Calcium Isotope Ratios Research Articles

A Comparative Study of Cave System Calcium Isotope Ratios: Implications for Quantitative Reconstruction of Paleorainfall From Speleothems

AbstractVariations in speleothem calcium isotope ratios (δ44Ca) are thought to be uniquely controlled by prior carbonate precipitation (PCP) above a drip site and, when calibrated with modern data, show promise as a semi‐quantitative proxy for paleorainfall. However, few monitoring studies have focused on δ44Ca in modern cave systems. We present a multi‐year comparative study of δ44Ca, carbon isotopes (δ13C), and trace elemental ratios from cave drip waters, modern calcite, and host rocks from two cave systems in California—White Moon Cave (WMC) and Lake Shasta Caverns (LSC). Drip water and calcite δ44Ca from both caves indicate PCP‐driven enrichment, and we used a simple Rayleigh fractionation model to quantify PCP variability over the monitoring period. Modern calcite trace element and δ44Ca data positively correlate at WMC, but not at LSC, indicating a shared PCP control on these proxies at WMC but not at LSC. At both WMC and LSC, we observe an inverse relationship between PCP and rainfall amounts, though this relationship is variable across individual drip sites. Our modeled data suggest that WMC experiences ∼20% more PCP than LSC, consistent with the fact that WMC receives less annual rainfall. This work supports speleothem δ44Ca as an independent constraint on PCP that can aid in the interpretation of other hydrologically sensitive proxies and provide quantitative estimates of paleorainfall. Additional, long‐term monitoring studies from a variety of climate settings will be key for understanding δ44Ca variability in cave systems more fully and better constraining the relationship between PCP and rainfall.

Changes in bone and mineral homeostasis after short-term androgen deprivation therapy with or without androgen receptor signalling inhibitor – substudy of a single-centre, double blind, randomised, placebo-controlled phase 2 trial

Prostate cancer (PCa) patients treated with androgen deprivation therapy (ADT) have an increased fracture risk. Exploring biomarkers for early bone loss detection is of great interest. Pre-planned substudy of the ARNEO-trial (NCT03080116): a double blind, randomised, placebo-controlled phase 2 trial performed in high-risk PCa patients without bone metastases between March 2019 and April 2021. Patients were 1:1 randomised to treatment with gonadotropin-releasing hormone antagonist (degarelix)+androgen receptor signalling inhibitor (ARSI; apalutamide) versus degarelix+matching placebo for 12 weeks prior to prostatectomy. Before and following ADT, serum and 24-h urinary samples were collected. Primary endpoints were changes in calcium-phosphate homeostasis and bone biomarkers. Of the 89 randomised patients, 43 in the degarelix+apalutamide and 44 patients in the degarelix+placebo group were included in this substudy. Serum corrected calcium levels increased similarly in both treatment arms (mean difference+0.04mmol/L, 95% confidence interval, 0.02; 0.06), and parathyroid hormone and 1,25-dihydroxyvitamin D3 levels decreased. Bone resorption markers increased, and stable calcium isotope ratios reflecting net bone mineral balance decreased in serum and urine similarly in both groups. This exploratory substudy suggests that 12 weeks of ADT in non-metastatic PCa patients results in early bone loss. Additional treatment with ARSI does not seem to more negatively influence bone loss in the early phase. Future studies should address if these early biomarkers are able to predict fracture risk, and can be implemented in clinical practice for follow-up of bone health in PCa patients under ADT. Research Foundation Flanders; KU Leuven; University-Hospitals-Leuven.

Calcimimetic AMG-416 induced short-term changes in calcium concentrations and calcium isotope ratios in rats

Calcium (Ca) isotopes (δ44/42Ca) in serum and urine have been suggested as novel sensitive markers of bone calcification. The response of δ44/42Ca to acute changes in Ca homeostasis, has not yet been demonstrated. We measured serum Ca and δ44/42Ca in rats maintained on a standard and a 50% Ca reduced diet for 4 weeks, and after injection of 1 mg/kg of the calcimimetic AMG-416, 24 h prior to sacrifice. AMG-416 decreased serum Ca by a maximum of 0.38 ± 0.10 and 0.53 ± 0.35 mmol/l after 12 and 6 h, respectively, in the standard and low-Ca diet groups (p = 0.0006/0.02), while serum δ44/42Ca did not change over 24 h in both groups. Urinary Ca concentrations were higher 24 h after AMG-416 injection in both groups (p = 0.03/0.06), urine δ44/42Ca was not different compared to the untreated control groups. Our data does not show acute changes in δ44/42Ca in response to a single dose of AMG-416 within 24 h after injection, possibly due to a lack of bone calcification.

