pickering emulsion was structured by calcium carbonate nanoparticles (CaCO 3 NPs; mean particle size: 80 nm) and medium-chain triglyceride (MCT) for delivery of lipophilic drugs and simultaneous oral supplementation of calcium. The microstructure of the as-made CaCO 3 NPs stabilized Pickering emulsion can be controlled by varying the particle concentration (c) and oil volume fraction (φ). The emulsification stabilizing capability of the CaCO 3 NPs also favored the formation of high internal phase emulsion at a high φ of 0.7-0.8 with excellent emulsion stability at room temperature and at 4 °C, thus protecting the encapsulated lipophilic bioactive, vitamin D3 (VD3), against degradation. Interestingly, the structured CaCO 3 NP-based Pickering emulsion displayed acid-trigged demulsification because of the disintegration of the CaCO 3 NPs into Ca 2+ in a simulated gastric environment, followed by efficient lipolysis of the lipid in simulated intestinal fluid. With the encapsulation and delivery of the emulsion, VD3 exhibited satisfying bioavailability after simulated gastrointestinal digestion., Conclusions: Taken together, the rationally designed CaCO 3 NP emulsion system holds potential as a calcium-fortified formulation for food, pharmaceutical and biomedical applications.

Developing chick embryos are a classical research tool in developmental biology. The whole embryo culture technique can be applied to various fields, such as embryo manipulation, toxicology, tumorigenesis, and basic research in regenerative medicine. When used for the generation of transgenic chickens, a high hatchability of genetically engineered embryos is essential to support normal embryonic development during culture. In this study, calcium carbonate, which is the main component of eggshells, was added as a calcium source in shell-less chick embryo cultures using a transparent plastic film as a culture vessel. In the absence of a calcium source in the shell-less culture system, embryogenesis ceased during culture, resulting in failed embryonic hatching. We found that the direct addition of calcium carbonate to the chorioallantoic membrane of the developing embryo was effective for the hatching of cultured chick embryos. The amount, timing, and location of calcium carbonate addition were investigated to maximize the hatchability of cultured embryos. Starting from the time of calcium carbonate supplementation, >40% hatchability was obtained with the optimal condition. This established method of shell-less chick embryo culture provides a useful tool in basic and applied fields of chick embryo manipulation., (Copyright © 2020 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Highly active antiretroviral therapy (HAART) has dramatically lowered rates of mother-to-child HIV transmission among patients with access to treatment. Barriers to complete viral suppression increase rates of transmission, even with only low levels of viral replication. Here, we present the case of a pregnant patient who developed a detectable viral load in pregnancy, thought to be related to calcium supplement consumption or emesis while using a dolutegravir-based HAART regimen. Ultimately, with adjustments, the patient again reached an undetectable viral load and had an uncomplicated perinatal and neonatal outcome. We discuss new data on the use of dolutegravir in pregnancy and precautions for maintaining viral suppression while on antiretroviral therapy in pregnancy., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Patients with advanced-stage ovarian cancer face a poor prognosis because of recurrent peritoneal cavity metastases following surgery and chemotherapy. Alpha-emitters may enable the efficient treatment of such disseminated diseases because of their short range and highly energetic radiation. Radium-224 is a candidate α-emitter due to its convenient 3.6-day half-life, with more than 90% of the decay energy originating from α-particles. However, its inherent skeletal accumulation must be overcome to facilitate intraperitoneal delivery of the radiation dose. Therefore, 224 Ra-labeled CaCO 3 microparticles have been developed., Objective: The antitumor effect of CaCO 3 microparticles as a carrier for 224 Ra was investigated, with an emphasis on the ratio of activity to mass dose of CaCO 3 , that is, specific activity., Methods: Nude athymic mice were inoculated intraperitoneally with human ovarian cancer cells (ES-2) and treated with a single intraperitoneal injection of 224 Ra-labeled CaCO 3 microparticles with varying combinations of mass and activity dose, or cationic 224 Ra in solution. Survival and ascites volume at sacrifice were evaluated., Results: Significant therapeutic effect was achieved for all tested specific activities ranging from 0.4 to 4.6 kBq/mg. Although treatment with a mean activity dose of 1305 kBq/kg of cationic 224 Ra prolonged the survival compared with the control, equivalent median survival could be achieved with 224 Ra-labeled microparticles with a mean dose of only 420 kBq/kg. The best outcome was achieved with the highest specific activities (2.6 and 4.6 kBq/mg)., Conclusion: Radium-224-labeled CaCO 3 microparticles present a promising therapy against cancer dissemination in body cavities., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Contrast-enhanced ultrasound (CEUS) is widely applied in cancer diagnosis clinically. However, the gas-filled contrast agents are unstable in the blood and exhibit shorter imaging time, which limit their clinical use. In this study, a diagnostic nanoparticle system was developed for dual-mode imaging (ultrasound and fluorescence), which after encapsulation with doxorubicin (DOX) demonstrated simultaneous therapeutic function towards cancer treatment. Thus, calcium carbonate (CaCO 3 ) nanoparticles were encapsulated with doxorubicin (DOX) to obtain CaCO 3 -DOX. Under acidic conditions, it produced carbon dioxide (CO 2 ) to enhance ultrasound imaging and increase the release of DOX. After intravenously injecting CaCO 3 -DOX to tumor-bearing mice, in the presence of an ultrasound field, CO 2 bubbles were sufficiently generated at the tumor tissues for echogenic reflectivity. Also, the indocyanine green (ICG) was encapsulated into CaCO 3 nanoparticles, to further detect the tumor with fluorescence. The resultant theranostic nanoparticle system exhibited therapeutic efficacy towards tumour-bearing mice. Overall, this investigation provides an attractive strategy for dual-mode cancer diagnostics., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We investigated effects of rearing cage type and dietary limestone particle size (LPS) on egg production, egg weight, eggshell, and bone quality in laying hens. The pullets were reared in conventional (CON; 20 chicks/cage, 270 cm 2 /chick) or furnished (FUR; 30 chicks/cage; 636 cm 2 /chick) cages and fed 3 LPS (fine, <0.595 mm; medium, 0.595 to <1.68 mm; and 1:1 mixture of F and M wt/wt) to 16 wk of age (woa). Pullets were transitioned to laying furnished cages and retained rearing treatment combination identities (n = 5, 20 hens/cage). Hens had free access to common commercial layer diet and water through to 40 woa. Eggs were recorded daily for calculation of hen day egg production (HDEP). Subsamples of eggs laid on the first day of 24, 28, 32, 36, and 40 woa were used for eggshell quality analyses. Two hens per cage were sacrificed on the last day of 24 and 40 woa for femur and tibia quality assessments. There was no interaction (P > 0.05) between rearing cage type and dietary LPS on response variables. At 19 and 20 woa, HDEP was higher (P < 0.01) for FUR than CON reared hens but was similar (P > 0.05) afterward. At 40 woa, FUR reared hens had higher (P < 0.05) body weight (BW), egg weight (EW), eggshell thickness, and eggshell weight and tended (P < 0.10) to have higher femur and tibia mineral density (BMD) and mineral content (BMC) than CON reared hens. Rearing dietary LPS had no effect (P > 0.05) on HDEP, BW, EW, and eggshell quality. Although, rearing dietary LPS did not affect (P > 0.05) femur and tibia BMD and BMC; at 24 woa, hens reared on medium LPS tended to have higher femur BMD (0.17 vs. 0.14 g/cm 2 ; P = 0.079) and BMC (0.99 vs.0.78 g; P = 0.088) than hens reared on fine LPS. In conclusion, hens reared in furnished cages had better eggshell quality but had marginal effects on femur and tibia quality, whereas rearing dietary LPS had no effects on eggshell and bone attributes in hens., (Copyright © 2020. Published by Elsevier Inc.)

