Oral supplementation with Scabiosa artropurperea L. aqueous extract improves the in vivo sexual behaviour, sperm quality and androgen level in rams.

The effects of an aqueous extract of Scabiosa atropurpurea L. (AES) on the reproduction potential of Queue Fine de l'Ouest rams were evaluated over 9 weeks. Eighteen mature (4-6 years old) rams (52.8 ± 2.6 kg) were divided into three groups. The control (C) group was fed oat hay ad libitum with 700 g of concentrate and the other two groups were fed the same diet supplemented with AES at 1 and 2 mg/kg body weight (AES1 and AES2, respectively). Ram sperm was collected with an artificial vagina (2 × 2 days/week) to evaluate sperm production and quality, antioxidant activity, the adenosine triphosphate (ATP) and calcium concentrations. Sexual behaviour and plasma testosterone concentrations were also investigated. The administration of AES improved sexual behaviour (the duration of contact and the number of lateral approaches). The addition of AES also improved individual spermatozoa motility (C: 71.7% ± 6.3%; AES1: 78.3% ± 4.9%; AES2: 83.8% ± 4.4%), the sperm concentration (C: 5.6 ± 0.36; AES1: 6.4 ± 0.81; AES2: 6.7 ± 0.52 × 109 spermatozoa/mL), the ATP ratio (C: 1 ± 0.08; AES1: 2.1 ± 0.08; AES2: 3.3 ± 0.08) and the calcium concentration (C: 5.6 ± 0.24; AES1: 7.7 ± 0.21; AES2: 8.1 ± 0.24 mmol/L). AES treatment decreased the percentage of abnormal sperm (C: 18.5% ± 1.2%; AES1: 16.2% ± 1.1%; AES2: 14.8% ± 0.94%) and DNA damage (C: 62%; AES1: 27%; AES2: 33%) and was associated with elevated seminal fluid antioxidant activity (C: 22 ± 0.27; AES1: 27.1 ± 1.08 and AES2: 27.5 ± 0.36 mmol Trolox equivalents/L) and plasma testosterone (C: 8.3 ± 0.7; AES1: 11.7 ± 0.4; AES2: 15 ± 0.7 ng/L). In conclusion, our study suggests that S. atropurpurea may be potentially useful to enhance libido and sperm production and quality in ram.

Coronary artery calcium score as independent risk factor of obstructive coronary artery disease.

OBJECTIVE: Aim: To prove an independence of CAC score comparatively to conventional risk factors such as age, and dyslipidemia especially in patients under forty years of age. PATIENTS AND METHODS: Materials and Methods: Thirty-four asymptomatic adult patients with no prior established atherosclerotic cardiovascular disease, diabetes mellitus or severe comorbidities, except of complex clinical examination, underwent CT scan with evaluation of coronary artery calcium score. RESULTS: Results: The average total cholesterol level in the group was (5.62±1.02) mmol/l, indicating the presence of dyslipidemia. The average HDL level was (1.26±0.24) mmol/l, suggesting an average risk of atherosclerosis. The average LDL levels were within the borderline range at (3.63±1.01) mmol/l. The average triglyceride level was within the safe range at (1.93±1.08) mmol/l. The atherogenicity coefficient indicated a moderate risk of atherosclerosis with an average value of 3.64±1.31. The average coronary artery calcium score was 56.71±143.85, indicating minor plaques and a moderate risk of coronary artery disease. Correlation analysis revealed no significant correlation between age and the CAC score (r=0.1, p>0.05). However, reliable direct correlation of weak strength was found between the CAC score and LDL level (r=0.35, p<0.05). Direct correlations of weak strength were also observed between age and the levels of total cholesterol, LDL and the atherogenicity coefficient (r=0.43, 0.49, 0.42 respectively, p<0.05). CONCLUSION: Conclusions: Coronary artery calcium score is a valuable screening tool for identifying potential obstructive coronary artery disease, not only for individuals aged forty and above, but also for younger asymptomatic patients.

Electron Microscopy for the Stability Assessment of Parenteral Nutrition Admixtures: Focus on Precipitation.

