Biomedical subjects

G You

Publications and source records attributed to G You.

At least 19 recordsLinked to original sources

PubMed2000

Regulation of mOAT-mediated organic anion transport by okadaic acid and protein kinase C in LLC-PK(1) cells.

Organic anion transporters in the kidney proximal tubule play an essential role in eliminating a wide range of organic anions including endogenous compounds, xenobiotics, and their metabolites, thereby preventing their potentially toxic effects within the body. We have previously cloned a cDNA encoding an organic anion transporter from mouse kidney (mOAT) (Lopez-Nieto, C. E., You, G., Bush, K. T., Barros, E. J. G., Beier, D. R., and Nigam, S. K. (1997) J. Biol. Chem. 272, 6471-6478; Kuze, K., Graves, P., Leahy, A., Wilson, P., Stuhlmann, H., and You, G. (1999) J. Biol. Chem. 274, 1519-1524). In the present study, we assessed the potential for regulation of this transporter by heterologous expression of mOAT in the pig proximal tubule-like cell line, LLC-PK(1). We report here that both protein phosphatase (PP1/PP2A) inhibitor, okadaic acid, and protein kinase C (PKC) activators down-regulate mOAT-mediated transport of para-aminohippuric acid (PAH), a prototypic organic anion, in a time- and concentrationdependent manner. However their mechanisms of action for this down-regulation are distinct. Okadaic acid modulated PAH transport, at least in part, through phosphorylation/dephosphorylation of mOAT; phosphoamino acid analysis indicated this phosphorylation occurs on serine. In contrast, PKC activation induced a decrease in the maximum transport velocity (V(max)) of PAH transport without direct phosphorylation of the transporter protein. Together these results provide the first demonstration that regulation of organic anion transport by mOAT is likely to be tightly controlled directly and indirectly by phosphatase PP1/PP2A and PKC. Our results also suggest that kinases other than PKC are involved in this process.

Animals↗

PubMed2000

[Clinical analysis of 106 cases of inhalation injury induced by gas explosion].

OBJECTIVE: To review the management of inhalation injury induced by gas explosion in our department, so as to draw some experience and lessons for future work. METHODS: One hundred and six cases of inhalation injury induced by gas explosion were analyzed in terms of the characteristics of and therapeutic methods for the disease. RESULTS: The injury induced by gas explosion was very complicated with high incidences of compound injuries, severe inhalation injury and systemic infection. In addition, respiratory dysfunction occurred very quickly and severely at early postburn stage, which could easily develop to respiratory failure. The latter could ultimately lead patients to death due to improper management. CONCLUSION: In order to raise cure rate of the inhalation injury induced by gas explosion, airway obstruction should be cleared immediately and fiberbronchoscopy and respirator should be employed as soon as possible to help remove endotracheal obstruction and to maintain effective gas exchange. In addition, correct pulmonary resuscitation and continuous blood gas analysis were advocated.

Adolescent↗

PubMed1999

Heterologous expression and functional characterization of a mouse renal organic anion transporter in mammalian cells.

Organic anion transporters play an essential role in eliminating a wide range of organic anions including endogenous compounds, xenobiotics, and their metabolites from kidney, thereby preventing their potentially toxic effects within the body. The goal of this study was to extend our previous study on the functional characterization and post-translational modification of a mouse kidney organic anion transporter (mOAT), in a mammalian cell system, COS-7 cells. The transporter-mediated p-aminohippurate (PAH) uptake was saturable, probenecid-sensitive, and inhibited by a wide range of organic anions including vitamins, anti-hypertensive drugs, anti-tumor drugs, and anti-inflammatory drugs. Tunicamycin, an inhibitor of asparagine-linked glycosylation, significantly inhibited the transport activity. Immunofluorescence provided evidence that most of the protein remained in the intracellular compartment in tunicamycin-treated cells. Diethyl pyrocarbonate (DEPC), a histidine residue-specific reagent, completely blocked PAH transport. The inhibitory effect by DEPC was significantly protected (90%) by pretreating the cells with excess unlabeled PAH, suggesting that the histidine residues may be close to the PAH binding sites. Finally, in situ mRNA localization was studied in postnatal mouse kidney. The expression was observed in proximal tubules throughout development. We conclude that COS-7 cells may be useful in pharmacological and molecular biological studies of this carrier. The carbohydrate moieties are necessary for the proper trafficking of mOAT to the plasma membrane, and histidine residues appear to be important for the transport function.

