Search Results (74)

Flunarizine is a calcium channel blocker widely used in neurological disorders; however, its low aqueous solubility may influence formulation stability and drug dispersion in polymer-based systems. The present study aimed to evaluate the compatibility of flunarizine with selected excipients and to investigate its incorporation into polymeric hydrogel matrices. Binary mixtures of flunarizine with excipients such as hydroxypropyl-β-cyclodextrin, polyethylene glycol (PEG 6000), Tween 20, gelatin, and citric acid were prepared and characterized using Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG/DTG), and high-performance liquid chromatography (HPLC). The FTIR spectra of the analyzed samples do not reveal the appearance of new absorption bands that may indicate chemical interactions; instead, minor spectral variations were observed due to weak intermolecular interactions within the polymer network. Thermal analysis revealed decomposition patterns consistent with those of the individual components, suggesting the absence of significant incompatibilities. A validated RP-HPLC method enabled sensitive and reliable quantification of flunarizine in the analyzed systems, with a limit of detection (LOD) of 0.05 µg/mL and a limit of quantitation (LOQ) of 0.16 µg/mL. Accuracy testing showed average recovery rates of 100% across 80–120% spiking levels. Overall, the results support the compatibility of flunarizine with the investigated excipients and the suitability of the studied hydrogels as potential drug delivery matrices. Full article

Neurogranin (Ng) is a postsynaptic calmodulin-binding protein highly enriched in forebrain neurons and widely implicated in synaptic plasticity. However, whether Ng contributes more broadly to neuronal network maturation and cellular homeostasis remains unclear. Here, we examined the consequences of silencing or restoring Ng to adult physiological levels in primary hippocampal neurons. Ng expression promoted dendritic expansion, increased synaptic number, and shifted the axon initial segment toward the soma, consistent with structural adaptations to enhanced connectivity. Calcium (Ca²⁺) imaging revealed a marked increase in spontaneous neuronal activity and network synchronization, which was confirmed by electrophysiological recordings showing enhanced burst firing and spike synchrony. At the molecular level, Ng altered Ca²⁺/calmodulin (CaM) signaling by increasing total CaM levels, reducing Ca²⁺/CaM-dependent protein kinase II (CaMKII) abundance while increasing its relative autophosphorylation, and downscaling specific ionotropic glutamate receptors. Despite elevated network activity, Ng expression enhanced neuronal metabolic competence and viability, reduced cellular stress signaling and induced modest caspase-3 activation without engagement of apoptotic pathways. Together, these results indicate that Ng promotes neuronal maturation and coordinated network activity while engaging compensatory mechanisms that preserve excitatory balance and neuronal resilience. Our findings identify Ng as a molecular integrator linking Ca²⁺/CaM signaling with the structural and functional maturation of neuronal networks. Full article

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is initiated by the viral spike proteins, which are key structural components that mediate host cell binding and entry and alter downstream signaling through multiple interactions with endothelial surface receptors. Endothelial dysfunction is a central consequence of COVID-19, contributing to vascular inflammation, barrier disruption, thrombosis, and multi-organ injury affecting the pulmonary, cardiovascular, cerebral, and renal systems. Emerging evidence demonstrates that spike protein-mediated effects, independent of productive viral infection, disrupt endothelial homeostasis through angiotensin-converting enzyme 2 (ACE2) dysregulation, integrin engagement, altered calcium signaling, junctional protein remodeling, oxidative stress, and pro-inflammatory and pro-apoptotic pathways. This review is intentionally focused on spike (S) protein-driven mechanisms of endothelial dysfunction; pathogenic vascular effects attributed to other SARS-CoV-2 structural proteins, including the nucleocapsid (N) protein, are beyond the scope of this discussion. In this review, we synthesize current experimental and translational data detailing the molecular mechanisms by which the SARS-CoV-2 spike protein drives endothelial dysfunction across multiple organ systems and discuss potential therapeutic strategies aimed at preserving endothelial integrity in acute COVID-19 and its long-term vascular sequela. Full article