Calcium isotopes as a biomarker for vascular calcification in chronic kidney disease.

Calcium balance is abnormal in adults with chronic kidney disease (CKD) and is associated with the development of vascular calcification. It is currently not routine to screen for vascular calcification in CKD patients. In this cross-sectional study, we investigate whether the ratio of naturally occurring calcium (Ca) isotopes, 44Ca and 42Ca, in serum could be used as a noninvasive marker of vascular calcification in CKD. We recruited 78 participants from a tertiary hospital renal center: 28 controls, 9 subjects with mild-moderate CKD, 22 undertaking dialysis and 19 who received a kidney transplant. For each participant, systolic blood pressure, ankle brachial index, pulse wave velocity, and estimated glomerular filtration rate were measured, along with serum markers. Calcium concentrations and isotope ratios were measured in urine and serum. While we found no significant association between urine Ca isotope composition (noted δ44/42Ca) between the different groups, δ44/42Ca values in serum were significantly different between healthy controls, subjects with mild-moderate CKD and those undertaking dialysis (P<0.01). Receiver operative characteristic curve analysis shows that the diagnostic utility of serum δ44/42Ca for detecting medial artery calcification is very good (AUC=0.818, sensitivity 81.8% and specificity 77.3%, P<0.01), and performs better than existing biomarkers. Although our results will need to be verified in prospective studies across different institutions, serum δ44/42Ca has the potential to be used as an early screening test for vascular calcification.

Calcium isotope ratios of malformed foraminifera reveal biocalcification stress preceded Oceanic Anoxic Event 2

Ocean acidification causes biocalcification stress. The calcium isotope composition of carbonate producers can archive such stress because calcium isotope fractionation is sensitive to precipitation rate. Here, we synthesize morphometric observations of planktic foraminifera with multi-archive calcium isotope records from Gubbio, Italy and the Western Interior Seaway spanning Cretaceous Ocean Anoxic Event 2 (~94 million years ago). Calcium isotope ratios increase ~60 thousand years prior to the event. The increase coincides with foraminiferal abnormalities and correlates with existing proxy records for carbon dioxide release during large igneous province volcanism. The results highlight Ocean Anoxic Event 2 as a geologic ocean acidification analog. Moreover, decreasing calcium isotope ratios during the event provide evidence for ocean alkalinization, which could have shifted air-sea carbon dioxide partitioning. These data offer an explanation for the Plenus Cold Event and further have implications for refining ocean alkalinity enhancement, a leading anthropogenic carbon dioxide removal strategy.

Calcium isotope ratio in kidney stones: preliminary exploration of mechanism from the geochemical perspective.

Stable calcium (Ca) isotope ratios are sensitive and radiation-free biomarkers in monitoring biological processes in human bodies. Recently, the Ca isotope ratios of bone, blood, and urine have been widely reported to study bone mineral balance. However, as a pure Ca crystallization product, there is no report on the Ca isotope ratios of kidney stones, even though the prevalence of kidney stones is currently on the rise. Here, we measured Ca isotope data of 21 kidney stone samples collected in Beijing, China. The δ44/42CaNIST 915a values ranged from 0.25‰ to 2.85‰ for calcium oxalate, and from 0.38‰ to 3.00‰ and 0.61‰ to 0.69‰ for carbonate apatite and uric acid, respectively. Kidney stones have heavier Ca isotope ratios than bone or blood, which is probably because complexed Ca contains more heavy Ca isotopes than free Ca2+. Ca isotope evidence suggests that magnesium (Mg) affects kidney stone formation, as the δ44/42CaNIST 915a value is inversely correlated with the Ca/Mg ratio. This study provides important preliminary reference values on the Ca isotopic composition of kidney stones and proposes a factor influencing Ca isotope fractionation in biological processes for future research.

