Shedding Light on Calcium Dynamics in the Budding Yeast: A Review on Calcium Monitoring with Recombinant Aequorin.

Recombinant aequorin has been extensively used in mammalian and plant systems as a powerful tool for calcium monitoring. While aequorin has also been widely applied in yeast research, a notable gap exists in the literature regarding comprehensive reviews of these applications. This review aims to address that gap by providing an overview of how aequorin has been used to explore calcium homeostasis, signaling pathways, and responses to stressors, heavy metals, and toxic compounds in .

Kcs1 and Vip1: The Key Enzymes behind Inositol Pyrophosphate Signaling in .

The inositol pyrophosphate pathway, a complex cell signaling network, plays a pivotal role in orchestrating vital cellular processes in the budding yeast, where it regulates cell cycle progression, growth, endocytosis, exocytosis, apoptosis, telomere elongation, ribosome biogenesis, and stress responses. This pathway has gained significant attention in pharmacology and medicine due to its role in generating inositol pyrophosphates, which serve as crucial signaling molecules not only in yeast, but also in higher eukaryotes. As targets for therapeutic development, genetic modifications within this pathway hold promise for disease treatment strategies, offering practical applications in biotechnology.

Formulation and Comprehensive Evaluation of Biohybrid Hydrogel Membranes Containing Doxycycline or Silver Nanoparticles.

Complicated wounds often require specialized medical treatments, and hydrogels have emerged as a popular choice for wound dressings in such cases due to their unique properties and the ability to incorporate and release therapeutic agents. Our focus was to develop and characterize a new optimized formula for biohybrid hydrogel membranes, which combine natural and synthetic polymers, bioactive natural compounds, like collagen and hyaluronic acid, and pharmacologically active substances (doxycycline or npAg). Dynamic (oscillatory) rheometry confirmed the strong gel-like properties of the obtained hydrogel membranes.

Nanobodies as Diagnostic and Therapeutic Tools for Cardiovascular Diseases (CVDs).

The aim of this review is to summarize some of the most recent work in the field of cardiovascular disease (CVD) diagnosis and therapy, focusing mainly on the role of nanobodies in the development of non-invasive imaging methods, diagnostic devices, and advanced biotechnological therapy tools. In the context of the increased number of people suffering from CVDs due to a variety of factors such as sedentariness, poor nutrition, stress, and smoking, there is an urgent need for new and improved diagnostic and therapeutic methods. Nanobodies can be easily produced in prokaryotes, lower eukaryotes, and plant and mammalian cells, and offer great advantages.

New Heterocyclic Compounds from Oxazol-5(4)-one and 1,2,4-Triazin-6(5)-one Classes: Synthesis, Characterization and Toxicity Evaluation.

This paper describes the synthesis of new heterocycles from oxazol-5(4)-one and 1,2,4-triazin-6(5)-one classes containing a phenyl-/4-bromophenylsulfonylphenyl moiety. The oxazol-5(4)-ones were obtained via condensation of 2-(4-(4-X-phenylsulfonyl)benzamido)acetic acids with benzaldehyde/4-fluorobenzaldehyde in acetic anhydride and in the presence of sodium acetate. The reaction of oxazolones with phenylhydrazine, in acetic acid and sodium acetate, yielded the corresponding 1,2,4-triazin-6(5)-ones.

Enantiomeric pairs of copper(II) complexes with tridentate Schiff bases derived from - and -methionine: the role of decorating organic groups of the ligand in crystal packing and biological activity.

Three enantiomeric pairs consisting of copper(II) complexes with tridentate Schiff bases have been synthesized for employing in biological assessments: 1∞[Cu(/-salmet)(HO)] (1-R/S·H2O), 1∞[Cu(/-3-HOMe-5-Me-salmet)] (2-R/S), and 1∞[Cu(/-3-MeO-salmet)] (3-R/S) (where /-salmetH, /-3-HOMe-5-Me-salmetH, and /-3-MeO-salmetH result from the condensation of /-methionine with salicylaldehyde, 2-hydroxy-3-(hydroxymethyl)-5-methylbenzaldehyde, and 3-methoxy-salicylaldehyde, respectively, in a 1 : 1 molar ratio). The crystal structures of 1-R·H2O and 2-R/S are reported. Moreover, the 1-R/S·H2O enantiomers have been subjected to a single-crystal-to-single-crystal (SC-SC) transformation by heating at 160 °C to afford their dehydrated forms, 1∞[Cu(/-salmet)] (1-R/S), whose structures have also been crystallographically determined.

