Facilitated Transport Research Articles

Edible film: A future thrust in packaging technology for horticultural crops

The significance of edible film in food packaging is increasingly recognized, especially in preserving perishable items such as fruits and vegetables. Packaging serves crucial functions, including containment, protection and facilitation of product transportation. However, conventional plastic packaging poses significant environmental challenges due to its non-biodegradable nature, contributing to pollution and heightened carbon footprints. In contrast, edible films, derived from natural ingredients like proteins, lipids and polysaccharides, offer a sustainable alternative. These films extend the shelf life of food products and improve food quality by reducing waste and minimizing environmental impact. Adopting edible films in the horticultural sector could significantly enhance post-harvest preservation, ensuring the freshness and quality of produce. As the food industry evolves, the development and application of edible film packaging present a promising avenue for future research, emphasizing sustainability and efficiency in food preservation.

Investigation of glucosuria in children.

Previous reports provided recommendations for familial renal glucosuria diagnosis without complex view on differential diagnosis of glucosuria. The aim of this review was to provide an overview of the causes of glucosuria and to create an evidence-based diagnostic approach for children with glucosuria. We searched the current literature with a focus to identify the possible etiology of glucosuria, gaining insight into the pathophysiology of glucosuria. Urinary glucose is completely reabsorbed in the proximal tubule of kidneys. It only appears in the urine if the plasma glucose concentration exceeds the renal threshold for glucose or in the case of insufficient renal glucose reabsorption. The proteins that provide glucose reabsorption are SGLT2 and SGLT1 - sodium-dependent co-transporters that transport glucose from the lumen into epithelial cells - and GLUT2 - a passive transporter providing facilitative glucose transport from epithelial cells to plasma. Renal glucose reabsorption is affected in case of acquired or inherited complex dysfunction of proximal tubule called Fanconi Syndrome or due to pathogenic variants of genes encoding glucose transporters. Prior to diagnosing any of these, diabetes mellitus must be excluded together with other conditions leading to hyperglycemia. In conclusion, glucosuria is always an abnormal finding. The review provides a simple evidence-based diagnostic approach to navigate the differential diagnosis of glucosuria.

Atmospherically Plasma-Sprayed Macro Porous Layers for Anion Exchange Membrane Water Electrolysis Operating with Alkaline Supporting Electrolyte

Green hydrogen produced via water electrolysis plays an important role in the decarbonisation of the global energy economy. Besides reducing electricity prices and increasing electrolyser efficiency, lowering electrolyser investment cost is an important avenue to make electrolysis technology more economically attractive. Anion exchange membrane water electrolysis (AEMWE) combines the advantages of proton exchange membrane water electrolysis (PEMWE), i.e., high current density while maintaining high efficiency and fast dynamic response with the advantages of alkaline water electrolysis (AWE), i.e., low-cost materials. However, an optimised porous transport layer (PTL) component for AEMWE is yet to be developed.Typically, PTLs in commercial electrolysers are metallic foams, felts, meshes, sintered sheets or graphitic fibre laminates, although these structures either perform poorly or they are extremely expensive. Furthermore, a high number of low-resistance contact points at the interface between the catalyst layer and the PTL is required to enable high catalyst utilisation. At the same time, operation at high current densities requires an especially efficient liquid/gas exchange at this interface. Both important functions, good electrical contact and mass transport facilitation, are fulfilled by a macro porous layer (MPL). Stiber et al designed and characterized a titanium/niobium MPL deposited via vacuum plasma spraying (VPS) on a stainless-steel mesh substrate, vastly improving the performance of a PEMWE [1]. Similarly, Razmjooei et al deposited a nickel layer via atmospheric plasma spraying (APS) and demonstrated the performance-increasing effect in AEMWE operating with pure water [2].In this work, within the frame of the German project AEM-Direkt, a Ni-based MPL was developed to improve high current density operation in AEMWE working with a supporting alkaline electrolyte. For this, a Ni-C composite powder was deposited on a stainless-steel multi-mesh PTL via APS (Fig. a). The carbon acts as a pore forming agent and is subsequently removed by an ex-situ oxidation step in air at 700°C followed by a reduction step in ammonia at 500°C. A highly porous, smooth and flat Ni-MPL is formed. Using the MPL on the anode, the AEMWE single cell achieves 2 V at 3 A cm-2 (Fig. b), which is comparable to the performance of PEMWE. As anode catalyst layer a novel 20 nm thickness sputtered nickel layer designed and manufactured by Siemens-Energy was used, deposited on a DURAION® AEM, manufactured by Evonik Operations GmbH. As cathode, a Pt/C based catalyst coated substrate (CCS) was employed.In continuation of this study, the designed MPL will be further optimised and characterised as cathode MPL as well. Acknowledgement This work is carried out within the project AEM-Direkt (Förderkennzeichen 03HY130) funded by the German Ministry of Science and Education (BMBF). References Stiber S, Sata N, Morawietz T, Ansar SA, Jahnke T, Lee JK, et al. A high-performance, durable and low-cost proton exchange membrane electrolyser with stainless steel components. Energy & Environmental Science. 2022;15(1):109-22.Razmjooei F, Morawietz T, Taghizadeh E, Hadjixenophontos E, Mues L, Gerle M, et al. Increasing the performance of an anion-exchange membrane electrolyzer operating in pure water with a nickel-based microporous layer. Joule. 2021;5(7):1776-99. Figure 1

