Sphingolipid Analysis in Clinical Research.

Sphingolipids are the most diverse class of lipids due to the numerous variations in their structural components. This diversity is also reflected in their extremely different functions. Sphingolipids are not only constituents of cell membranes but have emerged as key signaling molecules involved in a variety of cellular functions, such as cell growth and differentiation, proliferation and apoptotic cell death. Lipidomic analyses in clinical research have identified pathways and products of sphingolipid metabolism that are altered in several human pathologies. In this article, we describe how to properly design a lipidomic experiment in clinical research, how to handle plasma and serum samples for this purpose, and how to measure sphingolipids using liquid chromatography-mass spectrometry.

Quantifying Gq Signaling Using the IP1 Homogenous Time-Resolved Fluorescence (HTRF) Assay.

G protein-coupled receptors (GPCRs) play pivotal roles in cellular signaling and can regulate several cellular functions such as proliferation, secretion, protein expression, and cellular metabolism. Coupling of GPCRs to members of the Gq/11 protein family results in activation of inositol trisphosphate (IP3) and accumulation of calcium intracellularly. This protocol chapter outlines a step-by-step guide for utilizing the inositol phosphate-1 (IP1) accumulation assay, a time-resolved fluorescence resonance energy transfer (TR-FRET) method, to investigate Gq-IP3 signaling. The assay serves as a valuable tool for those conducting pharmacological investigations and compound screening targeting this critical cellular pathway. This protocol chapter covers experimental setup, sample preparation, and data analysis, providing researchers with an in-depth guide to explore the pharmacology of Gq-coupled receptors.

LiFe0.3Mn0.7PO4-on-MXene heterostructures as highly reversible cathode materials for Lithium-ion batteries.

Two-dimensional (2D) heterostructure materials, incorporating the collective strengths and synergetic properties of individual building blocks, have attracted great interest as a novel paradigm in electrode materials science. The family of 2D transition metal carbides and nitrides (e.g., MXenes) has become an appealing platform for fabricating functional materials with strong application performance. Herein, a 2D LiFe0.3Mn0.7PO4 (LFMP)-on-MXene heterostructure composite is prepared through an electrostatic self-assembly procedure. The functional groups on the surface of MXenes possess highly electronegative properties that facilitate the incorporation of LFMPs into MXenes to construct heterostructure composites. The special heterostructure of nanosized-LiFe0.3Mn0.7PO4 and MXene provides rapid Li+ and electron transport in the cathodes. This LiFe0.3Mn0.7PO4-3.0 wt% MXene composite can exhibit an excellent rate capability of 98.3 mAh g-1 at 50C and a very stable cycling performance with a capacity retention of 94.3 % at 5C after 1000 cycles. Furthermore, NaFe0.3Mn0.7PO4-3.0 wt% MXene with stable cyclability can be obtained by an electrochemical conversion method with LiFe0.3Mn0.7PO4-3.0 wt% MXene. Ex-situ XRD suggests that LiFe0.3Mn0.7PO4-on-MXene achieves a highly reversible structural evolution with a solid solution phase transformation (LFMP→LixFe0.3Mn0.7PO4 (LxFMP), LxFMP→LFMP) and a two-phase reaction (LxFMP←→Fe0.3Mn0.7PO4 (FMP)). This work provides a new direction for the use of MXenes to fabricate 2D heterostructures for lithium-ion batteries.

Porous nickel-cobalt phosphate with oxygen-rich vacancies in situ grown on dopamine-modified cellulose textiles as self-supporting high mass loadings supercapacitor electrode.

