Expression Profiles Of Transporters Research Articles

Reprogramming of Glutamine Amino Acid Transporters Expression and Prognostic Significance in Hepatocellular Carcinoma

Hepatocellular carcinoma (HCC) is the most prevalent primary liver malignancy and is a major cause of cancer-related mortality in the world. This study aimed to characterize glutamine amino acid transporter expression profiles in HCC compared to those of normal liver cells. In vitro and in vivo models of HCC were studied using qPCR, whereas the prognostic significance of glutamine transporter expression levels within patient tumors was analyzed through RNAseq. Solute carrier (SLC) 1A5 and SLC38A2 were targeted through siRNA or gamma-p-nitroanilide (GPNA). HCC cells depended on exogenous glutamine for optimal survival and growth. Murine HCC cells showed superior glutamine uptake rate than normal hepatocytes (p < 0.0001). HCC manifested a global reprogramming of glutamine transporters compared to normal liver: SLC38A3 levels decreased, whereas SLC38A1, SLC7A6, and SLC1A5 levels increased. Also, decreased SLC6A14 and SLC38A3 levels or increased SLC38A1, SLC7A6, and SLC1A5 levels predicted worse survival outcomes (all p < 0.05). Knockdown of SLC1A5 and/or SLC38A2 expression in human Huh7 and Hep3B HCC cells, as well as GPNA-mediated inhibition, significantly decreased the uptake of glutamine; combined SLC1A5 and SLC38A2 targeting had the most considerable impact (all p < 0.05). This study revealed glutamine transporter reprogramming as a novel hallmark of HCC and that such expression profiles are clinically significant.

Disruption of Long-Distance Transport Leads to Changes in Gene Expression Profiles of Sugar Transporters in Silver Birch

Disruption of Long-Distance Transport Leads to Changes in Gene Expression Profiles of Sugar Transporters in Silver Birch

Zinc transporters expression profile in professional handball players supplemented with zinc

IntroductionZinc (Zn) deficiency has been described not only on general human health but also within the sports context –as negatively affecting performance–. Thus, Zn status assessment is of great interest for athletes, especially in order to correct deficiency states of this mineral. ObjectiveThe overall objective of this work was to assess Zn status in professional handball players during the competitive period (through plasma levels, dietary intake and gene expression of the Zn transporters), as well as to determine the effect of Zn supplementation. MethodsA total of twenty-two participants were recruited, –twelve belonged to the Control Group (CG) and ten male handball players comprised the experimental group (ATH-G)–, being monitored over a 2-month period with 2 evaluation moments: baseline (i.e., initial conditions) and follow-up (i.e., after 8 weeks of training and competition). Zn intake, plasma Zn levels, and gene expression of Zn transporters were obtained. ResultsPlasma Zn levels were higher in ATH-G than in CG at the end of Zn intervention (p ≤ 0.010). Moreover, differences in the gene expression profile of Zn transporters were observed in ATH-G –with the down-regulation of several Zn transporters–, compared to the CG at baseline (p ≤ 0.05). Likewise, differences in the Zn transporters expression were observed in ATH-G at 8 weeks (all, p ≤ 0.001) –with ZnT2, ZnT5, ZIP3, ZIP5, ZIP11, ZIP13 and ZIP14 transporters being up-regulated–. ConclusionHandball players seemed to have different nutritional needs for Zn, with differences in the gene expression of Zn transporters compared to controls. Zn intervention in our athletes may have influenced the expression of Zn transporters, indicating a potential increase in Zn transporters expression to mobilize Zn at the cellular level at 8 weeks of Zn intervention.