Morpho-Molecular and Genomic Characterization of Penicillium mexicanum Isolates Retrieved from a Forsaken Gold Mine

During the ongoing studies designed to examine the fungal diversity present within the abandoned and flooded Escádia Grande gold mine (Góis, Portugal), we repeatedly isolated several specimens belonging to a Penicillium species. Molecular phylogenetic analysis, coupled with morphological observations, positioned this fungus within subgen. Penicillium sect. Paradoxa, series Atramentosa, pinpointing its identity as Penicillium mexicanum (the first record for mining soils and the country). Given the limited research conducted on Penicillia isolated from similar environments, the species genome was sequenced utilizing the Oxford Nanopore® MinION™ methodology and studied through bioinformatic analysis. The obtained genome has a size of 29.62 Mb, containing a 47.72% GC content, 10,156 genes, with 44 rRNAs and 178 tRNAs/tmRNAs, providing the first genomic resource for this microorganism. Bioinformatic analysis allowed us to identify multiple genomic traits that can contribute towards this species survival in these extreme environments, including the presence of high levels of major facilitator transporters (MFS), Zn (2)-C6 fungal-type DNA-binding domains, P-loop containing nucleoside triphosphate hydrolases, specific fungal transcription factors and sugar transporters. Furthermore, putative advantageous metabolic traits, such as methylotrophy, assimilatory nitrate and sulfate reduction abilities, were also detected. In addition, the results also highlighted a strong genomic and metabolic organization and investment towards arsenic detoxification (transport and oxidation). Lastly, thirty-two putative biosynthetic gene clusters were predicted, including some with high similarity values to monascorubrin, nidulanin A, histidyltryptophanyldiketopiperazine/dehydrohistidyltryptophanyldiketopiperazine/roquefortine D/roquefortine C/glandicoline A/glandicoline B/meleagrine, YWA1 and choline. Overall, this study expands the current Penicillia knowledge from mining environments while also enhancing our understanding regarding fungal arsenic resistance.

Probing into the low efficiency of CdS/SnS-based solar cells and remediation through surface treatments

Tin sulfide (SnS) is a highly promising photovoltaic absorption material in thin-film solar cells with abundant reserves, environmental friendliness, low cost and long-term stability. An efficiency of up to 4.8 % has been achieved for SnS absorber/CdS heterojunction solar cells where the CdS buffer layer is usually obtained by chemical bath deposition. However, current CdS/SnS devices often suffer from severe interface recombination loss, which deteriorates device performance. This report focuses on the CdS film surface to identify performance-degrading factors for CdS/SnS-based solar cells. XRD and XPS reveal numerous oxides on freshly fabricated CdS films increasing surface roughness and reducing conductivity. Herein, a method based on ammonium sulfide (AS) chemical treatment is proposed, which can effectively reduce the oxide content on the surface of CdS and optimize the band alignment, leading to the facilitation of charge carrier transport and the suppression of interface recombination. Consequently, the PCE of the FTO/CdS/SnS/Ag device is enhanced about three times (from 0.18 % to 0.47 %) with excellent moisture stability.

Glucose Transporter-Targeting Chimeras Enabling Tumor-Selective Degradation of Secreted and Membrane Proteins.