The Calcium Isotope (δ44/40Ca) Record Through Environmental Changes: Insights From the Late Triassic

AbstractCalcium isotopes (δ44/40Ca) are particularly useful in palaeo‐environmental studies due to the key role of carbonate minerals in continental weathering and their formation in seawater. The calcium isotope ratio can provide hints on past changes in the calcium fluxes, environmental shifts, ecological factors and alternatively diagenesis of carbonate rocks. The investigation of the Late Triassic calcium isotope record offers a great opportunity to evaluate such factors in a time interval that witnessed important environmental and ecological turnovers, such as the first appearance of calcareous nannoplankton, ocean acidification and periods of elevated extinction rates. In this study, we present a δ44/40Ca data set from the upper Norian (Upper Triassic) through the lower Hettangian (Lower Jurassic) interval. The isotope records reveal two globally significant signals: a ∼ 0.20‰ decrease through the early Rhaetian (Upper Triassic) and a small, negative (∼0.14‰) excursion corresponding to the emplacement of the Central Atlantic Magmatic Province, at the end of the Triassic. The possible explanations for these signals are changes in the isotopic ratio of the continental calcium influx to the ocean due to the high chemical weathering rate of carbonates and possibly ocean acidification, respectively. The considerable (∼0.15–0.30‰) offset in δ44/40Ca between study areas is likely the combined result of local differences in lithology and early marine diagenesis. The major evolutionary step represented by the first occurrence of calcareous nannoplankton did not have at this time a determining role on the calcium isotopic signature of the marine carbonates.

Natural stable calcium isotope ratios: a new gold standard for bone balance?

Bone calcium balance is the net gain, loss, or equilibrium of calcium moving to and from bone, which reflects bone balance. There are currently no clinically available tools for measuring real-time bone balance. In this issue, Shroff etal. demonstrate the use of natural stable calcium isotope ratios as a novel biomarker of bone balance in children with chronic kidney disease on dialysis that is highly repeatable and associated with radiological and biochemical markers of bone metabolism.

Controls on riverine calcium isotope ratios during basalt weathering in the Skagafjörður watershed, Iceland

This study examines the Ca isotope (δ44/40Ca) geochemistry of Icelandic rivers with the overall aims of improving the δ44/40Ca tracer, constraining solute sources, and addressing the hypothesis that basalt weathering disproportionately regulates Earth’s long-term climate. We report Ca isotope and elemental data for mainstem rivers, tributary streams, thermal and non-thermal springs, basalt, calcite, soil, vegetation, and sediment collected from the Skagafjörður region in North Iceland. Waters and calcite were further analyzed for their carbon isotope (δ13C) composition, and we conducted experiments to characterize δ44/40Ca values of riverine colloids, the clay-size fraction (<2 µm) of soil, and exchangeable leachates obtained from soil and bedload sediment. The dataset includes analyses of travertine and coexisting water samples collected from a CO2-rich spring (Hvanná river) in South Iceland.Samples show a ∼2‰ range, with groundwater, the ultrafiltered fraction of river water, and hydrothermal calcite producing among the highest δ44/40Ca values and travertine, soil exchangeable Ca, and vegetation the lowest. Riverine δ44/40Ca values are on average ∼0.20‰ higher than those for basalt, which shows minimal isotopic variability. Mainstem glacial-fed rivers entering the Skagafjörður valley from the soil- and vegetation-free highlands have higher δ44/40Ca values than direct-runoff tributaries draining catchments to the north where pedogenesis is more pervasive. Riverine δ44/40Ca values correlate with Sr/Ca and Na/Ca ratios, as well as the saturation index of calcite (SIcal). Riverine δ13C values increase from ∼−8‰ (atmospheric) to ∼−3‰ (calcite) as SIcal values increase from highly-undersaturated to near-equilibrium. One watershed (Svartá river) shows unique trends, where riverine δ44/40Ca values and Sr/Ca ratios do not correlate, and soil exchangeable leachates produced anomalously high amounts of Ca. Clay-size fractions isolated from three horizons composing a brown andosol show slightly lower δ44/40Ca values than bulk basalt, but the values overlap with those for primary minerals. Colloids appear to have lower δ44/40Ca values than truly dissolved Ca. The proxy could be developed as a tracer for colloidal contributions to riverine dissolved loads, especially in soil-mantled catchments where the inputs appear most pronounced. Groundwater δ44/40Ca values correlate with pH, temperature, distance from the coast, and Sr/Ca ratios, all consistent with fractionation control yielding varying degrees of geochemical evolution. The combined riverine patterns largely reflect three-component mixing between basalt weathering, calcite weathering, and hydrothermal water inputs. Subsurface Ca isotope fractionation indirectly impacts riverine δ44/40Ca values by elevating the δ44/40Ca values of groundwater and hydrothermal calcite. This study highlights the significance of hydrothermal water inputs of Ca and HCO3 to Icelandic rivers, which have been previously underappreciated. Mixing calculations suggest that a maximum of ∼20% of the Ca in mainstem rivers derives from surficial basalt weathering. Small tributaries draining the flanks of the valley show the clearest signals of basalt weathering by atmospheric CO2, but these waters have much lower solute concentrations than those employed in previous attempts to estimate basalt weathering rates and parameterize the climate sensitivity of the basalt weathering feedback.