Sex differences in renal acid-base regulation

Transport along the renal tubule differs between female and male rodents, possibly as a consequence of varying abundance and expression of transporters along the nephron. For several aspects such as renal Na+ transport and blood pressure regulation, this has been comprehensively studied; however, data on sex differences in renal acid-base regulation are less well understood. We hypothesized that there will be sex differences in transporter expression profiles in response to acid-base challenges. We used female and male C57Bl/6 mice (n=10 per sex and treatment), which were challenged by acid (NH4Cl, 0.28 mM) and base (NaHCO3, 0.28 mM) in drinking water for 8 days. Blood and urine samples were collected at baseline and at the end of the experimental period before kidneys were harvested under isoflurane anesthesia. Data were analyzed by 2-way ANOVA followed by Tukey multiple comparisons test. In response to NH4Cl challenge, urine pH significantly decreased in both sexes without sex differences (male baseline: 6.4±0.1 vs. NH4Cl: 5.7±0.1; female baseline: 6.2±0.2 vs NH4Cl: 5.6±0.1). In contrast to the urinary response, blood pH was not significantly affected in male mice (male baseline: 7.29±0.01 vs. NH4Cl: 7.26±0.02); however, significantly decreased in female mice (female baseline: 7.28±0.01 vs NH4Cl: 7.22±0.02). Despite differences in acid-base physiology, no differences were observed in the abundance of NHE3, pS552-NHE3, Npt2a, NKCC2, NCC, pT58-NCC or pendrin between sexes. In response to NaHCO3 challenge, urine pH significantly increased in both sexes; however, the increase was significantly smaller in female vs. male mice (male baseline: 6.3±0.1 vs. NaHCO3: 7.6±0.1; female baseline: 6.3±0.1 vs NaHCO3: 7.0±0.3). In contrast to the urinary response, blood pH significantly increased in male mice (male baseline: 7.28±0.01 vs. NaHCO3: 7.40±0.01); however, was not significantly affected in female mice (female baseline: 7.29±0.01 vs NaHCO3: 7.31±0.01). These differences in females compared to males were associated with a significantly greater abundance of Npt2a (~1.5-fold), pendrin (~1.4-fold) and H-ATPase (~1.5-fold). No differences were observed in the abundance of NHE3, pS552-NHE3, NKCC2, NCC, pT58-NCC, between sexes. In summary, our study demonstrates that acid-base regulation shows very specific sex differences: (i) female mice seem to be more prone to a NH4Cl disturbance compared with male mice, (ii) acid challenges under these conditions do not result in major changes in the abundance of the tested proteins, (iii) male mice are more prone to a NaHCO3 disturbance possibly because of (iv) lower abundances of Npt2a, pendrin and H-ATPase. This work was supported by a VA Merit Review Award IBX004968A (to Dr. Rieg) and a Pilot Project from the USF Microbiomes Institute (to T.R. and J.D.R). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Correlating sugar transporter expression and activities to identify transporters for an orphan sugar substrate

Filamentous fungi like Neurospora crassa are able to take up and metabolize important sugars present, for example, in agricultural and human food wastes. However, only a fraction of all putative sugar transporters in filamentous fungi has been characterized to date, and for many sugar substrates, the corresponding transporters are unknown. In N. crassa, only 14 out of the 42 putative major facilitator superfamily (MFS)–type sugar transporters have been characterized so far. To uncover this hidden potential for biotechnology, it is therefore necessary to find new strategies. By correlation of the uptake profile of sugars of interest after different induction conditions with the expression profiles of all 44 genes encoding predicted sugar transporters in N. crassa, together with an exhaustive phylogenetic analysis using sequences of characterized fungal sugar transporters, we aimed to identify transporter candidates for the tested sugars. Following this approach, we found a high correlation of uptake rates and expression strengths for many sugars with dedicated transporters, like galacturonic acid and arabinose, while the correlation is loose for sugars that are transported by several transporters due to functional redundancy. Nevertheless, this combinatorial approach allowed us to elucidate the uptake system for the disaccharide lactose, a by-product of the dairy industry, which consists of the two main cellodextrin transporters CDT-1 and CDT-2 with a minor contribution of the related transporter NCU00809. Moreover, a non-MFS transporter involved in glycerol transport was also identified. Deorphanization of sugar transporters or identification of transporters for orphan sugar substrates by correlation of uptake kinetics with transporter expression and phylogenetic information can thus provide a way to optimize the reuse of food industry by-products and agricultural wastes by filamentous fungi in order to create economic value and reduce their environmental impact.Key points• The Neurospora crassa genome contains 30 uncharacterized putative sugar transporter genes.• Correlation of transporter expression and sugar uptake profiles can help to identify transporters for orphan sugar substrates.• CDT-1, CDT-2, and NCU00809 are key players in the transport of the dairy by-product lactose in N. crassa.

