Increase In Calcium Research Articles

Neuronal Calcium Sensor-1 Protects Cortical Neurons from Hyperexcitation and Ca2+ Overload during Ischemia by Protecting the Population of GABAergic Neurons.

A defection of blood circulation in the brain leads to ischemia, damage, and the death of nerve cells. It is known that individual populations of GABAergic neurons are the least resistant to the damaging factors of ischemia and therefore they die first of all, which leads to impaired inhibition in neuronal networks. To date, the neuroprotective properties of a number of calcium-binding proteins (calbindin, calretinin, and parvalbumin), which are markers of GABAergic neurons, are known. Neuronal calcium sensor-1 (NCS-1) is a signaling protein that is expressed in all types of neurons and is involved in the regulation of neurotransmission. The role of NCS-1 in the protection of neurons and especially their individual populations from ischemia and hyperexcitation has not been practically studied. In this work, using the methods of fluorescence microscopy, vitality tests, immunocytochemistry, and PCR analysis, the molecular mechanisms of the protective action of NCS-1 in ischemia/reoxygenation and hyperammonemia were established. Since NCS-1 is most expressed in GABAergic neurons, the knockdown of this protein with siRNA led to the most pronounced consequences in GABAergic neurons. The knockdown of NCS-1 (NCS-1-KD) suppressed the basic expression of protective proteins without significantly reducing cell viability. However, ischemia-like conditions (oxygen-glucose deprivation, OGD) and subsequent 24-h reoxygenation led to a more massive activation of apoptosis and necrosis in neurons with NCS-1-KD, compared to control cells. The mass death of NCS-1-KD cells during OGD and hyperammonemia has been associated with the induction of a more pronounced network hyperexcitation symptom, especially in the population of GABAergic neurons, leading to a global increase in cytosolic calcium ([Ca2+]i). The symptom of hyperexcitation of neurons with NCS-1-KD correlated with a decrease in the level of expression of the calcium-binding protein-parvalbumin. This was accompanied by an increase in the expression of excitatory ionotropic glutamate receptors, N-methyl-D-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (NMDAR and AMPAR) against the background of suppression of the expression of glutamate decarboxylase (synthesis of γ-aminobutyric acid).

Chemosensory detection of aversive concentrations of ammonia and basic volatile amines in insects

Basic volatiles like ammonia are found in insect environments, and at high concentrations cause an atypical action potential burst, followed by inhibition in multiple classes of olfactory receptor neurons (ORNs) in Drosophila melanogaster. During the period of inhibition, ORNs are unable to fire action potentials to their ligands but continue to display receptor potentials. An increase in calcium is also observed in antennal cells of Drosophila and Aedes aegypti. In the gustatory system, ammonia inhibits sugar and salt responses in a dose-dependent manner. Other amines show similar effects in both gustatory and olfactory neurons, correlated with basicity. The concentrations that inhibit neurons reduce proboscis extension to sucrose in Drosophila. In Aedes, a brief exposure to volatile ammonia abolishes attraction to human skin odor for several minutes. These findings reveal an effect that prevents detection of attractive ligands in the olfactory and gustatory systems and has potential in insect control.

Activation of Piezo1 downregulates renin in juxtaglomerular cells and contributes to blood pressure homeostasis