Tumor-selective degradation of target proteins has the potential to offer superior therapeutic benefits with maximized therapeutic windows and minimized off-target effects. However, the development of effective lysosome-targeted degradation platforms for achieving selective protein degradation in tumors remains a substantial challenge. Cancer cells depend on certain solute carrier (SLC) transporters to acquire extracellular nutrients to sustain their metabolism and growth. This current study exploits facilitative glucose transporters (GLUTs), a group of SLC transporters widely overexpressed in numerous types of cancer, to drive the endocytosis and lysosomal degradation of target proteins in tumor cells. GLUT-targeting chimeras (GTACs) were generated by conjugating multiple glucose ligands to an antibody specific for the target protein. We demonstrate that the constructed GTACs can induce the internalization and lysosomal degradation of the extracellular and membrane proteins streptavidin, tumor necrosis factor-alpha (TNF-α), and human epidermal growth factor receptor 2 (HER2). Compared with the parent antibody, the GTAC exhibited higher potency in inhibiting the growth of tumor cells in vitro and enhanced tumor-targeting capacity in a tumor-bearing mouse model. Thus, the GTAC platform represents a novel degradation strategy that harnesses an SLC transporter for tumor-selective depletion of secreted and membrane proteins of interest.

Enhanced Local Ion Transport in High-Performance Anion Exchange Membrane Water Electrolyzers Via Quaternary Ammonium-Functionalized Metal-Organic Framework Electrocatalysts

Anion-exchange membrane water electrolyzers (AEMWEs) present a multitude of advantages, particularly in the utilization of non-platinum group (non-PGM) catalysts. While diverse catalysts have demonstrated notable electrocatalytic activity in alkaline conditions, the integration of the ionomer with the electrocatalyst at the active catalyst layer is crucial for enhancing overall AEMWE performance in practical device demonstrations by improving mass transport. However, the intrinsic instability of polymeric ionomers, leading to physical detachments or alkaline (chemical) degradation, poses a challenge that can potentially diminish performance. In this study, drawing inspiration from the aforementioned ionomer chemistry, we introduce non-PGM NiFe-based metal-organic frameworks (MOFs) incorporating quaternary ammonium (QA) cation moieties covalently functionalized to benzenedicarboxylic acid (BDC) ligands as promising electrocatalysts for the oxygen evolution reaction (OER) in alkaline environments. Three specifically designed functional groups with varying ion exchange capacities (IEC) and alkaline stability—trimethylamine (TMA), N-methylpyrrolidine (C4N), and N-methylpiperidine (C5N)—were directly tethered to BDC ligands. The alkaline OER activity of Ni2Fe1-QA-MOF was meticulously assessed using rotating disk electrodes and AEMWE devices. These directly tethered QA groups markedly facilitated improved ionic transport of OH- ions at the catalyst/electrolyte interfaces compared to BDC ligands, ultimately leading to a significant enhancement in half-cell activity and AEMWE performance. In-situ IR/RAMAN spectroscopy revealed that QA moieties highly promoted local ionic transport at the interfaces, as evidenced by characteristic peaks indicating a robust interaction between QA and OH- ions. Unexpectedly, QA moieties not only contributed to ionic transport in local environments but also induced oxidation states of Ni metal sites, while Fe sites exhibited no changes during the applied potential in the OER region, as comprehensively revealed by synchrotron X-ray absorption spectroscopy (XAS). In comparison with conventional NiFe layered double hydroxide (LDH) structures, we posit that the substantial structural transformation of Ni moieties can be attributed to an increased degree of M-O bond interaction, enhanced by the local interaction between QA and OH- ions. Density functional theory (DFT) calculations also validate the balanced adsorption of OER intermediates on QA-MOFs. Finally, QA-NiFe-MOF was directly synthesized on a porous transport layer (PTL), and its AEMWE performance was evaluated without the use of any ionomer. At 1.0 M KOH, C4N, C5N, and TMA-based NiFe MOFs exhibited current densities of 9.35 A/cm2, 9.11 A/cm2, and 7.6 A/cm2, respectively. These current densities surpassed those of bare BDC-NiFe MOF and commercial IrO2 anode catalysts, which exhibited only 7.39 A/cm2 and 6.4 A/cm2, respectively. Consequently, we emphasize that the facilitation of local ionic transport could yield remarkable AEMWE performance, even in the absence of conventional polymeric ionomers.

Localization of the MTP4 transporter to trans-Golgi network in pollen tubes of Arabidopsis thaliana.