Expression Profiles of Glucose Transporters Along the Nephron in Non-Diabetic and Diabetic Kidneys

Expression Profiles of Glucose Transporters Along the Nephron in Non-Diabetic and Diabetic Kidneys

Impact of hypoxia and reoxygenation on the extra/intracellular metabolome and on transporter expression in a human kidney proximal tubular cell line.

Ischemia-reperfusion injury (IRI) induces several perturbations that alter immediate kidney graft function after transplantation and may affect long-term graft outcomes. Given the IRI-dependent metabolic disturbances previously reported, we hypothesized that proximal transporters handling endo/exogenous substrates may be victims of such lesions. This study aimed to determine the impact of hypoxia/reoxygenation on the human proximal transport system through two semi-targeted omics analyses. Human proximal tubular cells were cultured in hypoxia (6 or 24h), each followed by 2, 24 or 48-h reoxygenation. We investigated the transcriptomic modulation of transporters. Using semi-targeted LC-MS/MS profiling, we characterized the extra/intracellular metabolome. Statistical modelling was used to identify significant metabolic variations. The expression profile of transporters was impacted during hypoxia (y + LAT1 and OCTN2), reoxygenation (MRP2, PEPT1/2, rBAT, and OATP4C1), or in both conditions (P-gp and GLUT1). The P-gp and GLUT1 transcripts increased (FC (fold change) = 2.93 and 4.11, respectively) after 2-h reoxygenation preceded by 24-h hypoxia. We observed a downregulation (FC = 0.42) of y+LAT1 after 24-h hypoxia, and of PEPT2 after 24-h hypoxia followed by 2-h reoxygenation (FC = 0.40). Metabolomics showed that hypoxia altered the energetic pathways. However, intracellular metabolic homeostasis and cellular exchanges were promptly restored after reoxygenation. This study provides insight into the transcriptomic response of the tubular transporters to hypoxia/reoxygenation. No correlation was found between the expression of transporters and the metabolic variations observed. Given the complexity of studying the global tubular transport systems, we propose that further studies focus on targeted transporters.

Short-chain fatty acids inhibit the activation of T lymphocytes and myeloid cells and induce innate immune tolerance.

The intestinal microbiota contributes to gut immune homeostasis, where short-chain fatty acids (SCFAs) function as the major mediators. We aimed to elucidate the immunomodulatory effects of acetate, propionate, and butyrate. With that in mind, we sought to characterise the expression of SCFA receptors and transporters as well as SCFAs' impact on the activation of different immune cells. Whereas all three SCFAs decreased tumour necrosis factor (TNF)-α production in activated T cells, only butyrate and propionate inhibited interferon (IFN)-γ, interleukin (IL)-17, IL-13, and IL-10 production. Butyrate and propionate inhibited the expression of the chemokine receptors CCR9 and CCR10 in activated T- and B-cells, respectively. Similarly, butyrate and propionate were effective inhibitors of IL-1β, IL-6, TNF-α, and IL-10 production in myeloid cells upon lipopolysaccharide and R848 stimulation. Acetate was less efficient at inhibiting cytokine production except for IFN-α. Moreover, SCFAs inhibited the production of IL-6 and TNF-α in monocytes, myeloid dendritic cells (mDC), and plasmacytoid dendritic cells (pDC), whereas acetate effects were relatively more prominent in pDCs. In monocytes and mDCs, acetate was a less efficient inhibitor, but it was equally effective in inhibiting pDCs activation. We also studied the ability of SCFAs to induce trained immunity or tolerance. Butyrate and propionate- but not acetate- prevented Toll-like receptor-mediated activation in SCFA-trained cells, as demonstrated by a reduced production of IL-6 and TNF-α. Our findings indicate that butyrate and propionate are equally efficient in inhibiting the adaptive and innate immune response and did not induce trained immunity. The findings may be explained by differential SCFA receptor and transporter expression profiles of the immune cells.

