Brain infection by the pathogenic free-living amoebae Naegleria fowleri is a life-threatening illness, with a mortality rate >95% in the US. One of the challenges faced in treating these infections is the lack of efficacious drugs. Here, we further characterize two leads. The first, a N. fowleri enolase inhibitor (HEX ((1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid)), emerged from a target-based approach while the other, BDGR-20191 (rac-(4aR,13bS)-10,11-dichloro-4-(oxetan-3-ylmethyl)-1,2,3,4,4a,5,6,13b-octahydro-8H-[1,6]naphthyridino[5,6-b]quinazolin-8-one), was identified in a phenotypic screen for anti-amoebic agents. Chronic low dose exposure to these agents led to cell lines that were tolerant to each. However, the tolerance phenotypes were reversed in the absence of selection with the agent. Differential gene expression analyses suggest that HEX tolerant cells upregulate cellular components that could impact levels of accumulated glycolytic intermediates, while BDGR-20191 tolerant cells could be targeting membrane biosynthesis. Unbiased metabolomics of the lines supported these possibilities. While HEX and BDGR-20191 are not synergistic with one another or with other standard-of-care drugs, their additive nature suggests they could be a useful addition to current therapeutic cocktails.

The induction period (IP, obtained via standard conditions and a commercial instrument) is nowadays one of the most common tools to investigate oxidative stability in lipid samples. Despite the advantages and widespread use of this methodology in industry, the development of algorithms to predict IP values has been impaired by the scarcity of data linking the composition of the sample with the corresponding IP value. Addressing this need and considering the persistent difficulties in bridging experimental and theoretical approaches related to edible fats, this paper reports on the development of a machine learning approach based on extreme gradient boosting (XGBoost) that was trained using a biodiesel database and then fine-tuned using a vegetable oils database, in both cases linking composition to their corresponding stability (via rancimat). The resulting model was able to predict accelerated oxidative stability of vegetable oils with a mean average error of only 1.37 h, one of the lowest ever reported. Beyond demonstrating the ability of our model to provide meaningful predictions, this report highlights the importance of incorporating oxidation-related chemical descriptors and the utility of using associated databases and transfer learning to support the development of accurate machine learning models.

Toxoplasma gondii infects approximately one-third of the human population, posing a severe and potentially fatal risk to individuals with compromised immune systems. Our previous studies demonstrated that modifying the arene in the herbicidal protoporphyrinogen oxidase (PPO) inhibitor, oxadiazon, yields analogs that potently inhibit T. gondii PPO, a key enzyme in the heme biosynthesis pathway. In this study, we further investigated the structure-activity relationship of oxadiazon analogs by introducing aliphatic chains with varying functionalities, resulting in 23 new derivatives. Some of these compounds exhibited significant intracellular inhibition of wild-type T. gondii, with IC50 values ranging from 2 to 3 μM. Biochemical analysis confirmed that their mode of action is mediated by potent PPO inhibition, which further blocked heme production and damaged mitochondrial health status in the parasites. These findings enhance our understanding of oxadiazon's structural optimization and highlight its derivatives as promising early-stage candidates for developing effective therapies against toxoplasmosis in humans and other animals.

The pathogenic free-living amoeba Naegleria fowleri causes primary amoebic meningoencephalitis (PAM), a highly fatal disease with limited treatment options, underscoring the urgent need for new therapeutics. Our previous studies identified (1-hydroxy-2-oxopiperidin-3-yl)phosphonic acid (HEX), an inhibitor of human enolase 2 (ENO2) involved in glucose metabolism, as a potent inhibitor of N. fowleri enolase (NfENO) with potent amoebicidal activity. In this study, we explored the structure-activity relationship (SAR) of HEX by modifying its hydroxamate and phosphonate functional groups, as well as introducing steric alterations to generate new analogs. Functional assays and computational-assisted SAR analysis provided insights into the impact of HEX modifications on N. fowleri agonism. Ultimately, the results of this study demonstrated that the activity of the HEX scaffold toward NfENO is rather sensitive to structural purturbations, confirming the necessity of both key functional groups - the hydroxamate and phosphonate - to maintain potency. Additionally, structural modifications of the parent compound into bicyclic analogs resulted in loss of biological activity ostensibly due to unfavorable steric interactions in the active site. These findings enhance our understanding of the activity of HEX's molecular architecture, and underscore potential limitations of further structural tuning efforts of the scaffold by means of SAR.