Excitable neurons are intrinsically capable of firing action potentials (AP), yet a state of hyperexcitability is prevented in the central nervous system by powerful GABAergic inhibition. For this inhibition to be effective, it must occur before excitatory signals can initiate runaway activity, implying the existence of a proactive control system. To test for such proactive inhibition, we used Ca²⁺ imaging and patch-clamp recording to measure how hippocampal neurons respond to depolarization and glutamatergic agonists. In mature hippocampal cultures (14 days in vitro (DIV)) and acute brain slices from two-month-old rats, neurons exhibited non-simultaneous responses to various excitatory stimuli, including KCl, NH₄Cl, forskolin, domoic acid, and glutamate. We observed that the Ca²⁺ rise occurred significantly earlier in GABAergic neurons than in glutamatergic neurons. This delay in glutamatergic neurons was abolished by GABA(A) receptor inhibitors, suggesting a mechanism of preliminary γ-aminobutyric acid (GABA) release. We further found that these early-responding GABAergic neurons express calcium-permeable kainate and AMPA receptors (CP-KARs and CP-AMPARs). Application of domoic acid induced an immediate Ca²⁺ increase in neurons expressing these receptors, but a delayed response in others. Crucially, when domoic acid was applied in the presence of the AMPA receptor inhibitors NBQX or GYKI-52466, the response delay in glutamatergic neurons was significantly prolonged. This confirms that CP-KARs on GABAergic neurons are responsible for the delayed excitation of glutamatergic neurons. In hippocampal slices from two-month-old rats, depolarization with 50 mM KCl revealed two distinct neuronal populations based on their calcium dynamics: a majority group (presumably glutamatergic) exhibited fluctuating Ca²⁺ signals, while a minority (presumably GABAergic) showed a steady, advancing increase in [Ca²⁺]i. This distinction was reinforced by the application of domoic acid. The “advancing-response” neurons reacted to domoic acid with a similar prompt increase, whereas the “fluctuating-response” neurons displayed an even more delayed and fluctuating reaction (80 s delay). Therefore, we identify a subgroup of hippocampal neurons—in both slices and cultures—that respond to depolarization and domoic acid with an early [Ca²⁺]_i signal. Consistent with our data from cultures, we conclude these early-responding neurons are GABAergic. Their early GABA release directly explains the delayed Ca²⁺ response observed in glutamatergic neurons. We propose that this proactive mechanism, mediated by CP-KARs on GABAergic neurons, is a primary means of protecting the network from hyperexcitation. Furthermore, the activity of these CP-KAR-expressing neurons is itself regulated by GABAergic neurons containing CP-AMPARs. Full article

The Gadouras Reservoir, Rhodes Island’s primary water source, experiences recurrent whiting events—milky turbidity from calcium carbonate precipitation—that challenge treatment operations, with impacts compounded by a major 2023 wildfire in this fire-prone Mediterranean setting. To elucidate these dynamics, a pragmatic, multi-source monitoring framework integrates archived Sentinel-2 and Landsat imagery with treatment-plant records (2017–mid-2025). Unitless spectral indices (e.g., AreaBGR) for whiting detection and chlorophyll-a proxies are combined with laboratory measurements of turbidity, pH, total organic carbon, manganese, and hydrological metrics, analyzed via spatiotemporal Hovmöller diagrams, Pearson correlations, and interrupted time-series models. Two seasonal whiting regimes are identified: a biogenic summer mode (southern origin; elevated chlorophyll-a; water temperature > 15 °C; pH > 8.5) and a non-biogenic winter mode (northern inflows). Following the wildfire, the system exhibits characteristics that could be related to possible hypolimnetic anoxia, prolonged whiting, a ~50% rise in organic carbon, and a manganese excursion to ~0.4 mg L⁻¹ at the deeper intake. Crucially, the post-fire period shows a decoupling of AreaBGR from turbidity (r ≈ 0.233 versus ≈ 0.859 pre-fire)—a key diagnostic finding that confirms a fundamental shift in the composition and optical properties of suspended particulates. The manganese spike is best explained by the confluence of a wildfire-induced biogeochemical predisposition (anoxia and Mn mobilization) and a consequential operational decision (relocation to a deeper, Mn-rich intake). This framework establishes diagnostic baselines and thresholds for managing fire-impacted reservoirs, supports the use of remote sensing in data-scarce systems, and informs adaptive operations under increasing climate pressures. Full article

