Calcium pantothenate 750mg modified-release tablets

In Mycobacterium tuberculosis (Mtb), proteins that are posttranslationally modified with a prokaryotic ubiquitin-like protein (Pup) can be degraded by bacterial proteasomes. A single Pup-ligase and depupylase shape the pupylome, but the mechanisms regulating their substrate specificity are incompletely understood. Here, we identified a depupylation regulator, a protein called CoaX, through its copurification with the depupylase Dop. CoaX is a pseudopantothenate kinase that showed evidence of binding to pantothenate, an essential nutrient Mtb synthesizes, but not its phosphorylation. In a ∆coaX mutant, pantothenate synthesis enzymes including PanB, a substrate of the Pup-proteasome system (PPS), were more abundant than in the parental strain. In vitro, CoaX specifically accelerated depupylation of Pup~PanB, while addition of pantothenate inhibited this reaction. In culture, media supplementation with pantothenate decreased PanB levels, which required CoaX. Collectively, we propose CoaX regulates PanB abundance in response to pantothenate levels by modulating its vulnerability to proteolysis by Mtb proteasomes.

Abstract licence: CC BY-NC-ND

AbstractConsiderable efforts have been devoted in recent years to enhancing the efficacy medicinal substance, leading to the discovery of innovative drug formulations and delivery techniques. The successful design of these processes necessitates a profound understanding at the molecular level of how these substances interact with biological membranes. Thorough thermodynamic investigations provide invaluable insights into these interactions and aid in selecting suitable compounds for pharmaceutical production. This study aims to determine the density and speed of sound for D-calcium pantothenate in mixtures of water and deep eutectic solvents (DESs), specifically choline chloride/sucrose, choline chloride/ glucose, and choline chloride/ fructose (with 2:1 molar ratio) over a temperature range of 288.15 K to 318.15 K under atmospheric pressure. In order to predict the behavior of molecules, COSMO model (the Conductor-Like Screening Model) offer complementary strengths in quantum chemistry. This approach allows for calculating solvation free energies, making it ideal for predicting properties like solubility, where understanding solvent-solute interactions is crucial. By correlating the measured parameters using standard relationships, important partial molar parameters such as apparent molar volumes and apparent molar isentropic compressibility are calculated. Additionally, apparent molar isobaric expansion, and Hepler’s constant are derived from the density and speed of sound data. The experimental apparent molar volumes, and apparent molar isentropic compressibility data is fitted to the Redlich-Meyer equation to obtain significant quantities such as standard partial molar volume, and partial molar isentropic compression. The comprehensive thermodynamic analysis of this studied system holds immense significance for advancements in the pharmaceutical industry.

Abstract licence: CC BY 4.0

Acute respiratory distress syndrome (ARDS) is a severe condition with complex pathogenesis, and emerging evidence highlights the potential role of metabolic factors, though the exact mechanisms are not fully understood. In this study, we used Mendelian randomisation (MR) and multi-omics approaches to investigate the causal relationship between plasma metabolites, immune cell profiles and ARDS risk. MR analysis of 1400 metabolites identified two causal metabolites linked to increased ARDS risk, primarily involved in pantothenate and CoA biosynthesis. Single-cell RNA sequencing of ARDS samples revealed that monocytes exhibited the highest levels of pantothenate synthesis. Intercellular communication and pseudotime analysis suggested that the pantothenate synthesis pathway influenced monocyte differentiation and interactions with other cell types. Gene set enrichment analysis showed that monocytes with high pantothenate synthesis were significantly enriched in phagocytosis-related pathways. Subsequent MR analysis demonstrated that CD33dim HLA DR+ CD11b+%CD33dim HLA DR+ were a risk factor against ARDS. Notably, monocytes with high pantothenate synthesis exhibited decreased expression of antigen presentation markers HLA-DRB5, HLA-DRB1 and HLA-DRA, suggesting that the high pantothenate synthesis monocytes exhibit attenuated antigen presentation and enhanced phagocytic function. Moreover, we developed a diagnostic model using machine learning algorithms. Shapley Additive explanation (SHAP) was leveraged to evaluate the model performance, with CALM2 identified as the most influential feature across the CatBoost and XGBoost models. In summary, this study integrates genetic, multi-omics and machine learning approaches to provide novel insights into the pathogenesis of ARDS and its potential therapeutic strategies targeting monocyte metabolism and function.

