Post-Translational Modifications Control Phase Transitions of Tau.

The self-assembly of Tau(297-391) into filaments, which mirror the structures observed in Alzheimer's disease (AD) brains, raises questions about the role of AD-specific post-translational modifications (PTMs) in the formation of paired helical filaments (PHFs). To investigate this, we developed a synthetic approach to produce Tau(291-391) featuring N-acetyllysine, phosphoserine, phosphotyrosine, and N-glycosylation at positions commonly modified in post-mortem AD brains, thus facilitating the study of their roles in Tau pathology. Using transmission electron microscopy (TEM), cryo-electron microscopy (cryo-EM), and a range of optical microscopy techniques, we discovered that these modifications generally hinder the in vitro assembly of Tau into PHFs. Interestingly, while acetylation's effect on Tau assembly displayed variability, either promoting or inhibiting phase transitions in the context of cofactor free aggregation, heparin-induced aggregation, and RNA-mediated liquid-liquid phase separation (LLPS), phosphorylation uniformly mitigated these processes. Our observations suggest that PTMs, particularly those situated outside the fibril's rigid core are pivotal in the nucleation of PHFs. Moreover, in scenarios involving heparin-induced aggregation leading to the formation of heterogeneous aggregates, most AD-specific PTMs, except for K311, appeared to decelerate the aggregation process. The impact of acetylation on RNA-induced LLPS was notably site-dependent, exhibiting both facilitative and inhibitory effects, whereas phosphorylation consistently reduced LLPS across all proteoforms examined. These insights underscore the complex interplay between site-specific PTMs and environmental factors in modulating Tau aggregation kinetics, enhancing our understanding of the molecular underpinnings of Tau pathology in AD and highlighting the critical role of PTMs located outside the ordered filament core in driving the self-assembly of Tau into PHF structures.

Peptide and Protein Desulfurization with Diboron Reagents.

In this Letter, we report a direct and robust desulfurization method employing water-soluble phosphine, specifically tris(2-carboxyethyl)phosphine hydrochloride (TCEP), and tetrahydroxydiboron (B2(OH)4), which serves as a radical initiator. This innovative reaction exhibits compatibility with a diverse array of substrates, including cysteine residues in chemically synthesized oligopeptides and cyclic peptides, alkyl thiols in bioactive molecules, disulfides in commercial proteins, and selenocysteine. We optimized the reaction conditions to minimize the formation of undesired oxidized and borylated byproducts. Furthermore, the refined desulfurization process is executed after native chemical ligation (NCL) in a single pot, streamlining the existing synthetic approaches. This demonstrates its potential applications in the synthesis of complex peptides and proteins, showcasing a significant advancement in the field.

Monocyte-to-lymphocyte ratio correlates with parathyroid hormone concentration in patients with severe symptomatic aortic stenosis.

PURPOSE: The normal healthy valve is devoid of inflammatory cells, however background of aortic stenosis (AS) may include inflammatory processes. Moreover, the link between hyperparathyroidism and heart failure is postulated. Simple whole blood analysis with indices is a beneficial tool in cardiovascular diseases' assessment. The purpose of the study was to evaluate correlation between parathyroid hormone (PTH) and simple blood parameters in severe AS. MATERIAL AND METHODS: The study included 62 patients with severe AS. Patients with inflammatory or autoimmune co-morbidities were excluded. Blood samples were collected, and clinical and demographic data were analyzed. RESULTS: The final study group comprised 55 patients (31 females, 56.4%; mean age 77.13 (SD 6.76)). In 23 patients (41.8%), PTH concentration was markedly increased. The study group was divided into two subgroups according to the PTH concentration. Patients from both groups did not differ significantly in terms of age and co-morbidities. PTH concentration correlated positively with monocyte-lymphocyte ratio (MLR) (p â€‹= â€‹0.008, Spearman rho 0.356) and platelet-lymphocyte ratio (PLR) (p â€‹= â€‹0.047, Spearman rho 0.269), creatinine level (p â€‹= â€‹0.001, Spearman rho 0.425) and glomerular filtration rate (GFR-MDRD) (p â€‹= â€‹0.009, Spearman rho -0.349). The multivariable logistic regression with backward analysis revealed MLR (p â€‹= â€‹0.029) and GFR (p â€‹= â€‹0.028) as independent significant predictors of abnormal PTH values. The receiver operator characteristics (ROC) curve was performed for the model of MLR and GFR-MDRD (AUC â€‹= â€‹0.777), yielding the sensitivity of 60.9% and specificity of 90.6%. CONCLUSIONS: PTH concentration correlates with monocyte-to-lymphocyte and platelet-to-lymphocyte ratios in calcified AS.