In this study, the role of CaCO 3 in n-butanol production was further investigated using corn straw hydrolysate (CSH) media by Clostridium acetobutylicum CICC 8016. CaCO 3 addition stimulated sugars utilization and butanol production. Further study showed that calcium salts addition to CSH media led to the increase in Ca 2+ concentration both intracellularly and extracellularly. Interestingly, without calcium salts addition, intracellular Ca 2+ concentration in the synthetic P2 medium was much higher than that in the CSH medium despite the lower extracellular Ca 2+ concentrations in the P2 medium. These results indicated that without additional calcium salts, Ca 2+ uptake by C. acetobutylicum CICC 8016 in the CSH medium may be inhibited by non-sugar biomass degradation compounds, such as furans, phenolics and organic acids. Comparative proteomics analysis results showed that most enzymes involved in glycolysis, redox balance and amino acids metabolism were up-regulated with CaCO 3 addition. This study provides further insights into the role of CaCO 3 in n-butanol production using real biomass hydrolysate.

The hypothesis was tested that an increased digestion of coarse compared with fine limestone can alleviate the negative effects of a low dietary Ca/P ratio on the growth performance and characteristics of tibia strength (CTS) in broilers. A total of 1,152 Ross 308 broiler chickens received a standard commercial starter feed from day 0 to 13. From day 14 onward, birds received 1 of 12 diets containing 1 of 6 Ca/P ratios (0.50, 0.75, 1.00, 1.25, 1.50, and 1.75) and 1 of 2 limestone particle sizes (<500 [fine] and 500 to 2,000 [coarse] μm) in a study with a 6 × 2 factorial arrangement of treatments. Total P content was fixed at 5.5 g/kg for all treatment diets. Each treatment was replicated 6 times with 16 birds per replicate pen. On day 20 and 21, twelve birds per pen were randomly selected from 4 of the 6 replicate pens for tibia analysis and digesta collection from different gut segments. The apparent Ca digestibility was higher for fine than coarse limestone in the jejunum (P = 0.043). However, this difference in Ca digestibility disappeared for the low, whereas it remained for the high Ca/P ratios in the proximal (P interaction = 0.067) and distal (P interaction = 0.052) ileum. In addition, coarse limestone improved apparent P digestibility in the proximal and distal ileum (P < 0.001) but not in the jejunum (P = 0.305). Regardless of limestone particle size, reducing dietary Ca/P ratio linearly improved apparent Ca and P digestibility in the proximal and distal ileum (P < 0.001). Moreover, decreasing dietary Ca/P ratio linearly (P < 0.001) and quadratically (P < 0.046) reduced the CTS. Reducing dietary Ca/P ratio linearly (P < 0.003) and quadratically (P ≤ 0.006) decreased body weight gain and increased feed conversion ratio. For both fine and coarse limestone, the optimal Ca/P ratio was 1.00 to 1.25 to optimize apparent Ca and P digestibility while maintaining growth performance and CTS. Reducing Ca/P ratio from 1.75 to 1.00 improved distal ileal Ca and P apparent digestibility from 36.6 to 53.7% and 48.0 to 58.3%, respectively. In conclusion, coarse limestone is equally digestible with fine limestone at a low Ca/P ratio but is less digestible at a high Ca/P ratio, and the optimal Ca/P ratio in the diet is 1.00 to 1.25 for both fine and coarse limestone., (Copyright © 2020. Published by Elsevier Inc.)

Effects of rearing cage type and dietary limestone particle size (LPS) on growth, apparent retention (AR) of nutrients, and bone quality were investigated. The treatments were arranged in a 2 × 3 factorial with cage (conventional, CON and furnished, FUR) and LPS (fine, < 0.595 mm, F; medium, 0.595 to < 1.68 mm, M; and 1:1 mixture of F and M wt/wt; FM). A total of 900-day-old Lohmann LSL-Lite chicks were placed in CON (20 chicks/cage) and FUR (30 chicks/cage) based on BW. The diets were formulated according to breeder's nutrient specifications for starter, grower, and developer phases. At the end of 4, 12, and 16 wk of age (woa), 2 pullets/cage were euthanized for samples. At 12 and 16 woa, 1 pullet/cage was transferred to metabolism cages for AR measurements. There was no interaction (P > 0.05) between cage type and LPS on response variables. At 4 woa, body (P = 0.002) and bone (P < 0.05) weight was higher for CON than FUR pullets, but this was reversed (P < 0.01) at 16 woa. Pullets fed M LPS had higher (P < 0.05) AR of Ca, whole body mineral density (BoMD), and whole body mineral content (BoMC) than pullets fed F LPS. However, pullets fed F LPS had higher (P < 0.05) femur bone mineral density (BMD) and tended (P = 0.059) to have higher tibia bone breaking strength (BBS) than pullets fed M LPS at 16 woa. Pullets reared in CON cages had higher (P < 0.05) AR of Ca than FUR pullets. At 4 woa, CON pullets had lower (P < 0.05) femur and tibia BMD but higher tibia (93 vs. 83 N P = 0.012) BBS than FUR pullets. However, at 16 woa, FUR pullets had higher (P < 0.05) BoMD, BoMC, and tibia BBS than CON pullets. In conclusions, cage type and dietary LPS had independent effects on Ca utilization and skeletal development. Despite poor Ca retention, FUR caged pullets showed improved bone quality at 16 woa. Finer LPS improved femur mineral density suggesting coarser LPS had limited effects on pullet bone quality., (Copyright © 2020. Published by Elsevier Inc.)

Background: The calcium supplement is a clinically approved approach for osteoporosis therapy but usually requires a large dosage without targetability and with poor outcome. This modality is not fully explored in current osteoporosis therapy due to the lack of proper calcium supplement carrier. Methods: In this study, we constructed a tetracycline (Tc) modified and simvastatin (Sim) loaded phospholipid-amorphous calcium carbonate (ACC) hybrid nanoparticle (Tc/ACC/Sim). Results: The resulted Tc/ACC/Sim was able to enhance its accumulation at the osteoporosis site. Most importantly, the combination of calcium supplement and Sim offered synergetic osteoblast promotion therapy of osteoporosis with advanced performance than non-targeted system or mono therapy. Conclusion: This platform provides an alternative approach to stimulate bone formation by synergetic promotion of osteoblast differentiation using calcium supplement and Sim., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Calcite processed particles (CaPPs, Megagreen ® ) elaborated from sedimentary limestone rock, and finned by tribomecanic process were found to increase photosynthetic CO 2 fixation grapevines and stimulate growth of various cultured plants. Due to their processing, the CaPPs present a jagged shape with some invaginations below the micrometer size. We hypothesised that CaPPs could have a nanoparticle (NP)-like effects on plants. Our data show that CaPPs spontaneously induced reactive oxygen species (ROS) in liquid medium. These ROS could in turn induce well-known cellular events such as increase in cytosolic Ca 2+ , biotic ROS generation and activation of anion channels indicating that these CaPPs could activate various signalling pathways in a NP-like manner.