Transition-metal phosphates/phosphides showcase significant promise for energy-related applications because of their high theoretical electrochemical characteristics. However, sluggish electro/ion transfer rates and kinetically unfavorable reaction sites hinder their application at high mass loading. Herein, a self-supporting electrode based on transition-metal phosphates was successfully fabricated via a one-step electrodeposition process. The nanosheet structure of transition-metal phosphates, formed by interconnecting nanoparticles, effectively mitigates the impact of stress and achieves a high mass-loading (21 mg cm-2) of the electrode. Additionally, the oxygen vacancy-rich and porous nanostructure of transition-metal phosphates endows the as-prepared electrodes with a significantly increased conductivity and fast ion migration rate for enhancing electrochemical kinetics. Consequently, the as-fabricated transition-metal phosphate electrode displays the highest areal specific capacity of 39.2F cm-2. Furthermore, the asymmetric supercapacitor achieves a maximum energy density of 0.79 mWh cm-2 and a high capacity retention of 93.0 % for 10000 cycles under 60 mA cm-2. This work provides an ideal strategy for fabricating flexible electrodes with high mass loading and synthesizing transition-metal phosphate electrodes rich in oxygen vacancies.

Oxalate modification enabled advanced phosphate removal of nZVI: In Situ formed surface ternary complex and altered multi-stage adsorption process.

Nano zero-valent iron (nZVI) is a promising phosphate adsorbent for advanced phosphate removal. However, the rapid passivation of nZVI and the low activity of adsorption sites seriously limit its phosphate removal performance, accounting for its inapplicability to meet the emission criteria of 0.1 mg P/L phosphate. In this study, we report that the oxalate modification can inhibit the passivation of nZVI and alter the multi-stage phosphate adsorption mechanism by changing the adsorption sites. As expected, the stronger anti-passivation ability of oxalate modified nZVI (OX-nZVI) strongly favored its phosphate adsorption. Interestingly, the oxalate modification endowed the surface Fe(III) sites with the lowest chemisorption energy and the fastest phosphate adsorption ability than the other adsorption sites, by in situ forming a Fe(III)-phosphate-oxalate ternary complex, therefore enabling an advanced phosphate removal process. At an initial phosphate concentration of 1.00 mg P/L, pH of 6.0 and a dosage of 0.3 g/L of adsorbents, OX-nZVI exhibited faster phosphate removal rate (0.11 g/mg/min) and lower residual phosphate level (0.02 mg P/L) than nZVI (0.055 g/mg/min and 0.19 mg P/L). This study sheds light on the importance of site manipulation in the development of high-performance adsorbents, and offers a facile surface modification strategy to prepare superior iron-based materials for advanced phosphate removal.

Electrochemical removal of nitrate in high-salt wastewater with low-cost iron electrode modified by phosphate.

Nitrate (NO3-) is a widespread pollutant in high-salt wastewater and causes serious harm to human health. Although electrochemical removal of nitrate has been demonstrated to be a promising treatment method, the development of low-cost electro-catalysts is still challenging. In this work, a phosphate modified iron (P-Fe) cathode was prepared for electrochemical removal of nitrate in high-salt wastewater. The phosphate modification greatly improved the activity of iron, and the removal rate of nitrate on P-Fe was three times higher than that on Fe electrode. Further experiments and density functional theory (DFT) calculations demonstrated that the modification of phosphoric acid improved the stability and the activity of the zero-valent iron electrode effectively for NO3- removal. The nitrate was firstly electrochemically reduced to ammonium, and then reacted with the anodic generated hypochlorite to N2. In this study, a strategy was developed to improve the activity and stability of metal electrode for NO3- removal, which opened up a new field for the efficient reduction of NO3- removal by metal electrode materials.

Circular economy strategies in sedimentary phosphate mine reclamation: Development of technosol from phosphate waste rock.