Animals↗

PubMed1999

Mitochondrial DNA analysis of Sporothrix schenckii clinical isolates from China.

Mitochondrial DNA (mtDNA) types based on restriction fragment length polymorphism (RFLP) patterns with HaeIII were investigated in clinical isolates of Sporothrix schenckii in China. In addition to 23 mtDNA types (Types 1-23) so far reported, a new mtDNA type (Type 24) was found in this study. Type 24 was divided into two subtypes, Subtype 24A and 24B based on RFLP with EcoRV. Sixty-seven isolates in China consisted of 58 isolates of Type 4, 5 of Type 6, 1 of Type 5, 1 of Type 20 and 2 of Type 24. Based on the phylogeny of the mtDNA types (Types 1-24) constructed by estimating sequence divergences of mtDNA, mtDNA types clustered into two groups: Group A (Types 1-3, Type 11, Types 14-19 and Types 22-23) and Group B (Types 4-10, Types 12-13, Types 20-21 and Type 24). These results suggest that most S. schenckii isolates in China belong to Group B.

China↗

PubMed1999

[The relationship between sleep related breathing disorders and stroke].

OBJECTIVE: To study the relationship between sleep related breathing disorder(SRBD) and cerebrovascular diseases (CVD). METHODS: Polysomnographic studies on 43 cases of CVD were performed and 43 sex-, age-, body mass index- compatible cases without CVD used as controls. RESULTS: The incidence of SRBD (mostly the obstructive one) was much more in CVD patients and both the obstructive and central apnea/hypopnea indices, deoxygenation indices were bigger and deoxygenation time were longer while the mean and lowest oxygen saturation were lower in CVD group than those in the controls (all P < 0.05). The blood pressure before sleep, after awakening and the mean BP during sleep were also higher in the CVD patients. In CVD group, the incidence of hypertension was also much higher in SRBD patients than that in non-SRBD ones (P < 0.01). SRBD was dangerous for CVD as OR value was 6.28. CONCLUSIONS: SRBD may be one important risk factor for CVD. An vicious circle might be formed between stroke and SRBD which inflences the outcome of stroke.

Blood Pressure↗

PubMed1999

[Changes of the interleukin-1 in dysfunction of central nervous system in myasthenia gravis].

OBJECTIVE: Recent studies have demonstrated that acetylcholine receptor antibodies (AChRab) present in the serum of the patients with myasthenia gravis (MG) not only block acetylcholine transmission at the neuromuscular junction, but also cause dysfunction of central nervous system (CNS). The mechanism remains unclear. In the present study the concentration of interleukin-1 in the brain, thymus and blood was investigated to reveal the mechanism of CNS dysfunction in MG. METHODS: The concentration of interleukin-1 in brain, thymus and blood was detected after establishing the experimental MG model of central nervous system dysfunction induced by injection of AChRab purified from MG sera into the rat cerebral ventricular system. RESULTS: Interleukin-1 level in brain, thymus and blood started to increase one week after injection of AChRab. The level of interleukin-1 in brain tissue showed a very significant increase, as compared to that in thymus and blood till the end of 2 weeks after injection, where as in thymus and blood the increasing rate was relatively slow. CONCLUSION: It is concluded that interleukin-1 increased in rat brain, thymus and blood may play an important role in generating dysfunction of CNS in MG.

Animals↗

PubMed1999

[A clinico-pathological and etiological study of Binswanger's Disease].