Several studies suggest the involvement of glutamatergic neurotransmission in obsessive-compulsive disorder (OCD). Some NMDA glutamatergic receptor antagonists, such as the general anesthetic ketamine, have shown anti-OCD effects in preclinical and clinical studies. Therefore, we investigated whether the inhalational anesthetics isoflurane and sevoflurane, which are general anesthetics acting as NMDA receptor antagonists, would induce the same effects. To test our hypothesis, adult male Swiss mice were exposed to different concentrations of isoflurane (0.5, 1.5 or 3 %) or sevoflurane (0.8, 2.5 or 4 %) for 20 min (short-time exposure) or 1 h (moderate-time exposure) and submitted to the open field test (OFT) and the marble-burying test (MBT) in the same day (acute effect) or 7 days (long-lasting effect) after anesthetics administration. We found that single short or moderate-time exposure to isoflurane or sevoflurane, at sub-anesthetic or anesthetic concentrations, did not affect marble-burying behavior acutely or even 7 days after their administration. The same treatment schedules with isoflurane or sevoflurane did not impair total distance travelled in the OFT. A single moderate-time exposure to isoflurane (3 %) reduced, acutely, the central exploration of the open field, suggesting an anxiogenic-like effect of isoflurane in mice. Our results suggest that isoflurane and sevoflurane may not be promising anti-compulsive drugs., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Inducing the osteogenic differentiation from bone marrow stromal cells (BMSCs) might be a potent strategy for treating bone loss and nonunion during fracture and improving fracture healing. Among several signaling pathways involved, mitogen-activated protein kinases (MAPKs) have been reported to play a critical role. Magnesium (Mg)-based alloys, including Mg-Zn alloy, have been used clinically as implants in the musculoskeletal field and could promote BMSC osteogenic differentiation. However, the underlying mechanisms remain unclear. In this study, we produced Mg-Zn alloy consists of Mg and low concentrations of Zn, calcium carbonate, and β-tricalcium phosphate (β-TCP; manifesting process not shown), prepared Mg, Zn, and Mg-Zn extracts, and investigated the specific effects of these extracts on human BMSC (hBMSC) osteogenic differentiation and MAPK signaling. Mg extracts and Mg-Zn extracts could significantly promote the osteogenic differentiation of hBMSCs as manifested as increased alkaline phosphatase levels, enhanced calcium nodules formation, and increased messenger RNA expression and protein levels of osteogenesis markers, including BMPs, Col-I, Runx2, and Osx; in the meantime, Mg culture medium (CM) and Mg-Zn CM both significantly enhanced the activation of MAPK signaling in hBMSCs. By adding ERK1/2 signaling, p38 signaling, or JNK signaling inhibitor to Mg-Zn CM, or conducting p38 MAPK silence in hBMSCs, we revealed that these extracts might promote hBMSC osteogenic differentiation via p38 MAPK signaling and MAPK-regulated Runx2/Osx. In conclusion, Mg 2+ in β-TCP/Mg-Zn extract promotes the osteogenic differentiation of hBMSCs via MAPK-regulated Runx2/Osx interaction., (© 2019 Wiley Periodicals, Inc.)

Calcium carbonate nanoparticles (CaCO 3 NPs) derived from CO 2 are promising materials for different industrial applications. It is imperative to understand their toxicological profile in biological systems as the human and environmental exposures to CaCO 3 NPs increases with growing production. Here, we analyse the cytotoxicity of CaCO 3 NPs synthesized from a CaO slurry on two cell lines, and in vivo on zebrafish (Danio Rerio). Our results demonstrate the CaCO 3 NPs in vitro safety as they do not cause cell death or genotoxicity. Moreover, zebrafish treated with CaCO 3 NPs develop without any abnormalities, confirming the safety and biocompatibility of this nanomaterial., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

There is a strong interest in cement additives that are able to prevent or mitigate the adverse effects of cracks in concrete that cause corrosion of the reinforcement. Inorganic polyphosphate (polyP), a natural polymer that is synthesized by bacteria, even those on cement/concrete, can increase the resistance of concrete to progressive damage from micro-cracking. Here we use a novel bioinspired strategy based on polyP-stabilized amorphous calcium carbonate (ACC) to give this material self-healing properties. Portland cement was supplemented with ACC nanoparticles which were stabilized with 10% ( w/w ) Na-polyP. Embedding these particles in the hydrated cement resulted in the formation of calcite crystals after a hardening time of 10 days, which were not seen in controls, indicating that the particles dissolve and then transform into calcite. While there was no significant repair in the controls without ACC, almost complete closure of the cracks was observed after a 10 days healing period in the ACC-supplemented samples. Nanoindentation measurements on the self-healed crack surfaces showed a similar or slightly higher elasticity at a lower hardness compared to non-cracked surfaces. Our results demonstrate that bioinspired approaches, like the use of polyP-stabilized ACC shown here, can significantly improve the repair capacity of Portland cement.

Lactobionic acid (LBA) was produced by fermentation of Pseudomonas taetrolens. First, to increase the production of LBA by P. taetrolens, we controlled the pH of culture medium by CaCO 3 addition (30 g/L) and then examined the initial lactose concentration ranging from 50 to 200 g/L and the growth temperature ranging from 20 to 37 °C. Both the LBA production titer (180 g/L) and the productivity (2.5 g/L h) were highest at 200 g/L lactose concentration and 25 °C of cell growth temperature in shake-flask culture. Although the production of LBA (178 g/L) was almost similar during the batch fermentation of P. taetrolens using 5 L bioreactor, the LBA productivity highly increased to 4.9 g/L h. The method using ethanol precipitation and ion-exchange chromatography was developed to recover the pure LBA from the fermentation broth. The optimum volume of ethanol and pH of culture medium for the precipitation of Ca 2+ salt form of LBA were six volume of ethanol and pH 6.5, respectively. The cation-exchange resin T42 finally showed the best recovery yield (97.6%) of LBA from the culture supernatant. The production titer (178 g/L) and the productivity (4.9 g/L h) of lactobionic acid in this study were highest among the previous studies ever reported using P. taetrolens as a production strain of LBA.

Evidence suggests that lipopolysaccharide (LPS) absorbed from the large intestine may contribute to the inflammatory response to high starch feeding in dairy cows. This work evaluated the impact of buffers or alkalinizing agents with expected large intestinal activity on faecal indicators of intestinal fermentation and LPS. Ten late-lactation cows were used in a replicated 5 × 5 Latin square design with 7-day periods. Cows were fed a diet containing 265 g/kg dry matter of starch and were abomasally infused with 1 g/kg body weight cornstarch daily. Treatments were control (CON), ration supplementation with 200 g/day sodium bicarbonate (FSB), 200 g/day calcium carbonate (FCC) or 125 g/day calcium carbonate plus 75 g/day of magnesium oxide (FCCM), or abomasal infusion of a lipid encapsulate providing 200 g/day sodium bicarbonate (ISB). The FCC, FCCM and ISB treatments were hypothesized to have large intestinal buffering effects, and FSB was included as a secondary control. Milk, feed, rumen and faecal samples were collected on day 7 of each period. Treatment did not affect intake, milk yield or milk composition. There were no effects of treatment on ruminal measures except that ISB tended to reduce and the post-ruminal treatments as a whole (FCC, FCCM and ISB) reduced rumen butyrate compared with CON. Faecal pH was greater for FCCM compared with all other treatments. Total faecal VFA tended to increase with FCC and FCCM compared with CON and was increased by the post-ruminal treatments as a whole compared with CON. Treatment did not affect faecal dry matter, lactate or LPS or apparent total tract nutrient digestibility. Although some treatments altered fermentation as evidenced by the change in faecal VFA, this was not accompanied by a decrease in faecal LPS. The strategies employed in this study had limited effects on large intestinal fermentation., (© 2020 Blackwell Verlag GmbH.)