Desulfurative Borylation of Small Molecules, Peptides, and Proteins.

We introduce a direct conversion of alkyl thiols into boronic acids, facilitated by a water-soluble phosphine, 1,3,5-triaza-7-phosphaadamantane (PTA), in conjunction with tetrahydroxydiboron (B2(OH)4), acting as both a radical initiator and a boron source. This desulfurative borylation reaction has been successfully applied to various substrates, including cysteine residues in oligopeptides and small proteins, primary alkyl thiols found in pharmaceutical compounds, disulfides, and selenocysteine. Optimization of reaction conditions was undertaken to reduce the formation of unwanted reactions, such as the reduction of alanyl or other primary radicals, and to prevent deleterious reactions between the phosphine and N-terminal amine that lead to methylene adducts by utilizing a buffer containing glycine-glycine (GG) dipeptide. The developed method is characterized by its operational simplicity and robustness. Moreover, its compatibility with various functional groups present in peptides and proteins makes it a promising tool for late-stage functionalization, extending its potential application across a broad spectrum of chemical and biological targets.

Chemical Synthesis of Microtubule-Associated Protein Tau.

Deposits of the microtubule-associated protein Tau (MAPT) serve as a hallmark of neurodegenerative diseases known as tauopathies. Numerous studies have demonstrated that in diseases such as Alzheimer's disease (AD), Tau undergoes extensive remodeling. The attachment of post-translational modifications distributed throughout the entire sequence of the protein correlates with clinical presentation. A systematic examination of these protein alterations can shed light on their roles in both healthy and diseased states. However, the ability to access these modifications in the entire protein chain is limited as Tau can only be produced recombinantly or through semisynthesis. In this article, we describe the first chemical synthesis of the longest 2N4R isoform of Tau, consisting of 441 amino acids. The 2N4R Tau was divided into 3 major segments and a total of 11 fragments, all of which were prepared via solid-phase peptide synthesis. The successful chemical strategy has relied on the strategic use of two cysteine sites (C291 and C322) for the native chemical ligations (NCLs). This was combined with modern preparative protein chemistries, such as mercaptothreonine ligation (T205), diselenide-selenoester ligation (D358), and mutations of mercaptoamino acids into native residues via homogeneous radical desulfurization (A40, A77, A119, A157, A246, and A390). The successful completion of the synthesis has established a robust and scalable route to the native protein in multimilligram quantities and high purity. In broader terms, the presented strategy can be applied to the preparation of other shorter isoforms of Tau as well as to introduce all post-translational modifications that are characteristic of tauopathies such as AD.

TMEM106B Fibrils from FTLD Patients and Healthy Controls.

Recent studies involving four research teams have revealed that amyloid fibrils in FTLD-TDP patients and cognitively healthy individuals primarily consist of TMEM106B, a protein previously identified as a risk factor for FTLD-TDP. Through cryogenic electron microscopy, the studies identified various protofilament structures of TMEM106B fibrils from individuals with several neurodegenerative diseases. These findings raise new questions and opportunities for future research, as they suggest that TMEM106B plays a central role in FTLD pathology. These discoveries also prompt the need for the development of specific antibodies for fibrillar TMEM106B and necessitate further investigation of the potential mechanistic link between TMEM106B and other filamentous aggregates. The power of cryo-EM techniques is underscored in these unexpected findings and may be a vital tool for gaining further molecular insights into neurodegenerative diseases characterized by amyloid deposits.

Stereoselective alkyl C-glycosylation of glycosyl esters via anomeric C-O bond homolysis: efficient access to C-glycosyl amino acids and C-glycosyl peptides.