(1) Background: parenteral nutrition (PN) is indispensable for patients unable to receive oral or enteral feeding. However, the complexity of PN solutions presents challenges regarding stability and compatibility. Precipitation reactions may occur. The most frequent is the formation of calcium phosphate (Ca-P). The different factors influencing these reactions must be considered to ensure patient safety. (2) Methods: eight paediatric PN solutions were prepared, following standard protocols. Samples were stored at room temperature and in a refrigerator. Electron microscopy, coupled with energy dispersive X-ray spectroscopy (EDS), was employed. Precipitates were analysed for composition and morphology. (3) Results: precipitates were observed in all samples, even at day 0. Crystalline structures, predominantly composed of calcium or magnesium, sometimes associated with chlorine or phosphorus, were detected. Additionally, amorphous precipitates, contained heterogeneous compositions, including unexpected elements, were identified. (4) Conclusions: various precipitates, primarily calcium- or magnesium-based, can form in PN solutions, although it is not expected that they can form under the real conditions of use. Calcium oxalate precipitation has been characterised, but the use of organic calcium and phosphate salts appears to mitigate calcium phosphate precipitation. Electron microscopy provides interesting results on NP precipitation, but sample preparation may present technical limitations that affect the interpretation of the results.

Pinecone derived hierarchical carbon nanostructure as a transducer in a solid-state ion-selective electrode for in vivo analysis of calcium ion.

Monitoring extracellular calcium ion (Ca2+) chemical signals in neurons is crucial for tracking physiological and pathological changes associated with brain diseases in live animals. Potentiometry based solid-state ion-selective electrodes (ISEs) with the assist of functional carbon nanomaterials as ideal solid-contact layer could realize the potential response for in vitro and in vivo analysis. Herein, we employ a kind of biomass derived porous carbon as a transducing layer to prompt efficient ion to electron transduction while stabilizes the potential drift. The eco-friendly porous carbon after activation (APB) displays a high specific area with inherit macropores, micropores, and large specific capacitance. When employed as transducer in ISEs, a stable potential response, minimized potential drift can be obtained. Benefiting from these excellent properties, a solid-state Ca2+ selective carbon fiber electrodes (CFEs) with a sandwich structure is constructed and employed for real time sensing of Ca2+ under electrical stimulation. This study presents a new approach to develop sustainable and versatile transducers in solid-state ISEs, a crucial way for in vivo sensing.

Time-of-day dependence of running averages for some components of metabolic panels at our hospital: Relation to strategy for patient-based quality control.

BACKGROUND AND OBJECTIVES: We assessed properties of running averages for our hospital's most common chemistry analytes, for use in real-time patient-based quality control (PBQC). We determined whether there was dependence of any running averages on 24-h clock time (time-of-day, TOD). MATERIALS AND METHODS: We analyzed 3-months' data for measurements of 13 metabolic panel components. Running averages for 20 consecutive results (20-mers) were computed for data restricted to results within reference intervals. This produced an overall mean (X) and standard-deviation (SD) of 20-mers for each analyte. We then computed the average 20-mer result (Y) reported within 1-h bins across 24-hour clock time (t). Y(t) was regarded as having TOD-dependence if either nadir or apex values for |Y-X| exceeded 0.5 SD, occurring within a contiguous series of at least 4 Y(t) values on one side of the mean. RESULTS: Seven analytes (albumin, aspartate aminotransferase, calcium, chloride, CO2, potassium, total protein) demonstrated TOD-dependence of running means for 20-mers. CONCLUSIONS: At our hospital, TOD-dependence of running means was identified for 7 of 13 metabolic panel analytes. TOD-dependence is likely to be hospital-specific. Utilization of TOD-dependent targets for PBQC, rather than fixed targets, would be appropriate in these cases.

Fluorescent Protein-Based Sensors for Detecting Essential Metal Ions across the Tree of Life.