Aims: This study aimed to explore whether pearl could help prevent cognitional morbidity and improve the metabolic processes of hippocampus., Methods: Rats were divided into group of control (CTL), sleep deprivation (SD) and pearl powder (PP). The sleeplessness was introduced to all rats except control. Before and after administration with vehicle or pearl powder, cognition was evaluated by Morris water maze (MWM). The protein expression in hippocampus among all groups was examined using iTRAQ-based global proteomic analysis., Results: Morris water maze tests revealed improvements of insomnia-induced cognitive deficit in both PP- and ES-treated rats, as compared to SD rats. However, proteomic analysis indicates that the pharmacological impact on gene expression of these two medicines is quite different: pearl is more capable of correcting aberrant gene expression caused by SD than estazolam. Therefore, pearl is more suitable for treatment of insomnia. These data, together with protein-protein interaction analysis, indicate that several pathways, affected by sleep deprivation, may be rescued by pearl powder: retrograde endocannabinoid signalling pathway, and the protein interaction or network enrich in oxidative phosphorylation Parkinson's disease and Huntington disease, etc CONCLUSIONS: Sleep deprivation can mimic cognition decline caused by insomnia with altered protein expression in the hippocampus; such behavioural and pathological changes can be significantly ameliorated by pearl powder., (© 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.)

Porous calcium carbonate (CaCO 3 ) particles have been shown to be highly advantageous for biological applications, mainly due to their large surface area and their stability in physiological media. Also, developing appropriate antibacterial materials presenting the benefits of non-formation of harmful compounds is of major interest. Two characteristics of CaCO 3 particles were investigated herein: (i) antibiotic-loading capacity and (ii) the possibility of using CaCO 3 particles as a template for the fabrication of biocapsules presenting inherent antibacterial capacity. The particles were tested against two representative pathogenic bacteria (Staphylococcus aureus and Escherichia coli). On one hand, a method for antibiotic (namely penicillin, ampicillin and ciprofloxacin) loading inside calcium carbonate particles was developed and antibacterial activity was investigated. Encapsulation efficiency and loading content were 95% and 5%, respectively. We showed that antibiotics prevented bacterial growth within 2 h, with no evidence of bacterial regrowth within 16 h; bactericidal effects were also observed. On the other hand, the self-assembly of charged polysaccharides, namely chitosan (chi + ) and dextran sulfate (dex - ), were assessed on calcium carbonate microparticles used as a sacrificial matrix. During bacterial growth in a liquid medium, an inhibitory effect of these particles was observed, i.e. Staphylococcus aureus (Gram-positive) (from 16.3% to 48.8% for (chi + /dex - ) n -chi + coated CaCO 3 materials and from 41.9% to 93.0% for (chi + /dex - ) n -chi + capsules) and Escherichia coli (Gram-negative) (from 18.2% to 45.5% for (chi + /dex - ) n -chi + coated CaCO 3 materials and from 40.0% to 89.1% for (chi + /dex - ) n -chi + capsules). Staining with acridine orange highlighted the bactericidal effect of the designed particles. These findings demonstrate the excellent potential of using calcium carbonate particles in antibiotic therapy as a starting point for the development of smart materials., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Bacteria that are resistant to high temperatures and alkaline environments are essential for the biological repair of damaged concrete. Alkaliphilic and halotolerant Bacillus sp. AK13 was isolated from the rhizosphere of Miscanthus sacchariflorus . Unlike other tested Bacillus species, the AK13 strain grows at pH 13 and withstands 11% (w/v) NaCl. Growth of the AK13 strain at elevated pH without urea promoted calcium carbonate (CaCO 3 ) formation. Irregular vateritelike CaCO 3 minerals that were tightly attached to cells were observed using field-emission scanning electron microscopy. Energy-dispersive X-ray spectrometry, confocal laser scanning microscopy, and X-ray diffraction analyses confirmed the presence of CaCO 3 around the cell. Isotope ration mass spectrometry analysis confirmed that the majority of CO 3 2- ions in the CaCO 3 were produced by cellular respiration rather than being derived from atmospheric carbon dioxide. The minerals produced from calcium acetate-added growth medium formed smaller crystals than those formed in calcium lactate-added medium. Strain AK13 appears to heal cracks on mortar specimens when applied as a pelletized spore powder. Alkaliphilic Bacillus sp. AK13 is a promising candidate for self-healing agents in concrete.

Background and Objective: Climate change affecting on weather in many different ways especially drought and temperature rise. This will drastically down plant production, if not start looking for another source to increase water productivity to cope up with water stress conditions. In this study efforts were conducted to interpret the use of anti-transpirants to conserving irrigation water, aiding plant survival under dry conditions and protecting plant against drought stress., Materials and Methods: Two field experiments were carried out during 2017 and 2018 successive growing summer seasons at the experimental farm of National Research Centre, Nubaria, El-Beheira Governorate, Egypt. Anti-transparent ,i.e., chitosan (100 and 150 mg L-1), calcium carbonate (5 and 10 g L-1), salicylic acid (200 and 300 mg L-1) were foliar sprayed on sunflower plants grown under two water levels (normal 100% and deficit 50%)., Results: The results showed that water stress decreased growth criteria, photosynthetic pigments, osmoprotectants, yield components, oil and carbohydrate (%) as compared to 100% of irrigation requirements. Meanwhile, water deficit induced significant increases in (proline). Foliar treatments of sunflower plant with chitosan, calcium carbonate, salicylic acid increased growth criteria, yield components, photosynthetic pigments, total soluble sugars, proline and free amino acid as compared to control plant. Data also illustrated that, all used treatment improved seed yield, oil and carbohydrate% of sunflower plants., Conclusion: Generally, it could be concluded that 10 g L-1 CaCO3 and 300 mg L-1 SA at 50% level of water irrigation could be recommended for sunflower plant grown under similar field conditions in order to get optimum yield and to save irrigation water.

Ovary fragments from six sexually mature cats were vitrified in the presence or absence of betaine or ascorbic acid, loaded (7.4 or 74µM betaine; 20 or 200µM ascorbic acid) or not (1mM betaine or 0.3mM ascorbic acid) into CaCO3 microparticles, and assessed for follicular morphology, oxidative stress and mitochondrial activity Feline ovarian tissue was successfully preserved after vitrification in the presence of 74µM betaine loaded in CaCO3 microparticles, as confirmed by morphological analysis and the density of preantral follicles and stromal cells, as well as by the increased mitochondrial activity and decreased production of reactive oxygen species.