Insights into Structure and Biological Activity of Copper(II) and Zinc(II) Complexes with Triazolopyrimidine Ligands.

In an attempt to increase the biological activity of the 1,2,4-triazolo[1,5-]pyrimidine scaffold through complexation with essential metal ions, the complexes -[Cu(mptp)Cl] (), [Zn(mptp)Cl(DMSO)] () (mptp: 5-methyl-7-phenyl-1,2,4-triazolo[1,5-]pyrimidine), [Cu(dmtp)Cl]·2HO () and [Zn(dmtp)Cl] () (dmtp: 5,7-dimethyl-1,2,4-triazolo[1,5-]pyrimidine), were synthesized and characterized as new antiproliferative and antimicrobial species. Both complexes () and () crystallize in the 2/n monoclinic space group, with the tetrahedral surroundings generating a square-planar stereochemistry in the Cu(II) complex and a tetrahedral stereochemistry in the Zn(II) species. The mononuclear units are interconnected in a supramolecular network through π-π interactions between the pyrimidine moiety and the phenyl ring in () while supramolecular chains resulting from C-H∙∙∙π interactions were observed in ().

Biological Activity of Triazolopyrimidine Copper(II) Complexes Modulated by an Auxiliary N-N-Chelating Heterocycle Ligands.

Novel complexes of type [Cu(N-N)(dmtp)(OH)](ClO)·dmtp (() N-N: 2,2'-bipyridine; () L: 1,10-phenantroline and dmtp: 5,7-dimethyl-1,2,4-triazolo[1,5-]pyrimidine) were designed in order to obtain biologically active compounds. Complexes were characterized as mononuclear species that crystallized in the space group P-1 of the triclinic system with a square pyramidal geometry around the copper (II). In addition to the antiproliferative effect on murine melanoma B16 cells, complex () exhibited low toxicity on normal BJ cells and did not affect membrane integrity.

Antiproliferative and antibacterial properties of biocompatible copper(II) complexes bearing chelating N,N-heterocycle ligands and potential mechanisms of action.

In an attempt to propose new applications for the biomedical field, complexes with mixed ligands {[Cu(bpy)(μOClO)]ClO}n (1) and [Cu(phen)(OH)](ClO) (2) (bpy: 2,2'-biyridine; phen and 1,10-phenantroline) were evaluated for their antibacterial and cytotoxicicity features and for the elucidation of some of the mechanisms involved. Complex (2) proved to be a very potent antibacterial agent, exhibing MIC and MBEC values 2 to 54 times lower than those obtained for complex (1) against both susceptible or resistant Gram-positive and Gram-negative strains, in planktonic or biofilm growth state. In exchange, complex (1) exhibited selective cytotoxicity against melanoma tumor cells (B16), proving a promising potential for developing novel anticancer drugs.

Concentrates Subtoxic Copper onto Cell Wall from Solid Media Containing Reducing Sugars as Carbon Source.

Copper is essential for life, but it can be deleterious in concentrations that surpass the physiological limits. Copper pollution is related to widespread human activities, such as viticulture and wine production. To unravel aspects of how organisms cope with copper insults, we used as a model for adaptation to high but subtoxic concentrations of copper.

Interaction between Polyphenolic Antioxidants and Cells Defective in Heavy Metal Transport across the Plasma Membrane.

Natural polyphenols are compounds with important biological implications which include antioxidant and metal-chelating characteristics relevant for their antimicrobial, antitumor, or antiaging potential. The mechanisms linking polyphenols and heavy metals in their concerted actions on cells are not completely elucidated. In this study, we used the model eukaryotic microorganism to detect the action of widely prevalent natural polyphenols on yeast cells defective in the main components involved in essential heavy metal transport across the plasma membrane.

cells lacking transcription factors Skn7 or Yap1 exhibit different susceptibility to cyanidin.