BackgroundThe synthesis and secretion of renin in juxtaglomerular (JG) cells are closely regulated by the blood pressure. To date, however, the molecular identity through which JG cells respond to the blood pressure remains unclear.ResultsHere we discovered that Piezo1, a mechanosensitive ion channel, was colocalized with renin in mouse kidney as well as As4.1 cells, a commonly used JG cell line. Activation of Piezo1 by its agonist Yoda1 induced an intracellular calcium increase and downregulated the expression of renin in these cells, while knockout of Piezo1 in JG cells abolished the effect of Yoda1. Meanwhile, mechanical stress using microfluidics also induced an intracellular calcium increase in wildtype but not Piezo1 knockout JG cells. Mechanistically, we demonstrated that activation of Piezo1 upregulated the Ptgs2 expression via the calcineurin-NFAT pathway and increased the production of Ptgs2 downstream molecule PGE2 in JG cells. Surprisingly, we discovered that increased PGE2 could decreased the renin expression through the PGE2 receptor EP1 and EP3, which inhibited the cAMP production in JG cells. In mice, we found that activation of Piezo1 significantly downregulated the renin expression and blood pressure in wildtype but not adeno-associated virus (AAV)-mediated kidney specific Piezo1 knockdown mice.ConclusionsIn summary, these results revealed that activation of Piezo1 could downregulate the renin expression in JG cells and mice, subsequently a reduction of blood pressure, highlighting its therapeutic potential as a drug target of the renin-angiotensin system.

OR2AT4 and OR1A2 counterregulate molecular pathophysiological processes of steroid-resistant inflammatory lung diseases in human alveolar macrophages

BackgroundTherapeutic options for steroid-resistant non-type 2 inflammation in obstructive lung diseases are lacking. Alveolar macrophages are central in the progression of these diseases by releasing proinflammatory cytokines, making them promising targets for new therapeutic approaches. Extra nasal expressed olfactory receptors (ORs) mediate various cellular processes, but clinical data are lacking. This work investigates whether ORs in human primary alveolar macrophages could impact pathophysiological processes and could be considered as therapeutic targets.MethodsHuman primary alveolar macrophages were isolated from bronchoalveolar lavages of 50 patients with pulmonary diseases. The expression of ORs was validated using RT-PCR, immunocytochemical staining, and Western blot. Changes in intracellular calcium levels were analyzed in real-time by calcium imaging. A luminescent assay was used to measure the cAMP concentration after OR stimulation. Cytokine secretion was measured in cell supernatants 24 h after stimulation by ELISA. Phagocytic ability was measured by the uptake of fluorescent-labeled beads by flow cytometry.ResultsWe demonstrated the expression of functional OR2AT4 and OR1A2 on mRNA and protein levels. Both ORs were primarily located in the plasma membrane. Stimulation with Sandalore, the ligand of OR2AT4, and Citronellal, the ligand of OR1A2, triggered a transient increase of intracellular calcium and cAMP. In the case of Sandalore, this calcium increase was based on a cAMP-dependent signaling pathway. Stimulation of alveolar macrophages with Sandalore and Citronellal reduced phagocytic capacity and release of proinflammatory cytokines.ConclusionThese are the first indications for utilizing olfactory receptors as therapeutic target molecules in treating steroid-resistant lung diseases with non-type 2 inflammation.

Rapid astrocyte-dependent facilitation amplifies multi-vesicular release in hippocampal synapses

Synaptic facilitation is a major form of short-term plasticity typically driven by an increase in residual presynaptic calcium. Using near-total internal reflection fluorescence (near-TIRF) imaging of single vesicle release in cultured hippocampal synapses, we demonstrate a distinctive, release-dependent form of facilitation in which probability of vesicle release is higher following a successful glutamate release event than following a failure. This phenomenon has an onset of ≤500ms and lasts several seconds, resulting in clusters of successful release events. The release-dependent facilitation requires neuronal contact with astrocytes and astrocytic glutamate uptake by EAAT1. It is not observed in neurons grown alone or in the presence of astrocyte-conditioned media. This form of facilitation dynamically amplifies multi-vesicular release. Facilitation-evoked release events exhibit spatial clustering and have a preferential localization toward the active zone center. These results uncover a rapid astrocyte-dependent form of facilitation acting via modulation of multi-vesicular release and displaying distinctive spatiotemporal properties.