Genetically encoded fluorescent metal ion sensors are powerful tools for elucidating metal dynamics in living systems. Over the last 25 years since the first examples of genetically encoded fluorescent protein-based calcium indicators, this toolbox of probes has expanded to include other essential and non-essential metal ions. Collectively, these tools have illuminated fundamental aspects of metal homeostasis and trafficking that are crucial to fields ranging from neurobiology to human nutrition. Despite these advances, much of the application of metal ion sensors remains limited to mammalian cells and tissues and a limited number of essential metals. Applications beyond mammalian systems and in vivo applications in living organisms have primarily used genetically encoded calcium ion sensors. The aim of this Perspective is to provide, with the support of historical and recent literature, an updated and critical view of the design and use of fluorescent protein-based sensors for detecting essential metal ions in various organisms. We highlight the historical progress and achievements with calcium sensors and discuss more recent advances and opportunities for the detection of other essential metal ions. We also discuss outstanding challenges in the field and directions for future studies, including detecting a wider variety of metal ions, developing and implementing a broader spectral range of sensors for multiplexing experiments, and applying sensors to a wider range of single- and multi-species biological systems.

Machine learning reveals the control mechanics of an insect wing hinge.

Insects constitute the most species-rich radiation of metazoa, a success that is due to the evolution of active flight. Unlike pterosaurs, birds and bats, the wings of insects did not evolve from legs1, but are novel structures that are attached to the body via a biomechanically complex hinge that transforms tiny, high-frequency oscillations of specialized power muscles into the sweeping back-and-forth motion of the wings2. The hinge consists of a system of tiny, hardened structures called sclerites that are interconnected to one another via flexible joints and regulated by the activity of specialized control muscles. Here we imaged the activity of these muscles in a fly using a genetically encoded calcium indicator, while simultaneously tracking the three-dimensional motion of the wings with high-speed cameras. Using machine learning, we created a convolutional neural network3 that accurately predicts wing motion from the activity of the steering muscles, and an encoder-decoder4 that predicts the role of the individual sclerites on wing motion. By replaying patterns of wing motion on a dynamically scaled robotic fly, we quantified the effects of steering muscle activity on aerodynamic forces. A physics-based simulation incorporating our hinge model generates flight manoeuvres that are remarkably similar to those of free-flying flies. This integrative, multi-disciplinary approach reveals the mechanical control logic of the insect wing hinge, arguably among the most sophisticated and evolutionarily important skeletal structures in the natural world.

Sizeable net export of base cations from a Carpathian flysch catchment indicates their geogenic origin while the 26Mg/24Mg, 44Ca/40Ca and 87Sr/86Sr isotope ratios in runoff are indistinguishable from atmospheric input.

Nutrient imbalances may negatively affect the health status of forests exposed to multiple stress factors, including drought and bark beetle calamities. We studied the origin of base cations in runoff from a small Carpathian catchment underlain by base-poor flysch turbidites using magnesium (Mg), calcium (Ca) and strontium (Sr) isotope composition of 10 ecosystem compartments. Our objective was to constrain conclusions drawn from long-term hydrochemical monitoring of inputs and outputs. Annual export of Mg, Ca and Sr exceeds 5-to-15 times their atmospheric input. Mass budgets per se thus indicate sizeable net leaching of Mg, Ca and Sr from bedrock sandstones and claystones. Surprisingly, δ26Mg, δ44Ca and 87Sr/86Sr isotope ratios of runoff were practically identical to those of atmospheric deposition and soil water but significantly different from bedrock isotope ratios. We did not find any carbonates in the studied area as a hypothetical, easily dissolvable source of base cations whose isotope composition might corroborate the predominance of geogenic base cations in the runoff. Marine carbonates typically have lower δ26 Mg and 87Sr/86Sr ratios, and silicate sediments often have higher δ26Mg and 87Sr/86Sr ratios than runoff at the study site. Mixing of these two sources, if confirmed, could reconcile the flux and isotope data.

Improvement of the Green-Red Förster Resonance Energy Transfer-Based Ca2+ Indicator by Using the Green Fluorescent Protein, Gamillus, with a Trans Chromophore as the Donor.

