Physiological and Molecular Investigation of Urea Uptake Dynamics in Cucumis sativus L. Plants Fertilized With Urea-Doped Amorphous Calcium Phosphate Nanoparticles

At present, the quest for innovative and sustainable fertilization approaches aiming to improve agricultural productivity represents one of the major challenges for research. In this context, nanoparticle-based fertilizers can indeed offer an interesting alternative with respect to traditional bulk fertilizers. Several pieces of evidence have already addressed the effectiveness of amorphous calcium phosphate-based nanoparticles as carriers for macronutrients, such as nitrogen (N), demonstrating increase in crop productivity and improvement in quality. Nevertheless, despite N being a fundamental nutrient for crop growth and productivity, very little research has been carried out to understand the physiological and molecular mechanisms underpinning N-based fertilizers supplied to plants via nanocarriers. For these reasons, this study aimed to investigate the responses of Cucumis sativus L. to amorphous calcium phosphate nanoparticles doped with urea (U-ACP). Urea uptake dynamics at root level have been investigated by monitoring both the urea acquisition rates and the modulation of urea transporter CsDUR3, whereas growth parameters, the accumulation of N in both root and shoots, and the general ionomic profile of both tissues have been determined to assess the potentiality of U-ACP as innovative fertilizers. The slow release of urea from nanoparticles and/or their chemical composition contributed to the upregulation of the urea uptake system for a longer period (up to 24 h after treatment) as compared to plants treated with bulk urea. This prolonged activation was mirrored by a higher accumulation of N in nanoparticle-treated plants (approximately threefold increase in the shoot of NP-treated plants compared to controls), even when the concentration of urea conveyed through nanoparticles was halved. In addition, besides impacting N nutrition, U-ACP also enhanced Ca and P concentration in cucumber tissues, thus having possible effects on plant growth and yield, and on the nutritional value of agricultural products.

Water transport mediated by murine urea transporters: implications for urine concentration mechanisms

ABSTRACTUrea transporters (UTs) facilitate urea diffusion across cell membranes and play an important role in the urinary concentration mechanisms in the kidney. Herein, we injected cRNAs encoding for c-Myc-tagged murine UT-B, UT-A2 or UT-A3 (versus water-injected control) in Lithobates oocytes and evaluated oocyte surface protein expression with biotinylation and immunoblotting, urea uptake using [14C] counts and water permeability (Pf) by video microscopy. Immunoblots of UT-injected oocyte membranes revealed bands with a molecular weight consistent with that of a UT monomer (34 kDa), and UT-injected oocytes displayed significantly increased and phloretin-sensitive urea uptake and Pf when compared to day-matched control oocytes. Subtracting the water-injected urea uptake or Pf values from those of UT-injected oocytes yielded UT-dependent values*. We demonstrate for the first time that UT-A2 and UT-A3 can transport water, and we confirm that UT-B is permeable to water. Moreover, the [14C] urea*/Pf* ratios fell in the sequence mUT-B>mUT-A2>mUT-A3, indicating that UTs can exhibit selectivity to urea and/or water. It is likely that specific kidney regions with high levels of UTs will exhibit increased urea and/or water permeabilities, directly influencing urine concentration. Furthermore, UT-mediated water transport activity must be considered when developing UT-inhibitors as novel diuretics.This article has an associated First Person interview with the first author of the paper.

Urea uptake and urease activity in the Chesapeake Bay

The importance of urea in supplying the nitrogen (N) required by planktonic communities has long been recognized, notably by James J. McCarthy in studies as early as the 1970s. Utilization of urea involves a two-step enzymatic process in phytoplankton, with urea first entering the cell via transport (i.e., urea uptake), followed by the conversion of urea into ammonium by the enzyme urease. This article describes a series of field observations and experiments conducted in the Chesapeake Bay, USA, from 2001 through 2018, aimed at understanding the relationship between urea uptake and urease activity and the role of environmental factors on that relationship. Principal component analysis revealed a few patterns. Urea uptake, for example, was consistently positively related to combined variables that included urea concentrations. Similarly, urease activity was consistently positively related to combined variables that included temperature. Contrary to findings in culture studies, however, relationships with environmental factors within different phytoplankton taxa in the field were not clear. This suggests that factors other than those examined may be involved in the regulation of urea uptake and urease activity. New insights into the role of the urea cycle in phytoplankton nitrogen dynamics suggest that the regulation of urease may not be directly impacted by environmental factors, but indirectly regulated by different metabolic pathways responding to nutrient availability, light, and temperature conditions.

Artificial cell microcapsules containing live bacterial cells and activated charcoal for managing renal failure creatinine: preparation and in-vitro analysis

Activated charcoal was microencapsulated with Lactobacillus acidophilus 314 previously adapted for urea uptake. The creatinine removal capacity of this combination microcapsule was evaluated in-vitro in media simulating the small intestine. Results show that microcapsules containing both activated charcoal and L. acidophilus 314 demonstrated potential for decreasing creatinine. Interestingly, when co-encapsulating both activated charcoal and L. acidophilus 314 a smaller decrease in creatinine was observed than when encapsulating them separately. However, co-encapsulated microcapsules were more stable in various parts of the gastrointestinal system and survived longer in storage. These results suggest the feasibility of using microcapsules containing activated charcoal and probiotic bacteria as oral adjuvants for creatinine removal and provides a theoretical model for the use of these microcapsules to remove any unwanted metabolite.

Alternative channels for urea in the inner medulla of the rat kidney

The ascending thin limbs (ATLs) and lower descending thin limbs (DTLs) of Henle's loop in the inner medulla of the rat are highly permeable to urea, and yet no urea transporters have been identified in these sections. We hypothesized that novel, yet-unidentified transporters in these tubule segments could explain the high urea permeability. cDNAs encoding for Na+-glucose transporter 1a (SGLT1a), Na+-glucose transporter 1 (NaGLT1), urea transporter (UT)-A2c, and UT-A2d were isolated and cloned from the Munich-Wistar rat inner medulla. SGLT1a is a novel NH2-terminal truncated variant of SGLT1. NaGLT1 is a Na+-dependent glucose transporter primarily located in the proximal tubules and not previously described in the thin limbs. UT-A2c and UT-A2d are novel variants of UT-A2. UT-A2c is truncated at the COOH terminus, and UT-A2d has one exon skipped. When rats underwent water restriction for 72 h, mRNA levels of SGLT1a increased in ATLs, NaGLT1 levels increased in both ATLs and DTLs, and UT-A2c increased in ATLs. [14C]urea uptake assays performed on Xenopus oocytes heterologously expressing these proteins revealed that despite having structural differences from their full-length versions, SGLT1a, UT-A2c, and UT-A2d enhanced urea uptake. NaGLT1 also facilitated urea uptake. Uptakes were Na+ independent and inhibitable by phloretin and/or phloridzin. Our data indicate that there are several alternative channels for urea in the rat inner medulla that could potentially contribute to the high urea permeabilities in thin limb segments.