Naturally occurring stable calcium isotope ratios are a novel biomarker of bone calcium balance in chronic kidney disease

Dysregulated calcium homeostasis is common in chronic kidney disease and causally associated with disorders of bone mineralization. However, radiological measures and biomarkers do not allow accurate evaluation of bone calcium balance. Non-radioactive calcium isotopes, 42Ca and 44Ca, are present in our diet and sequestered into body compartments following principles of kinetic isotope fractionation. Isotopically light 42Ca is preferentially incorporated into bone, while heavier 44Ca is excreted. The ratio (44/42Caserum) increases when bone formation exceeds resorption and vice versa, reflecting bone calcium balance. We measured these calcium isotopes by inductively coupled plasma mass-spectrometry in blood, urine and feces of 42 children with chronic kidney disease and 92 receiving dialysis therapy. We compared the isotope ratios with bone biomarkers and determined total bone mineral content by dual-energy x-ray absorptiometry and peripheral quantitative CT expressed as age-adjusted z-scores. The 44/42Caserum ratio positively correlated with serum calcium, 25-hydroxyvitamin D and alkaline phosphatases and inversely with serum parathyroid hormone and other bone resorption markers. The 44/42Caserum ratio positively correlated with age-adjusted z-scores of tibial trabecular bone mineral density and total bone mineral content measured by peripheral quantitative CT, and hip bone mineral density measured by dual-energy X-ray absorptiometry. Significant and independent predictors of total bone mineral content, measured by, were the 44/42Caserum ratio and parathyroid hormone. The44/42Caserum ratio, repeated after four weeks, highly correlated with baseline values. When adjusted for calcium-containing medications and kidney impairment, the 44/42Caserum ratio in patients receiving dialysis was 157% lower than that of age-matched children and 29% lower than levels in elderly women with osteoporosis, implying significantly lower bone mineral content. Thus, calcium isotope ratios may provide a novel, sensitive and non-invasive method of assessing bone calcium balance in chronic kidney disease.

Adapting automated instrumentation for high-throughput calcium isotope measurements by multi-collector inductively coupled plasma mass spectrometry.

