High Calcium Research Articles

Unleashing Customer Empathy in the Circular Economy: Development of A High-Calcium Fish Sausage Prototype from Fermented Fish Residue

In a circular economy, fermented fish residue could be converted into high-calcium fish bone powder for fish sausages to reduce fish waste. To create product concepts for such sausages, this study aimed to (1) identify customer behaviors, needs, and problems associated with purchasing fish sausages; (2) select high-value consumers of high-calcium fish sausages from fermented fish residues; and (3) propose product prototypes of high-calcium fish from fermented fish residues. We interviewed 50 sausage purchasers and a particular producer of calcium fish bone powder. Then, thematic analyses were performed to evaluate the minimum viable product (MVP) for the final high calcium fish sausage prototype. The research suggested that household consumers were the target market for this novel food. This result emphasized nutrition and sensory characteristics as the primary reason for purchasing sausages and indicated common problems with the inaccessibility of healthy sausages. Based on the findings, fish sausage should contain nutritional value and healthy ingredients to address the consumers’ pain points. This circular food system was estimated to make the food product more sustainable and resource-efficient when fully applied.

#1834 Prevalence of nephrolithiasis in patients with idiopathic hypercalciuria after two decades of follow-up

Abstract Background and Aims Idiopathic hypercalciuria (IH) is the main metabolic abnormality that causes kidney stones. It is linked to high intestinal calcium absorption and osteopenia. However, not all patients diagnosed with IH develop kidney stones. In this study, we examined a cohort of patients diagnosed of IH during childhood to determine the prevalence of lithiasis in adulthood. Method Longitudinal study that evaluated the prevalence of lithiasis in 35 patients (12 female, 23 male) diagnosed of IH during childhood (mean age of 7.7 ± 3 years). The patients were followed until adulthood (mean age 28.2 ± 3.1 years) with a median follow-up of 18.2 years (IQR: 12.2-19.3). IH diagnosis was made by urinary calcium excretion higher than 4 mg/Kg/day in two consecutive samples. In both periods, clinical and demographic data were collected, including urinalysis and urinary ratios (calcium/creatinine, calcium/citrate, oxalate/creatinine, and citrate/creatinine) in an isolated sample of the first morning urine, as well as the performance of renovesical ultrasound. Results In pediatric age, 8 patients (26%) presented nephrolithiasis on ultrasound, and 14 (43.8%) in adulthood, 6 of them experiencing episodes of nephritic colic. During the study, we observed a decrease in urinary calcium excretion, as expressed by the calcium/creatinine ratio (adult age 0.16 ± 0.9 mg/dl vs. childhood 0.28 ± 0.1 mg/dl, p = 0.000), and an hypocitraturia increase (citrate/creatinine ratio in childhood 887 ± 349 mg/dl and in adulthood 357.7 ± 363 mg/dl, p = 0.000). A decreased citrate/creatinine ratio in adulthood was associated with nephrolithiasis (p < 0.007). Only 4 patients (50%) had nephrolithiasis at both ages. Conclusion These results suggest that hypercalciuria tends to correct spontaneously in adulthood, and although the prevalence of nephrolithiasis increased, it is more related to hypocitraturia.

Combined full-length transcriptomic and metabolomic analysis reveals the molecular mechanisms underlying nutrients and taste components development in Primulina juliae