Proteomics-based comparison of transporter expression profiles in BeWo and JEG-3 cells

Proteomics-based comparison of transporter expression profiles in BeWo and JEG-3 cells

Fetal membranes exhibit similar nutrient transporter expression profiles to the placenta

IntroductionDuring pregnancy, the growth of the fetus is supported by the exchange of nutrients, waste, and other molecules between maternal and fetal circulations in the utero-placental unit. Nutrient transfer, in particular, is mediated by solute transporters such as solute carrier (SLC) and adenosine triphosphate-binding cassette (ABC) proteins. While nutrient transport has been extensively studied in the placenta, the role of human fetal membranes (FM), which was recently reported to have a role in drug transport, in nutrient uptake remains unknown. ObjectivesThis study determined nutrient transport expression in human FM and FM cells and compared expression with placental tissues and BeWo cells. MethodsRNA sequencing (RNA-Seq) of placental and FM tissues and cells was done. Genes of major solute transporter groups, such as SLC and ABC, were identified. Proteomic analysis of cell lysates was performed via nano-liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS) to confirm expression at a protein level. ResultsWe determined that FM tissues and cells derived from the fetal membrane tissues express nutrient transporter genes, and their expression is similar to that seen in the placenta or BeWo cells. In particular, transporters involved in macronutrient and micronutrient transfer were identified in both placental and FM cells. Consistent with RNA-Seq findings, carbohydrate transporters (3), vitamin transport-related proteins (8), amino acid transporters (21), fatty acid transport-related proteins (9), cholesterol transport-related proteins (6) and nucleoside transporters (3) were identified in BeWo and FM cells, with both groups sharing similar nutrient transporter expression. ConclusionThis study determined the expression of nutrient transporters in human FMs. This knowledge is the first step in improving our understanding of nutrient uptake kinetics during pregnancy. Functional studies are required to determine the properties of nutrient transporters in human FMs.

Renal Transporter Alterations in Patients with Chronic Liver Diseases: Nonalcoholic Steatohepatitis, Alcohol-Associated, Viral Hepatitis, and Alcohol-Viral Combination.

Alterations in hepatic transporters have been identified in precirrhotic chronic liver diseases (CLDs) that result in pharmacokinetic variations causing adverse drug reactions (ADRs). However, the effect of CLD on the expression of renal transporters is unknown despite the overwhelming evidence of kidney injury in CLD patients. This study determines the transcriptomic and proteomic expression profiles of renal drug transporters in patients with defined CLD etiology. Renal biopsies were obtained from patients with a history of CLD etiologies, including nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), alcohol-associated liver disease (ALD), viral hepatitis C (HCV), and combination ALD/HCV. A significant decrease in organic anion transporter (OAT)-3 was identified in NASH, ALD, HCV, and ALD/HCV (1.56 ± 1.10; 1.01 ± 0.46; 1.03 ± 0.43; 0.86 ± 0.57 pmol/mg protein) relative to control (2.77 ± 1.39 pmol/mg protein). Additionally, a decrease was shown for OAT4 in NASH (24.9 ± 5.69 pmol/mg protein) relative to control (43.8 ± 19.9 pmol/mg protein) and in urate transporter 1 (URAT1) for ALD and HCV (1.56 ± 0.15 and 1.65 ± 0.69 pmol/mg protein) relative to control (4.69 ± 4.59 pmol/mg protein). These decreases in organic anion transporter expression could result in increased and prolonged systemic exposure to drugs and possible toxicity. Renal transporter changes, in addition to hepatic transporter alterations, should be considered in dose adjustments for CLD patients for a more accurate disposition profile. It is important to consider a multiorgan approach to altered pharmacokinetics of drugs prescribed to CLD patients to prevent ADRs and improve patient outcomes. SIGNIFICANCE STATEMENT: Chronic liver diseases are known to elicit alterations in hepatic transporters that result in a disrupted pharmacokinetic profile for various drugs. However, it is unknown if there are alterations in renal transporters during chronic liver disease, despite strong indications of renal dysfunction associated with chronic liver disease. Identifying renal transporter expression changes in patients with chronic liver disease facilitates essential investigations on the multifaceted relationship between liver dysfunction and kidney physiology to offer dose adjustments and prevent adverse drug reactions.

Significance of differential expression profiles of ABC transporters in azole susceptibility between Cryptococcus gattii VGI and VGII strains.

Azoles were used as the primary antifungal agents for treating Cryptococuss gattii infection. Since subtypes of C. gattii respond differently to azoles. We analyzed mRNA expression profiles of different subtypes and identified four ABC transporters that could be potential genes regulating azole sensitivity.

Atmospheric Reactive Oxygen Species and Some Aspects of the Antiviral Protection at the Respiratory Epithelium.