Anthocyanidins - the aglycone moiety of anthocyanins - are responsible for the antioxidant traits and for many of the health benefits brought by the consumption of anthocyanin-rich foods, but whether excessive anthocyanidins are deleterious to living organisms is still a matter of debate. In the present study we used the model eukaryotic microorganism to evaluate the potential toxicity of cyanidin, one of the most prevalent anthocyanidins found in berries, grapes, purple vegetables, and red wine. We found that yeast cells lacking the transcription factors responsible for regulating the response to oxidative stress - Skn7 and Yap1 - exhibited different sensitivities to cyanidin.

Cytotoxicity of Oleandrin Is Mediated by Calcium Influx and by Increased Manganese Uptake in Cells.

Oleandrin, the main component of L. extracts, is a cardiotoxic glycoside with multiple pharmacological implications, having potential anti-tumoral and antiviral characteristics. Although it is accepted that the main mechanism of oleandrin action is the inhibition of Na/K-ATPases and subsequent increase in cell calcium, many aspects which determine oleandrin cytotoxicity remain elusive.

Copper(II) Complexes with Mixed Heterocycle Ligands as Promising Antibacterial and Antitumor Species.

Complexes with mixed ligands [Cu(N-N)(pmtp)](ClO) (() N-N: 2,2'-bipyridine; () L: 1,10-phenanthroline and pmpt: 5-phenyl-7-methyl-1,2,4-triazolo[1,5-]pyrimidine) were synthesized and structurally and biologically characterized. Compound () crystallizes into space group and () in -1. Both complexes display an intermediate stereochemistry between the two five-coordinated ones.

and Caffeine Implications on the Eukaryotic Cell.

Caffeine-a methylxanthine analogue of the purine bases adenine and guanine-is by far the most consumed neuro-stimulant, being the active principle of widely consumed beverages such as coffee, tea, hot chocolate, and cola. While the best-known action of caffeine is to prevent sleepiness by blocking the adenosine receptors, caffeine exerts a pleiotropic effect on cells, which lead to the activation or inhibition of various cell integrity pathways. The aim of this review is to present the main studies set to investigate the effects of caffeine on cells using the model eukaryotic microorganism , highlighting the caffeine synergy with external cell stressors, such as irradiation or exposure to various chemical hazards, including cigarette smoke or chemical carcinogens.

Dietary Anthocyanins and Stroke: A Review of Pharmacokinetic and Pharmacodynamic Studies.

Cerebrovascular accidents are currently the second major cause of death and the third leading cause of disability in the world, according to the World Health Organization (WHO), which has provided protocols for stroke prevention. Although there is a multitude of studies on the health benefits associated with anthocyanin (ACN) consumption, there is no a rigorous systematization of the data linking dietary ACN with stroke prevention. This review is intended to present data from epidemiological, , , and clinical studies dealing with the stroke related to ACN-rich diets or ACN supplements, along with possible mechanisms of action revealed by pharmacokinetic studies, including ACN passage through the blood-brain barrier (BBB).

Anthocyanins and Anthocyanin-Derived Products in Yeast-Fermented Beverages.

The beverages obtained by yeast fermentation from anthocyanin-rich natural sources (grapes, berries, brown rice, etc.) retain part of the initial pigments in the maturated drink. During the fermentation and aging processes anthocyanins undergo various chemical transformations, which include reactions with glycolytic products (especially pyruvate and acetaldehyde) or with other compounds present in the complex fermentation milieu (such as vinylphenols obtained from cinnamic acids by means of a yeast decarboxylase) yielding pigments which can be more stable than the initial anthocyanins.

Manganese Suppresses the Haploinsufficiency of Heterozygous Cells and Stimulates the TRPY1-Dependent Release of Vacuolar Ca under H₂O₂ Stress.

Transient potential receptor (TRP) channels are conserved cation channels found in most eukaryotes, known to sense a variety of chemical, thermal or mechanical stimuli. The TRPY1 is a TRP channel with vacuolar localization involved in the cellular response to hyperosmotic shock and oxidative stress. In this study, we found that diploid cells with heterozygous deletion in gene are haploinsufficient when grown in synthetic media deficient in essential metal ions and that this growth defect is alleviated by non-toxic Mn surplus.