Proper management of mine waste plays a crucial role in minimizing environmental impacts. One potential solution to tackle this problem involves transforming mine waste rock into soil to facilitate the process of mine restoration. The aim of this study was to assess the mineralogical, chemical, and physical characteristics of technosol derived from phosphate mine waste dumps. Following this evaluation, a novel rehabilitation strategy was proposed. For this purpose, a total of 32 samples were systematically collected across a 4 ha area of technosols, which had been established in accordance with the waste rock soil rehabilitation strategy involving geomorphic reshaping. According to the findings, phosphate mining left the soil with a sandy texture, resulting in a degraded soil structure with severely unfavorable crop growth conditions, notably poor stability, and low water retention. The chemistry of the studied soils was characterized by the dominance of CaO (29.02 wt%± 1.01) > SiO2 (27.61 wt% ± 0.61) > P2O5 (11.34 wt% ± 0.23) > MgO (5.97 wt%±0.16). Mineralogically, the samples were mainly formed by quartz, dolomite, calcite, apatite, and clay minerals. The prevalence of dolomite played a significant role in enhancing the accessibility of Mg as an essential nutrient and the occurrence of apatite in the soil resulted in the presence of P2O5. However, the abundance of Ca was linked to three major minerals: calcite, apatite, and dolomite. X-ray fluorescence analyses demonstrated that the concentrations of Fe2O3, K2O, and SO3 did not exceed 2 wt%.Organic matter, represented by SOC <0.2% and N < 0.02%, demonstrated an extraordinary deficiency in the study area. The analysis of element bioavailability confirmed that the soil was rich in Ca (10383,26 mg/kg), Mg (278,47 mg/kg), Zn (12,82 mg/kg), and Cu (3,7 mg/kg) but deficient in other essential nutrients such as P, K, S, Mn, and Fe. Our research results provide a set of recommendations aimed at enhancing existing mine rehabilitation practices applicable to both pre- and post-rehabilitation phases, leveraging automated mineralogy and circular economy principles. Notably, we propose a rehabilitation strategy to be implemented prior to the geomorphic reshaping phase, which is intended to reduce costs and efforts associated with soil reconstitution.

Preclinical/clinical trials of thrice-weekly administration of a combination of tegafur/gimeracil/oteracil (TS-1) and toceranib phosphate in dogs with intranasal tumors.

Intranasal tumors in dogs are malignant solid tumors that are primarily treated with radiotherapy and often recur post-treatment. Combination therapy is pivotal in cancer therapy. Effective drugs include fluoropyrimidine 5-fluorouracil (5-FU) and toceranib phosphate. TS-1, an oral formulation containing the 5-FU prodrug tegafur and enzyme modulators gimeracil and oteracil, is proven to be safe in dogs with solid tumors. While the oral drug toceranib phosphate (Palladia®) is safely administered, the combined toxicity with TS-1 is unknown. We aimed to determine the dosage of this combination in dogs. In the preclinical/clinical trials conducted here, we used a standard 3+3 cohort design with fixed doses of toceranib phosphate (2.4 mg/kg) administered thrice weekly. TS-1 administration was initiated at a dose of 0.5 mg/kg (upper limit 2.0 mg/kg) thrice weekly. Four cohorts were included to confirm the safety of TS-1 and toceranib phosphate. Each cohort was followed up for 1 month. The intranasal tumor types included in the clinical trial (n=13) were adenocarcinoma (n=7), squamous cell carcinoma (n=1), non-epithelial malignancy (n=2), undifferentiated carcinoma (n=1), and transitional carcinoma (n=2). The TS-1 dosage could be increased up to its dose limit in the preclinical/clinical trials. The TS-1 dose to combine with toceranib phosphate thrice weekly was 2.0 mg/kg. This regimen was well-tolerated in dogs. Thus, combined TS-1 and toceranib phosphate therapy is safe for dogs with intranasal tumors.

Phosphate promotes Arabidopsis root skewing and circumnutation through reorganisation of the microtubule cytoskeleton.