Large, efficiently generated datasets of calcium isotope ratios offer tremendous utility in an increasing number of scientific disciplines. Modern analytical capabilities in mass spectrometry present unique challenges to previously established sample preparation techniques, the extent of which must be thoroughly examined before these data are interpreted and reported. This study addresses key methodological challenges in the determination of calcium isotopes using state-of-the-art, commercially available instruments. Automated ion chromatography was used to isolate calcium from carbonate- and seawater-like samples prior to analysis by multi-collector inductively coupled plasma mass spectrometry (MC-ICP-MS). A new, membrane-bearing desolvator (Apex-Ω) is implemented and achieves enhanced sensitivity, yet necessitates an update to established sample preparation techniques due to previously unobserved matrix effects. Performance of this method was assessed through analyses of multiple standard reference materials (SRM 915b, USGS EN-1, and seawater) and several in-house standards using a Neptune MC-ICP-MS instrument. The enhanced sensitivity afforded by the Apex-Ω also yields pronounced matrix effects during mass spectrometry, resulting specifically from heightened sample Na and Sr content and deviations in sample [Ca] from their bracketing standards. While the latter can be addressed by a concentration correction, the first two matrix effects are mitigated using a modified chromatography technique that implements unique rinsing protocols. This precise (0.14‰ 2σ), high-throughput method is very reproducible for small sample amounts and, at optimal efficiency, can generate approximately 140 sample δ44/40 Ca values per week with 5h of in-person effort per day. Documented matrix effects are successfully mitigated through corrections and modified chromatographic techniques. Additionally, this method may be permuted to accommodate most major cations.

Application of calcium isotopes to the study of calcareous materials on Chinese glaze: A new methodological approach

AbstractIn this work, we propose an innovative application of the calcium isotope technique for the study of calcareous materials in Chinese glazes, supported by simulation experiments. The δ44/42Ca values have been determined for limestone, plant ash and glazed ceramic samples collected from the Ru kiln in Henan Province, the Lauding kiln in Hangzhou city, Zhejiang Province, and Jingdezhen in Jiangxi Province. The results show that the δ44/42Ca values in limestone in Henan and Zhejiang are all between 0.15‰ and 0.41‰, whereas the δ44/42Ca values of plant ash from the Ru and Laohudong kilns are all between −0.23‰ and −0.04‰. This means that the calcium isotope technique can effectively identify whether calcium levels in the glazes were supplied by plant ash or by limestone materials. More importantly, the calcium isotope technique can evaluate the proportions of plant ash and limestone if the local glaze materials have been studied thoroughly. According to the calcium isotope ratios established for this study, it is speculated that for the Jingdezhen kilns plant ash was used from the Late Tang to Five Dynasties, and limestone and plant ash were used for glazes in the Song and Yuan Dynasties. The calcium isotope technique can therefore offer an approach to identifying both the calcareous materials and the origins of ancient Chinese glazes, and provides a potentially powerful tool in their interpretation.

A reactive transport approach to modeling cave seepage water chemistry I: Carbon isotope transformations

The majority of Critical Zone research has emphasized silicate lithologies, which are typified by relatively slow rates of reactivity and incongruent weathering. However, the relatively simpler weathering of carbonate-dominated lithology can result in secondary mineral deposits, such as speleothems, which provide a long-term archive for Critical Zone processes. In particular, carbon isotopic variability in speleothems has the potential to provide records of changes in vegetation, soil respiration, carbon stabilization in deep soils, and/or chemical weathering in the host rock. Despite this opportunity to reconstruct many Critical Zone processes, multiple influences can also make interpretion of these speleothem carbon isotope records challenging. The integration of observational data and simulations specific to karst systems offers an interpretive framework for these unique time-averaged records accumulated through the evolution of carbonate landscapes. Here, we present a forward and process-based reactive transport simulation based on a multi-year monitoring study of Blue Spring Cave in central Tennessee, USA. The simulations describe the fluid-driven weathering of limestone including explicit tracking of dissolved calcium, stable carbon, and radiocarbon isotope ratios based on reaction rates calibrated through laboratory batch reaction data. We find that calcium concentrations and radiocarbon isotope ratios are strongly influenced by the combination of fluid flow rate and soil CO2 content, and require rapid gas phase communication between the overlying soil boundary condition and interior karst to sustain both elevated limestone weathering rates and relatively modern radiocarbon signatures. Stable carbon isotopes are largely dictated by temperature-dependent equilibrium fractionation among contemporaneous species. These simulations are extended to a wide range of parameter space to demonstrate the environmental factors that these isotope proxies record.