C-Glycosyl peptides possess excellent metabolic stability and therapeutic properties and thus play critical roles in biological studies as well as drug discoveries. However, the limited accessibility of C-glycosyl amino acids has significantly hindered the broader research of their structural features and mode of action. Herein, for the first time we disclose a novel visible-light-driven radical conjugate addition of 1,4-dihydropyridine (DHP)-derived glycosyl esters with dehydroalanine derivatives, generating C-glycosyl amino acids and C-glycosyl peptides in good yields with excellent stereoselectivities. Redox-active glycosyl esters, as readily accessible and bench-stable radical precursors, could be easily converted to glycosyl radicals via anomeric C(sp3)-O bond homolysis under mild conditions. Importantly, the generality and practicality of this transformation were fully demonstrated in >40 examples including 2-dexosugars, oligosaccharides, oligopeptides, and complex drug molecules. Given its mild reaction conditions, robust sugar scope, and high anomeric control and diastereoselectivity, the method presented herein could find widespread utility in the preparation of C(sp3)-linked sugar-based peptidomimetics.

Catalytic Amide Activation with Thermally Stable Molybdenum(VI) Dioxide Complexes.

Oxazolines and thiazolines are important constituents of bioactive natural products and pharmaceuticals. Here, we report the development of an effective and practical method of oxazoline and thiazoline formation, which can facilitate the synthesis of natural products, chiral ligands, and pharmaceutical intermediates. This method capitalized on a Mo(VI) dioxide catalyst stabilized by substituted picolinic acid ligands, which is tolerant to many functional groups that would otherwise be sensitive to highly electrophilic alternative reagents.

Glomus sp. and Bacillus sp. strains mitigate the adverse effects of drought on maize (Zea mays L.).

Maize (Zea mays L.) is an economically important source of food and feed. This species is highly sensitive to drought, which is the most limiting factor for the biomass yield of a crop. Thus, maize cultivation methods should be improved, especially by environment-friendly agricultural practices, such as microorganisms. Here, we provide evidence that Glomus sp. and Bacillus sp. modulate maize response to drought. Inoculation of maize seeds by these microorganisms restored the proper photosynthetic activity of the plant under drought and stabilized the osmoprotectant content of the leaf. The beneficial effect of Glomus sp. and Bacillus sp. was also related to the stabilization of cell redox status reflected by hydrogen peroxide content, antioxidant enzymes, and malondialdehyde level in leaves. As we revealed by several methods, shaping maize response to drought is mediated by both microorganism-mediated modifications of cell wall composition and structure of leaves, such as downregulating pectin, affecting their methylation degree, and increasing hemicellulose content. Overall, we provide new information about the mechanisms by which Glomus sp. and Bacillus sp. induce drought tolerance in maize, which is a promising approach for mitigating abiotic stresses.

Umpolung AlaB Reagents for the Synthesis of Non-Proteogenic Amino Acids, Peptides and Proteins.

Non-proteogenic amino acids and functionalized peptides are important motifs in modern drug discovery. Here we report that AlaB can serve as universal building blocks in the synthesis of a diverse collection of modified amino acids, peptides, and proteins. First, we develop the synthesis of AlaB from redox-active esters of aspartic acid resulting in a series of ß-boronoalanine derivatives. Next, we show that AlaB can be integrated into automated oligopeptide solid-phase synthesis. AlaB is compatible with common transformations used in preparative peptide chemistry such as native chemical ligation and radical desulfurization as showcased by total synthesis of AlaB -containing ubiquitin. Furthermore, AlaB reagents participate in Pd-catalyzed reactions, including C-C cross-couplings and macrocyclizations. Taken together, AlaB synthons are practical reagents to access modified peptides, proteins, and in the synthesis of cyclic/stapled peptides.

Current Landscape of Coccidioidomycosis.

Coccidioidomycosis, also known as Valley fever, is an endemic fungal infection commonly found in the southwestern parts of the United States. However, the disease has seen an increase in both in its area of residency and its prevalence. This review compiles some of the latest information on the epidemiology, current and in-development pharmaceutical approaches to treat the disease, trends and projections, diagnostic concerns, and the overlapping dynamics of coccidioidomycosis and COVID-19, including in special populations. This review provides an overview of the current diagnostic and therapeutic strategies and identifies areas of future development.