Zinc (Zn) is an essential element for plants. Numerous proteins in different cellular compartments require Zn for their structure and function. Zn can be toxic when it accumulates in high levels in the cytoplasm. Therefore, Zn homeostasis at tissue, cell, and organelle levels is vital for plant growth. A part of the metal tolerance protein (MTP) / Cation Diffusion Facilitator (CDF) transporters functions as Zn transporters, exporting Zn from the cytosol to various membrane compartments. In Arabidopsis thaliana, MTP1, MTP2, MTP3, MTP4, MTP5, and MTP12 are classified as Zn transporters (Zn-CDF). In this study, we systematically analyzed the localization of GFP-fused Zn-CDFs in the leaf epidermal cells of Nicotiana benthamiana. As previously reported, MTP1 and MTP3 were localized to tonoplast, MTP2 to endoplasmic reticulum, and MTP5 to Golgi. In addition, we identified the localization of MTP4 to trans-Golgi Network (TGN). Since MTP4 is specifically expressed in pollen, we analyzed the localization of MTP4-GFP in the Arabidopsis pollen tubes and confirmed that it is in the TGN. We also showed the Zn transport capability of MTP4 in yeast cells. We then analyzed the phenotype of an mtp4 T-DNA insertion mutant under both limited and excess Zn conditions. We found that their growth and fertility were not largely different from the wild-type. Our study has paved the way for investigating the possible roles of MTP4 in metallating proteins in the secretory pathway or in exporting excess Zn through exocytosis. In addition, our system of GFP-fused MTPs will help study the mechanisms for targeting transporters to specific membrane compartments.

Glucose transporters in lymphocytes: expression and effects on lymphocyte functions

Glucose uptake by lymphocytes is dependent on the facilitative glucose transporters (GLUT1, GLUT3, GLUT4, and GLUT6) of the GLUT family and the Na+-coupled glucose transporter SGLT1. GLUTs and SGLTs are widely expressed in mammals, and their expression and functions may affect cell development, homeostasis, activation, and differentiation. This article details the important functions of several GLUTs and SGLTs in lymphocytes and points out that glucose transporters play a key role in supplying energy for lymphocytes, maintaining intracellular glucose homeostasis, and improving the efficiency of immune responses, which reflect their key roles in signal transduction. Probing into the effects of glucose transporters on lymphocyte functions will help to decipher the functioning mechanisms of lymphocytes in diseases. Furthermore, this paper prospects the application values of glucose transporters in lymphocytes from molecular biology, aiming to provide better strategies for the clinical treatment of lymphocyte-related diseases and promote the research and development of targeted therapeutic drugs.

Neurodevelopment Is Dependent on Maternal Diet: Placenta and Brain Glucose Transporters GLUT1 and GLUT3.

Glucose is the primary energy source for most mammalian cells and its transport is affected by a family of facilitative glucose transporters (GLUTs) encoded by the SLC2 gene. GLUT1 and GLUT3, highly expressed isoforms in the blood-brain barrier and neuronal membranes, respectively, are associated with multiple neurodevelopmental disorders including epilepsy, dyslexia, ADHD, and autism spectrum disorder (ASD). Dietary therapies, such as the ketogenic diet, are widely accepted treatments for patients with the GLUT1 deficiency syndrome, while ameliorating certain symptoms associated with GLUT3 deficiency in animal models. A ketogenic diet, high-fat diet, and calorie/energy restriction during prenatal and postnatal stages can also alter the placental and brain GLUTs expression with long-term consequences on neurobehavior. This review focuses primarily on the role of diet/energy perturbations upon GLUT isoform-mediated emergence of neurodevelopmental and neurodegenerative disorders.

Genome-Scale Screening of Saccharomyces cerevisiae Deletion Mutants to Gain Molecular Insight into Tolerance to Mercury Ions.