We describe a new one-pot synthesis to access richly functionalized tetrahydroindoles and related fused pyrroles. The key to this transformation is a robust cascade cyclization between ene-yne sulfides and azodicarboxylates involving diazacyclobutene formation primed for rapid electrocyclic ring opening to an α-iminothioimidate intermediate, followed by cyclization and aromatization to yield the N-heterocycles in good yields. This tactic represents a significant departure from conventional approaches to the tetrahydroindole motif, which typically involve either staging the 140-year-old Paal-Knorr cyclization or leveraging transition metal catalysis. Further synthetic applications demonstrated that this method can be used for the late-stage functionalization of complex bioactive molecules or natural product-like structures in an efficient manner. The mechanism of the cyclization event leading to the heterocyclic products was probed computationally and is more consistent with a polar process instead of a concerted aza-Nazarov cyclization. The products can be leveraged in a mechanistically interesting desulfination reaction or converted to highly substituted indoles under mild conditions.

A convenient, metal-free one-pot synthesis has been developed to access 1,3,4-oxadiazol-2(3H)-ones using hydrazines, carboxylic acids, and 1,1 '-carbonyldiimidazole (CDI) as an activating agent in the presence of triethylamine (TEA). This method can access the desired heterocycles in moderate to excellent yields in a one-pot transformation from commercially available starting materials, representing a significantly simpler path to access these compounds compared with existing tactics. The substrate scope investigation demonstrated that this method is broadly applicable and scalable.

We report an efficient two-step strategy for the synthesis of 1,2-diazetidin-3-ones (or aza-β-lactams) from thiolated diazacyclobutene intermediates. The transformation involves acid-promoted ring scission, followed by base-induced intramolecular nucleophilic cyclization, affording the desired diazetidinones in good yields. This method offers straightforward access to a structurally unique and underexplored four-membered heterocycle, expanding the synthetic toolbox for strained nitrogen-containing ring systems.

Rationale: X-ray computed tomography (CT) is crucial in precision medicine for diagnostic and therapeutic guidance. However, current small molecule CT contrast agents pose risks such as nephrotoxicity, short blood circulation time, limited scan durations, potential thyroid impact, and immune responses. These challenges necessitate the development of kidney-safe nanoparticle (NP)-based contrast agents (CAs). Methods: We developed safe intravenous blood pool NP-based CT CAs at a clinical-equivalent dose of 300 mgI/kg, suitable for vascular and hepatic imaging. Our iodinated lipid nanoemulsions (ILNEs) were optimized for shelf-life stability, osmolarity, and viscosity for excellent injectability. The ILNEs were designed to offer high contrast and were tested for minimal protein interaction, prolonged blood circulation, and hepatic clearance. In vitro studies, along with tests in mice and porcine models, were conducted to confirm safety, cytocompatibility, and absence of tissue damage. Results: The ILNEs demonstrated high x-ray attenuation, improved contrast enhancement, extended stability, and batch-to-batch consistency. They exhibited minimal protein interaction, prolonged blood residency of about 4 h, and hepatic clearance within three days, avoiding nephrotoxicity. Blood and thyroid-stimulating hormone (TSH) analyses, along with kidney and liver function tests, confirmed the safety of ILNEs. Conclusion: Our ILNEs offer a promising alternative to current CT contrast agents, with improved safety and efficacy profiles. The results support further toxicity evaluations for clinical translation, highlighting the potential of ILNEs in vascular and hepatic imaging without the associated risks of nephrotoxicity.