Phosphate (Pi) plays a key role in plant growth and development. Hence, plants display a range of adaptations to acquire it, including changes in root system architecture (RSA). Whether Pi triggers directional root growth is unknown. We investigated whether Arabidopsis roots sense Pi and grow towards it, that is whether they exhibit phosphotropism. While roots did exhibit a clear Pi-specific directional growth response, it was, however, always to the left, independent of the direction of the Pi gradient. We discovered that increasing concentrations of KH2PO4, trigger a dose-dependent skewing response, in both primary and lateral roots. This phenomenon is Pi-specific - other nutrients do not trigger this - and involves the reorganisation of the microtubule cytoskeleton in epidermal cells of the root elongation zone. Higher Pi levels promote left-handed cell file rotation that results in right-handed, clockwise, root growth and leftward skewing as a result of the helical movement of roots (circumnutation). Our results shed new light on the role of Pi in root growth, and may provide novel insights for crop breeding to optimise RSA and P-use efficiency.

Repairing Dehiscence Defects at Implant Sites using ß-Tricalcium Phosphate/Calcium Sulfate versus Xenograft Combined With Membrane: A Randomized Clinical Trial.

Guided bone regeneration (GBR) typically involves bone grafts and a membrane to enhance bone formation. Beta-tricalcium phosphate calcium sulfate (ß-TCP/CS) is a novel material with self-hardening and tissue growth inhibition properties and can potentially replace the need for a membrane. This study compares ß-TricalciumPhosphate/CalciumSulfate with deproteinized bovine bone mineral and a collagen membrane (DBBM/CM) to repair bone defects at implant sites over six months. Sixteen implant defects were divided into ß-TCP/CS (n = 8) and DBBM/CM (n = 8). The results showed no significant differences in vertical and horizontal defect fill in millimeters between ß-TCP/CS (2.87±1.25 and 2.37±1.06 mm., respectively) and DBBM/CM (3.5±0.92 and 2.87±1.12 mm. , respectively). Buccal bone thickness (BT) alterations at the implant platform levels (BT0) were similar for both materials. However, ß-TCP/CS exhibited greater bone alteration at the 2-mm level (BT2: -1.85 mm vs. -0.47 mm) and 4-mm level (BT4: -1.79 mm vs. 0.12 mm) apical to the implant platform compared to DBBM/CM. When assessing volume alteration, ß-TCP/CS showed a significantly greater reduction at the platform to the 2 mm level (-61.98% vs. -23.76%) than DBBM/CM. In conclusion, ß-TCP/CS demonstrated promise for treating buccal bone defects around implants but exhibited higher graft reduction. This suggests that while ß-TricalciumPhosphate/CalciumSulfate may offer clinical benefits, its potential for greater graft reduction should be considered. Further research and evaluation are warranted to fully understand the long-term implications of using ß-TCP/CS in guided bone regeneration procedures.

Mitigating harmful cyanobacterial blooms in drinking water reservoirs through in-situ sediment resuspension.

Mitigating harmful cyanobacterial blooms is a global challenge, particularly crucial for safeguarding source water. Given the limitations of current technologies for application in drinking water reservoirs, we propose an innovative strategy based on in-situ sediment resuspension (SR). This method's effectiveness in cyanobacterial control and its potential impacts on water quality were assessed through laboratory culture experiments and further validated via field applications in five drinking water reservoirs. The results revealed that SR could significantly mitigate cyanobacterial growth, evidenced by the treated sets (removal rate: 3.82×106 cells L-1d-1) compared to the control set (growth rate: 2.22×107 cells L-1d-1) according to the laboratory experiments. The underlying mechanisms identified included underwater light reduction (2.38× increase in extinction coefficient) and flocculation and entrainment of cells by resuspended particles (30 % reduction per operation). Additional contributions were noted in the reduction of bioavailable phosphate and remediation of anaerobic sediment characterized by increased redox potential. This facilitated the oxidation of iron, which in turn promoted the co-precipitation of phosphate (removal rate: 46 µg L-1d-1) and inhibited its release from the sediment. The SR operation, devoid of importing extra substances, represents a safe and economical technology for controlling harmful cyanobacteria in drinking water reservoirs.

The concentration of free glycerol in goat milk increases during feed restrictions.