Abstract licence: CC BY

Toxoplasma gondii is an obligate intracellular pathogen that infects humans and many animals. It harbors unique secretory organelles to facilitate its parasitic lifestyle, including dense granules that secrete diverse proteins to different destinations. Yet the biological functions of these secretory proteins are mostly unknown. Here, we examined the roles of GRA1, the first dense granule protein discovered. GRA1 is secreted to the parasitophorous vacuole (PV), and its conditional depletion led to growth arrest of tachyzoites, suggesting a crucial role of GRA1 for parasite propagation during acute infection. Furthermore, GRA1 inactivation resulted in severe metabolic defects, including reduced glycolysis and tricarboxylic acid cycle activities and a drop in cellular ATP levels. GRA1 depletion also disrupted the integrity of the mitochondrion and apicoplasts. Mechanistically, disruption of GRA1 altered the structures of membranous tubules in the PV, called the intravacuolar network (IVN), which are thought to mediate the transport of molecules in and out of the parasites. Consistently, GRA1 inactivation impaired the secretion of various GRA proteins to the PV membrane (PVM) or host cells, including GRA17 that is involved in nutrient uptake from host cells. In agreement with the mislocalization of GRA proteins, mutants lacking GRA1 had a strong defect in absorbing nutrients like glucose and pantothenate from host cells. Consequently, the parasites displayed slower growth and a higher tendency to differentiate into bradyzoites. Together, these results suggest that GRA1 plays a vital role in maintaining the structural integrity of the IVN, thus contributing to the scavenging of host nutrients.ImportanceToxoplasma gondii critically relies on host nutrients for growth, but the underlying mechanisms are largely unknown. Here, we discovered that the secretory protein GRA1 in Toxoplasma played a crucial role in scavenging host nutrients to support parasite proliferation. GRA1 is the first reported GRA protein, but its function remains enigmatic since its discovery in 1989. GRA1 is secreted to the PV from the dense granules, which harbor many proteins that traffic to PV, PVM, and even host cells to perform diverse functions, including forming channels on the PVM to take up host nutrients to establish parasitism. GRA1 disruption impaired the structure of IVN and abolished the targeting of other GRA proteins to their destinations, which compromised the parasite's ability to import nutrients from the hosts. These findings reveal the functional mode of GRA1 in T. gondii and highlight its potential as a target for developing new interventions against toxoplasmosis.

Abstract licence: CC BY

Recombinant adeno associated virus (rAAV) vectors have become popular delivery vehicles for in vivo gene therapies, but demand for rAAVs continues to outpace supply. Platform processes for rAAV production are being developed by many manufacturers, and transient chemical transfection of human embryonic kidney 293 (HEK293) cells is currently the most popular approach. However, the cutting edge nature of rAAV process development encourages manufacturers to keep cell culture media formulations, plasmid sequences, and other details proprietary, which creates hurdles for small companies and academic labs seeking to innovate in this space. To address this problem, we leveraged the resources of an academic-industry consortium (Advanced Mammalian Biomanufacturing Innovation Center, AMBIC) to develop an rAAV production system based on transient transfection of suspension HEK293 cells adapted to an in-house, chemically defined medium. We found that balancing iron and calcium levels in the medium were crucial for maintaining transfection efficiency and minimizing cell aggregation, respectively. A design of experiments approach was used to optimize the transient transfection process for batch rAAV production, and PEI:DNA ratio and cell density at transfection were the parameters with the strongest effects on vector genome (VG) titer. When the optimized transient process was transferred between two university sites, VG titers were within a twofold range. Analytical characterization showed that purified rAAV from the AMBIC process had comparable viral protein molecular weights versus vector derived from commercial processes, but differences in transducing unit (TU) titer were observed between vector preps. The developed media formulation, transient transfection process, and analytics for VG titer, capsid identity, and TU titer constitute a set of workflows that can be adopted by others to study fundamental problems that could improve product yield and quality in the nascent field of rAAV manufacturing.

Abstract licence: CC BY-NC