A regio- and stereoselective debenzylative cycloetherification (DBCE) reaction of protected hexoses to form stereodefined hydroxylated tetrahydrofurans of synthetic utility has been investigated under very mild room temperature reaction conditions. This study revealed the potential application of the DBCE reaction of simple, abundant starting materials to access stereochemically rich tetrahydrofuran compounds under mild reaction conditions.

ABSTRACT Infections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is very important for the infectious trophozoite lifecycle stage and inhibitors of glucose metabolism have been found to be toxic to the pathogen. Recently, human enolase 2 (ENO2) phosphonate inhibitors have been developed as lead agents to treat glioblastoma multiforme (GBM). These compounds, which cure GBM in a rodent model, are well-tolerated in mammals because enolase 1 (ENO1) is the predominant isoform used systemically. Here, we describe findings that demonstrate that these agents are potent inhibitors of N. fowleri ENO ( Nf ENO) and are lethal to amoebae. In particular, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX) was a potent enzyme inhibitor (IC 50 value of 0.14 ± 0.04 µM) that was toxic to trophozoites (EC 50 value of 0.21 ± 0.02 µM) while the reported CC 50 was >300 µM. Molecular docking simulation revealed that HEX binds strongly to the active site of Nf ENO with a binding affinity of –8.6 kcal/mol. Metabolomic studies of parasites treated with HEX revealed a 4.5 to 78-fold accumulation of glycolytic intermediates upstream of Nf ENO. Last, nasal instillation of HEX increased longevity of amoebae-infected rodents. Two days after infection, animals were treated for 10 days with 3 mg/kg HEX, followed by one week of observation. At the conclusion of the experiment, eight of 12 HEX-treated animals remained alive (resulting in an indeterminable median survival time) while one of 12 vehicle-treated rodents remained, yielding a median survival time of 10.9 days. Brains of six of the eight survivors were positive for amoebae, suggesting the agent at the tested dose suppressed, but did not eliminate, infection. These findings suggest that HEX is a promising lead for the treatment of PAM.

Previously, we described the synthesis of stable, bicyclic examples of the rather rare diazacyclobutene (DCB) motif by means of a cycloaddition between triazolinediones and electron-rich thiolated alkynes. Here, we report the investigation of the cycloaddition of triazolinediones with related electron-rich yne-carbamates and carbazole-alkynes. Bicyclic DCBs arising from yne-carbamates were isolated in 8-65% yield, while those arising from carbazole-alkynes were isolated in 28-59% yield. Mechanistic studies and characterization of isolable byproducts shed light on the underlying issues leading to poor to moderate yields.

Infections with the pathogenic free-living amoebae Naegleria fowleri can lead to life-threatening illnesses including catastrophic primary amoebic meningoencephalitis (PAM). Efficacious treatment options for these infections are lacking and the mortality rate remains >95% in the US. Glycolysis is very important for the infectious trophozoite lifecycle stage and inhibitors of glucose metabolism have been found to be toxic to the pathogen. Recently, human enolase 2 (ENO2) phosphonate inhibitors have been developed as lead agents to treat glioblastoma multiforme (GBM). These compounds, which cure GBM in a rodent model, are well-tolerated in mammals because enolase 1 (ENO1) is the predominant isoform used systemically. Here, we describe findings that demonstrate these agents are potent inhibitors of N. fowleri ENO (NfENO) and are lethal to amoebae. In particular, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX) was a potent enzyme inhibitor (IC50 = 0.14 ± 0.04 μM) that was toxic to trophozoites (EC50 = 0.21 ± 0.02 μM) while the reported CC50 was >300 μM. Molecular docking simulation revealed that HEX binds strongly to the active site of NfENO with a binding affinity of -8.6 kcal/mol. Metabolomic studies of parasites treated with HEX revealed a 4.5 to 78-fold accumulation of glycolytic intermediates upstream of NfENO. Last, nasal instillation of HEX increased longevity of amoebae-infected rodents. Two days after infection, animals were treated for 10 days with 3 mg/kg HEX, followed by one week of observation. At the end of the one-week observation, eight of 12 HEX-treated animals remained alive (resulting in an indeterminable median survival time) while one of 12 vehicle-treated rodents remained, yielding a median survival time of 10.9 days. However, intranasal HEX delivery was not curative as brains of six of the eight survivors were positive for amoebae. These findings suggest that HEX requires further evaluation to develop as a lead for treatment of PAM.