This Research Communication introduces a novel enzymatic-fluorometric analytical procedure for glycerol and glycerol 3-phosphate in milk. Milk from thirty-seven goats was analysed during 9 consecutive days during which a two-day feed restriction was introduced. Fractional milk triacylglyceride and free glycerol increased significantly while glycerol 3-phosphate reacted more moderately. The energy status of the mammary cell is discussed.

A Room-Temperature Lithium-Restocking Strategy for the Direct Reuse of Degraded LiFePO4 Electrodes.

The sustainable development of lithium iron phosphate (LFP) batteries calls for efficient recycling technologies for spent LFP (SLFP). Even for the advanced direct material regeneration (DMR) method, multiple steps including separation, regeneration, and electrode refabrication processes are still needed. To circumvent these intricacies, new regeneration methods that allow direct electrode reuse (DER) by rejuvenating SLFP electrodes without damaging its structure are desired. Here, a 0.1 M lithium triethyl borohydride/tetrahydrofuran solution, which has the proper reductive capability to reduce Fe3+ in SLFP to Fe2+ without alloying with the aluminum current collector, is selected as the lithiation/regeneration reagent to restock the Li loss and regenerate SLFP electrodes. By soaking the SLFP electrodes in the lithiation solution, we successfully rejuvenated the crystal structure and electrochemical activity of SLFP electrodes with structural integrity within only 6 minutes at room temperature. When being directly reused, the regenerated LFP electrodes deliver a high specific capacity of 162.6 mAh g-1 even after being exposed to air for 3 months. The DER strategy presents significant economic and environmental benefits compared with the DMR method. This research provides a timely and innovative solution for recycling spent blade batteries using large-sized LFP electrodes, boosting the closed-loop development of LFP batteries.

Functional analysis of ZmPHR1 and ZmPHR2 under low-phosphate stress in maize.

The PHOSPHATE STARVATION RESPONSE REGULATOR (PHR) plays a crucial regulatory role in plants during the process of responding to phosphate starvation. In this study, we combined reverse genetics and biotechnology to investigate the function of ZmPHR1 and ZmPHR2, including proteins containing the Myb_DNA_banding and Myb_CC-LHEQLE structural domains, in maize seedlings. Phylogenetic analysis revealed that ZmPHR1 and ZmPHR2 have high homology with AtPHR1 and OsPHR2, and share the characteristic features of nuclear localisation and transcriptional self-activation. Real-time quantitative PCR analysis showed that low phosphate (Pi) stress significantly induced the expression of ZmPHR1 and ZmPHR2 in maize seedling stage, and candidate gene association analysis further revealed the close association of these two genes with root traits under Pi stress conditions. Transgenic plants overexpressing ZmPHR1 and ZmPHR2 in Arabidopsis show a significant increase in lateral root number, fresh weight and total phosphorus accumulation under low-Pi stress. Besides, CHIP-PCR experiments identified target genes involved in hormone regulation, metal ion transport and homeostasis, phosphatase encoding, and photosynthesis, providing new insights into the biological functions of ZmPHR1 and ZmPHR2. Furthermore, our study showed that ZmPHR1 interacts with six SPX domain-only proteins (ZmSPXs) in maize, while ZmPHR2 interacts with five of these proteins. ZmPHR1 and ZmPHR2 expression was repressed in low Pi conditions, but was up-regulated in ZmSPX1 knockout material, according to our study of transgenic seedlings overexpressing ZmSPX1 in maize. We identified downstream target genes involved in the phosphorus signaling pathway, which are mainly involved in plant-pathogen interactions, ascorbic acid and arabinose metabolism, and ABC transporter proteins, by RNA-seq analysis of transgenic seedlings grown under low Pi stress for 7 days. Collectively, these results provide important clues to elucidate the role and functional significance of ZmPHR1 and ZmPHR2 under low Pi stress and also provide insights into understand the molecular mechanism of phosphorus homeostasis in maize. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-024-01508-2.

Synthesis of high-efficient low-cost fertilizer carriers based on biodegradable lignin hydrogels.