BackgroundPrimulina juliae has recently emerged as a novel functional vegetable, boasting a significant biomass and high calcium content. Various breeding strategies have been employed to the domestication of P. juliae. However, the absence of genome and transcriptome information has hindered the research of mechanisms governing the taste and nutrients in this plant. In this study, we conducted a comprehensive analysis, combining the full-length transcriptomics and metabolomics, to unveil the molecular mechanisms responsible for the development of nutrients and taste components in P. juliae.ResultsWe obtain a high-quality reference transcriptome of P. juliae by combing the PacBio Iso-seq and Illumina sequencing technologies. A total of 58,536 cluster consensus sequences were obtained, including 28,168 complete protein coding transcripts and 8,021 Long Non-coding RNAs. Significant differences were observed in the composition and content of compounds related to nutrients and taste, particularly flavonoids, during the leaf development. Our results showed a decrease in the content of most flavonoids as leaves develop. Malate and succinate accumulated with leaf development, while some sugar metabolites were decreased. Furthermore, we identified the different accumulation of amino acids and fatty acids, which are associated with taste traits. Moreover, our transcriptomic analysis provided a molecular basis for understanding the metabolic variations during leaf development. We identified 4,689 differentially expressed genes in the two developmental stages, and through a comprehensive transcriptome and metabolome analysis, we discovered the key structure genes and transcription factors involved in the pathways.ConclusionsThis study provides a high-quality reference transcriptome and reveals molecular mechanisms associated with the development of nutrients and taste components in P. juliae. These findings will enhance our understanding of the breeding and utilization of P. juliae as a vegetable.

Key Roles of CACNA1C/Cav1.2 and CALB1/Calbindin in Prefrontal Neurons Altered in Cognitive Disorders.

The risk of mental disorders is consistently associated with variants in CACNA1C (L-type calcium channel Cav1.2) but it is not known why these channels are critical to cognition, and whether they affect the layer III pyramidal cells in the dorsolateral prefrontal cortex that are especially vulnerable in cognitive disorders. To examine the molecular mechanisms expressed in layer III pyramidal cells in primate dorsolateral prefrontal cortices. The design included transcriptomic analyses from human and macaque dorsolateral prefrontal cortex, and connectivity, protein expression, physiology, and cognitive behavior in macaques. The research was performed in academic laboratories at Yale, Harvard, Princeton, and the University of Pittsburgh. As dorsolateral prefrontal cortex only exists in primates, the work evaluated humans and macaques. Outcome measures included transcriptomic signatures of human and macaque pyramidal cells, protein expression and interactions in layer III macaque pyramidal cells using light and electron microscopy, changes in neuronal firing during spatial working memory, and working memory performance following pharmacological treatments. Layer III pyramidal cells in dorsolateral prefrontal cortex coexpress a constellation of calcium-related proteins, delineated by CALB1 (calbindin), and high levels of CACNA1C (Cav1.2), GRIN2B (NMDA receptor GluN2B), and KCNN3 (SK3 potassium channel), concentrated in dendritic spines near the calcium-storing smooth endoplasmic reticulum. L-type calcium channels influenced neuronal firing needed for working memory, where either blockade or increased drive by β1-adrenoceptors, reduced neuronal firing by a mean (SD) 37.3% (5.5%) or 40% (6.3%), respectively, the latter via SK potassium channel opening. An L-type calcium channel blocker or β1-adrenoceptor antagonist protected working memory from stress. The layer III pyramidal cells in the dorsolateral prefrontal cortex especially vulnerable in cognitive disorders differentially express calbindin and a constellation of calcium-related proteins including L-type calcium channels Cav1.2 (CACNA1C), GluN2B-NMDA receptors (GRIN2B), and SK3 potassium channels (KCNN3), which influence memory-related neuronal firing. The finding that either inadequate or excessive L-type calcium channel activation reduced neuronal firing explains why either loss- or gain-of-function variants in CACNA1C were associated with increased risk of cognitive disorders. The selective expression of calbindin in these pyramidal cells highlights the importance of regulatory mechanisms in neurons with high calcium signaling, consistent with Alzheimer tau pathology emerging when calbindin is lost with age and/or inflammation.

Efficiency of cavitary varnishes containing experimental bioglass particles in the occlusion of dentinal tubules.