A stable 2,2,6,6-tetramethylpiperidine-1-yl)oxyl (TEMPO)-oxidized cellulose nanofibril (TCNF)/graphene oxide (GO)/polyethylenimine (PEI) composite hydrogel was synthesized by self-assembly instead of chemical crosslinking. Their chemical, morphology, surface, and mechanical properties were characterized and adsorption behavior for methyl blue (–) was systematically investigated in terms of the optimal GO content, pH effect, kinetics, and isotherm models. Additionally, to assess the adsorption capability of the TCNF/GO/PEI hydrogel for various contaminants, its effectiveness was also tested for methylene blue (+), Cu (II), and soybean oils. The maximum adsorption capability for the methyl blue (−) dyes increased from 3125 to 3962 mg/g when 13.3 % of GO was added. The adsorption capability for Cu (II) and soybean oils rose from 205.3 to 218.5 and 2.1 to 7.2 mg/g, respectively. The adsorption capability of optimized TCNF/GO/PEI hydrogel for a variety of contaminants was improved overall based on the increase of surface area, electrostatic interactions, and hydrophobic domains. Moreover, adding GO did not impact the adsorption mechanism but increased the external diffusion rate in the intraparticle diffusion model. This work provides a self-assembling route to TCNF/GO/PEI hydrogels with great potential for the removal of multiple water pollutants.

Current clinical small molecule x-ray CT agents are effective but pose risks such as nephrotoxicity, short blood circulation time, limiting scan durations, potential thyroid impact, and immune responses. These challenges drive the development of kidney-safe x-ray nanoparticle (NP)-based contrast agents (CAs), though translation to clinical practice is hindered by chemical complexities and potential toxicity. We have engineered an intravenous, injectable, and safe blood pool NP-based CT CAs at a clinical-equivalent dose of ∼300 mgI/kg (∼2 mL/kg), ideal for vascular and hepatic imaging which are limited by clinical agents. Our iodinated lipid nanodroplet emulsions (ILNEs) contrast agent offers high x-ray attenuation thus improved contrast enhancement, extended stability, and exceptional batch-to-batch consistency. It also boasts a straightforward and scalable manufacturing process with minimal protein interaction, prolonged blood residency (∼4h), and hepatic clearance within 3 days, avoiding nephrotoxicity. Studies in vitro, in mice, and 16.6kg porcine animal model studies confirm its safety, cytocompatibility, and absence of tissue damage. Blood, and thyroid-stimulating hormone (TSH) analyses, and kidney and liver function tests, also support further toxicity evaluations for clinical translation.

Cellulose nanomaterial (CNM) and polyethylenimine (PEI) composites have attracted growing attention due to their multifunctional characteristics, which have been applied in different fields. In this work, soybean hulls were valorized into carboxyl cellulose nanofibrils (COOH-CNFs), and composited into hydrogels with PEI by combining them with cationic chelating and physical adsorption strategies. Cellulose nanofibrils (CNFs) were produced from soybean hulls prior to oxidation by a TEMPO mediated reaction to obtain COOH–CNFs; then drops of zinc chloride were added to 1.5% aqueous COOH–CNF dispersions, which were left for 24 h to form COOH-CNF hydrogels. Finally, the hydrogels were functionalized using different concentration of PEI solutions over a range of pH values. Elemental analysis results showed that 20% aq. PEI at pH 11.6 is the optimum condition to synthesize the COOH–CNF/PEI hydrogels. Additionally, the adsorption efficiency for the removal of anionic methyl blue dyes and Cu(II) ions from water was tested, reaching 82.6% and 69.8%, respectively, after 24 h. These results demonstrate the great potential of COOH–CNF/PEI hydrogels as adsorbent materials for water remediation. Graphical abstract