Conventional fertilizers face environmental and economic challenges due to their high solubility, leading to significant losses via runoff and leachate. This study presents a biodegradable hydrogel, synthesized from lignin and polyvinyl alcohol (PVA), designed as an eco-friendly carrier for struvite (fertilizer) with controlled phosphate release. The hydrogel was analysed through scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), Thermogravimetric analysis (TGA) and Differential scanning calorimetry (DSC). Furthermore, the prepared hydrogels demonstrated high water absorption capacities (963.4 %, 706.4 %, and 410 % for LH4, LH8, and LH12, respectively) and exhibited Fickian diffusion behaviour. Phosphate release studies showed a gradual release over 6-8 h with concentrations of 20.5 ppm, 19.45 ppm, and 17.85 ppm for St-LH4, St-LH8, and St-LH12. These lignin-based hydrogels offer a promising, cost-effective solution for slow-release fertilizers with high efficiency.

Phage-Loaded Biomimetic Apatite Powder With Antibiofilm Activity to Treat Bone-Associated Infections.

For decades, calcium phosphate (CaP)-based ceramics have been used for coating of bone and joint substitutes after arthroplasty due to their biocompatible properties. Infections following orthopedic replacement occur in 1%-5% of cases, causing serious complications. Biofilm formation either on the biomaterial's surface or on patient's tissues greatly enhances the resistance against antibiotic treatments and can induce a chronic infection, emphasizing the need for novel antimicrobial delivery systems. In this study, we established a protocol enabling bacteriophage loading during the synthesis of a CaP-based powder. The resulting biomaterial proved to be noncytotoxic against human osteoblastic cells and able to significantly inhibit 24-h matured S. aureus biofilm cultures or even completely eradicate it after 5 days of contact. Additional S. aureus biofilm assays with a freeze-dried material using two different excipients showed that sucrose had a protective role against Remus bacteriophage treatment of S. aureus biofilms, whereas lactose-freeze-dried powder maintained the antibiofilm activity.

Obesity Promotes the Ceramide-Mediated NADPH Oxidase in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is a type of blood cancer of the myeloid cell lineage. Obesity is characterized by an increase in body weight that results in excessive fat accumulation. Obesity has been associated with an increased incidence of many cancers, including blood cancers. This study evaluated the role obesity in AML progression in a novel transgenic mouse model developed by crossing Flt3ITD mice with Lepob/ob mice. Leukemia burden was augmented in obese AML mice. In addition, it was determined that obesity upregulated the ceramide-mediated and ceramide-1-phosphate-mediated NADPH oxidase 2 (NOX2). Notably, increased oxidative pathways has been attributed to disease progression in AML. Taken together, this study demonstrates a direct link between obesity and the progression of AML in part by augmenting the ceramide mediated NOX2.

Crystal structure of l-threonine-O-3-phosphate decarboxylase CobC from Sinorhizobium meliloti involved in vitamin B12 biosynthesis.

Vitamin B12 is involved in many important biochemical reactions for humans, and its deficiency can lead to serious diseases. The industrial production of vitamin B12 is achieved through microbial fermentation. In this work, we determine the crystal structures of the l-threonine-O-3-phosphate (Thr-P) decarboxylase CobC from Sinorhizobium meliloti (SmCobC), an industrial vitamin B12-producing bacterium, in apo form and in complex with a reaction intermediate. Our structures supported the Thr-P decarboxylase activity of SmCobC and revealed that the positively charged substrate-binding pocket between the large and small domains determines its substrate selectivity for Thr-P. Moreover, our results provided evidence for the proposition that the AP-P linker is formed by direct incorporation of AP-P in the biosynthetic pathway of vitamin B12 in S.meliloti.

Medical dissolution of presumptive upper urinary tract struvite uroliths in 6 dogs (2012-2018).