This research aims to evaluate the efficiency of cavitary varnishes containing experimental bioglasses in the occlusion of dentinal tubules. One hundred and sixty-eight cervical buccal dentin samples were obtained from bovine teeth. Samples were randomized into the following groups: I. Distilled Water (DW); II. Cavity Varnish (CV); III. Colgate® Sensitive Pro-Relief™ (CS); IV. 45S5 Bioglass (45S5); V. KSr Bioglass strontium potassium (KSr); VI. P Bioglass phosphorus (P); and VII. PSi Bioglass phosphorus silica (PSi). The treatments were applied to the surfaces of the samples, which were then subjected to simulated brushing. The samples were analyzed for a) characterization of bioactive glasses; b) surface roughness; c) descriptive analysis of the dentin surface; d) total versus occluded number of dentinal tubules; e) diameter of the dentinal tubules; f) chemical composition of the dentin surfaces, and g) dentin permeability. All groups treated with biomaterials without the brushing challenge showed an increase in roughness and (total or partial) occlusion of the dentinal tubules. The PSi group had the best values for occlusion, while the KSr group had the highest calcium and phosphorus concentrations. After the brushing challenge the roughness was controlled by the presence of biomaterials; 45S5, KSr, and PSi showed occlusion of the dentin tubules. All bioactive glasses showed reduced tooth permeability compared to distilled water. The PSi group had the smallest tubule diameter and highest phosphorus concentration. KSr and PSi bioglasses are promising materials for dentin occlusion and remineralization and are promising new biomaterials for the treatment of dentin hypersensitivity.

Volcanic Rock of Slamet Volcano as the Potential of Soil Ameliorant

Mount Slamet is an active volcano in Java Island, Indonesia. Slamet volcanic rocks comprise various igneous and pyroclastic rocks, including basaltic lava, andesitic lava, pyroclastic rocks, and intrusions. Geochemical analysis of rocks in the studied area (301300 mE - 303300 mE and 9189400 mN - 9191400 mN) showed the presence of high calcium and iron elements. This geological study aims to determine the potential of material resources contained to be used for agricultural needs. The potential nutrients to be found such as P, K, Mg, Ca, Fe, Ti, Na, Mn, and Si with a DHL conductivity value of 0.0473 - 0.1318 mmhos /cm are classified as non-salinity, which is safe for soil improvement. Then the neutralization value relative to calcite is between 15.45 - 27.27 %, and the abrasion pH value is between 8.05 - 8.91. The agrogeological analysis shows that the Slamet volcanic rock in Baturraden area has good prospects as an ameliorant for highly weathered (acid) soils.

Development of gluten free crackers using water chestnut (Trapa natans) and Pearl Millet (Pennisetum glaucum)

Baked goods are favored by the public due to their widespread availability, convenience, ready-to-eat nature, and excellent storage capabilities. The study focused on developing high-quality gluten-free crackers by incorporating pearl millet (Bajra) and water chestnut flour which enhances nutritional value and is gluten free, catering to individuals with celiac disease.To Formulate Crackers with Pearl Millet and water chestnut flour. To determine the Sensory quality of the developed product. To Estimate the Proximate composition and cost of the developed product.Pearl millet flour, water chestnut flour, whole wheat flour,salt, jaggery, garlic powder, ajwain, mixed herbs, oil, and baking powder were procured from local grocery stores of Mysore and standard A.O.A.C (2005) methods for Proximate analysis and 9-point hedonic scale was used for sensory analysis.: The sensory evaluation revealed that PMC4 was highly acceptable on par with control. Proximate analysis showed that PMC4 had high phosphorous and calcium content.The study successfully developed superior gluten-free crackers with incorporation of Pearl millet and water chestnut flour. Among all the variations PMC4 was rich in phosphorous and calcium and is gluten free which caters to individuals with celiac disease.

Investigating the Influence of Spinach Sauce in Modulating Serum Calcium in Osteoporotic Females