Potassium (K) is a critical macronutrient for sugarcane (Saccharum spp.), playing a vital role in metabolic processes, sucrose accumulation, and yield formation. Herein, this study systematically evaluated the effects of potassium oxide (K₂O) application on sugarcane (cultivar YZ1696) growth at the seedling and tillering stages. Hydroponic experiments demonstrated that 6 mmol/L K₂O optimally promoted seedling growth, whereas field trials revealed that 150 kg/ha K₂O maximized growth rate, yield, and sucrose content. Sugarcane growth exhibited a biphasic response—stimulation followed by inhibition—with increasing K₂O dosage at both developmental stages. Transcriptomic profiling of sugarcane roots under low-potassium (K-deficient), optimal potassium, and high-potassium conditions identified 10,266 differentially expressed genes (DEGs), with the most pronounced transcriptional shifts occurring under K deficiency. Functional enrichment analysis identified DEGs associated with potassium transport, calcium signaling, and carbohydrate metabolism. Notably, potassium uptake was mediated by distinct mechanisms: Shaker family channels (AKT1, AKT2, SPIKE) and the TPK family member KCO1 were induced under optimal K supply, whereas HAK/KUP/KT transporters (HAK1/5/10/21/25) exhibited broad activation across K concentrations, underscoring their key role in K homeostasis. Furthermore, calcium signaling genes (e.g., CIPK23) displayed K-dependent expression patterns. Weighted gene co-expression network analysis identified key gene modules that correlated strongly with agronomic traits, including plant height, yield, and sucrose content. Optimal K conditions favored the expression of yield- and sucrose-associated genes, suggesting a molecular basis for K-mediated productivity enhancement. Our findings revealed the genetic and physiological mechanisms underlying K-dependent sugarcane improvement, providing actionable insights for precise potassium fertilization to maximize the yield and sugar content. Full article

With increasing global challenges related to water scarcity and phosphorus depletion, the recovery and reuse of wastewater-derived nutrients offer a sustainable path forward. This study evaluates the dual role of lanthanides (Ce³⁺ and La³⁺) in recovering phosphorus from municipal wastewater and supporting corn (Zea mays) cultivation through lanthanide phosphate (Ln-P) and lanthanide-reclaimed wastewater (LRWW, wastewater spiked with lanthanide). High-purity precipitates of CePO₄ (98%) and LaPO₄ (92%) were successfully obtained without pH adjustment, as confirmed by X-ray photoelectron spectroscopy (XPS) and energy-dispersive spectroscopy (EDS). Germination assays revealed that lanthanides, even at concentrations up to 2000 mg/L, did not significantly alter germination rates compared to traditional coagulants, though root and shoot development declined above this threshold—likely due to reduced hydrogen peroxide (H₂O₂) production and elevated total dissolved solids (TDSs), which induced physiological drought. Greenhouse experiments using desert-like soil amended with Ln-P and irrigated with LRWW showed no statistically significant differences in corn growth parameters—including plant height, stem diameter, leaf number, leaf area, and biomass—when compared to control treatments. Photosynthetic performance, including stomatal conductance, quantum efficiency, and chlorophyll content, remained unaffected by lanthanide application. Metal uptake analysis indicated that lanthanides did not inhibit phosphorus absorption and even enhanced the uptake of calcium and magnesium. Minimal lanthanide accumulation was detected in plant tissues, with most retained in the root zone, highlighting their limited mobility. These findings suggest that lanthanides can be safely and effectively used for phosphorus recovery and agricultural reuse, contributing to sustainable nutrient cycling and aligning with the United Nations’ Sustainable Development Goals of zero hunger and sustainable cities. Full article

The search for novel compounds with anticonvulsant properties remains a key focus in neuropharmacology. Recently, the diazepine-benzimidazole derivative, DAB-19, has emerged as a promising candidate due to its demonstrated anxiolytic and analgesic effects. In this study, we investigate the mechanisms underlying DAB-19’s activity, focusing on its impact on glutamatergic transmission, a key target in the pathophysiology of various central nervous system disorders. Intriguingly, while DAB-19 suppressed evoked glutamatergic transmission in rat brain slices, it simultaneously enhanced spontaneous neurotransmission. Further experiments on glutamatergic neuromuscular synapses in fly larvae revealed two distinct mechanisms: calcium-dependent potentiation of glutamate release and inhibition of spike propagation via blockade of voltage-gated sodium channels. The latter effect was directly confirmed in rat brain neurons. Given its action on sodium channels, we tested DAB-19 in the pentylenetetrazole model, where it delayed seizure onset but did not prevent seizures. These findings position DAB-19 as a multifaceted compound with significant therapeutic potential. Full article