Accumulation of Ag(I) by Saccharomyces cerevisiae Cells Expressing Plant Metallothioneins.

The various applications of Ag(I) generated the necessity to obtain Ag(I)-accumulating organisms for the removal of surplus Ag(I) from contaminated sites or for the concentration of Ag(I) from Ag(I)-poor environments. In this study we obtained Ag(I)-accumulating cells by expressing plant metallothioneins (MTs) in the model . The cDNAs of seven MTs (AtMT1a, AtMT1c, AtMT2a, AtMT2b, AtMT3, AtMT4a and AtMT4b) and four MTs (NcMT1, NcMT2a, NcMT2b and NcMT3) fused to myrGFP displaying an -terminal myristoylation sequence for plasma membrane targeting were expressed in and checked for Ag(I)-related phenotype.

Epigallocatechin-3-O-gallate, the main green tea component, is toxic to Saccharomyces cerevisiae cells lacking the Fet3/Ftr1.

Epigallocatechin-3-O-gallate (EGCG), the main green tea component, is intensively studied for its anti-oxidant, anti-inflammatory, anti-microbial and anti-cancer effects. In the present study, a screen on a Saccharomyces cerevisiae gene deletion library was performed to identify conditions under which EGCG had deleterious rather than beneficial effects. Two genes were identified whose deletion resulted in sensitivity to EGCG: FET3 and FTR1, encoding the components of the Fet3/Ftr1 high-affinity iron uptake system, also involved in Cu(I)/Cu(II) balance on the surface of yeast cells.

Heavy metal accumulation by Saccharomyces cerevisiae cells armed with metal binding hexapeptides targeted to the inner face of the plasma membrane.

Accumulation of heavy metals without developing toxicity symptoms is a phenotype restricted to a small group of plants called hyperaccumulators, whose metal-related characteristics suggested the high potential in biotechnologies such as bioremediation and bioextraction. In an attempt to extrapolate the heavy metal hyperaccumulating phenotype to yeast, we obtained Saccharomyces cerevisiae cells armed with non-natural metal-binding hexapeptides targeted to the inner face of the plasma membrane, expected to sequester the metal ions once they penetrated the cell. We describe the construction of S.

Anchoring plant metallothioneins to the inner face of the plasma membrane of Saccharomyces cerevisiae cells leads to heavy metal accumulation.

In this study we engineered yeast cells armed for heavy metal accumulation by targeting plant metallothioneins to the inner face of the yeast plasma membrane. Metallothioneins (MTs) are cysteine-rich proteins involved in the buffering of excess metal ions, especially Cu(I), Zn(II) or Cd(II). The cDNAs of seven Arabidopsis thaliana MTs (AtMT1a, AtMT1c, AtMT2a, AtMT2b, AtMT3, AtMT4a and AtMT4b) and four Noccaea caerulescens MTs (NcMT1, NcMT2a, NcMT2b and NcMT3) were each translationally fused to the C-terminus of a myristoylation green fluorescent protein variant (myrGFP) and expressed in Saccharomyces cerevisiae cells.

Calcium signaling and copper toxicity in Saccharomyces cerevisiae cells.

To respond to metal surpluses, cells have developed intricate ways of defense against the excessive metallic ions. To understand the ways in which cells sense the presence of toxic concentration in the environment, the role of Ca in mediating the cell response to high Cu was investigated in Saccharomyces cerevisiae cells. It was found that the cell exposure to high Cu was accompanied by elevations in cytosolic Ca with patterns that were influenced not only by Cu concentration but also by the oxidative state of the cell.

Interaction between lanthanide ions and Saccharomyces cerevisiae cells.

Lanthanides are a group of non-essential elements with important imaging and therapeutic applications. Although trivalent lanthanide ions (Ln³⁺) are used as potent blockers of Ca²⁺ channels, the systematic studies correlating Ln³⁺ accumulation and toxicity to Ca²⁺ channel blocking activity are scarce. In this study, we made use of the eukaryotic model Saccharomyces cerevisiae to investigate the correlation between Ln³⁺ accumulation, their toxicity and their capacity to block the exogenous stress-induced Ca²⁺ influx into the cytosol.