Germline deletion of the mitochondrial calcium uniporter (MCU) does not affect mitochondrial calcium increases during ischemia/reperfusion injury in ex vivo perfused mouse hearts

Germline deletion of the mitochondrial calcium uniporter (MCU) does not affect mitochondrial calcium increases during ischemia/reperfusion injury in ex vivo perfused mouse hearts

Medicinal treatment of primary hyperparathyroidism

Primary hyperparathyroidism (PHPT) is a hyperproduction of parathyroid hormone that is not provoked by a decrease in the blood calcium level, which leads to a violation of calcium homeostasis. The prevalence of PHPT has been increasing over the past 10 years due to improvements in its diagnosis. Currently, it ranks third by this indicator after diabetes and thyroid pathology. The leading clinical manifestations of PHPT are demineralization and resorption of bone tissue, as well as nephrolithiasis and the development of renal failure. It was found that a decrease in the glomerular filtration rate is inversely correlated with the level of parathyroid hormone and is associated with a greater decrease in bone mineral density. In addition, the frequency of left ventricular hypertrophy increases against the background of a significant persistent increase in calcium. Patients with the indicated symptoms due to an unestablished PHPT are under the supervision of orthopedists, cardiologists, and family doctors. Monitoring of PHPT is not a fully addressed issue today. This, in particular, is due to the fact that the functioning organizational system for the diagnosis of this pathology has significantly increased both the number of identified patients and the proportion of asymptomatic PHPT among them. Some people refuse surgery, which is currently recognized as the best option for the treatment of PHPT. Other patients have contraindications to surgery due to severe comorbid pathology. These factors increase the relevance for the development of the principles of medicinal treatment of pathology, which requires a serious analysis of existing drugs, their therapeutic potential, and the risk of complications in order to develop clinical recommendations. These questions became the basis of this review article. The base of scientific publications on PHPT and pharmacological aspects of the action of drugs in patients with this pathology were analyzed. It was found that among the full-text articles with the analysis of therapy effectiveness, the most common drugs of choice were bisphosphonates, calcimimetic cinacalcet, vitamin D and its analogues. Their prospects for the impact on the main symptoms of PHPT are carefully considered. The results of the analysis will allow clinicians to choose individual treatment approach to a particular patient.

Biogeochemical shifts and zooplankton responses in post-CAAA northeastern lakes: The success of acid recovery, complexity of biological recovery, and value of long-term monitoring

Lakes in the northeastern U.S., as in other regions, experienced significant declines in water quality and ecosystem health in the mid to late 20th century as atmospheric deposition from industrial emissions caused surface waters to acidify. After the Clean Air Act Amendments of 1990 enhanced regulations to limit emissions of sulfur and nitrogen, many lakes in the Northeast recovered from acidification. The organisms that inhabit lakes are susceptible to these shifts, and zooplankton can serve as valuable indicators of lake ecosystem responses to shifts due to their position as primary consumers in the food web. We evaluated a large historical dataset from the U.S. Environmental Protection Agency (EPA) Eastern Lake Survey (ELS) of 143 lakes throughout the northeastern U.S. to understand how recovery from acidification and subsequent chemical changes influenced zooplankton body size and community composition. We observed significant changes in chemistry; surface water sulfate concentrations decreased by 22% in the ELS lakes between the two sample years of 1986 and 2004. Increasing concentrations of DOC are common from the combined influence of deacidification and climate change, and the ELS lakes experienced a 26% increase in DOC. Notably, in the more developed regions of our study area, chloride concentration in lakes tripled, likely due to increased use of road deicers, and this is related to a subsequent increase in calcium and magnesium base cations. Zooplankton body size increased significantly between 1986 and 2004 across all taxa, and body size of Daphnia species, which require higher amounts of calcium than other zooplankton taxa, was positively associated with high calcium lakes. Shifts in zooplankton community structure, however, were most strongly influenced by variation in ANC, sulfate, and DOC. Our findings indicate that, while surface water acidity has a strong influence on zooplankton community structure, zooplankton populations are likely on a different trajectory than a simple recovery due to multiple chemical changes, such as increased chloride, occurring in lakes undergoing acid recovery. These, along with changes in climate and land-use, elicit multiple simultaneous responses in zooplankton communities.