Influence of Fermentation Beetroot Juice Process on the Physico-Chemical Properties of Spray Dried Powder.

Picking vegetables is, along with salting and drying, one of the oldest ways to preserve food in the world. This is the process of decomposition of simple sugars into lactic acid with the participation of lactic bacteria. The aim of the study was to obtain powders from fermented red beet juice with the highest possible amount of lactic acid bacteria (LAB) and active ingredients. For the analysis, juices were squeezed from the vegetables and two types of fermentation were used: a spontaneous fermentation and a dedicated one. After inoculation, samples were taken for analysis on a daily basis. Extract, pH, total acidity, pigments, and color were measured. In addition, microbiological tests were also carried out. The juices from the fifth day of fermentation was also spray dried, to obtain fermented beetroot powder. Juices from 3-5th day were characterized by a high content of LAB and betanin, had also a low pH, which proves that the lactic fermentation is working properly. The exception was the juice from spontaneous fermentation. According to the observations, the fermentation process did not run properly, and further analysis is needed. The powders were stable; however, results obtained from the pigment content and the LAB content are not satisfactory and require further analysis.

Identification of Ca2+ oscillations with the 0-1 test for periodic and chaotic time-series: One- and two-parameter bifurcations.

This paper examines the oscillatory responses (periodic and chaotic) of a biosystem store model for bursting and complex Ca2+ oscillations in which three compartments have been taken into consideration: the cytosol, endoplasmic reticulum (ER) and mitochondria. The oscillatory model is used to examine the reliability of the 0-1 test for chaos in the bifurcation analysis of continuous signals obtained when the frequencies of oscillatory responses vary significantly with a relatively small changes of the bifurcation parameters. The illustrative examples in both the one- and two-parameter cases are designed to show that for a periodic time-series the test's reliability may be questioned when a periodic series is classified as a chaotic one - the 'false-positive' case. To prevent the incorrect result an additional computational work is needed to examine the frequency spectrum of the periodic time-series. The illustrative examples utilize an autonomous dynamical model of cytosolic calcium oscillations with three dynamical variables and sixteen parameters. The dynamical model is such that the frequency of oscillations may change by the factor of about 200, when a certain dynamical system's parameter changes from its minimum to maximum values, making selection of the parameters in the 0-1 test extremely difficult. The extra computational work improves the test's reliability and eliminates the 'false-positive' outcomes of the test. The paper is focused on the computational aspects of the 0-1 test for periodic and chaotic oscillations rather than on the properties of the store model for bursting and complex Ca2+ oscillations.

Site-Selective Amide Functionalization by Catalytic Azoline Engrafting.

Direct peptide and protein activation is a challenging transformation because of the stabilizing effect of the amide group. While enzymes can be considered as prototypical systems that have evolved to achieve high selectivity and specificity, small-molecule catalysts that functionalize the amide group may accommodate a much larger selection of substrates but currently remain scarce. Here, by combining the desired features from both catalytic regimes we designed an artificial cyclodehydratase, a catalytic system for the site-selective modification of peptides and natural products by engrafting heterocycles into their scaffolds. The catalytic system features a molybdenum(VI) center that was decorated with a sterically congested tripod ligand. The optimized catalyst can introduce azolines into small molecules, natural products, and oligopeptides with high efficiency and minimal waste. We further demonstrate the utility of the new protocol in the direct functionalization of a single amide group in the presence of up to seven other chemically similar positions and in the direct conversion of these groups into amines and thioamides. This new mechanistic paradigm may address an unmet need for a general method for the selective and sustainable functionalization of peptides and natural products.

Organometallic AlaM Reagents for Umpolung Peptide Diversification.