The role of calcium, an essential secondary messenger, in cell division remains an outstanding question in cell biology despite several significant findings over the past few decades. Among them is the landmark discovery of intracellular calcium waves during cytokinesis, the last stage of cell division, in fish cells. Nevertheless, subsequent studies have been largely unable to determine the underlying molecular mechanism of these cytokinetic transients. At the center of this stalemate stands two challenging questions, how these calcium transients rise and what they do during cytokinesis. Yeast, despite its proven prowess as a model organism to study cell cycle, has not drawn much interest in addressing these questions. However, the recent discovery of cytokinetic calcium spikes in the fission yeast Schizosaccharomyces pombe has provided novel insights into how calcium regulates cytokinesis. In this review, I will primarily focus on our current understanding of the molecular mechanism of cytokinetic calcium transients in yeast cells. First, I will briefly recount the discovery of cytokinetic calcium transients in animal cells. This will be followed by an introduction to the intracellular calcium homeostasis. Next, I will discuss yeast cytokinetic calcium spikes, the ion channel Pkd2 that promotes these spikes, and the potential molecular targets of these spikes. I will also compare the calcium regulation of cytokinesis between yeast and animal cells. I will conclude by presenting a few critical questions in our continued quest to understand how calcium regulates cytokinesis. Full article

Arbuscular mycorrhizal (AM) symbiosis, a mutually beneficial interaction between plant roots and AM fungi, plays a key role in plant growth, nutrient acquisition, and stress tolerance, which make it a major focus for sustainable agricultural strategies. This intricate association involves extensive transcriptional reprogramming in host plant cells during the formation of arbuscules, which are specialized fungal structures for nutrient exchange. The symbiosis is initiated by molecular signaling pathways triggered by fungal chitooligosaccharides and strigolactones released by plant roots, which act as chemoattractants and signaling molecules to promote fungal spore germination, colonization, and arbuscule development. Calcium spiking, mediated by LysM domain receptor kinases, serves as a critical second messenger in coordinating fungal infection and intracellular accommodation. GRAS transcription factors are key components that regulate the transcriptional networks necessary for arbuscule development and maintenance, while small RNAs (sRNAs) from both plant and fungi, contribute to modifications in gene expression, including potential bidirectional sRNA exchange to modulate symbiosis. Understanding the molecular mechanisms related to AM symbiosis may provide valuable insights for implementation of strategies related to enhancing plant productivity and resilience. Full article

Diabetes mellitus is a metabolic disorder that is rapidly growing across the world. Our laboratory has recently demonstrated that photobiomodulation (PBM) can couple to its metabolic pathways by modulating calcium dynamics in islet cells, including α- and β-cells. Using computer vision algorithms, changes in PBM-induced calcium dynamics can be verified, and, more importantly, this led us to propose hypotheses that will likely advance our understanding of photostimulatory effects in islet cells. In our previous paper, we determined changes in calcium spiking in response to PBM at 810 nm by manually segmenting the cells and the calcium spiking patterns. We have since developed a computer vison pipeline to automate cell segmentation and subsequent image analyses. By using automated methods for segmentation, registration, tracking, and statistical analysis, we were able to improve the accuracy of previously observed changes in calcium spiking in response to PBM in both cell types. Moreover, this pipeline was applied to elucidate the wavelength-dependent modulation of calcium dynamics at 1064 nm. The extent of increase in calcium spiking appears to have been overestimated by manual analysis, and the machine learning pipeline was able to capture and segment nearly 3-fold more cells, suggesting improved accuracy in the analysis of calcium spiking in islet cells. Detailed calcium analysis also indicates a biphasic dose response among α- and β-cells in response to PBM therapy at different wavelengths. The current findings offer a novel hypothesis and may facilitate the use of translational PBM as a potential therapy for diabetes mellitus. Full article

Background: The Precision ID mitochondrial (mt) DNA Whole Genome system is a fully automated massively parallel sequencing (MPS) solution for the whole mitochondrial genome. While extremely sensitive, the Precision ID system is susceptible to inhibitors and microbial DNA that are often co-extracted from human remains. Methods: DNA templates spiked with varying amounts of hematin, humic acid, and calcium, along with bones containing degraded and non-human DNA, were sequenced using the Precision ID system with and without the addition of bovine serum albumin (BSA). Results: BSA added to the initial PCR reaction successfully improved the robustness of the Precision ID system while not negatively impacting the sequencing success of uninhibited samples. The success of BSA is inhibitor-concentration dependent and is effective for templates containing at least 50 ng/μL humic acid, 50 μM hematin, and 1500 μM calcium ions. Furthermore, the presence of microbial DNA in addition to an inhibitor, results in non-specific adaptor ligation to the non-human DNA; BSA can alleviate the inhibition, allowing the human mtDNA to be amplified and sequenced. Conclusions: The addition of BSA to the Precision ID mtDNA system can yield successful sequencing results from challenging case samples that would otherwise fail. Full article