Review of techniques, challenges, and new developments for calcium isotope ratio measurements

Calcium isotope meaurements apply to problems spanning the ‘cosmos to benthos.’ Fractionation, source mixing, radioactive decay, and nucleosynthetic processes cause variations in Ca isotope ratios, which can be measured using different types of mass spectrometers. Early measurements employed thermal ionization mass spectrometry (TIMS), which continues to be the most common and precise reference technique. An increasing number of studies use multi-collector inductively coupled plasma mass spectrometry (MC-ICPMS). Both TIMS and MC-ICPMS can yield high-quality data for δ44/40Ca and δ44/42Ca, respectively, but wide ranges in precision exist. Sample size requirements are lower for TIMS, but MC-ICPMS offers faster throughput. Both techniques require purifying Ca by ion-exchange chromatography when analyzing samples with complex matrices. In-situ techniques, like laser ablation MC-ICPMS (LA-MC-ICPMS) and secondary ion mass spectrometry (SIMS), do not require purification. In-situ techniques provide high spatial resolution, but LA-MC-ICPMS and (nano)-SIMS suffer from relatively poor precision compared to TIMS and MC-ICPMS, and the methods lack well-characterized standards, which are essential for comparing datasets.The past few years have seen advances in Ca purification techniques, including automated methods that accelerate sample throughput. The next generation of Ca purification techniques should focus on complete purification and quantitative separation of low-Ca samples. Application of collision-cell technology for suppressing Ar-related molecular isobaric interferences will improve quantification of 40Ca using certain MC-ICPMS models. Similarly, development of new amplifier technology will foster high-precision Faraday cup measurements of low abundance Ca isotopes. Adopting common reference materials, including newly developed standards with complex matrices, would signficantly aid inter-laboratory comparisons, method development, and data compilation efforts. In general, at least two widely-used standards should be measured along with unknown samples to confirm data quality, as well as improve utilization of published Ca isotope datasets by a broader community.

Calcium isotope fractionation by osteoblasts and osteoclasts, across endothelial and epithelial cell barriers, and with binding to proteins.

Timely and accurate diagnosis of osteoporosis is essential for adequate therapy. Calcium isotope ratio (δ44/42Ca) determination has been suggested as a sensitive, noninvasive, and radiation-free biomarker for the diagnosis of osteoporosis, reflecting bone calcium balance. The quantitative diagnostic is based on the calculation of the δ44/42Ca difference between blood, urine, and bone. The underlying cellular processes, however, have not been studied systematically. We quantified calcium transport and δ44/42Ca fractionation during in vitro bone formation and resorption by osteoblasts and osteoclasts and across renal proximal tubular epithelial cells (HK-2), human vein umbilical endothelial cells (HUVECs), and enterocytes (Caco-2) in transwell systems and determined transepithelial electrical resistance characteristics. δ44/42Ca fractionation was furthermore quantified with calcium binding to albumin and collagen. Calcified matrix formed by osteoblasts was isotopically lighter than culture medium by -0.27 ± 0.03‰ within 5 days, while a consistent effect of activated osteoclasts on δ44/42Ca could not be demonstrated. A transient increase in δ44/42Ca in the apical compartment by 0.26‰ occured across HK-2 cells, while δ44/42Ca fractionation was small across the HUVEC barrier and absent with Caco-2 enterocytes, and with binding of calcium to albumin and collagen. In conclusion, δ44/42Ca fractionation follows similar universal principles as during inorganic mineral precipitation; osteoblast activity results in δ44/42Ca fractionation. δ44/42Ca fractionation also occurs across the proximal tubular cell barrier and needs to be considered for in vivo bone mineralization modeling. In contrast, the effect of calcium transport across endothelial and enterocyte barriers on blood δ44/42Ca should be low and is absent with physiochemical binding of calcium to proteins.

Semiquantitative Estimates of Rainfall Variability During the 8.2 kyr Event in California Using Speleothem Calcium Isotope Ratios

AbstractA multiproxy record from a fast‐growing stalagmite reveals variable hydroclimate on the California coast across the 8.2 kyr event and a precursor event likely caused by initial drainage of proglacial Lake Agassiz. Using speleothem δ44Ca, we develop the first semiquantitative estimates of paleorainfall variability for California through calibration with measurements of the modern climate and cave environment. We find that the magnitude of rainfall variability during the 8.2 kyr event approached the multiyear variability observable in the recent past (1950–2019) and the magnitude of variability during the precursor event likely exceeded this range. Additionally, we observe other instances of multidecadal variability comparable in magnitude to the precursor event during the record. Our work suggests that speleothem calcium isotope ratios are a powerful semiquantitative means to reconstruct paleorainfall, although numerous factors must be assessed in each cave system before applying this approach.