Background: The modulation of serum calcium levels through diet is a critical aspect of managing osteoporosis, particularly in postmenopausal women. Spinach, known for its high calcium content, offers potential benefits for bone health due to its rich nutrient profile. This study aimed to develop a spinach sauce, evaluate its consumer acceptability, and investigate its impact on serum calcium levels in osteoporotic females. Objective: To develop a nutritionally rich spinach sauce, assess its sensory acceptability, and evaluate its effectiveness in modulating serum calcium levels among osteoporotic females. Methods: The spinach sauce was formulated using various recipes to maximize calcium bioavailability. The preparation process involved controlled cooking techniques to preserve nutrients, followed by proximate composition analysis. A sensory evaluation was conducted with osteoporotic females and a control group using standard hedonic scales. Participants were instructed to incorporate the spinach sauce into their daily diet for a specified period. Serum calcium levels were measured before and after the intervention. Statistical analysis was conducted using SPSS version 25, controlling for covariates such as age, baseline dietary habits, and baseline serum calcium levels. Ethical approval was obtained, and informed consent was provided by all participants. Results: Treatment T2, containing 40% spinach and 60% mint, scored highest in sensory evaluation with color (8.94 ± 0.26), flavor (8.92 ± 0.30), taste (8.98 ± 0.27), texture (8.22 ± 0.29), and overall acceptability (8.94 ± 0.31). The proximate composition of the spinach sauce showed moisture (79.84%), crude protein (6.27%), crude fat (1.91%), fiber (0.57%), ash (4.21%), and nitrogen-free extract (10.27%). Serum calcium levels increased significantly in the spinach sauce group from 0.84 ± 0.01 to 1.03 ± 0.04, and in the combined spinach sauce and calcium supplement group from 0.69 ± 0.01 to 0.91 ± 0.07. ANOVA revealed significant effects of treatments (F = 1.99, p ≤ 0.01) and days (F = 41.96, p ≤ 0.01) on bone mineral density. Conclusion: The developed spinach sauce was nutritionally rich and well-received, particularly in treatment T2. It demonstrated potential in improving serum calcium levels among osteoporotic females, suggesting that dietary interventions with spinach could be a viable strategy for managing osteoporosis. Further research with larger sample sizes and longer durations is recommended to substantiate these findings.

Interaction of extreme drought and insect herbivores on grassland community is affected by drought pattern

Abstract Extreme drought and insect herbivores can affect plant community non-independently, and their interaction may be influenced by drought pattern. However, few studies have explored the interaction on plant community structure, and no study has investigated whether the interaction is affected by drought pattern. We explored the interaction of different extreme drought patterns and insect herbivores with a manipulated experiment in a semiarid grassland. There were three treatments for drought: control – ambient precipitation; chronic drought – 66% precipitation reduction in growing season (May – August); intense drought – completely exclusion rain events from June to July. Herbivores removal and present treatments were imposed within each drought treatments. We found that although herbivores had no significant effects on the impacts of droughts on total species richness, it alleviated the impacts of chronic drought on total cover, and the alleviating role decreased under intense drought. Comparing with intense drought, more increasing of grass cover led to more alleviation under chronic drought associated with the decrease of forb which have low leaf carbon and dry matter content, along with high starch, calcium, magnesium and manganese concentration. These results indicate that the interaction of drought and herbivores is affected by drought pattern, and the changes of leaf traits that reduce herbivores feeding in grass might alleviate the impact of drought on grassland.

Prevotella copri promotes vascular calcification via lipopolysaccharide through activation of NF-κB signaling pathway

ABSTRACT Emerging evidence indicates that alteration of gut microbiota plays an important role in chronic kidney disease (CKD)-related vascular calcification (VC). We aimed to investigate the specific gut microbiota and the underlying mechanism involved in CKD-VC. We identified an increased abundance of Prevotella copri (P. copri) in the feces of CKD rats (induced by using 5/6 nephrectomy followed by a high calcium and phosphate diet) with aortic calcification via amplicon sequencing of 16S rRNA genes. In patients with CKD, we further confirmed a positive correlation between abundance of P. copri and aortic calcification scores. Moreover, oral administration of live P. copri aggravated CKD-related VC and osteogenic differentiation of vascular smooth muscle cells in vivo, accompanied by intestinal destruction, enhanced expression of Toll-like receptor-4 (TLR4), and elevated lipopolysaccharide (LPS) levels. In vitro and ex vivo experiments consistently demonstrated that P. copri-derived LPS (Pc-LPS) accelerated high phosphate-induced VC and VSMC osteogenic differentiation. Mechanistically, Pc-LPS bound to TLR4, then activated the nuclear factor κB (NF-κB) and nucleotide-binding domain, leucine-rich–containing family, pyrin domain–containing-3 (NLRP3) inflammasome signals during VC. Inhibition of NF-κB reduced NLRP3 inflammasome and attenuated Pc-LPS-induced VSMC calcification. Our study clarifies a novel role of P. copri in CKD-related VC, by the mechanisms involving increased inflammation-regulating metabolites including Pc-LPS, and activation of the NF-κB/NLRP3 signaling pathway. These findings highlight P. copri and its-derived LPS as potential therapeutic targets for VC in CKD.