Sensitivity of Pathogenic Bacteria Strains to Treated Mine Water

Mine water as a result of meteoric and/or underground water's contact with tailings and underground workings could have an elevated content of metals associated with sulfate, often acidic, due to the bio-oxidation of sulfides. When entering aquatic ecosystems, the mine water can cause significant changes in the species' trophic levels, therefore a treatment is required to adjust the alkalinity and to remove the heavy metals and metalloids. The conventional mine water treatment removes metals, but in many cases it does not reduce the sulfate content. This paper aimed to predict the impact of conventionally treated mine water on the receiving river by assessing the genotoxic activity on an engineered Escherichia coli and by evaluating the toxic effects generated on two Gram-negative bacterial strains, Pseudomonas aeruginosa and Escherichia coli. Although the main chemical impact is the severe increases of calcium and sulfate concentrations, no significant genotoxic characteristics were detected on the Escherichia coli strain and on the cell-viability with a positive survival rate higher than 80%. Pseudomonas aeruginosa was more resistant than Escherichia coli in the presence of 1890 mg SO42-/L. This paper reveals different sensitivities and adaptabilities of pathogenic bacteria to high concentrations of sulfates in mine waters.

Piezo1 in endothelial cells is involved in vitamin D-induced vascular calcification

The relationship between the Piezo1 channel of vascular endothelial cells and vascular calcification is unknown. In this study, after subcutaneous injection of vitamin D for 10 consecutive days, the mice showed an increase in serum calcium, aortic calcium content, vascular tension and pulse wave velocity. Piezo1channel antagonist, GsMTx4 alleviated arteriosclerosis and decreased the aortic calcium content, while Piezo1 agonist Yoda1 produced opposite effect. In addition, activation of Piezo1 by Yoda1 impaired the function of human umbilical vein endothelial cells (HUVECs), as evidenced by further decreased production of NO, reduction in expression levels of eNOS, MMP-2, PCNA and VEGFA. When co-culture of HUVECs and vascular smooth muscle cells (VSMCs), activation of Piezo1 in HUVECs enhanced expression levels of calcification-related SOX9 and Runx2 genes, increased ALP activity and calcium deposition in VSMCs. We concluded that Piezo1 in endothelial cells is involved in the pathogenesis of vascular calcification. This study provides a new experimental basis for the prevention and treatment of vascular calcification.

Autophagy Mediates Escherichia Coli-Induced Cellular Inflammatory Injury by Regulating Calcium Mobilization, Mitochondrial Dysfunction, and Endoplasmic Reticulum Stress.

Bovine endometritis is a reproductive disorder that is induced by mucus or purulent inflammation of the uterine mucosa. However, the intracellular control chain during inflammatory injury remains unclear. In the present study, we found that E. coli activated the inflammatory response through the assembly of the NLRP3 inflammasome and activation of the NF-κB p65 subunit in primary bovine endometrial epithelial cells (bEECs). Infection with E. coli also led to an abnormal increase in cytoplasmic calcium and mitochondrial dysfunction. Additionally, live-cell imaging of calcium reporters indicated that the increase in cytosolic calcium mainly was caused by the release of Ca2+ ions stored in the ER and mitochondria, which was independent of extracellular calcium. Cytoplasmic calcium regulates mitochondrial respiratory chain transmission, DNA replication, and biogenesis. Pretreatment with NAC, BAPTA-AM, or 2-APB reduced the expression of IL-1β and IL-18. Moreover, ERS was involved in the regulation of bovine endometritis and cytosolic calcium was an important factor for regulating ERS in E. coli-induced inflammation. Finally, activation of autophagy inhibited the release of IL-1β and IL-18, cytochrome c, ATP, ERS-related proteins, and cytoplasmic calcium. Collectively, our findings demonstrate that autophagy mediated E. coli-induced cellular inflammatory injury by regulating cytoplasmic calcium, mitochondrial dysfunction, and ERS.

Chemical-Physical Properties and Bioactivity of New Premixed Calcium Silicate-Bioceramic Root Canal Sealers.