Small calcium isotope fractionation at slow precipitation rates in methane seep authigenic carbonates

Natural calcium carbonate minerals express a range of calcium isotope fractionations, with the precipitated mineral typically enriched in the lighter isotopes of calcium relative to source fluids. Experimental and theoretical evidence shows a strong dependence on precipitation rate, although this relationship has not been well quantified over the range of precipitation rates observed in natural settings. Endmember cases show that average marine carbonate precipitation expresses a large fractionation (δ44/40Ca values lower than seawater by approximately 1‰), while diagenetic carbonate phases assumed to have precipitated or recrystallized at very slow rates show negligible fractionation. The limited examples of quantified precipitation rates in natural settings with measurable, non-zero fractionation represents a barrier for applying mechanistic models of calcium isotope fractionation to geological applications. This study examines a methane seep system in the northern Barents Sea south of Svalbard where authigenic carbonate minerals are precipitating, driven by anaerobic oxidation of methane, and where the apparent calcium isotope fractionation factor and precipitation rate can be constrained by measuring properties of the pore fluids. Pore fluid profiles are analyzed in two shallow cores, and authigenic carbonate nodules are analyzed in one of these cores. The pore fluid profiles point to a transitional, non-steady state which approximates a closed system, where the elevation of pore fluid calcium isotope ratios through carbonate precipitation can be modeled as a Rayleigh distillation system. The apparent fractionation factors for 44Ca/40Ca ratios at these sites are α = 0.99985 and 0.9996, although the carbonate nodules suggest a different calcium isotope fractionation factor may have been expressed under past conditions. Precipitation rates at the two sites are estimated to be 1.4 and 3.5 μmol/m2/h, intermediate between those of typical laboratory experiments and the much slower rates of marine diagenesis. Trace element analyses of the nodules (Mg/Ca and Sr/Ca ratios) suggest that both precipitation rate and mineralogy affect nodule composition. These results provide new constraints for the relationship between precipitation rate and calcium isotope fractionation and can inform modeling efforts leading towards mechanistic understanding of calcium isotope fractionation and trace element distributions in carbonate minerals.

Isotope Microscopic Observation of Osteogenesis Process Forming Robust Bonding of Double Network Hydrogel to Bone.

Tough double network (DN) hydrogels are promising substitutes of soft supporting tissues such as cartilage and ligaments. For such applications, it is indispensable to robustly fix the hydrogels to bones with medically feasible methods. Recently, robustly bonding the DN hydrogels to defected bones of rabbits in vivo has been proved successful. The low crystalline hydroxyapatite (HAp) of calcium-phosphate-hydroxide salt coated on the surface layer of the DN hydrogels induced spontaneous osteogenesis penetrating into the semi-permeable hydrogels to form a gel/bone composite layer. In this work, the 44 Ca isotope-doped HAp/DN hydrogel is implanted in a defect of rabbit femoral bone and the dynamic osteogenesis process at the gel/bone interface is analyzed by tracing the calcium isotope ratio using isotope microscopy. The synthetic HAp hybridized on the surface layer of DN gel dissolves rapidly in the first two weeks by inflammation, and then the immature bone with a gradient structure starts to form in the gel region, reutilizing the dissolved Ca ions. These results reveal, for the first time, that synthetic HAp is reutilized for osteogenesis. These facts help to understand the lifetime of bone absorbable materials and to elucidate the mechanism of spontaneous, non-toxic, but strong fixation of hydrogels to bones.

Naturally Occurring Stable Calcium Isotope Ratios in Body Compartments Provide a Novel Biomarker of Bone Mineral Balance in Children and Young Adults.