Optimizing nitrogen (N) sources has the potential to improve wheat tillering, nitrogen use efficiency (NUE), and grain yield, yet the underlying mechanisms remain unclear. This study hypothesizes that combining specific N sources can increase zeatin riboside + zeatin (ZR + ZT) content in tiller nodes and maintain a higher ZR + ZT/gibberellin A7 (GA₇) ratio, thereby promoting tiller development, enhancing NUE, and increasing yield. The effects of N source treatments on two wheat cultivars, the multi-spike Shannong 28 (SN28) and the large-spike Tainong 18 (TN18), were investigated. A total of seven N treatments were tested: no nitrogen (N0), urea (N1), calcium nitrate (N2), ammonium chloride (N3), and equal doses of urea and calcium nitrate (N4), urea and ammonium chloride (N5), and calcium nitrate and ammonium chloride (N6). The results showed that treatment N4 significantly increased the levels of ZR and ZT in tiller nodes, while maintaining a higher ZR + ZT to GA₇ ratio. This hormonal shift promoted tiller formation and biomass accumulation. Under N4, both cultivars exhibited the highest number of effective spikes and biomass in higher-order tillers. N4 also enhanced N accumulation in the grains, N absorption efficiency, and N translocation, while reducing N loss. Compared to N1, effective spike numbers increased by 7.8% in SN28 and 5.6% in TN18, resulting in a 6.4% increase in grain yield for SN28 and a 2.2% increase for TN18. In conclusion, the combined application of urea and calcium nitrate optimizes hormonal regulation, improves NUE, and significantly enhances wheat tillering and grain yield, providing a promising strategy for enhancing wheat productivity. Full article

According to the FAO, 828 million people were facing acute food insecurity in 2021. Fertilization is a critical input factor in crop production and food security achievement. Therefore, fertilization is a critical input factor in crop production and food security achievement. However, there is room for improvement in the application of fertilizers in certain regions. Thus, new fertilizers with a relatively low cost could enhance farmers’ access to these essential inputs. Phosphogypsum (PG) is used as fertilizer because it contains many nutrients essential for plant growth, including calcium, sulfur, and phosphorus. A two-year field experiment was conducted using two Moroccan PG products (PG1 and PG2, obtained from two different industrial sites), applied at four rates (0, 1.5, 3, and 4.5 t/ha). The aim was to assess the impact of PG source and rate on barley crops, including yield component, nutrients uptake, and heavy metals content. The study’s findings revealed that as the rate of PG application increased, there were significant enhancements in the number of spikes, tillers, grains, total biomass, grain yield, and thousand-grain weight. In fact, when compared to the control, the application of 1.5, 3, and 4.5 t/ha of PG led to a remarkable increase in grain yield by 21%, 34%, and 39%, respectively. Furthermore, the uptake of nutrients (N, P, K, Ca, Mg, and S) by the shoots and grains was significantly influenced by the PG application rates, with higher rates resulting in greater nutrient uptake. Notably, the application of PG had no discernible impact on the heavy metal content in shoots, grains, or soil. Full article

Acidic stress is a formidable environmental factor that exerts adverse effects on plant growth and development, ultimately leading to a potential reduction in agricultural productivity. A low pH triggers Ca²⁺ influx across the plasma membrane (PM), eliciting distinct responses under various acidic pH levels. However, the underlying mechanisms by which Arabidopsis plant cells generate stimulus-specific Ca²⁺ signals in response to acidic stress remain largely unexplored. The experimentally induced stimulus may elicit spikes in cytosolic free Ca²⁺ concentration ([Ca²⁺]_i) spikes or complex [Ca²⁺]_i oscillations that persist for 20 min over a long-term of 24 h or even several days within the plant cytosol and chloroplast. This study investigated the increase in [Ca²⁺]_i under a gradient of low pH stress ranging from pH 3.0 to 6.0. Notably, the peak of [Ca²⁺]_i elevation was lower at pH 4.0 than at pH 3.0 during the initial 8 h, while other pH levels did not significantly increase [Ca²⁺]_i compared to low acidic stress conditions. Lanthanum chloride (LaCl₃) can effectively suppress the influx of [Ca²⁺]_i from the apoplastic to the cytoplasm in plants under acid stress, with no discernible difference in intracellular calcium levels observed in Arabidopsis. Following 8 h of acid treatment in the darkness, the intracellular baseline Ca²⁺ levels in Arabidopsis were significantly elevated when exposed to low pH stress. A moderately low pH, specifically 4.0, may function as a spatial-temporal input into the circadian clock system. These findings suggest that acid stimulation can exert a continuous influence on intracellular calcium levels, as well as plant growth and development. Full article