Based on the concept of starving tumor therapy, in this study we put forward a new idea that the pH-sensitive Ca 2+ delivery of calcium carbonate nanoparticles (CaCO 3 NPs) induced blood coagulation of tumor vessels, and first explored the effect of CaCO 3 NPs on the in vitro and in vivo blood coagulation by acid stimulus. CaCO 3 NPs with a size of about 100 nm and a porous structure of several nanometers were synthesized in an emulsion system, which showed a high loading capacity (49%) of doxorubicin hydrochloride (DOX) with an encapsulation efficiency of 98% and a pH-sensitive drug delivery. The hemolysis test showed that CaCO 3 NPs were blood compatible. The in vitro Ca 2+ delivery and blood clotting tests indicated that CaCO 3 NPs pH-sensitively released Ca 2+ , and caused rapid blood coagulation at pH 5.0 but no thrombus at pH 7.4. Confocal laser scanning microscopy showed that after uptake by MCF-7 or MDA-MB-231 breast cancer cells, CaCO 3 NPs mainly distributed in endosomes/lysosomes within the initial 2 h and then decomposed by acid stimulus, leading to the intracellular delivery of Ca 2+ that subsequently migrated outside the cells. CaCO 3 NPs were nontoxic to NIH3T3 mouse fibroblasts, but highly toxic to both MCF-7 and MDA-MB-231 cells after loading DOX. After topical administration into the breast tumors of mice, CaCO 3 NPs evoked significant thrombosis and hemorrhage of tumor vasculature by hematoxylin-eosin and Masson's trichrome staining. These results indicated that CaCO 3 NPs could induce blood coagulation via acid stimulus, showing potential applications in blocking tumor vessels for starving tumor therapy.

Background: Both falls and fractures are increased in older persons living with HIV (PLWH). Low serum total 25-hydroxyvitamin D (25-OHD) levels have been associated with falls, fractures and poor muscle strength. We hypothesized that vitamin D (VitD) supplementation would improve muscle strength in postmenopausal PLWH., Methods: In a 12-month prospective, randomized, double-blind, study of 69 African American and Hispanic postmenopausal PLWH on antiretroviral therapy with 25-OHD ≥10 ng/ml and ≤32 ng/ml, we investigated the effects of daily low (1,000 IU; n=31) and moderate (3,000 IU; n=38) cholecalciferol doses on lean mass and strength. Change in lean body mass was assessed by dual-energy X-ray absorptiometry (DXA), and isometric and isokinetic muscle strength in the dominant lower extremity was assessed using the Biodex System 4 Pro., Results: Mean age was 56 ±5 years, median CD4 + T-cell count 722 cells/mm 3 and 74% had HIV RNA≤50 copies/ml. Serum 25-OHD did not differ at baseline, but was higher in the moderate than low VitD group at 6 and 12 months. In both groups, there were significant increases in lower extremity isokinetic torque, work and power at 12 months, with no change in lean mass., Conclusions: VitD supplementation was associated with a modest increase in lower extremity strength in postmenopausal PLWH, without a concomitant increase in muscle mass. Magnitude of increase in strength were similar with 3,000 IU and 1,000 IU daily. Future larger studies will be required to determine the optimal dose of VitD to improve muscle strength and to determine whether supplementation reduces the risk of falls and fractures in PLWH.

Polymeric coatings with positive surface charge offer potential antimicrobial activity, which they owe to a simple electrostatic attraction with negatively charged bacterial walls and membranes. We describe synthesis and characterization of poly(2-aminoethyl methacrylate) and its copolymers with methyl methacrylate and butyl acrylate, as potential binders for antimicrobial solvent-cast paints. TiO 2 and CaCO 3 mineral particles were employed as model pigments/fillers, as they are used in most real-life paint formulations. Electrokinetic (ζ) potential and antimicrobial activity of thin films made of the (co)polymers in the absence and presence of TiO 2 and CaCO 3 nanopowders were assessed using streaming current measurements and microbial growth inhibition tests, respectively. Independently of the structure of the monomers used for the synthesis, the films showed positive ζ-potential values (up to +95 mV) in the pH range 3.5-8.0. The presence of mineral particles at 50% dry weight of the films did not affect significantly the ζ(pH) curves. The films made of the mixed dispersions remained positively charged and inhibited growth of both Gram-negative (E. coli) and Gram-positive (S. aureus) bacteria, as well as yeast (C. albicans). The mixed polymeric-mineral films described in this study seem to be promising potential candidates for designing antimicrobial coatings aimed to prevent spreading of bacterial infections., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Introduction: Recent clinical studies demonstrated the favorable effects of calcium-free phosphate binders on mortality and vascular calcification in hemodialysis (HD) patients. The aim of the present study was to investigate the effects of a calcium-free phosphate binder, lanthanum carbonate (LC), on bone metabolic markers and bone mineral density (BMD), compared with those of calcium carbonate (CC), in subjects new to HD., Materials and Methods: The present study included 65 subjects from our previous randomized controlled trial (LC group, N = 31; CC group, N = 34). We investigated the effects of LC on serum intact parathyroid hormone (iPTH), osteocalcin (OC), bone-specific alkaline phosphatase (BAP), tartrate-resistant acid phosphatase 5b (TRACP-5b), sclerostin levels, and BMD, compared with those of CC in patients new to HD at baseline and at 12 and 18 months., Results: Serum OC levels at 18 months were significantly higher in the LC group than in the CC group. During the study period, serum BAP and TRACP-5b and iPTH levels tended to be higher in the LC group than in the CC group. At 18 months, the percentage of low bone turnover, based on a serum BAP cutoff value, was significantly lower in the LC group than in the CC group. There were no significant differences in the lumbar and femoral BMD between the two groups., Conclusions: The results of the present study suggest that LC has potential in preventing low bone turnover, in comparison to CC, in patients new to HD.