The aim of the study was to analyze the chemical-physical properties and bioactivity (apatite-forming ability) of three recently introduced premixed bioceramic root canal sealers containing varied amounts of different calcium silicates (CaSi): a dicalcium and tricalcium silicate (1-10% and 20-30%)-containing sealer with zirconium dioxide and tricalcium aluminate (CERASEAL); a tricalcium silicate (5-15%)-containing sealer with zirconium dioxide, dimethyl sulfoxide and lithium carbonate (AH PLUS BIOCERAMIC) and a dicalcium and tricalcium silicate (10% and 25%)-containing sealer with calcium aluminate, tricalcium aluminate and tantalite (NEOSEALER FLO). An epoxy resin-based sealer (AH PLUS) was used as control. The initial and final setting times, radiopacity, flowability, film thickness, open pore volume, water absorption, solubility, calcium release and alkalizing activity were tested. The nucleation of calcium phosphates and/or apatite after 28 days aging in Hanks balanced salt solution (HBSS) was evaluated by ESEM-EDX, vibrational IR and micro-Raman spectroscopy. The analyses showed for NeoSealer Flo and AH Plus the longest final setting times (1344 ± 60 and 1300 ± 60 min, respectively), while shorter times for AH Plus Bioceramic and Ceraseal (660 ± 60 and 720 ± 60 min, respectively). Radiopacity, flowability and film thickness complied with ISO 6876/12 for all tested materials. A significantly higher open pore volume was observed for NeoSealer Flo, AH Plus Bioceramic and Ceraseal when compared to AH Plus (p < 0.05), significantly higher values were observed for NeoSealer Flo and AH Plus Bioceramic (p < 0.05). Ceraseal and AH Plus revealed the lowest solubility. All CaSi-containing sealers released calcium and alkalized the soaking water. After 28 days immersion in HBSS, ESEM-EDX analyses revealed the formation of a mineral layer that covered the surface of all bioceramic sealers, with a lower detection of radiopacifiers (Zirconium for Ceraseal and AH Plus Bioceramic, Tantalum for NeoSealer Flo) and an increase in calcium, phosphorous and carbon. The calcium phosphate (CaP) layer was more evident on NeoSealer Flo and AH Plus Bioceramic. IR and micro-Raman revealed the formation of calcium carbonate on the surface of all set materials. A thin layer of a CaP phase was detected only on AH Plus Bioceramic and NeoSealer Flo. Ceraseal did not show CaP deposit despite its highest calcium release among all the tested CaSi-containing sealers. In conclusion, CaSi-containing sealers met the required chemical and physical standards and released biologically relevant ions. Slight/limited apatite nucleation was observed in relation to the high carbonation processes.

Dynamic Balance between PTH1R-Dependent Signal Cascades Determines Its Pro- or Anti-Osteogenic Effects on MSC.

Parathyroid hormone (PTH) is one of the key regulators of calcium and phosphate metabolism in the body, controlling bone metabolism and ion excretion by the kidneys. At present, attempts to use PTH as a therapeutic agent have been associated with side-effects, the nature of which is not always clear and predictable. In addition, it is known that in vivo impairment of PTH post-receptor signaling is associated with atypical differentiation behavior not only of bone cells, but also of connective tissues, including adipose tissue. In this work, we studied the functional responses of multipotent mesenchymal stromal cells (MSCs) to the action of PTH at the level of single cells. We used MSCs isolated from the periosteum and subcutaneous adipose tissue to compare characteristics of cell responses to PTH. We found that the hormone can activate three key responses via its receptor located on the surface of MSCs: single transients of calcium, calcium oscillations, and hormone-activated smooth increase in intracellular calcium. These types of calcium responses led to principally different cellular responses of MSCs. The cAMP-dependent smooth increase of intracellular calcium was associated with pro-osteogenic action of PTH, whereas phospholipase C dependent calcium oscillations led to a decrease in osteogenic differentiation intensity. Different variants of calcium responses are in dynamic equilibrium. Suppression of one type of response leads to increased activation of another type and, accordingly, to a change in the effect of PTH on cell differentiation.