To monitor the Ca2+ dynamics in cells, various genetically encoded Ca2+ indicators (GECIs) based on Förster resonance energy transfer (FRET) between fluorescent proteins are widely used for live imaging. Conventionally, cyan and yellow fluorescent proteins have been often used as FRET pairs. Meanwhile, bathochromically shifted indicators with green and red fluorescent protein pairs have various advantages, such as low toxicity and autofluorescence in cells. However, it remains difficult to develop them with a similar level of dynamic range as cyan and yellow fluorescent protein pairs. To improve this, we used Gamillus, which has a unique trans-configuration chromophore, as a green fluorescent protein. Based on one of the best high-dynamic-range GECIs, Twitch-NR, we developed a GECI with 1.5-times higher dynamic range (253%), Twitch-GmRR, using RRvT as a red fluorescent protein. Twitch-GmRR had high brightness and photostability and was successfully applied for imaging the Ca2+ dynamics in live cells. Our results suggest that Gamillus with trans-type chromophores contributes to improving the dynamic range of GECIs. Therefore, selection of the cis-trans isomer of the chromophore may be a fundamental approach to improve the dynamic range of green-red FRET indicators, unlimited by GECIs.

Serum and tissue metallome of pancreatic ductal adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) patients have late presentation at the time of diagnosis and a poor prognosis. Metal dyshomeostasis is known to play a role in cancer progression. However, the blood and tissue metallome of PDAC patients has not been assessed. This study aimed to determine the levels of essential and toxic metals in the serum and pancreatic tissue from PDAC patients. Serum samples were obtained from PDAC patients before surgical resection. Tissue (tumor and adjacent normal pancreas) were obtained from the surgically resected specimen. Inductively coupled plasma-mass spectrometry (ICP-MS) analysis was performed to quantify the levels of 10 essential and 3 toxic metals in these samples. Statistical analysis was performed to identify dysregulated metals in PDAC and their role as potential diagnostic and prognostic biomarkers. Significantly decreased serum levels of magnesium, potassium, calcium, iron, zinc, selenium, arsenic, and mercury and increased levels of molybdenum were shown to be associated with PDAC. There were significantly decreased levels of zinc, manganese and molybdenum, and increased levels of calcium and selenium in the pancreatic tumor tissue compared with the adjacent normal pancreas. Notably, lower serum levels of calcium, iron, and selenium, and higher levels of manganese, were significantly associated with a poor prognosis (i.e., overall survival) in PDAC patients. In conclusion, this is the first study to comprehensively assess the serum and tissue metallome of PDAC patients. It identified the association of metals with PDAC diagnosis and prognosis.

Trace metals and nutrient analysis of marine fish species from the Gwadar coast.

Trace metals are naturally occurring metals found in very small concentrations in the environment. In the context of fish flesh, metals such as copper, calcium, potassium, sodium, zinc, iron, and manganese are absorbed by fish and play vital roles in various physiological functions. However, if these metals exceed the recommended limits set by WHO/FAO, they are termed 'toxic metals' due to their harmful impacts on both the fish and its consumers. Therefore, the present study aims to analyze the levels of protein, lipids, and certain metals-Aluminum (Al), Sodium (Na), Zinc (Zn), Titanium (Ti), Iron (Fe), Copper (Cu), Potassium (K), and Calcium (Ca) in three commercially important marine fishes i.e. Rastrelliger kanagurta, Sardinella abella, and Otolithes ruber. The study also aims to assess their potential impact on human health. The macro-Kjeldhal method and Soxhlet apparatus were used to estimate protein and lipid contents, while atomic absorption spectroscopy (AAS) was used to estimate trace metals found in fishes. The study found that these fish species are valuable sources of protein, lipids, and certain essential minerals. The protein content (CP) in these three species ranged from 63.35 to 86.57%, while lipid content was from 21.05 to 23.86%. The overall results of the trace metal concentrations analyzed in the present study revealed that Aluminum (Al), Sodium (Na), Zinc (Zn), Titanium (Ti), Copper (Cu), Potassium (K), and Calcium (Ca) were found in low concentration or traces and also within suitable ranges as set by WHO/FAO. However, Iron (Fe) was absent in all three species. Moreover, both copper and potassium were found in all three species, while Zinc was present in Rastrelliger kanagurta and Sardinella abella, calcium in Sardinella abella, and sodium in Otolithes ruber only. Titanium was recorded for the first time in S. abella. However, the total health risk assessment associated with these fish food consumption was measured by THQ and TTHQ and found to be less than 1, which shows no potential risk related to trace metals found in these fishes on human health upon their consumption. In conclusion, these commercially important marine fish species were found valuable sources of protein, lipids, and essential trace minerals that are necessary for human health. Thus, the current study provides useful information for the local population to make informed decisions about their daily diets and highlights the importance of sustainable fishing practices to maintain these valuable marine resources by periodical monitoring of their ecosystem.