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

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

Utilization of waste high calcium oxide fly ash as hybrid activator for vulcanization of natural rubber/carbon black composites: Reducing zinc oxide requirement

A high calcium oxide fly ash (FA), a by-product waste of electric power generation has been used as hybrid activator in natural rubber (NR) composites filled with carbon black (CB). The aim of using FA is to reduce the environmental contamination by zinc oxide (ZnO) needed as activator for sulfur-vulcanization/crosslinking in final NR products. The FA was varied by adding in the composites with and without ZnO, and it was observed that chemical crosslinking could be generated although no ZnO was used anymore. The synergistic FA/ZnO-blend with FA 9 phr and ZnO 3 phr presented the best fit for the NR/CB composites, where it showed a proper crosslink density, reinforcement efficiency, cure characteristics, and mechanical properties in terms of modulus, tensile strength, elongation at break and tear resistance. This relates to the inorganic composition of the FA, in particular calcium oxide (CaO), which induces the thermal conductivity throughout the NR matrix and activates crosslinking between the NR molecules. The results showed effective use of FA in the composites, which can be applied to the CB-based formulations used in commercial NR applications like tires, barriers, and elastic stopper.

Bond Model for High Calcium Fly Ash Geopolymer Concrete

Concrete industries and researchers have devoted great efforts to promoting sustainability in the construction sector. This includes finding innovative alternatives to OPC concrete to ensure environmental sustainability and prevent the depletion of natural resources. The present research explained the steps to develop a bond stress-slip model for high calcium fly ash (HCFA) geopolymer concrete based on the experimental results in the literature. This model is based on the concrete compressive strength (fc), concrete cover (Cc), and bar diameter (db), which are considered the most influential parameters on the bond response between the reinforcement and surrounding concrete. The regressed model shows a quite accurate prediction of the bond behaviour of HCFA, particularly those with high strength. The slight difference was attributed to the difficulty in identifying the exact values of Slips (δr1 and δr2).

Analisis Kadar Kalsium (Ca) Pada Lansia Di Panti Perlindungan Dan Rehabilitasi Sosial Lanjut Usia Provinsi Kalimantan Selatan

The elderly, or senior citizens, are a group of individuals who have reached old age. People in this age group are usually more susceptible to health problems. Calcium is a nutrient that is essential for the body and is the most abundant mineral found in the body, accounting for approximately 1.5-2% of an adult's body weight or about 1kg. Insufficient or excessive calcium intake can lead to abnormal metabolism. Calcium deficiency can result in osteoporosis and hypocalcemia, while excess calcium can cause hypercalcemia. The purpose of this study is to determine the calcium (Ca) levels in the elderly at the "Budi Sejahtera" Social Protection and Rehabilitation Home for the Elderly in South Kalimantan Province. The research design used in this study is a descriptive survey, which is a method used to describe or analyze research findings but not to draw broader conclusions. The calcium levels in the elderly were measured using a cross-sectional design, which observes population or sample data only once at the same time. The sampling was conducted on June 10, 2023. The sample used was blood from the elderly at the "Budi Sejahtera" Social Protection and Rehabilitation Home for the Elderly in South Kalimantan Province, with purposive sampling technique, which is a sampling technique based on specific criteria, resulting in 30 participants out of 69. The calcium levels (Ca) of the 30 respondents were calculated to have a mean of 6.81mg/dl, a maximum of 10.80 mg/dl, and a minimum of 3.30 mg/dl. The results showed that 21 respondents had low calcium levels, 8 respondents had normal levels, and 1 respondent had high calcium levels. Based on the characteristics of the 30 respondents, there were 18 male and female respondents in the younger age group (60-74 years), and 3 respondents in the older age group (75-95 year).