Serum calcium (Ca), bone biomarkers, and radiological imaging do not allow accurate evaluation of bone mineral balance (BMB), a key determinant of bone mineral density (BMD) and fracture risk. We studied naturally occurring stable (non-radioactive) Ca isotopes in different body pools as a potential biomarker of BMB. 42 Ca and 44 Ca are absorbed from our diet and sequestered into different body compartments following kinetic principles of isotope fractionation; isotopically light 42 Ca is preferentially incorporated into bone, whereas heavier 44 Ca preferentially remains in blood and is excreted in urine and feces. Their ratio (δ44/42 Ca) in serum and urine increases during bone formation and decreases with bone resorption. In 117 healthy participants, we measured Ca isotopes, biomarkers, and BMD by dual-energy X-ray absorptiometry (DXA) and tibial peripheral quantitative CT (pQCT). 44 Ca and 42 Ca were measured by multi-collector ionization-coupled plasma mass-spectrometry in serum, urine, and feces. The relationship between bone Ca gain and loss was calculated using a compartment model. δ44/42 Caserum and δ44/42 Caurine were higher in children (n = 66, median age 13 years) compared with adults (n = 51, median age 28 years; p < 0.0001 and p = 0.008, respectively). δ44/42 Caserum increased with height in boys (p < 0.001, R2 = 0.65) and was greatest at Tanner stage 4. δ44/42 Caserum correlated positively with biomarkers of bone formation (25-hydroxyvitaminD [p < 0.0001, R2 = 0.37] and alkaline phosphatase [p = 0.009, R2 = 0.18]) and negatively with bone resorption marker parathyroid hormone (PTH; p = 0.03, R2 = 0.13). δ44/42 Caserum strongly positively correlated with tibial cortical BMD Z-score (n = 62; p < 0.001, R2 = 0.39) but not DXA. Independent predictors of tibial cortical BMD Z-score were δ44/42 Caserum (p = 0.004, β = 0.37), 25-hydroxyvitaminD (p = 0.04, β = 0.19) and PTH (p = 0.03, β = -0.13), together predicting 76% of variability. In conclusion, naturally occurring Ca isotope ratios in different body compartments may provide a novel, non-invasive method of assessing bone mineralization. Defining an accurate biomarker of BMB could form the basis of future studies investigating Ca dynamics in disease states and the impact of treatments that affect bone homeostasis. © 2020 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

Early effects of androgen deprivation on bone and mineral homeostasis in adult men: a prospective cohort study.

Long-term androgen deprivation therapy (ADT) negatively influences bone. The short-term effects on bone and mineral homeostasis are less known. Therefore, we aimed to investigate the early effects of ADT on calcium/phosphate homeostasis and bone turnover. Prospective cohort study. Eugonadal adult, male sex offenders, who were referred for ADT to the endocrine outpatient clinic, received cyproterone acetate. Changes in blood markers of calcium/phosphate homeostasis and bone turnover between baseline and first follow-up visit were studied. Of 26 screened patients, 17 were included. The median age was 44 (range 20-75) years. The median time interval between baseline and first follow-up was 13 (6-27) weeks. Compared to baseline, an 81% decrease was observed for median total testosterone (to 3.4 nmol/L (0.4-12.2); P < 0.0001) and free testosterone (to 0.06 nmol/L (0.01-0.18); P < 0.0001). Median total estradiol decreased by 71% (to 17.6 pmol/L (4.7-35.6); P < 0.0001). Increased serum calcium (P < 0.0001) and phosphate (P = 0.0016) was observed, paralleled by decreased PTH (P = 0.0156) and 1,25-dihydroxyvitamin D3 (P = 0.0134). The stable calcium isotope ratio (δ44/42Ca) decreased (P = 0.0458), indicating net calcium loss from bone. Bone-specific alkaline phosphatase and osteocalcin decreased (P < 0.0001 and P = 0.0056, respectively), periostin tended to decrease (P = 0.0500), whereas sclerostin increased (P < 0.0001), indicating suppressed bone formation. Serum bone resorption markers (TRAP, CTX) were unaltered. In adult men, calcium release from the skeleton occurs early following sex steroid deprivation, reflecting early bone resorption. The increase of sclerostin and reduction of bone formation markers, without changes in resorption markers, suggests a dominant negative effect on bone formation in the acute phase.