—The review summarizes literature data on molecular and biochemical mechanisms of nonspecific protection of respiratory epithelium. The special attention is paid to comprehensive analysis of up-to-date data on the activity of the lactoperoxidase system expressed on the surface of the respiratory epithelium which provides the generation of hypothiocyanate and hypoiodite in the presence of locally produced or inhaled hydrogen peroxide. Molecular mechanisms of production of active compounds with antiviral and antibacterial effects, expression profiles of enzymes, transporters and ion channels involved in the generation of hypothiocyanite and hypoiodite in the mucous membrane of the respiratory system in physiological and pathological conditions (inflammation) are discussed. A hypothesis about the effect of atmospheric air composition on the efficiency of hypothiocyanate and hypoiodite generation in the respiratory epithelium in the context of its antibacterial and antiviral protection is presented. The causes and consequences of insufficiency of the lactoperoxidase system caused by the action of atmospheric factors are discussed in the context of controlling the sensitivity of the epithelium to the action of bacterial agents and viruses. Good evidence exists that restoration of the lactoperoxidase system activity can be achieved by application of pharmacological agents aimed to compensate for the deficit of halides in tissues, and by the control of chemical composition of the inhaled air.

Cover Image

The front cover image is based on the Original Article Visualization of phosphorus re-translocation and phosphate transporter expression profiles in a shortened annual cycle system of poplar by Yuko Kurita et al., https://doi.org/10.1111/pce.14319.

Quantitative Expression of Drug, Phospholipid and Nucleoside Transporters in the Lung Tissues across Species

ObjectiveIt has been proven that transporters play a pivotal role in organ distribution of drugs, and subsequently in drug efficacy and toxicity. Active transport processes may affect the local pulmonary disposition of drugs administered directly to the lung by the inhaled route as well as the uptake of drugs to the lung from the systemic circulation. While drug transporter expressions and their roles in drug disposition and elimination in the major elimination organs including intestine, liver and kidney are well‐characterized, the expression of drug transporters and the comparison across species in the lung remain overall poorly understood. For example, azithromycin levels in pulmonary tissue remained to be high for up to 5 days after a single oral dose, in contrast to much lower plasma levels. However, the mechanisms associated with azithromycin lung loading and efficacy remain unknown. The aim of this study was to comprehensively determine the mRNA and protein expression levels of drug transporters in the lung in human and preclinical species using quantitative targeted proteomics approaches. The findings can be useful to further explore potential mechanisms in lung specific disposition.MethodsTotal cellular mRNA was isolated from human lower lung tissues using a Qiagen mRNA isolation kit. qRT‐PCR was performed, and β‐actin mRNA expression was used to normalize target gene expression. The mRNA levels were expressed as a fold change using the 2‐ΔΔCt method. The total membrane proteins of lung tissues from human, monkey, dog and rat were extracted by Native Membrane Protein Extraction Kit, and subsequently digested by trypsin followed by the treatment of DTT and iodoacetamide (IAA). The levels of transporter proteins were quantified using a Sciex 7500 triple‐quadruple mass spectrometer coupled to a Shimadzu LC (SLC‐30A) system with optimized ionization conditions. The Na K‐ATPase level in the lung was measured as the membrane protein marker.Results and conclusionIn the current investigation, thirteen ABC efflux transporters and twenty‐three solute carriers (SLC) were characterized. Among those, mRNAs of eleven ABC transporters and fifteen SLC transporters were detected by the RT‐PCR method. In addition, eight ABC transporter and eleven SLC transporter proteins were quantified using the targeted proteomics LC‐MS/MS method. The mRNA expression pattern agrees with the protein expression; however, no quantitative correlation can be drawn between mRNA and protein expression. While OATP2B1 was only detected in human lung tissues, OATP2A1 and 3A1 are expressed in the lungs of human, monkey dog and rat. The expressions of three equilibrative nucleoside transporters (ENT1/2/3) in human lung tissues were higher than those in monkey. The expression of ABCA1, ABCA3 and ABCG1, the ABC transporters that are the regulators of cellular cholesterol and phospholipid homeostasis, were detected in all tested species, and found to be high in rat. Collectively, the comparison of transporter expression profiles can contribute to a better understanding of drug distribution and transport in the lung and aid pulmonary drug discovery and development.