Microbial pathogens use proteases for their infections, such as digestion of proteins for nutrients and activation of their virulence factors. As an obligate intracellular parasite, Toxoplasma gondii must invade host cells to establish its intracellular propagation. To facilitate invasion, the parasites secrete invasion effectors from microneme and rhoptry, two unique organelles in apicomplexans. Previous work has shown that some micronemal invasion effectors experience a series of proteolytic cleavages within the parasite’s secretion pathway for maturation, such as the aspartyl protease (TgASP3) and the cathepsin L-like protease (TgCPL), localized within the post-Golgi compartment and the endolysosomal system, respectively. Furthermore, it has been shown that the precise maturation of micronemal effectors is critical for Toxoplasma invasion and egress. Here, we show that an endosome-like compartment (ELC)-residing cathepsin C-like protease (TgCPC1) mediates the final trimming of some micronemal effectors, and its loss further results in defects in the steps of invasion, egress, and migration throughout the parasite’s lytic cycle. Notably, the deletion of TgCPC1 completely blocks the activation of subtilisin-like protease 1 (TgSUB1) in the parasites, which globally impairs the surface-trimming of many key micronemal invasion and egress effectors. Additionally, we found that Toxoplasma is not efficiently inhibited by the chemical inhibitor targeting the malarial CPC ortholog, suggesting that these cathepsin C-like orthologs are structurally different within the apicomplexan phylum. Collectively, our findings identify a novel function of TgCPC1 in processing micronemal proteins within the Toxoplasma parasite’s secretory pathway and expand the understanding of the roles of cathepsin C protease. IMPORTANCE Toxoplasma gondii is a microbial pathogen that is well adapted for disseminating infections. It can infect virtually all warm-blooded animals. Approximately one-third of the human population carries toxoplasmosis. During infection, the parasites sequentially secrete protein effectors from the microneme, rhoptry, and dense granule, three organelles exclusively found in apicomplexan parasites, to help establish their lytic cycle. Proteolytic cleavage of these secretory proteins is required for the parasite’s optimal function. Previous work has revealed that two proteases residing within the parasite’s secretory pathway cleave micronemal and rhoptry proteins, which mediate parasite invasion and egress. Here, we demonstrate that a cathepsin C-like protease (TgCPC1) is involved in processing several invasion and egress effectors. The genetic deletion of TgCPC1 prevented the complete maturation of some effectors in the parasites. Strikingly, the deletion led to a full inactivation of one surface-anchored protease, which globally impaired the trimming of some key micronemal proteins before secretion. Therefore, this finding represents a novel post-translational mechanism for the processing of virulence factors within microbial pathogens.

Nanocellulose/polyethylenimine composites have attracted growing attention due to their versatility as new materials for application in different fields. Water remediation is one of the traditional applications of these composites and their investigation as adsorbents for single water pollutants is well established. However, most water resources such as rivers, lakes, and even oceans contain complex mixtures of pollutants. Despite several recently published reviews on water purification technology, they only focused on these material as single pollutant removers and hardly mentioned their capacity to simultaneously recover multiple pollutants. There-fore, there is still a gap in the archived literature considering nanocellulose/polyethylenimine composites tar-geting water remediation with multiple water pollutants. In this review, methods for synthesizing such composites are classified and compared according to the mechanism of reactions, such as chemical crosslinking and physical adsorption, while outlining advantages and limitations. Then, the water pollutants mainly targeted by those composites are discussed in detail to expound the relationship between the synthesis method and the type and adsorption capacity. Finally, the last section presents challenges and opportunities of these nano-cellulose/polyethylenimine composites as emerging sorbents for sustainable multiple water pollutants purifi-cation technologies. This review aims to lay out the basis for future developments of these composites for multiple water pollutants.