Evaluation of Dentin Remineralization with Zinc Oxide and Calcium Fluoride Nanoparticles - An In vitro Study.

Partially demineralized dentin is remineralizable when mineral ions are made available in the near vicinity. Nanoparticles (NPs) have wide applications in remineralization process. Zinc promotes remineralization and has a synergistic effect when combined with fluoride. Hence, zinc oxide and calcium fluoride NPs were considered for dentin remineralization. The aim of this study was to evaluate the remineralizing ability of zinc oxide and calcium fluoride NPs on demineralized dentin individually and in combined form. This was an in vitro study. Forty-eight dentin disks were prepared from the crowns of 12 extracted human molars and were allocated into four groups of 12 each, i.e., Group I - nano zinc oxide (nZnO), Group II - nano calcium fluoride (nCaF2), Group III - combined group (nZnO + nCaF2), and Group IV - artificial saliva (AS, control group). The specimens were demineralized with 37% phosphoric acid and placed in the respective remineralizing solutions for 24 h and 1 month. Calcium (Ca) and phosphorous (P) uptake was measured using energy-dispersive X-ray spectrometry, and structural changes were analyzed using scanning electron microscopy (SEM). One-way analysis of variance, Student's t-test, and post hoc Tukey's test were used for statistical analysis. At 1-month interval, all the groups showed an increase in Ca/P ratio, with highest being the combined group (4.24), followed by nCaF2 (3.30), nZnO (1.71), and AS (1.31) groups, and these differences were statistically significant (P = 0.000). On SEM analysis of dentinal samples at 1 month, depositions were evident in intertubular regions, wherein the highest deposits were observed in the nZnO group, followed by nZnO + nCaF2 and nCaF2 groups. Dentin samples subjected to remineralization with aqueous solutions of nZnO and nCaF2 showed an increase in calcium and phosphorous uptake and also dense granular depositions were evident in intertubular regions of dentin.

A mathematical model of calcium signals around laser-induced epithelial wounds.

Cells around epithelial wounds must first become aware of the wound's presence in order to initiate the wound-healing process. An initial response to an epithelial wound is an increase in cytosolic calcium followed by complex calcium-signaling events. While these calcium signals are driven by both physical and chemical wound responses, cells around the wound will all be equipped with the same cellular components to produce and interact with the calcium signals. Here we have developed a mathematical model in the context of laser ablation of the Drosophila pupal notum that integrates tissue-level damage models with a cellular calcium-signaling toolkit. The model replicates experiments in the contexts of control wounds as well as knockdowns of specific cellular components, but it also provides new insights that are not easily accessible experimentally. The model suggests that cell-cell variability is necessary to produce calcium-signaling events observed in experiments; it quantifies calcium concentrations during wound-induced signaling events, and it shows that intercellular transfer of the molecule IP3 is required to coordinate calcium signals across distal cells around the wound. The mathematical model developed here serves as a framework for quantitative studies in both wound signaling and calcium signaling in the Drosophila system.

PMON55 Calcium Imaging in the Intact Mouse Pituitary: A Novel Design for Understanding the Formation and Modulation of the Gonadotrope Network