Unveiling 4500 years of environmental dynamics and human activity at Songo Mnara, Tanzania

Coastal East Africa has undergone massive transformations through the Late Holocene, with a combination of changes in sea level, increasing human settlement, and ensuing use of coastal resources. A comprehensive multi-proxy analysis, including pollen, phytolith, charcoal, stratigraphy, particle size, and geochemical data from sedimentary cores extracted from mangrove ecosystems combined with soils from archaeological contexts, provided valuable insights into vegetation dynamics, environmental changes, and human interactions within the mangrove ecosystem of Songo Mnara Island, Tanzania over the last 2590 BCE (4540 cal yr BP). The bottommost layers indicate a lack of vegetation, as deduced from the presence of coral rags and high calcium and carbonate content, possibly due to high mid-Holocene sea-level. Evidence of mangrove taxa suggests a decrease in sea level, enabling the establishment of mangroves from around 2590 BCE. A brief period of sea-level rise occurred between 90 BCE and 320 CE before sea-level fell until 1570 CE. Significant evidence of human activity is recorded from around 1400 CE indicated by increased charcoal, crop phytoliths, and evidence of marine resource utilisation. The timing of this human-environment interaction is also linked to the time of lower sea level. However, there was evidence suggesting human abandonment of the island from around 1500 CE. This coincided with a subsequent rise in sea levels and potentially prolonged drought conditions spanning from 1570 to 1700 CE. These factors likely contributed to a shortage of food resources in the area, impacting both agricultural practices due to the scarcity of natural freshwater and the accessibility of marine food resources. From 1700 CE to the present, fluctuations in sea level have been observed, with a signal of recent sea-level rise in tandem with shifts in mangrove, terrestrial herbaceous taxa and fire activity.The low sedimentation rates within mangrove areas suggest that the mangroves on Songo Mnara Island may not keep pace with the current rate of sea-level rise.

Process intensification of the electrodialysis metathesis for calcium bromide preparation: Effects of key influencing factors and phenomena of water migration

Calcium bromide, a high-value bromine resource, is widely used in the industrial synthesis processes and mercury removal from flue gas. However, there are some problems in traditional preparation methods for calcium bromide, such as long processing time, environmentally unfriendly and so on. To address these challenges, an electrodialysis metathesis (EDM) process was developed in this study to prepare high value-added calcium bromide using sodium bromide and calcium chloride as the raw materials. In order to obtain calcium bromide with high purity and improve the economic efficiency of the EDM process, the key influencing factors were investigated. Furthermore, the water migration during EDM was analyzed, and potential prospects of reducing apparent water flux volume by lowering temperature, current density, and the concentration difference between the dilute chamber and the concentrated chamber within an appropriate range. As a result, a calcium bromide product with a purity higher than 99 % was obtained with no Cl− detected. Mass fraction of Na+ was found to be 0.016 %, meeting the standards required by the industry. This method provides a new perspective and a good guide for the efficient preparation of highly valued bromine resources.

Dietary potassium effects on renal calcium transport at single nucleus resolution