Mercury (Hg) is a global pollutant and a bioaccumulative toxin that seriously affects the environment. Though increasing information has been obtained on the mechanisms involved in mercury toxicity, there is still a knowledge gap between the adverse effects and action mechanisms, especially at the molecular level. In the current study, we screened a diploid library of Saccharomyces cerevisiae single-gene deletion mutants to identify the nonessential genes associated with increased sensitivity to mercury ions. By genome-scale screening, we identified 64 yeast single-gene deletion mutants. These genes are involved in metabolism, transcription, antioxidant activity, cellular transport, transport facilitation, transport routes, and the cell cycle, as well as in protein synthesis, folding, modification, and protein destination. The concentration of mercury ions was different in the cells of yeast deletion mutants. Moreover, the disruption of antioxidant systems may play a key role in the mercurial toxic effects. The related functions of sensitive genes and signal pathways were further analyzed using bioinformatics-related technologies. Among 64 sensitive genes, 37 genes have human homologous analogs. Our results may provide a meaningful reference for understanding the action mode, cellular detoxification, and molecular regulation mechanisms of mercury toxicity.

A COMPREHENSIVE STUDY OF MURRAYA KOENIGII (LINN.) ON RESTORATION OF CONSCIOUSNESS BY AUGMENTING GLUCOSE UPTAKE IN THE NEURONS: A REVIEW

Fainting, also known as syncope or passing out, is caused by decreased cerebral blood flow leading to loss of consciousness and muscle strength. It is caused by lack of blood supply to the brain leading to decrease blood sugar supply to the neurons. Symptoms like dizziness, sweating, pale complexion, blurred vision, nausea, vomiting or a warm feeling precede the loss of consciousness[1]. For the brain, glucose serves as its main energy source. In order for glucose to be transported from the blood into the brain, it must pass through the endothelial cells that line the blood-brain barrier and enter the neurons and glia. These processes are mediated by facilitative Glucose transporter protein. Kaidarya is an Ayurvedic drug from the ancient samhitas, commonly known as curry leaves. Due to its grahi properties it increases the glucose uptake activity. When glucose uptake activity increases in the brain, it supplies sufficient glucose to the brain cells, which helps in management of fainting or syncope.

Galacturonic acid-capsaicin prodrug for prolonged nociceptive-selective nerve blockade

There is an urgent clinical need to develop nerve-blocking agents capable of inducing long duration sensory block without muscle weakness or paralysis to treat post-operative and chronic pain conditions. Here, we report a galacturonic acid-capsaicin (GalA-CAP) prodrug as an effective nociceptive-selective axon blocking agent. Capsaicin selectively acts on nociceptive signaling without motor nerve blockade or disruption of proprioception and touch sensation, and the galacturonic acid moiety enhance prodrug permeability across the restrictive peripheral nerve barriers (PNBs) via carrier-mediated transport by the facilitative glucose transporters (GLUTs). In addition, following prodrug transport across PNBs, the inactive prodrug is converted to active capsaicin through linker hydrolysis, leading to sustained drug release. A single injection of GalA-CAP prodrug at the sciatic nerves of rats led to nociceptive-selective nerve blockade lasting for 234 ± 37 h, which is a sufficient duration to address the most intense period of postsurgical pain. Furthermore, the prodrug markedly mitigated capsaicin-associated side effects, leading to a notable decrease in systemic toxicity, benign local tissue reactions, and diminished burning and irritant effects.

Drought response revealed by chromatin organization variation and transcriptional regulation in cotton

BackgroundCotton is a major world cash crop and an important source of natural fiber, oil, and protein. Drought stress is becoming a restrictive factor affecting cotton production. To facilitate the development of drought-tolerant cotton varieties, it is necessary to study the molecular mechanism of drought stress response by exploring key drought-resistant genes and related regulatory factors.ResultsIn this study, two cotton varieties, ZY007 (drought-sensitive) and ZY168 (drought-tolerant), showing obvious phenotypic differences under drought stress, were selected. A total of 25,898 drought-induced genes were identified, exhibiting significant enrichment in pathways related to plant stress responses. Under drought induction, At subgenome expression bias was observed at the whole-genome level, which may be due to stronger inhibition of Dt subgenome expression. A gene co-expression module that was significantly associated with drought resistance was identified. About 90% of topologically associating domain (TAD) boundaries were stable, and 6613 TAD variation events were identified between the two varieties under drought. We identified 92 genes in ZY007 and 98 in ZY168 related to chromatin 3D structural variation and induced by drought stress. These genes are closely linked to the cotton response to drought stress through canonical hormone-responsive pathways, modulation of kinase and phosphatase activities, facilitation of calcium ion transport, and other related molecular mechanisms.ConclusionsThese results lay a foundation for elucidating the molecular mechanism of the cotton drought response and provide important regulatory locus and gene resources for the future molecular breeding of drought-resistant cotton varieties.