Abstract The reproductive cells of the pituitary, the gonadotropes, develop after e12.75 in mice and continue to differentiate and migrate throughout puberty. In response to physiological signals, these cells change in morphology to alter connections with other cells. Previous studies have indicated that gonadotropes have functional relationships in an interconnected, organized network. However, little work has been done to determine (1) what developmental cues are needed to organize this network, or (2) how this network functions to promote gonadotropin secretion. Activation of gonadotropin-releasing hormone receptors (GnRHRs) leads to an increase in intracellular calcium; therefore, this pathway can be used for visualizing the activation of gonadotropes. We have developed a novel system to analyze the gonadotrope network which will allow us to study the influence of physiological signals to this interconnected population. To visualize calcium signals specifically in gonadotropes, we created a line of Gonadotrope-GCaMP mice using Gnrhr-driven Cre to express the GCaMP calcium indicator. For present studies, intact pituitary glands were taken from adult Gonadotrope-GCaMP mice (males and proestrous females). Whole pituitaries were mounted in a chamber, and gonadotrope network responses to GnRH (1 or 5nM, at least 4 pituitaries/dose) were recorded using confocal microscopy. Calcium signals from an average of 177 ± 8 gonadotropes per pituitary were analyzed. MATLAB software was used to quantify network connections of gonadotropes for each treatment. Correlation coefficients were determined by their patterns of calcium signaling. Gonadotrope connectivity increases following GnRH stimulation in a dose-dependent manner in female pituitaries (17% following 1nM GnRH, p<0.05; 44% following 5nM GnRH, p<0.05). While male gonadotropes also increase connectivity following stimulation with GnRH, the effect is not enhanced by the higher dose (27% following 1nM GnRH, p<0.05; 20% following 5nM GnRH, NS). Although the strength of gonadotrope correlations is not changed by 1nM GnRH in females, higher dosage of GnRH doubles the strength of correlations over pre-stimulation levels (Pre: 0.08, Post: 0.16, p<0.05). The strength of correlations in males was not significantly altered by GnRH at either dose. In conclusion, the data from whole mouse pituitaries confirm that gonadotropes are capable of high levels of coordinated activity. All pituitaries showed baseline levels of coordination among gonadotropes, confirming the presence of the network. Network activation is significantly increased by GnRH stimulation in both males and females. In females, coordination of gonadotropes is also strengthened by GnRH in a dose-dependent manner. Using a novel gonadotrope-specific calcium indicator mouse model, we have shown GnRH-mediated changes in gonadotrope network connectivity in the intact mouse pituitary. Future studies in our lab will determine how other physiological signals are involved in development and/or maintenance of this coordinated activation. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Salivary Calcium Level and Its Correlation with Salivary pH, Salivary Volume, and Calcium Intake in Hypertensive Female Patients with Different Blood Sugar Levels.

This study aimed to evaluate the salivary calcium level and its correlation with salivary pH, salivary volume, and calcium intake in hypertensive female patients with different blood sugar levels. This cross-sectional study included 49 female subjects with hypertension. Subjects were divided into three groups based on the HbA1c test: normal, prediabetes, and diabetes. Unstimulated whole saliva was collected using the spitting method. Salivary calcium levels were evaluated using an Atomic Absorption Spectrophotometer. Salivary pH was obtained using a universal indicator pH paper test. Calcium intake was assessed by a semi-quantitative food frequency questionnaire. The Kruskal-Wallis test was used to compare salivary parameters and total calcium intake within study groups. Spearman rank correlation and multiple regression analysis were used to evaluate the correlation between salivary calcium levels and all variables in the study. No significant difference in salivary calcium, volume, pH, and total calcium intake was observed within the study groups. However, a significant correlation was found between salivary calcium levels and salivary pH in hypertensive females with normal blood glucose levels. Moreover, salivary calcium levels have a significant correlation with systolic blood pressure in prediabetes and diabetes groups. This study found a decrease in all salivary parameters (calcium, pH, and volume) as well as a low calcium intake in hypertensive females, despite no significant difference found in groups with different blood glucose levels. Blood glucose levels appeared to be a confounder in the relationship between salivary calcium with salivary pH and systolic blood pressure.

Towards a better understanding of arterial calcification disease progression in CKD: investigation of early pathological alterations.