Calcium homeostasis is maintained by the coordination of the intestines, bones, and kidneys. Dietary calcium is absorbed in the intestines, enters the bloodstream, and eventually finds its place in the bone reservoir or undergoes filtration by the glomerulus before being reabsorbed throughout the nephron. The late distal convoluted tubule (DCT2) and connecting tubule (CNT) determine the final rate of urinary calcium excretion, since no calcium reabsorption takes place beyond the connecting tubule (CNT). When excess calcium is excreted in the urine, also known as hypercalciuria, it contributes to the development of osteoporosis and the formation of kidney stones. High dietary potassium intake is strongly associated with a lower risk of kidney stone formation. Yet, the precise mechanism by which potassium intake modulates calcium excretion is not clear. To examine the transcriptional changes linking high dietary potassium intake and calcium transport, we applied single-nucleus RNA-sequencing (snRNA-Seq) in enriched distal nephron cells. To achieve the enrichment, we crossed a Calbindin 1 (Calb1)-driven Cre mouse line with the INTACT (for Isolation of Nuclei TAgged in Specific Cell Types) reporter line, which allows the GFP reporter to be expressed at the nuclear envelope of all calbindin 1-expressing cells. As the Calb1-Cre is constitutively expressed, all cells that have calbindin 1 expression at any time point during development will express the reporter. We examined the GFP expression by immunofluorescence, and found that it is expressed along the entire distal nephron from the distal convoluted tubule (DCT), CNT, to the collecting duct (CD). Male Calb1-Cre-INTACT mice were provided either a normal (NK, 1.05%) or a high (HK, 2%) potassium diet for 4 days and kidneys were snap-frozen for targeted snRNA-Seq using 10X Chromium (n=3 mice per group, targeting 10,000 nuclei per mouse). Our snRNA-seq dataset showed 6 major clusters, including DCT (DCT1 and DCT2), CNT (3 subclusters), CD principal cells (3 subclusters), ɑ-intercalated cells (2 subclusters), β-intercalated cells (2 subclusters), and proliferating cells. We curated a calcium score from the expression of known calciotropic genes ( Slc8a1, Vdr, Trpv5, Calb1, S100g, Ryr2, Trpv6) and used it as an index of calcium handling capacity. The calcium score is the highest in DCT2 and CNT along the distal nephron. In all CNT subclusters of the HK-treated animals, the calcium score is higher compared to the NK-treated animals, suggesting more calcium transport in CNT. Given that HK stimulates aldosterone and causes CNT hypertrophy, we compared the results with mice subjected to low dietary potassium (LK) treatment and metolazone (thiazide) treatment. The aim was to delineate the effects of dietary potassium and aldosterone A comprehensive summary of plasma potassium levels, urinary calcium excretion, aldosterone levels, calcium scores, and CNT morphology has been presented in the table below. Notably, the comparative analysis reveals a significant correlation between aldosterone levels and CNT size, both of which exhibit an inverse relationship with urinary calcium excretion: [Formula: see text] Our results suggest that DCT2 and CNT are the major sites for calcium transport. Yet, CNT exhibit more heightened adaptability in response to physiological or pharmacological perturbations at both transcriptional and morphological levels. Calcium homeostasis coincides with the regulation of aldosterone levels and CNT remodeling. DK51496 and DK133220 to DHE; K01DK121737 and AHA 20CDA35320169 to JWN. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

The A986S calcium sensing receptor variant supports the adaptation of breast cancer cells to high calcium — a novel paradigm in breast cancer progression

Breast cancer and other solid tumors that metastasize to bone are frequently accompanied by a progressive increase in circulating calcium as the disease becomes more malignant. The increase in circulating calcium not only disrupts systemic calcium homeostasis but also alters the ability of cells to sense slight changes in extracellular Ca2+ via the ubiquitous calcium sensing receptor (CaSR). Of the CASR polymorphisms at rs1801725 (p.A986S), rs1801726 (p.Q1011E), and rs1042636 (p.R990G) in exon 7 of the receptor, previous studies have shown that only the A986S CaSR variant is associated with calcium in breast, prostate and skin cancers. In this study, we assessed if exon 7 inactivating A986S (AS-CaSR) and Q1011E (QE-CaSR) variants of the receptor influence the survival of breast tumor cells at higher Ca2+. We generated isogenic HEK293T and BT-549 cells expressing these CaSR variants and carried out bulk RNA sequencing of high Ca2+ adapted control and breast cancer cells. CaSR expression was detected by western blotting using anti-flag M2 (Sigma) antibody. By using phospho-ERK1/2 as the readout, the activation of AS-CaSR required higher concentrations of Ca2+ (EC50 ~5.28 mM) compared to the QE-CaSR (EC50 of ~3.81 mM) and the wild type (WT-CaSR; EC50 ~3.69 mM) receptor. When the CaSR variants transfected cells initially cultured at 7.5 mM for 6 days, and then sub-cultured in medium containing 7.5 to 20 mM Ca2+, the viability of empty vector control and WT-CaSR transfected cells strongly decreased at Ca2+ levels beyond 7.5 mM and these cells were mostly necrotic at ≥15 mM Ca2+. The QE-CaSR variant transfected cells on the other hand exhibited limited cell death beyond 10 mM Ca2+, while the surviving colonies of the AS-CaSR variant were viable at up to 15 mM Ca2+ with invasive tendencies. Analysis of differentially expressed genes in high Ca2+ adapted mesenchymal-like BT-549 and epithelial MDA-MB-468 breast cancer cells compared to MCF10A control cells revealed a small set of differentially expressed genes that were significantly associated with relapse free survival of basal-like breast cancer patients. Among these genes are ligands and negative regulators of the WNT/FZD pathway. Our data suggest that the A986S-CaSR variant with reduced sensitivity to Ca2+ supported adaptation of TNBC cells at high Ca2+, and that high Ca2+ adaptation of breast cancer cells may be mediated via a panel of high Ca2+ inducible genes with implications in bone metastasis. This study is supported by NIH SC1GM139814, T32AI007281, T32GM144927, T32HL007737; and the American Cancer Society. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Innovative use of shrimp shell powder in carbon paste electrode for the electrochemical detection of dopamine and paracetamol: Valorization, characterization and application