BACKGROUND: Minimally invasive approaches are the standard for treatment of upper urinary tract uroliths in humans. OBJECTIVE: To describe the medical dissolution of upper urinary tract uroliths in a series of dogs and report clinical outcomes. ANIMALS: 6 female dogs (9 kidneys). METHODS: Retrospective case series. A review of medical records in dogs that underwent medical dissolution of upper urinary tract uroliths utilizing diet, administration of antibiotics, and double-pigtail ureteral stent(s) placement, when indicated, was performed. Medical management was generally continued for 4 weeks beyond urolith dissolution. Information on biochemical, microbiological, imaging, and clinical outcomes before and after dissolution were recorded. RESULTS: Six dogs (9 kidneys) were included with bilateral (3) or unilateral (3) nephrolithiasis, ureterolithiasis, or a combination. A ureteral stent(s) was placed endoscopically in 5/6 dogs (6/9 kidneys) for obstructive ureterolithiasis (n = 5) or a nonobstructive massive nephrolith (n = 1). All dogs had a positive urine culture of Staphylococcus pseudintermedius with a median urine pH of 7.25 (range, 6.5-8) and 4/5 had pyonephrosis. All dogs had initial evidence of urolith dissolution at a median of 1.1 months (range, 0.42-5.9), with complete dissolution of ureteroliths at a median of 3.9 months (range, 1.5-7.6), nephroliths at 5.3 months (range, 1.5-7.6), and lower urinary tract uroliths at 0.87 months (range, 0.42-5.9). Stents were removed in 3/6 once dissolution was documented. The median follow-up time was 519 days (range, 177-2492 days). CONCLUSION AND CLINICAL IMPORTANCE: Medical dissolution and decompression of upper urinary tract struvite uroliths should be considered a minimally invasive treatment for dogs before more invasive options.

Improving Oral Absorption of a Rapidly Crystallizing Parent Drug Using Prodrug Strategy: Comparison of Phosphate Versus Glycine Based Prodrugs.

With an increasing number of Biopharmaceutical Classification System (BCS) II/IV pipeline compounds, solubilizing and supersaturating formulation strategies are becoming prevalent. Beyond formulation and solid form strategies, prodrugs are also employed to overcome solubility-limited absorption of poorly water-soluble compounds. Prodrugs can potentially yield supersaturated systems upon conversion to the parent drug intraluminally and thus enhance absorption. However, supersaturation also increases the driving force for crystallization, resulting in low solution concentrations, which can potentially negate the advantage of prodrugs. In this work, two unique solubility-enhancing prodrugs, phosphate and glycine esters, were investigated for a rapidly crystallizing parent drug. Ex vivo absorption studies using rat tissue and in vivo studies in dogs were performed. Conversion rate of the phosphate prodrug to the parent was dependent on the milieu and increased ∼24-fold in the presence of intestinal contents as medium and tissue relative to neat buffer. In contrast, conversion of the glycine prodrug was minimal under any conditions tested, suggesting that the conversion occurs after absorption into the enterocytes. Phosphate prodrug showed a non-linear increase in parent drug absorptive flux across rat intestinal tissue with concentration when intestinal contents were used as donor media. This was attributed to rapid conversion and high supersaturation of the parent drug which subsequently resulted in crystallization at high doses in the donor chamber. Glycine prodrug did not undergo complete conversion at high doses and was absorbed unchanged on the basolateral side, indicating saturation of the converting enzymes in the enterocytes. The combined flux (parent drug and glycine) showed a linear increase with dose and crystallization was not observed. Under physiological conditions, glycine prodrug that is absorbed unchanged from the intestine can potentially undergo complete conversion in hepatocytes after absorption and make the parent drug systemically available. Thus, glycine prodrug provided overall higher absorption compared to phosphate prodrug. The observed flux levels for both the prodrugs were higher compared to the parent drug alone, highlighting an advantage to use of a prodrug strategy to improve absorption of such compounds. Oral dosing in a dog PK study revealed that the bioavailability using the phosphate prodrug was ∼50% whereas, it was ∼100% with glycine prodrug, supporting the in vitro observations.