Dihydroartemisinin (DHA) has attracted increasing attention as an anticancer agent. However, using DHA to treat cancer usually depends on the synergistic effects of exogenous components, and the loss of DHA during delivery reduces its effectiveness in cancer therapy. Reported herein is a programmed release nanoplatform of DHA to synergistically treat cancer with a Fe-TCPP [(4,4,4,4-(porphine-5,10,15,20-tetrayl) tetrakis(benzoic acid)] NMOF (nanoscale MOF) having a CaCO 3 mineralized coating, which prevents DHA leakage during transport in the bloodstream. When the nanoplatform arrives at the tumor site, the weakly acidic microenvironment and high concentration of glutathione (GSH) trigger DHA release and TCPP activation, enabling the synergistic Fe 2+ -DHA-mediated chemodynamic therapy, Ca 2+ -DHA-mediated oncosis therapy, and TCPP-mediated photodynamic therapy. In vivo experiments demonstrated that the nanoplatform showed enhanced anticancer efficiency and negligible toxicity., (© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The present work reports the very first hydrothermal synthesis of 100% triclinic phase pure aragonite (A1) with microdumbbell microstructural architecture and Au Nanoparticle-decorated (AuNP-decorated) aragonites (A2, A3 and A4) with spherical, pentagonal/hexagonal and agglomerated AuNP-decorated microdumbbells having triclinic aragonite phase as the major and cubic AuNPs as the minor phase. Even in dark the AuNP-decorated aragonites (especially A2) show efficacies as high 90% against gram-negative e.g., Pseudomonas putida (P. putida) bacteria. Further the AuNP-decorated aragonites (A3) show anti-biofilm capability of as high as about 20% against P. putida. Most importantly the AuNP-decorated aragonites (A3) offer anti-cancer efficacy of as high as 53% while those of A1, A2, and A4 are e.g., 26%, 46% and 37%, respectively. For the very first time, based on detailed investigations, the mechanisms behind such advance antibiofilm and anticancer activities are linked to the generation of excess labile toxic reactive oxygen species (ROS). Thus, these materials show enormous potential as futuristic, multi-functional biomaterials for anti-bacterial, anti-biofilm and anti-cancer applications., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Purpose: Lusutrombopag is a novel, orally active thrombopoietin receptor agonist. This report describes 3 studies aimed at assessing the effects of food and calcium carbonate on the pharmacokinetic parameters of lusutrombopag in healthy subjects., Methods: Three single-dose, open-label crossover studies were conducted. In study 1, eighteen healthy subjects were administered a single 2-mg dose of lusutrombopag as a single tablet in the fasted or fed state or as a 2-mg solution in the fasted state. In study 2, fifteen healthy subjects were administered a single 0.75-mg dose of lusutrombopag as three 0.25-mg tablets in the fasted or fed state, or in the fasted state with coadministration of 4000-mg calcium carbonate. In study 3, fifteen healthy subjects were administered 4-mg lusutrombopag as a single tablet in the fasted or fed state. Pharmacokinetic parameters were estimated from plasma lusutrombopag concentrations., Findings: Mean fed versus fasted state ratios (90% CIs) of C max and AUC 0-∞ , respectively, were: 0.904 (0.864-0.945) and 0.920 (0.886-0.956) (study 1); 0.972 (0.864-1.09) and 1.02 (0.945-1.11) (study 2); and 0.917 (0.842-0.999) and 0.908 (0.855-0.964) (study 3). The respective ratios for calcium carbonate versus no calcium carbonate (fasted state) were 1.08 (0.959-1.21) and 0.989 (0.913-1.07) (study 2). Lusutrombopag exposure remained unaffected, except for a slight decrease in exposure with food. Lusutrombopag exposure did not change with the coadministration of calcium carbonate. These findings suggest that there was no clinically significant effect of food or calcium carbonate on the bioavailability of lusutrombopag. Each treatment regimen was well tolerated., Implications: According to the present findings, no specific restrictions are required for lusutrombopag administration with regard to meals (including those with dairy products), mineral supplements, or coadministration of antacids., Clinical Trial Registration: JapicCTI-No.: JapicCTI-194690, JapicCTI-194689. ClinicalTrials.gov identifier: NCT03897413., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)

Doxorubicin (DOX) is an effective and commonly used anthracycline anticancer drug for the treatment of osteosarcoma (OS). However, its antitumor effect is hampered by the nonspecific distribution and significant adverse effects. Nanoparticles based drug delivery systems are promising approaches to maximize the anticancer efficacy while decrease the side effects. In this study, biogenic aragonite nanoparticles (ANPs) were developed from cockle shells and loaded with DOX. An orthotopic rat OS model was induced by UMR-106 cells tibia cavity injection. The anticancer efficacy study included five groups: normal control group, OS model group, free DOX group (2 mg/kg), DOX-ANPs 1 group (2 mg of equivalent DOX/kg) and DOX-ANPs 2 group (1.5 mg of equivalent DOX/kg). This study demonstrates that the DOX-ANPs treatment groups can significantly reduce the tumor volume and increase the surviving ratio as compared to the OS model group. In addition, these two DOX-ANPs groups showed less toxicity to the normal organs compared to the free DOX group. Furthermore, DOX-ANPs 2 group showed the similar anticancer efficacy as DOX-ANPs 1 group, which suggested that DOX loaded onto the ANPs may allow the reduction of chemotherapy doses. These results highlight the promising application of ANPs derived from cockle shells as an effective drug delivery system for a successful chemotherapy against OS. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 1898-1907, 2019., (© 2018 Wiley Periodicals, Inc.)

Foliar fungicide application in grapevines increases the content of heavy metals such as copper (Cu) in vineyard soils, which may reach phytotoxic levels. The application of soil amendments such as limestone and vermicompost may reduce Cu availability and phytotoxicity. The study aimed to assess growth, physiological status and changes in root morphology in young grapevines grown for 12 months in Cu-contaminated soil with and without the application of soil amendments. Samples of a Typic Hapludalf soil were collected in a vineyard with more than 30 years of cultivation. The following treatments were used: 1) control (without amendment), 2) application of organic vermicompost (86.7 g kg -1 ) and 3) application of limestone (3 Mg ha -1 ). Grapevines (Paulsen 1103 rootstock) were transplanted and grown for 12 months in PVC soil columns. We assessed parameters of growth, photosynthesis and root morphology. Grapevines grown in soil treated with limestone showed increased growth, dry matter yield and photosynthetic efficiency. The highest Cu concentrations in root tissue were found in grapevines grown in control soil. The application of vermicompost in this study did not alleviate Cu toxicity. Grapevines grown in soil treated with vermicompost showed high manganese (Mn) concentration in shoots before the winter pruning, reflecting the high Mn concentrations in soil solution that caused Mn phytotoxicity, resulting in plant death after the winter pruning. The vermicompost used in this study is not suitable for agronomic use. The use of limestone was an effective strategy to reduce Cu availability and phytotoxicity., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Two clinical studies were performed in healthy volunteers to investigate food and antacid effects on lesinurad, a novel selective uric acid reabsorption inhibitor approved for treatment of hyperuricemia associated with gout in combination with xanthine oxidase inhibitors. Study 1 evaluated a high-fat, high-calorie meal or high doses of antacids (3000 mg calcium carbonate or 1600 mg magnesium hydroxide/1600 mg aluminum hydroxide) on the pharmacokinetics (PK) and pharmacodynamics (PD) of 400 mg oral lesinurad. Study 2 evaluated low doses of antacids (1250 mg calcium carbonate or 800 mg magnesium hydroxide/800 mg aluminum hydroxide) on the PK and PD of 400 mg lesinurad. Food did not alter the plasma AUC of lesinurad and only reduced its C max by 18%. In the fasted conditions, high-dose calcium carbonate reduced the C max and AUC of lesinurad by 54% and 38%, respectively, whereas high-dose magnesium hydroxide/aluminum hydroxide reduced C max and AUC by 36% and 31%, respectively. Food enhanced the maximum serum urate (sUA)-lowering effect of lesinurad by approximately 20% despite reducing the C max of lesinurad. High-dose calcium carbonate decreased the urate-lowering effect approximately 20% in the first 6 hours, whereas high-dose magnesium hydroxide/aluminum hydroxide reduced the effect by 26%. Low-dose calcium carbonate or magnesium hydroxide/aluminum hydroxide in the presence of food did not significantly affect plasma lesinurad C max and AUC or the sUA lowering and renal handling of uric acid. In summary, study results suggest food did not meaningfully alter lesinurad PK and PD. High doses of antacids reduced lesinurad AUC up to 40% and reduced the lesinurad uric acid-lowering effect., (© 2019, The American College of Clinical Pharmacology.)