The present study outlines the design of an innovative, cost-effective, and environmentally friendly electrochemical sensor, based on a carbon paste electrode (CPE) modified by shrimp shell waste (SSW). The developed electrode was used to facilitate the electrocatalytic detection of two compounds, namely dopamine (DA) and paracetamol (PAR). The use of SSW as an electrode material is ground breaking. The results have shown that the addition of shrimp shell (SS) in the CPE has led to a significant increase in the active surface area (0.504 cm2) and has improved electron transfer by decreasing the charge transfer resistance (17.45 Ω.cm2) at the electrode/solution interface. The SSW was characterized by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), X-ray diffractometry (XRD), and fourier transform infra-red (FTIR). The results show a fibrous and porous structure with high calcium content and other chemical elements including carbon, oxygen, sulphur, phosphorus and nitrogen. The electrochemical behavior of DA and PAR on the CPE-SS electrode was studied using cyclic voltammetry (CV), chronoamperometry (CA), electrochemical impedance spectroscopy (EIS), and square wave voltammetry (SWV) techniques. Additionally, CPE and CPE-SS electrodes were examined by SEM/EDS and FTIR. The effect of scan rate shows that the electrochemical process is diffusion controlled for DA and PAR. The CPE-SS electrode provide a wide linear range, good selectivity, repeatability, precision (≤5%), and low detection limits 3.6 µM and 5.54 µM for DA and PAR, respectively. In addition, the developed sensor was applied to detect DA and PAR in urine, tap water, and pharmaceutical tablets with apparent recoveries.

Pengaruh Pemberian Pupuk Organik Jenis Bokashi Terhadap Produktivitas Tanaman Kacang Tanah (Arachis hypogaea L.)

Peanuts are rich in fat, contain high protein, iron, vitamin E and calcium, vitamin B complex and phosphorus, vitamins A and K, lecithin, choline and calcium. Problems faced by local communities include low peanut production due to poor growth systems due to less than optimal fertilization. This is because the price of chemical fertilizers is high and difficult to obtain. This type of research is experimental research (true experiment research). This research aims to determine the effect of providing bokashi type organic fertilizer on the productivity of peanut plants. One way ANOVA test results obtained: 1) calculated F value of 4.915 with a significant value of 0.019 for the number of fruits, 2) calculated F value of 4.936 with a significant value of 0.019 for gross weight, 3) calculated F value of 6.149 with a significant value of 0.009 for net weight. Based on these data, it is known that the significant value for the number of fruit is 0.019, the gross weight is 0.019, and the net weight is 0.009, which is smaller than 0.05, so it can be concluded that the application of bokashi fertilizer has a real effect on the productivity of peanut plants (A hypogaea L.), this shows that Ha is accepted and Ho is rejected, then in this case it is necessary to proceed to the Duncan test.