Hydrogels are promising candidates for wound healing bandages because they can mimic the native skin microenvironment. Additionally, there is increasing growth in the use of naturally derived materials and plant-based biomaterials to produce healthcare products with healing purposes because of their biocompatibility and biodegradation properties. In this study, cellulose extracted from biodiverse sources in Ecuador was used as the raw material for the fabrication of hydrogels with enhanced antifouling properties. Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and scanning electron microscopy (SEM) were used to characterize the cellulose and hydrogels. In vitro and ex vivo tests were performed to evaluate the antimicrobial activity of hydrogels against Gram-negative bacteria as a model. Finally, the hydrogel synthesized with cellulose extracted from pitahaya showed improved antibacterial activity when applied over pigskin as a proof of concept for wound dressing. Therefore, the present results suggest that cellulose-based hydrogels are good candidates for application as wound dressings.

Meatpacking and poultry operations produce an enormous amount of co-products including offal, fat, blood, feathers etc. that are collected and processed by the rendering industry into value-added materials such as various protein meals and rendered fat products. Rendered fats (mainly composed of triglycerides from the adipose tissue of animals or used cooking oil from the restaurant industry) are sold for a variety of applications including animal feed formulations. Nonetheless, in the current context of energy scarcity, their use as feedstocks for the generation of renewable fuels including biodiesel and renewable diesel represents a growing market. The diverse composition of the source material can impose significant challenges in terms of compliance, requiring the control (and reduction) of the concentration of elements such as phosphorus, sulfur, calcium, magnesium, sodium, potassium, and other undesirable metals that can otherwise interfere with critical aspects of the refining process or contaminate the renewable fuel products. To address this critical need, we describe the application of poly(ethylenimine)-modified cellulose nanocrystals as a low-cost material for the removal of unwanted metal/inorganic cations from rendered fat. A total of 28 real samples including poultry, white pork grease, and beef tallow were analyzed. Test results showed that the approach can effectively decrease the concentration of the target elements by 95 ± 2%, suggesting that this treatment protocol could dramatically improve the application of rendered fat products for renewable fuel refining.

Glucose metabolism is critical for the African trypanosome, Trypanosoma brucei, serving as the lone source of ATP production for the bloodstream form (BSF) parasite in the glucose-rich environment of the host blood. Recently, phosphonate inhibitors of human enolase (ENO), the enzyme responsible for the interconversion of 2-phosphoglycerate (2-PG) to phosphoenolpyruvate (PEP) in glycolysis or PEP to 2-PG in gluconeogenesis, have been developed for the treatment of glioblastoma multiforme (GBM). Here, we have tested these agents against T. brucei ENO (TbENO) and found the compounds to be potent enzyme inhibitors and trypanocides. For example, (1-hydroxy-2-oxopyrrolidin-3-yl) phosphonic acid (deoxy-SF2312) was a potent enzyme inhibitor (IC50 value of 0.60 ± 0.23 µM), while a six-membered ring-bearing phosphonate, (1-hydroxy-2-oxopiperidin-3-yl) phosphonic acid (HEX), was less potent (IC50 value of 2.1 ± 1.1 µM). An analog with a larger seven-membered ring, (1-hydroxy-2-oxoazepan-3-yl) phosphonic acid (HEPTA), was not active. Molecular docking simulations revealed that deoxy-SF2312 and HEX had binding affinities of −6.8 and −7.5 kcal/mol, respectively, while the larger HEPTA did not bind as well, with a binding of affinity of −4.8 kcal/mol. None of these compounds were toxic to BSF parasites; however, modification of enzyme-active phosphonates through the addition of pivaloyloxymethyl (POM) groups improved activity against T. brucei, with POM-modified (1,5-dihydroxy-2-oxopyrrolidin-3-yl) phosphonic acid (POMSF) and POMHEX having EC50 values of 0.45 ± 0.10 and 0.61 ± 0.08 µM, respectively. These findings suggest that HEX is a promising lead against T. brucei and that further development of prodrug HEX analogs is warranted.