Fetal Hypoglycemia Induced by Placental SLC2A3-RNA Interference Alters Fetal Pancreas Development and Transcriptome at Mid-Gestation.

Glucose, the primary energy substrate for fetal oxidative processes and growth, is transferred from maternal to fetal circulation down a concentration gradient by placental facilitative glucose transporters. In sheep, SLC2A1 and SLC2A3 are the primary transporters available in the placental epithelium, with SLC2A3 located on the maternal-facing apical trophoblast membrane and SLC2A1 located on the fetal-facing basolateral trophoblast membrane. We have previously reported that impaired placental SLC2A3 glucose transport resulted in smaller, hypoglycemic fetuses with reduced umbilical artery insulin and glucagon concentrations, in addition to diminished pancreas weights. These findings led us to subject RNA derived from SLC2A3-RNAi (RNA interference) and NTS-RNAi (non-targeting sequence) fetal pancreases to qPCR followed by transcriptomic analysis. We identified a total of 771 differentially expressed genes (DEGs). Upregulated pathways were associated with fat digestion and absorption, particularly fatty acid transport, lipid metabolism, and cholesterol biosynthesis, suggesting a potential switch in energetic substrates due to hypoglycemia. Pathways related to molecular transport and cell signaling in addition to pathways influencing growth and metabolism of the developing pancreas were also impacted. A few genes directly related to gluconeogenesis were also differentially expressed. Our results suggest that fetal hypoglycemia during the first half of gestation impacts fetal pancreas development and function that is not limited to β cell activity.

Tanycytes release glucose using the glucose-6-phosphatase system during hypoglycemia to control hypothalamic energy balance

ObjectiveThe liver releases glucose into the blood using the glucose-6-phosphatase (G6Pase) system, a multiprotein complex located in the endoplasmic reticulum (ER). Here, we show for the first time that the G6Pase system is also expressed in hypothalamic tanycytes, and it is required to regulate energy balance. MethodsUsing automatized qRT-PCR and immunohistochemical analyses, we evaluated the expression of the G6Pase system. Fluorescent glucose analogue (2-NBDG) uptake was evaluated by 4D live-cell microscopy. Glucose release was tested using a glucose detection kit and high-content live-cell analysis instrument, Incucyte s3. In vivo G6pt knockdown in tanycytes was performed by AAV1-shG6PT-mCherry intracerebroventricular injection. Body weight gain, adipose tissue weight, food intake, glucose metabolism, c-Fos, and neuropeptide expression were evaluated at 4 weeks post-transduction. ResultsTanycytes sequester glucose-6-phosphate (G6P) into the ER through the G6Pase system and release glucose in hypoglycaemia via facilitative glucose transporters (GLUTs). Strikingly, in vivo tanycytic G6pt knockdown has a powerful peripheral anabolic effect observed through decreased body weight, white adipose tissue (WAT) tissue mass, and strong downregulation of lipogenesis genes. Selective deletion of G6pt in tanycytes also decreases food intake, c-Fos expression in the arcuate nucleus (ARC), and Npy mRNA expression in fasted mice. ConclusionsThe tanycyte-associated G6Pase system is a central mechanism involved in controlling metabolism and energy balance.

Phosphate and illite colloid pose a synergistic risk of enhanced uranium transport in groundwater: A challenge for phosphate immobilization remediation of uranium contaminated environmental water

The phosphorus-containing reagents have been proposed to remediate the uranium contaminated sites due to the formation of insoluble uranyl phosphate mineralization products. However, the colloids, including both pseudo and intrinsic uranium colloids, could disturb the environmental fate of uranium due to its nonnegligible mobility. In this work, the transport pattern and micro-mechanism of uranium coupled to phosphate and illite colloid (IC) were investigated by combining column experiments and micro-spectroscopic evidences. Results showed that uranium transport was facilitated in granular media by forming the intrinsic uranyl phosphate colloid (such as Na-autunite) when the pH > 3.5 and CNa+ < 10 mM. Meanwhile, the mobility of uranium depended greatly on the typical water chemistry parameters governing the aggregation and deposit of intrinsic uranium colloids. However, the attachment of phosphate on illite granule increased the repulsive force and enhanced the dispersion stability of IC in the IC-U(VI)-phosphate ternary system. The non-preequilibrium transport and retention profiles, HRTEM-mapping, as well as TRLFS spectra revealed that the IC enhanced uranium mobility by forming the ternary IC-uranyl phosphate hybrid, and acted as the coagulation preventing agent for uranyl phosphate particles. This observed facilitation of uranium transport resulted from the formation of intrinsic uranyl phosphate colloids and IC-uranyl phosphate hybrids should be taken into consideration when evaluating the potential risk of uranium migration and optimizing the in-situ mineralization remediation strategy for uranium contaminated environmental water.