Selective modifications of peptides and proteins have emerged as a promising strategy to develop novel mechanistic probes and prepare compounds with translational potentials. Here, we report alanine carbastannatranes AlaSn as a universal synthon in various C-C and C-heteroatom bond-forming reactions. These reagents are compatible with peptide manipulation techniques and can undergo chemoselective conjugation in minutes when promoted by Pd(0). Despite their increased nucleophilicity and propensity to transfer the alkyl group, C(sp3)-C(sp2) coupling with AlaSn can be accomplished at room temperature under buffered conditions (pH 6.5-8.5). We also show that AlaSn can be easily transformed into several canonical L- and D-amino acids in arylation, acylation, and etherification reactions. Furthermore, AlaSn can partake in macrocyclizations exemplified by the synthesis of medium size cyclic peptides with various topologies. Taken together, metalated alanine AlaSn demonstrates unparalleled scope and represents a new type of umpolung reagents suitable for structure-activity relationship studies and peptide diversification.

Characterization of Collagen/Beta Glucan Hydrogels Crosslinked with Tannic Acid.

Hydrogels based on collagen/ß-glucan crosslinked with tannic acid were obtained by neutralization using dialysis. The presence of tannic acid allowed obtaining stable hydrogel materials with better mechanical properties. Tannic acid was released from matrices gradually and not rapidly. The antioxidant properties of the obtained hydrogels increased over the course of their incubation in culture media and were dependent on the concentration of tannic acid in the matrices. The obtained materials influenced dehydrogenase activity and the ATP level of pathogens. Additionally, the materials' extracts improved the HaCaT cells' viability. Therefore, the obtained hydrogels seem to be promising biocompatible materials which display antimicrobial properties.

Computational Analysis of Ca2+ Oscillatory Bio-Signals: Two-Parameter Bifurcation Diagrams.

Two types of bifurcation diagrams of cytosolic calcium nonlinear oscillatory systems are presented in rectangular areas determined by two slowly varying parameters. Verification of the periodic dynamics in the two-parameter areas requires solving the underlying model a few hundred thousand or a few million times, depending on the assumed resolution of the desired diagrams (color bifurcation figures). One type of diagram shows period-n oscillations, that is, periodic oscillations having n maximum values in one period. The second type of diagram shows frequency distributions in the rectangular areas. Each of those types of diagrams gives different information regarding the analyzed autonomous systems and they complement each other. In some parts of the considered rectangular areas, the analyzed systems may exhibit non-periodic steady-state solutions, i.e., constant (equilibrium points), oscillatory chaotic or unstable solutions. The identification process distinguishes the later types from the former one (periodic). Our bifurcation diagrams complement other possible two-parameter diagrams one may create for the same autonomous systems, for example, the diagrams of Lyapunov exponents, Ls diagrams for mixed-mode oscillations or the 0-1 test for chaos and sample entropy diagrams. Computing our two-parameter bifurcation diagrams in practice and determining the areas of periodicity is based on using an appropriate numerical solver of the underlying mathematical model (system of differential equations) with an adaptive (or constant) step-size of integration, using parallel computations. The case presented in this paper is illustrated by the diagrams for an autonomous dynamical model for cytosolic calcium oscillations, an interesting nonlinear model with three dynamical variables, sixteen parameters and various nonlinear terms of polynomial and rational types. The identified frequency of oscillations may increase or decrease a few hundred times within the assumed range of parameters, which is a rather unusual property. Such a dynamical model of cytosolic calcium oscillations, with mitochondria included, is an important model in which control of the basic functions of cells is achieved through the Ca2+ signal regulation.

Evaluation of the Mechanical and Biocidal Properties of Lapacho from Tabebuia Plant as a Biocomposite Material.

The aim of this article is to discuss in detail the physicochemical properties of polylactide (PLA) reinforced by cortex fibers, which may cause bacterial mortality and increased biodegradation rates. PLA biocomposites containing cortex Lapacho fibers from Tabebuia (1-10 wt%) were prepared by extrusion and injection moulding processes. The effects of Lapacho on the mechanical and biocidal properties of the biocomposites were studied using tensile and impact tests, dynamic mechanical analysis (DMA), differential scanning calorimetry (DSC), thermogravimetry (TG), and the method of evaluating the antibacterial activity of antibacterial treated according to the standard ISO 22196:2011. It also presented the effects of Lapacho on the structural properties and biodegradation rates of biocomposites. This research study provides very important results complementing the current state of knowledge about the biocidal properties of Lapacho from Tabebuia plants and about cortex-reinforced biocomposites.