Cardiovascular disease remains the leading cause of death in chronic kidney disease (CKD) patients, especially in those undergoing dialysis and kidney transplant surgery. CKD patients are at high risk of developing arterial media calcifications (AMC) and arterial stiffness. We hypothesized that investigation of disease progression at an early stage could provide novel insights in understanding AMC etiology. An adenine diet was administered to male Wistar rats to induce AMC. Rats were sacrificed after 2, 4 and 8 weeks. AMC was measured by assessment of aortic calcium and visualized using histology. Arterial stiffness was measured in vivo by ultrasound and ex vivo by applying cyclic stretch of physiological magnitude on isolated arterial segments, allowing us to generate the corresponding pressure-diameter loops. Further, ex vivo arterial reactivity was assessed in organ baths at 2 and 4 weeks to investigate early alterations in biomechanics/cellular functionality. CKD rats showed a time-dependent increase in aortic calcium which was confirmed on histology. Accordingly, ex vivo arterial stiffness progressively worsened. Pressure-diameter loops showed a gradual loss of arterial compliance in CKD rats. Additionally, viscoelastic properties of isolated arterial segments were altered in CKD rats. Furthermore, after 2 and 4 weeks of adenine treatment, a progressive loss in basal, nitric oxide (NO) levels was observed, which was linked to an increased vessel tonus and translates into an increasing viscous modulus. Our observations indicate that AMC-related vascular alterations develop early after CKD induction prior to media calcifications being present. Preventive action, related to restoration of NO bioavailability, might combat AMC development.

A comparative study on the antifungal efficacy of cold atmospheric plasma at low and high surface density on Aspergillus chevalieri and mechanisms of action

Cold atmospheric plasma (CAP) is a promising innovative technology for microbial inactivation on food surfaces. In this study, we evaluated the potential of CAP at low (CAP-O3) and high (CAP-NOx) power using Aspergillus chevalieri as a fungal model, one of the most xerophilic and xerotolerant molds causing food spoilage, We observed a strong reduction in mycelial growth (60%) and conidial germination (90%) after 30 min of CAP-NOx, while CAP-O3 was less effective showing no reduction in mycelia growth and only 36% of spore germination reduction. Furthermore, cell death was accompanied by a loss of cellular and mitochondrial potential membrane, a significant (p < 0.05) increase in intracellular ROS, RNS, calcium, and DNA damage. For the first time, we reported that trehalose, glycerol, chitin and glucan accumulation are some adaptive mechanisms of A. chevalieri to counteract CAP stress, with CAP-NOx treatments inducing a major accumulation of these two osmolites. Our results suggested that Cold atmospheric plasma-induced cell death can be explained by oxidative stress-dependent through a cascade of reactions initiated by membrane depolarization.

Cannabidiol as a modulator of α7 nicotinic receptors.

Cannabidiol (CBD), an important terpenoid compound from marijuana with no psychoactive effects, has become of great pharmaceutical interest for several health conditions. As CBD is a multitarget drug, there is a need to establish the molecular mechanisms by which CBD may exert therapeutic as well as adverse effects. The α7 nicotinic acetylcholine receptor (α7 nAChR) is a cation-permeable ACh-gated channel present in the nervous system and in non-neuronal cells. It is involved in different pathological conditions, including neurological and neurodegenerative disorders, inflammation, and cancer. By high-resolution single-channel recordings and confocal microscopy, we here reveal how CBD modulates α7 nAChR ionotropic and metabotropic functions. CBD leads to a profound concentration-dependent decrease of α7 nAChR single-channel activity with an IC50 in the sub-micromolar range. The inhibition of α7 nAChR activity, which takes place through a membrane pathway, is neither mediated by receptor phosphorylation nor overcome by positive allosteric modulators and is compatible with CBD stabilization of resting or desensitized α7 nAChR conformational states. CBD modulation is complex as it also leads to the later appearance of atypical, low-frequency α7 nAChR channel openings. At the cellular level, CBD inhibits the increase in intracellular calcium triggered by α7 nAChR activation, thus decreasing cell calcium responses. The modulation of α7 nAChR is of pharmacological relevance and should be considered in the evaluation of CBD potential therapeutic uses. Thus, our study provides novel molecular information of CBD multiple actions and targets, which is required to set the basis for prospective applications in human health.