Abstract 5883: Role of major facilitative folate transporters SLC19A1 and SLC46A1 in cyclic dinucleotide transport and STING signaling

Abstract Activation of stimulator of interferon genes (STING) activates the transcription factor pIRF3, promoting gene transcription encoding type I interferons including IFNβ that stimulate broader immune responses. Targeting the STING pathway for cancer is especially compelling given its importance to the malignant phenotype. Cyclic dinucleotides (CDNs) function as STING agonists. Inspired by preclinical studies, two phase I clinical trials with CDNs including ADU-S100 were initiated but concluded early due to lack of anti-tumor efficacy. The reduced folate carrier (RFC; SLC19A1) has been identified as one of the primary means to transport CDNs through the cell membrane, although a study suggested that CDN and folate binding to RFC was as least partly distinct. While depletion of the proton-coupled folate transporter (PCFT; SLC46A1) in THP-1 cells had nominal effects on CDN stimulation, overexpressing PCFT increased STING signaling. Limited clinical responses may reflect the impact of physiologic variables governing RFC and PCFT not previously considered. Understanding the mechanisms of CDN uptake by RFC and PCFT under physiologic conditions may elucidate the role of CDNs on STING signaling. To study the impact of RFC levels on CDN transport and STING signaling, we used an engineered R1-11/Tet-on-RFC HeLa cell model induced with doxycycline. In R1-11/Tet-on-RFC cells, ADU-S100 showed a low affinity for binding to RFC, as measured by direct competition for transport with [3H]methotrexate. RFC dose-dependence of ADU-S100 uptake and STING activation was confirmed by monitoring IFN-β (qPCR) and pIRF3 (Westerns). Maximum induction of pIRF3/IFNβ was seen in THP-1 and R1-11/Tet-on-RFC cells by ADU-S100 treatment at pH 7.2 (RFC pH optimum) in the presence of 25 nM leucovorin. Induction of pIRF3/IFNβ by ADU-S100 in THP-1 and R1-11/Tet-on-RFC cells were partially abolished by treatment with 200 μM of leucovorin or 10 μM of PT523 (RFC-specific inhibitor), suggesting the transport of folates and CDNs may share common albeit distinct mechanisms. ADU-S100 treatment showed no pIRF3 or IFN-β production in an engineered R1-11/Tet-on-PCFT HeLa model expressing high levels of PCFT, suggesting no direct transport by PCFT, even under optimal pH conditions. Interestingly, pIRF3/IFN-β induction was enhanced in R1-11/Tet-on-RFC/PCFT cells expressing constitutive PCFT in addition to RFC, compared to R1-11/Tet-on-RFC cells that express comparable RFC without PCFT. This may reflect previously unrecognized functional interactions between RFC and PCFT. In conclusion, RFC mediates ADU-S100 uptake and STING activation under physiologic conditions and this process is enhanced by concomitant expression of PCFT. Additional characterization of key molecular and biochemical determinants of CDN-based cancer therapeutics may lead to improved approaches for CDN therapy based on enhanced cellular uptake. Citation Format: Holly McQuithey, Madelyn Brzezinski, Carrie O’Connor, Larry Matherly, Zhanjun Hou. Role of major facilitative folate transporters SLC19A1 and SLC46A1 in cyclic dinucleotide transport and STING signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 5883.