Association between the Composition of Drinking Water and Cognitive Function in the Elderly: A Systematic Review.

The prevalence of dementia increases with nearly 10 million new cases each year, with Alzheimer's disease contributing to 60-70% of cases. Environmental factors such as drinking water have been evaluated to determine if a relationship exists between trace elements in drinking water and the risk of developing cognitive disorders in the elderly. The purpose of the current systematic review was to evaluate an association between the composition of drinking water and cognitive function in the elderly. In accordance with the Preferred Reporting Items for Systematic Review and Meta-Analyses guidelines, a literature search was conducted using PubMed and CINAHL databases. A total of 10 studies were included in the current systematic review. Aluminum is the most commonly evaluated trace element in studies (n = 8), followed by silica (n = 5), calcium (n = 4), and fluoride (n = 4). Aluminum exposure showed an increased risk of cognitive decline in four studies, with no association reported in the other studies. Higher silica and pH levels were shown to be protective against a decline in cognitive function. A similar protective effect of calcium was found in two studies. Future research should measure multiple trace mineral levels in all water sources to evaluate the impact on cognitive function.

Geochemical, mineralogical records, and statistical approaches in establishing sedimentary in the environment of a Mediterranean coastal system: case of Sebkha El-Guettiate (southeastern Tunisia).

The current study was conducted within the context of the Holocene era in Sebkha El-Guettiate, located in southeastern Tunisia. The aim was to determine the factors influencing the geochemical and mineralogical composition of sediments and to elucidate the sedimentary characteristics of the Holocene within the Sebkha core. We examined a sediment core extending 100 cm from this Sebkha, subjecting it to comprehensive analysis to uncover its sedimentological, mineralogical, and geochemical properties. Several techniques were employed to strengthen and validate the connections between geochemical and mineralogical analyses, including X-ray diffraction (XRD), X-ray fluorescence (XRF), scanning electron microscopy (SEM), and infrared (IR) spectroscopy, among others. Furthermore, statistical analyses utilizing principal component analysis (PCA) were applied to the results of the geochemical and mineralogical studies, aiding in the identification of patterns and relationships. A comprehensive mineralogical assessment of the core's sediments revealed the presence and interpretation of carbonate minerals, evaporite minerals, and detrital minerals. Through the application of infrared (IR) spectrometer techniques to all sediment samples, we gained insight into the mineralogical components and the distribution of key elements such as quartz, kaolinite, calcite, feldspar, and organic carbon. The geochemical composition demonstrated a clear dominance of silica (SiO2), accompanied by fluctuations in carbonate percentages (CaCO3). The prominent major elements, primarily magnesium (Mg) and calcium (Ca) originating from dolomitization, sodium (Na) and chlorine (Cl) from halite, and calcium (Ca) from gypsum, exhibited varying levels. Results from Rock-Eval 6 pyrolysis indicated that the organic matter within the sediments is generally a mixture of terrestrial and aquatic origins. This study provides practical information that underscores the diverse origins contributing to Sebkha sediment formation, often influenced by saline systems.

Subicular neurons encode concave and convex geometries.