Elaboration of novel biocomposites providing simultaneously both biodegradability and stimulated bone tissue repair is essential for regenerative medicine. In particular, piezoelectric biocomposites are attractive because of a possibility to electrically stimulate cell response. In the present study, novel CaCO 3 -mineralized piezoelectric biodegradable scaffolds based on two polymers, poly[( R)3-hydroxybutyrate] (PHB) and poly[3-hydroxybutyrate- co-3-hydroxyvalerate] (PHBV), are presented. Mineralization of the scaffold surface is carried out by the in situ synthesis of CaCO 3 in the vaterite and calcite polymorphs using ultrasound (U/S). Comparative characterization of PHB and PHBV scaffolds demonstrated an impact of the porosity and surface charge on the mineralization in a dynamic mechanical system, as no essential distinction was observed in wettability, structure, and surface chemical compositions. A significantly higher (4.3 times) piezoelectric charge and a higher porosity (∼15%) lead to a more homogenous CaCO 3 growth in 3-D fibrous structures and result in a two times higher relative mass increase for PHB scaffolds compared to that for PHBV. This also increases the local ion concentration incurred upon mineralization under U/S-generated dynamic mechanical conditions. The modification of the wettability for PHB and PHBV scaffolds from hydrophobic (nonmineralized fibers) to superhydrophilic (mineralized fibers) led to a pronounced apatite-forming behavior of scaffolds in a simulated body fluid. In turn, this results in the formation of a dense monolayer of well-distributed and proliferated osteoblast cells along the fibers. CaCO 3 -mineralized PHBV surfaces had a higher osteoblast cell adhesion and proliferation assigned to a higher amount of CaCO 3 on the surface compared to that on PHB scaffolds, as incurred from micro-computed tomography (μCT). Importantly, a cell viability study confirmed biocompatibility of all the scaffolds. Thus, hybrid biocomposites based on the piezoelectric PHB polymers represent an effective scaffold platform functionalized by an inorganic phase and stimulating the growth of the bone tissue.

Transdermal administration via skin appendages enables both localized and systemic drug delivery, as well as minimizes incidental toxicity. However, the design of an appropriate effective method for clinical use remains challenging. Here, we introduce calcium carbonate-based carriers for the transdermal transportation of bioactive substances. The proposed system presents easily manufacturable biodegradable particles with a large surface area enabling a high payload ability. Topical application of submicron porous CaCO 3 particles in rats followed by the therapeutic ultrasound treatment results in their deep penetration through the skin along with plentiful filling of the hair follicles. Exploiting the loading capacity of the porous particles, we demonstrate efficient transportation of a fluorescent marker along the entire depth of the hair follicle down the bulb region. In vivo monitoring of the carrier degradation reveals the active dissolution/recrystallization of CaCO 3 particles, resulting in their total resorption within 12 days. The proposed particulate system serves as an intrafollicular depot for drug storage and prolonged in situ release over this period. The urinary excretion profile proves the systemic absorption of the fluorescent marker. Hence, the elaborated transdermal delivery system looks promising for medical applications. The drug delivery to different target regions of the hair follicle may contribute to regenerative medicine, immunomodulation, and treatment of various skin disorders. In the meantime, the systemic uptake of the transported drug opens an avenue for prospective delivery routes beyond the scope of dermatology.

Carbonate apatite (CO 3 Ap) block can be used as a bone substitute because it can be remodeled to new natural bone in a manner conforming with the bone remodeling process. Among the many porous structures available, honeycomb (HC) structure is advantageous for rapid replacement of CO 3 Ap to bone. In this study, the feasibility to fabricate a CO 3 Ap HC was studied, along with its initial tissue response in rabbit femur bone defect. First, a mixture of Ca(OH) 2 and a wax-based binder was extruded from a HC mold. Then the fabricated HC was heated for binder removal and carbonation at 450°C in a mixed O 2 -CO 2 atmosphere, forming a CaCO 3 HC. When the CaCO 3 HC was immersed in 1 mol/L Na 3 PO 4 solution at 80°C for 7 days, its composition changed from CaCO 3 to CO 3 Ap, maintaining the structure of the original CaCO 3 HC. Compressive strengths of the CaCO 3 and CO 3 Ap HCs were 65.2 ± 7.4 MPa and 88.7 ± 4.7 MPa, respectively. When the rabbit femur bone defect was reconstructed with the CO 3 Ap HC, new bone penetrated the CO 3 Ap HC completely. Osteoclasts and osteoblasts were found on the surface of the newly formed bone and osteocytes were also found in the newly formed bone, indicating ongoing bone remodeling. Furthermore, blood vessels were formed inside the pores of CO 3 Ap HC. Therefore, CO 3 Ap HC has good potential as an ideal bone substitute. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1014-1020, 2019., (© 2019 Wiley Periodicals, Inc.)

Purpose: To evaluate the effect on dentin of chondroitin sulfate and L-arginine on dentin tubule occlusion., Methods: The dentin samples were activated by submersion in an aqueous ( aq. ) solution of chondroitin sulfate ( ChS) or L-arginine prior to application of a commercial or custom-made toothpaste. After rinsing with water and ultrasonication, adhesion to dentin and occlusion of dentin tubules were evaluated by scanning electron microscopy and the elemental composition of the deposits was evaluated by energy-dispersive x-ray spectroscopy., Results: Rinsing a dentin sample with a solution of ChS resulted in an increase in the adherence of dentifrices containing either titanium dioxide (TiO₂ ) or calcium-based nanoparticles [ hydroxyapatite ( HA\ or calcium carbonate( to the dentin surface. ChS does not appear to enhance the adherence of dentifrices lacking TiO₂. Pretreatment by L-arginine improved adherence of calcium carbonate nanoparticles, but less efficiently than ChS. Addition of nanoparticles of hydroxyapatite or calcium citrate to dentifrices improved their adherence to dentin without any pre-treatment., Clinical Significance: The significant increase in adherence to the dentin surface of dentifrices of either TiO₂ or calcium-supplying nanoparticles to the dentin surface following pre-treatment with ChS or L-arginine opens the door to the development of two-step dental treatments, which accomplish dentin tubule occlusion and help to deliver active dentifrice components to the dentin surface. The ability of the aqueous pastes of nanoparticles of hydroxyapatite or calcium citrate to occlude dentin tubules enables the formulation of desensitizing dentifrices, which also supply the mineral and organic nutrients to the tooth surface., Competing Interests: The authors declared no conflict of interest. The USD Chemistry Department, the NSF-MRI grant CHE-1337707 (SEM/EDS instruments) and the South Dakota Surface Engineering Research Center provided financial support of this work., (Copyright©American Journal of Dentistry.)