Abstract 4730: Pharmacodynamic determinants of anti tumor activity of SHMT2 inhibitors

Abstract These studies investigate factors that impact the antitumor activity of SHMT2 targeting including drug accumulation, metabolism, expression of one carbon (C1) related enzymes, and tumor hypoxia. Mitochondrial C1 metabolism is upregulated in a variety of cancer types including leukemias and solid tumors. The mitochondrial C1 enzymes serine hydroxymethyltransferase (SHMT) 2 and 5,10-methylene tetrahydrofolate (THF) dehydrogenase 2 (MTHFD2) are among the top 5 overexpressed metabolic enzymes in tumors vs normal tissues in a wide range of tumor types. Thus, SHMT2 and MTHFD2 are potential therapeutic targets for cancer, sparing normal tissues that are less reliant on mitochondrial C1 metabolism. SHMT2 metabolizes serine and THF to produce glycine and 5,10-methylene THF; MTHFD2 metabolizes 5,10-methylene THF to NADH and 10-formyl THF which is converted to formate by MTHFD1L. Thus, mitochondrial C1 metabolism is the principal source of C1 units for cellular biosynthesis and provides the majority of glycine for protein, purine and glutathione synthesis. We previously discovered novel 5-substituted pyrrolo[3,2-d]pyrimidine antifolate compounds which exhibit primary inhibition at mitochondrial SHMT2 along with cytosolic SHMT1 and de novo purine biosynthesis at glycinamide ribonucleotide and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferases. The lead compound AGF347 is taken up into cells by the facilitative transporters reduced folate carrier (SLC19A1) and the proton coupled folate transporter (SLC46A1) and like folate cofactors is metabolized to polyglutamate (PG) forms by folylpolyglutamate synthase (FPGS) which promotes retention in cytosolic and mitochondrial compartments and target enzyme engagement. We investigated the impact of folate transporter expression on metabolism to PGs for SHMT2 targeted antifolates. Folate transporter expression inversely correlated with anti-tumor efficacy of SHMT2 targeted antifolates and overexpression of FPGS resulted in dramatically increased sensitivities. However, for pyrazolopyran inhibitors of SHMT2 (e.g. SHIN1) which enter cells by diffusion and are not metabolized to polyglutamates, increased transporter and FPGS levels dramatically decreased anti-tumor activity. As SHMT2 is a target of HIF1α, we studied the impact of multitargeting SHMT2 and purine biosynthesis by pyrrolo[3,2-d]pyrimidine antifolates under hypoxic conditions. Our results establish that multi-targeting SHMT2 and purine biosynthesis preserves anti-tumor activity under hypoxia. Collectively, our results demonstrate that SHMT2 targeted pyrrolo[3,2-d]pyrimidine antifolates exhibit increased antitumor efficacies in cells expressing low folate transporter expression and is further enhanced by increased FPGS activity. Additionally, we find multitargeting of SHMT2 and de novo purine biosynthesis manifests as antitumor activity even under hypoxic conditions. Citation Format: Mathew Joseph Schneider, Carrie O'Connor, Xun Bao, Md. Junayed Nayeen, Zhanjun Hou, Jing Li, Aleem Gangjee, Larry Matherly. Pharmacodynamic determinants of anti tumor activity of SHMT2 inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4730.

The Impact of SLC2A8 RNA Interference on Glucose Uptake and the Transcriptome of Human Trophoblast Cells.

While glucose is the primary fuel for fetal growth, the placenta utilizes the majority of glucose taken up from the maternal circulation. Of the facilitative glucose transporters in the placenta, SLC2A8 (GLUT8) is thought to primarily function as an intracellular glucose transporter; however, its function in trophoblast cells has not been determined. To gain insight into the function of SLC2A8 in the placenta, lentiviral-mediated RNA interference (RNAi) was performed in the human first-trimester trophoblast cell line ACH-3P. Non-targeting sequence controls (NTS RNAi; n = 4) and SLC2A8 RNAi (n = 4) infected ACH-3P cells were compared. A 79% reduction in SLC2A8 mRNA concentration was associated with an 11% reduction (p ≤ 0.05) in ACH-3P glucose uptake. NTS RNAi and SLC2A8 RNAi ACH-3P mRNA were subjected to RNAseq, identifying 1525 transcripts that were differentially expressed (|log2FC| > 1 and adjusted p-value < 0.05), with 273 transcripts derived from protein-coding genes, and the change in 10 of these mRNAs was validated by real-time qPCR. Additionally, there were 147 differentially expressed long non-coding RNAs. Functional analyses revealed differentially expressed genes involved in various metabolic pathways associated with cellular respiration, oxidative phosphorylation, and ATP synthesis. Collectively, these data indicate that SLC2A8 deficiency may impact placental uptake of glucose, but that its likely primary function in trophoblast cells is to support cellular respiration. Since the placenta oxidizes the majority of the glucose it takes up to support its own metabolic needs, impairment of SLC2A8 function could set the stage for functional placental insufficiency.