Evaluation of Potential Herb-Drug Interactions Between Shengmai Injection and Losartan Potassium in Rat and In Vitro

Aim: The present study aimed to explore the potential herb-drug interactions (HDI) between Shengmai injection (SMI) and losartan potassium (LOS) based on the expression profiles of cytochromes P450 (CYP450) and drug transporters in rat and in vitro. Methods: Different concentrations of SMI were used to explore the influence of SMI on the antihypertensive efficacy of LOS in the hypertension rat model established by N (omega)-nitro-L-arginine methyl ester (L-NAME) for 4 weeks. Subsequently, the serum concentration levels of LOS and losartan carboxylic acid (EXP3174) were determined by Liquid Chromatography Mass Spectrometry (LC-MS) and pharmacokinetic analysis. Human liver microsomes, human multidrug resistance protein 1 (MDR1/P-gp), and breast cancer resistance protein (BCRP) vesicles, human embryonic kidney 293 cell line with stable expression of the organic anion transporting polypeptide 1B1 (HEK293-OATP1B1 cells) and mock-transfected HEK293 (HEK293-MOCK) cells were used to verify the effects of SMI on CYP450 enzymes and drug transporters in vitro. Results: Low, medium, and high concentrations of SMI increased the antihypertensive efficacy of LOS to varying degrees. The high dose SMI increased the half-life (t 1/2 ), the maximum plasma concentration (C max), the area under the plasma concentration-time curve (AUC) from time zero to the time of the last measurable plasma concentration (AUC 0-t ), AUC from time zero to infinity (AUC 0-∞ ), and mean residence time (MRT) values of LOS and decreased its apparent volume of distribution (Vd) and clearance (CL) values. The AUC 0-t , AUC 0-∞ , and MRT of LOS were increased, whereas the CL was decreased by the medium concentration of SMI. In addition, the high, medium, and low doses of SMI increased the relative bioavailability (Frel) of LOS. SMI exhibited no significant effects on the pharmacokinetics of EXP3174. In vitro, SMI exhibited different suppressive effects on the enzyme activity levels of CYP1A2 (6.12%), CYP2B6 (2.72%), CYP2C9 (14.31%), CYP2C19 (12.96%), CYP2D6 (12.26%), CYP3A4 (3.72%), CYP2C8 (10.00–30.00%), MDR1 (0.75%), OATP1B1(2.03%), and BCRP (0.15%). Conclusion: In conclusion, SMI improved the antihypertensive efficacy of LOS in the L-NAME-induced hypertension rat model by increasing the concentration of LOS, while leaving the concentration of EXP3174 intact. SMI affected the pharmacokinetic properties of LOS by decreasing the elimination of LOS. These effects might partly be attributed to the inhibition of the activities of CYP3A4, CYP2C9, and of the drug transporters (P-gp, BCRP, and OATP1B1) by SMI, which need further scrutiny.

Intracellular pH affects mitochondrial homeostasis in cultured human corneal endothelial cells prepared for cell injection therapy

This study aimed to uncover the mechanism responsible for the clinical efficacy of cell injection therapy with fully differentiated cultured cells. Analysis of polarized expression of ion transporters on cultured human corneal endothelial cells (CECs) subpopulations (SPs) was performed. The intracellular pH (pHi) between two CEC SPs, distinct in the proportion of differentiated cells, was measured, and the association with mitochondrial respiration homeostasis was investigated. The effects of the ion transporter inhibition by their selective inhibitors or siRNA transfection were also explored. Na+/K+-ATPase, Aquaporin 1, SLC4A11, NBCe1, NHE1 as transporters, and ZO-1, were all selectively expressed in differentiated SPs, but were almost null in the cell-state-transitioned SPs. We also confirmed that the pHi of CEC SPs affected their mitochondrial respiration by modulating the expression of these ion transporters via inhibitors or siRNA transfection. Ion and water transporters might participate in the maintenance of pHi and mitochondria homeostasis in differentiated SPs, which may contribute, combined with integral barrier functions, to efficient water efflux. The differences in intracellular pH between the two SPs is attributed to variations in the expression profile of specific ion transporters and mitochondrial functions, which may associate with the efficacy of the SPs in cell injection therapy.