Animals in the natural world constantly encounter geometrically complex landscapes. Successful navigation requires that they understand geometric features of these landscapes, including boundaries, landmarks, corners and curved areas, all of which collectively define the geometry of the environment1-12. Crucial to the reconstruction of the geometric layout of natural environments are concave and convex features, such as corners and protrusions. However, the neural substrates that could underlie the perception of concavity and convexity in the environment remain elusive. Here we show that the dorsal subiculum contains neurons that encode corners across environmental geometries in an allocentric reference frame. Using longitudinal calcium imaging in freely behaving mice, we find that corner cells tune their activity to reflect the geometric properties of corners, including corner angles, wall height and the degree of wall intersection. A separate population of subicular neurons encode convex corners of both larger environments and discrete objects. Both corner cells are non-overlapping with the population of subicular neurons that encode environmental boundaries. Furthermore, corner cells that encode concave or convex corners generalize their activity such that they respond, respectively, to concave or convex curvatures within an environment. Together, our findings suggest that the subiculum contains the geometric information needed to reconstruct the shape and layout of naturalistic spatial environments.

The dual function of calcium ion in fruit edible coating: Regulating polymer internal crosslinking state and improving fruit postharvest quality.

The edible coating is proved to be a convenient approach for fruit preservation. Among these published explorations, naturally sourced macromolecules and green crosslinking strategies gain attention. This work centers on edible coatings containing Ca2+ as crosslinker for the first time, delving into crosslinking mechanisms, include alginate, chitosan, Aloe vera gel, gums, etc. Additionally, the crucial functions of Ca2+ in fruit's quality control are also elaborated in-depth, involving cell wall, calmodulin, antioxidant, etc. Through a comprehensive review, it becomes evident that Ca2+ plays a dual role in fruit edible coating. Specifically, Ca2+ constructs a three-dimensional dense network structure with polymers through ionic bonding. Moreover, Ca2+ acts directly with cell wall to maintain fruit firmness and serve as a second messenger to participate secondary physiological metabolism. In brief, coatings containing Ca2+ present remarkable effects in preserving fruit and this work may provide guidance for Ca2+ related fruit preservation coatings.

Breastmilk calcium concentrations in Palestinian lactating women.

AIM: Breastmilk calcium concentrations can vary between lactating women and over the lactation period. This study assessed breastmilk calcium concentrations among Palestinian lactating women. METHODS: The demographic and dietary variables of the lactating women were collected using a questionnaire. The women provided a sample of about 5 mL of breastmilk using hand expression. Breastmilk calcium concentrations were quantified using an inductively coupled plasma-mass spectrometric method. RESULTS: Breastmilk samples were taken from 240 lactating women. The mean breastmilk calcium concentration was 285.4 ± 115.1 mg/L. Lower breastmilk calcium concentrations were associated with age, lactating period, unemployment, dissatisfaction with income and insufficient consumption of vitamins and minerals. CONCLUSION: Breastmilk calcium concentrations were affected by demographic variables of the lactating women and insufficient consumption of vitamins and minerals. The findings reported in this study are informative to healthcare providers and decision makers who might be interested in improving the health of lactating women and their infants.

Non-destructive monitoring and prediction of fouling by organic matters and residual anionic coagulant during membrane process.

Physical fouling characteristics on silicon carbide (SiC) membranes induced by various organic matter compounds vary depending on the presence of calcium ions (Ca2+). Both destructive techniques (morphological surface analysis) and non-destructive techniques (fouling properties monitoring) were used to determine the fouling mechanisms and behavior during the membrane filtration systems. Destructive analysis and a modified Hermia model were employed to assess the fouling mechanisms. Fouling behavior was also analyzed through non-destructive monitoring techniques including optical coherence tomography (OCT) and three-dimensional laser scanning confocal microscopy (3D-LSM). At concentrations of 10, 30, and 100 mg/L without Ca2+, the flux decreased by 57-95% for humic acid (HA) and anionic polyacrylamide (APAM). APAM exhibited a notable removal rate of up to 56% without Ca2+. At concentration of 10, 30, and 100 mg/L in the absence of Ca2+, the flux decreased by 6-8% for sodium alginate (SA). However, the addition of Ca2+ led to a reduction in the flux for SA by up to 91% and resulted in a removal rate of 40%. Furthermore, addition of Ca2+ led to an alteration of the fouling characteristics of HA and SA. In the case of HA, higher concentrations resulted in elevated thickness and roughness with correlation coefficients of 0.991 and 0.992, respectively. For SA, increased SA concentration led to a thicker (correlation coefficient of 0.999) but smoother surfaces (correlation coefficients of 0.502). Monitoring of these physical characteristics of the fouling layer through non-destructive analysis is crucial for effective fouling management, optimization of the system performance and extending the lifespan of the membrane. By continuously assessing the fouling layer thickness and surface roughness, we expect to be able to provide insights on the fouling behavior, identify trends, that can help scientists and engineers to make informed decisions regarding fouling control strategies in future.