Conventional photodynamic therapy (PDT) is limited by its penetration depth due to the photosensitizer and light source. In this study, we developed X-ray induced photodynamic therapy that applied X-ray as the light source to activate Ce-doped CaCO₃ (CaCO₃:Ce) to generate an intracellular reactive oxygen species (ROS) for killing cancer cells. The A549 cell line was used as the in vitro and in vivo model to evaluate the efficacy of X-ray-induced CaCO₃:Ce. The cell viability significantly decreased and cell cytotoxicity obviously increased with CaCO₃:Ce exposure under X-ray irradiation, which is less harmful than radiotherapy in tumor treatment. CaCO₃:Ce produced significant ROS under X-ray irradiation and promoted A549 cancer cell death. CaCO₃:Ce can enhance the efficacy of X-ray induced PDT, and tumor growth was inhibited in vivo. The blood analysis and hematoxylin and eosin stain (H & E) stain fully supported the safety of the treatment. The mechanisms underlying ROS and CO₂ generation by CaCO₃:Ce activated by X-ray irradiation to induce cell toxicity, thereby inhibiting tumor growth, is discussed. These findings and advances are of great importance in providing a novel therapeutic approach as an alternative tumor treatment.

Context: Sex-specific effects of pregnancy calcium carbonate supplementation have been reported in 8-12 year old Gambian children, indicating faster growth in boys but slower growth in girls born to calcium-supplemented mothers., Objective: To determine whether the pregnancy supplement resulted in sex-specific effects on offspring IGF1 and other growth-related indices in mid-childhood., Design: Analysis of archived data obtained in mid-childhood from the children of rural Gambian mothers who had been randomised to 1500 mgCa/d (Ca) or placebo (P) from 20 weeks pregnancy to delivery (ISRCTN96502494)., Participants and Methods: Of the 526 children born and followed in infancy, 290 had early-morning, fasting plasma assayed for IGF1, IGFBP3, leptin, insulin and calcium-related indices and had anthropometry performed at age 7.5 (SD1.2) years (N/group: Males(M)-Ca = 64, Females(F)-Ca = 77; M-P = 76, F-P = 73). Sex-specific effects of maternal supplementation were considered using regression with sexes separated and together to test for sex ∗ supplement interactions., Results: Boys had lower IGF1, IGFBP3, leptin and insulin than girls (P ≤ 0.004). IGF1 was higher in M-Ca than M-P (+14.2 (SE7.7)%, P = 0.05) but lower in F-Ca than F-P (-17.8 (SE7.4)%, P = 0.01); sex ∗ supplement interaction P = 0.001. IGF1 concentrations (ng/ml, geometric mean [-1SE,+1SE]) were M-Ca = 78.1[4.3,4.5], M-P = 67.8[3.4,3.6]; F-Ca = 99.5[4.8,5.1], F-P = 118.9[6.4,6.8]. Similar sex ∗ supplement interactions were seen for IGFBP3 and IGF1-adjusted-for-IGFBP3 but group differences were smaller. There were no significant supplement effects on the other biochemical indices., Conclusions: Calcium carbonate supplementation of pregnant Gambian mothers resulted in higher IGF1 in boys and lower IGF1 in girls during mid-childhood, consistent with the reported maternal supplement effects on growth of the offspring in later childhood., (Copyright © 2018. Published by Elsevier Inc.)

Rodent marble-burying behavior in the marble-burying test (MBT) is employed as a model or measure to study anxiety- and compulsive-like behaviors or anxiolytic and anticompulsive drug action. However, the test responds variably to a range of pharmacological interventions, and little consensus exists regarding specific methodologies for its execution. Regardless, the test is widely applied to investigate the effects of pharmacological, genetic, and behavioral manipulations on purported behaviors related to the said neuropsychiatric constructs. Therefore, in the present review we attempt to expound the collective translational significance of the MBT. We do this by (1) reviewing burying behavior as a natural behavioral phenotype, (2) highlighting key aspects of anxiety and obsessive-compulsive disorder from a translational perspective, (3) reviewing the history and proof of concept of the MBT, (4) critically appraising potential methodological confounds in execution of the MBT, and (5) dissecting responses of the MBT to various pharmacological interventions. We conclude by underlining that the collective translational value of the MBT will be strengthened by contextually valid experimental designs and objective reporting of data.

Aim: Hyperphosphataemia is associated with increased mortality and morbidity in end stage renal disease. Despite phosphate binder therapy, a large proportion of patients do not reach the treatment target. In five contemporary binders we explored the influence of pH and phosphate concentration on phosphate binding. This interaction could be of relevance in clinical practice., Methods: Phosphate binding was quantified in vitro in 25 mL of purified water containing phosphate concentrations of 10, 15 and 20 mM and baseline pH values of 3.0 or 6.0, with a binder over 6 h. Lanthanum carbonate, calcium acetate/magnesium carbonate, sevelamer carbonate, calcium carbonate and sucroferric oxyhydroxide, 67 mg of each, were used. The experiments were performed in duplicate. The primary outcome was the difference in the amount of bound phosphate for each binder after 6 h in solutions at two different pH values. Secondary outcomes were the influence of phosphate concentration on phosphate binding, next to binding patterns and phosphate binder saturation., Results and Conclusion: In this specific in vitro setting, lanthanum carbonate, sevelamer carbonate, calcium carbonate and sucroferric oxyhydroxide bound more phosphate in the solution with baseline pH of 3.0. Differences however were small except for lanthanum carbonate. Calcium acetate/magnesium carbonate was most effective in a solution with baseline pH of 6.0. All phosphate binders bound more phosphate in solutions with higher concentrations of phosphate. Sevelamer carbonate, calcium acetate/magnesium carbonate and sucroferric oxyhydroxide bound most phosphate in the first hour and reached maximum binding capacity in less than 6 h., (© 2018 The Authors Nephrology published by John Wiley & Sons Australia, Ltd on behalf of Asian Pacific Society of Nephrology.)

Carbonate apatites are sought as a bone substitute due to their biocompatibility and excellent resorbability. The present study deals with Cowrie's shell derived powder (CSDP) as natural biomaterial for bone regenerative medicine. Structural and physicochemical analysis showed that Cowrie's shells, presenting brick and mortar microstructures, were mainly composed of aragonite crystals, which were converted into poorly crystalline B-type carbonate apatite once soaked, at 37 °C, in simulated body fluid for 7 days, reflecting bioactive features. Cytotoxic assays showed that CSDP boosted human stem cell proliferation over the study time compared to nacre derived powder (NDP), used as positive control. Human stem cells adopted a flattened morphology and established physical contact with CSDP, signature of a good biocompatibility. Thus, these results suggested that CSDP presents a great interest for bone regenerative medicine, and could be a useful and versatile carrier/scaffold for bone tissue engineering or a raw material for 3D printed orthopedic devices., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Aim: CaCO 3 nanoparticles (nano-CaCO 3 ) can neutralize the acidic pHe of solid tumors, but the lack of intrinsic imaging signal precludes noninvasive monitoring of pH-perturbation in tumor microenvironment. We aim to develop a theranostic version of nano-CaCO 3 to noninvasively monitor pH modulation and subsequent tumor response., Materials & Methods: We synthesized ferromagnetic core coated with CaCO 3 (magnetite CaCO 3 ). Magnetic resonance imaging (MRI) was used to determine the biodistribution and pH modulation using murine fibrosarcoma and breast cancer models., Results: Magnetite CaCO 3 -MRI imaging showed that nano-CaCO 3 rapidly raised tumor pHe, followed by excessive tumor-associated acid production after its clearance. Continuous nano-CaCO 3 infusion could inhibit metastasis., Conclusion: Nano-CaCO 3 exposure induces tumor metabolic reprogramming that could account for the failure of previous intermittent pH-modulation strategies to achieve sustainable therapeutic effect.