Visualization of phosphorus re‐translocation and phosphate transporter expression profiles in a shortened annual cycle system of poplar

Phosphorus (P) is an essential macronutrient for plant growth. In deciduous trees, P is remobilized from senescing leaves and stored in perennial tissues during winter for further growth. Annual internal recycling and accumulation of P are considered an important strategy to support the vigorous growth of trees. However, the pathways of seasonal re-translocation of P and the molecular mechanisms of this transport have not been clarified. Here we show the seasonal P re-translocation route visualized using real-time radioisotope imaging and the macro- and micro-autoradiography. We analysed the seasonal re-translocation P in poplar (Populus alba. L) cultivated under 'a shortened annual cycle system', which mimicked seasonal phenology in a laboratory. From growing to senescing season, sink tissues of 32 P and/or 33 P shifted from young leaves and the apex to the lower stem and roots. The radioisotope P re-translocated from a leaf was stored in phloem and xylem parenchyma cells and redistributed to new shoots after dormancy. Seasonal expression profile of phosphate transporters (PHT1, PHT5 and PHO1 family) was obtained in the same system. Our results reveal the seasonal P re-translocation routes at the organ and tissue levels and provide a foothold for elucidating its molecular mechanisms.

The WWOX/HIF1A Axis Downregulation Alters Glucose Metabolism and Predispose to Metabolic Disorders.

Recent reports indicate that the hypoxia-induced factor (HIF1α) and the Warburg effect play an initiating role in glucotoxicity, which underlies disorders in metabolic diseases. WWOX has been identified as a HIF1α regulator. WWOX downregulation leads to an increased expression of HIF1α target genes encoding glucose transporters and glycolysis’ enzymes. It has been proven in the normoglycemic mice cells and in gestational diabetes patients. The aim of the study was to determine WWOX’s role in glucose metabolism regulation in hyperglycemia and hypoxia to confirm its importance in the development of metabolic disorders. For this purpose, the WWOX gene was silenced in human normal fibroblasts, and then cells were cultured under different sugar and oxygen levels. Thereafter, it was investigated how WWOX silencing alters the genes and proteins expression profile of glucose transporters and glycolysis pathway enzymes, and their activity. In normoxia normoglycemia, higher glycolysis genes expression, their activity, and the lactate concentration were observed in WWOX KO fibroblasts in comparison to control cells. In normoxia hyperglycemia, it was observed a decrease of insulin-dependent glucose uptake and a further increase of lactate. It likely intensifies hyperglycemia condition, which deepen the glucose toxic effect. Then, in hypoxia hyperglycemia, WWOX KO caused weaker glucose uptake and elevated lactate production. In conclusion, the WWOX/HIF1A axis downregulation alters glucose metabolism and probably predispose to metabolic disorders.

Fe3O4-urea nanocomposites as a novel nitrogen fertilizer for improving nutrient utilization efficiency and reducing environmental pollution

Almost 81% of nitrogen fertilizers are applied in form of urea but most of it is lost due to volatilization and leaching leading to environmental pollution. In this regard, slow-release nano fertilizers can be an effective solution. Here, we have synthesized different Fe3O4-urea nanocomposites with Fe3O4 NPs: urea ratio (1:1, 1:2, 1:3) ie. NC-1, 2, and 3 respectively, and checked their efficacy for growth and yield enhancement. Oryza sativa L. cv. Swarna seedlings were treated with different NCs for 14 days in hydroponic conditions and significant up-regulation of photosynthetic efficiency and nitrogen metabolism were observed due to increased availability of nitrogen and iron. The discriminant functional analysis confirmed that the NC3 treatment yielded the best results so further gene expression studies were performed for NC-3 treated seedlings. Significant changes in expression profiles of ammonia and nitrate transporters indicated that NC-3 treatment enhanced nitrogen utilization efficiency (NUE) due to sustained slow release of urea. From pot experiments, we found significant enhancement of growth, grain nutrient content, and NUE in NC supplemented sets. 1.45 fold increase in crop yield was achieved when 50% N was supplemented in form of NC-3 and the rest in form of ammonium nitrate. NC supplementation can also play a vital role in minimizing the use of bulk N fertilizers because, when 75% of the recommended N dose was supplied in form of NC-3, 1.18 fold yield enhancement was found. Thus our results highlight that, slow-release NC-3 can play a major role in increasing the NUE of rice.