Evidence from experiments, modeling, and field observations for effects of increased salinization on re-distribution of sediment base cations in Taihu Lake, China.

Taihu Lake, the third largest freshwater lake in China, has experienced rapid salinization in the past decades; however, little is known about the impact of sodium (Na) on ion exchange in the lake environment. To explore the potential effect of increased Na on the migration of base cations (Ca and Mg) and resulting redistribution between the water and sediment, we used the adsorption-exchange experiment, MINTEQ modeling to explore the cation exchange induced by high Na input, and its impact on the redistribution of Ca and Mg in Taihu different media. The results indicated that exchanged quantity of Ca and Mg increased with time, and the exchange process reached 90% during 0-4 h and reached equilibrium after 24 h under 100 mg/L Na (the maximum Na concentration in Taihu sediment pore water). Our MINTEQ modeled result indicated that the exchanged quantity of Ca and Mg increased with the increasing Na concentration, with Ca being preferably exchanged over Mg at the same Na concentration. The MINTEQ model further predicted that, in the Taihu lake environment, the exchange adsorption would reach the equilibrium at the concentration of 6000 mg/L Na, with exchanged Ca2+ and Mg2+ accounting for 47% and 55% of the total exchangeable Ca and Mg in the sediment, respectively. Although current Na-induced exchange in the Taihu lake has been far from the equilibrium, the MINTEQ result confirmed the existence of this reaction and predicted the potential redistribution of base cations or Ca/Mg ratio in the lake sediment and water phase with further Na increase. Furthermore, our field observations not only confirmed the existence of Na-induced cation exchange in this lake environment but also were generally in agreement with our experimental and modeled results. The increased salinization-induced ion exchange would alter the re-distribution of base cations and the resulting potential ecosystem consequences should be given close attention in this large freshwater lake.

Live Calcium Imaging of Virus-Infected Human Intestinal Organoid Monolayers Using Genetically Encoded Calcium Indicators.

Calcium signaling is an integral regulator of nearly every tissue. Within the intestinal epithelium, calcium is involved in the regulation of secretory activity, actin dynamics, inflammatory responses, stem cell proliferation, and many other uncharacterized cellular functions. As such, mapping calcium signaling dynamics within the intestinal epithelium can provide insight into homeostatic cellular processes and unveil unique responses to various stimuli. Human intestinal organoids (HIOs) are a high-throughput, human-derived model to study the intestinal epithelium and thus represent a useful system to investigate calcium dynamics. This paper describes a protocol to stably transduce HIOs with genetically encoded calcium indicators (GECIs), perform live fluorescence microscopy, and analyze imaging data to meaningfully characterize calcium signals. As a representative example, 3-dimensional HIOs were transduced with lentivirus to stably express GCaMP6s, a green fluorescent protein-based cytosolic GECI. The engineered HIOs were then dispersed into a single-cell suspension and seeded as monolayers. After differentiation, the HIO monolayers were infected with rotavirus and/or treated with drugs known to stimulate a calcium response. An epifluorescence microscope fitted with a temperature-controlled, humidified live-imaging chamber allowed for long-term imaging of infected or drug-treated monolayers. Following imaging, acquired images were analyzed using the freely available analysis software, ImageJ. Overall, this work establishes an adaptable pipeline for characterizing cellular signaling in HIOs.