OBJECTIVE: To study the pathology, incidence and etiology of Binswanger's disease. METHODS: Autopsied brains from 9 cases of Binswanger's disease and 13 cases of non-demented elderly individuals were studied by clinico-pathological, immunohistochemical and micrometer methods. RESULTS: The lesions of Binswanger's disease were mainly located in the subcortical white matter, periventrical regions, brainstem and cerebellum. The pathological changes were demyelination, lacunar infarction, gliosis and dilatation of perivascular space (Virchow-Robin space, VR space, P < 0.01). The wall thickness of deeply penetrating arteries in the white matter was significantly increased (P < 0.01). CONCLUSION: Binswanger's disease is not a rare entity. The findings of this study suggest that arteriosclerosis is a primary factor in the pathogenesis of Binswanger's disease.

Aged↗

PubMed1997

Molecular cloning and characterization of NKT, a gene product related to the organic cation transporter family that is almost exclusively expressed in the kidney.

We have identified a gene product (NKT) encoding an apparently novel transcript that appears to be related to the organic ion transporter family and is expressed almost exclusively in the kidney. Analysis of the deduced 546-amino acid protein sequence indicates that NKT is a unique gene product which shares a similar transmembrane domain hydropathy profile as well as transporter-specific amino acid motifs with a variety of bacterial and mammalian nutrient transporters. Nevertheless, the overall homology of NKT to two recently cloned organic ion transport proteins (NLT and OCT-1) is significantly greater; together these three gene products may represent a new subgroup of transporters. The NKT was characterized further with respect to its tissue distribution and its expression during kidney development. A 2.5-kilobase transcript was found in kidney and at much lower levels in brain, but not in a number of other tissues. Studies on the embryonic kidney indicate that the NKT transcript is developmentally regulated with significant expression beginning at mouse gestational day 18 and rising just before birth, consistent with a role in differentiated kidney function. Moreover, in situ hybridization detected specific signals in mouse renal proximal tubules. NKT was mapped by linkage disequilibrium to mouse chromosome 19, the same site to which several mouse mutations localize, including that for osteochondrodystrophy (ocd). Although initial experiments in a Xenopus oocyte expression system failed to demonstrate transport of known substrates for OCT-1, the homology to OCT-1 and other transporters, along with the proximal tubule localization, raise the possibility that this gene may play a role in organic solute transport or drug elimination by the kidney.

Amino Acid Sequence↗

PubMed1997

[Preliminary study on method for predicting lower body negative pressure tolerance].

To find out a method to predict lower body negative pressure (LBNP) tolerance with noninvasive physiological measurements, 11 healthy young subjects were tested in an LBNP chamber in sitting position. Incremental negative pressure was used for evaluating LBNP tolerance time. End points of the test were onset of presyncopal symptoms or completion of 15 min test. According to their reactions, the 11 subjects were divided into good tolerance group (group A, n = 5) and poor tolerance group (group B, n = 6); At the 11th minute of LBNP, mean arterial pressure (MAP), magnitude of decrease of regional cerebral oxygen saturation (delta SrO2) and cardiac output (delta CO) showed significant difference between group A and B(P <0.05); Mean error was 0.54 +/- 0.42 min when LBNP tolerance was predicted using MAP and delta SrO2 at this moment by regression analysis.

Aerospace Medicine↗

PubMed1997

Changes of cerebral oxygen saturation under lower body negative pressure.

The purpose of this study was to observe and analyse the changes of regional cerebral oxygen saturation (SrO2) under lower body negative pressure (LBNP). 12 healthy young subjects were tested in an LBNP chamber in sitting position. Incremental negative pressure was used and end points of test were onset of presyncopal symptoms or completion of the 15min test. The results were: (1) SrO2 showed a significant decrease under LBNP; (2) The magnitude of decrease of SaO2 showed significant differences among subjects with different reactions at the termination of LBNP; (3) Under LBNP the arterial oxygen saturation (SrO2) kept constant. The changes of SrO2 correlated strongly with the percentage changes of superaorbitalis arterial blood flow (r = 0.59-0.86, P < 0.05 or 0.01). It was suggested that the changes of SrO2 were related to the decrease of cerebral blood flow under LBNP. It was possible to use SrO2 to detect presyncopal and syncopal symptoms. So the monitoring of SrO2 could be used in studies of G-LOC.

Adult↗

PubMed1996

Structure, regulation and physiological roles of urea transporters.

Urea is the major constituent of the urine and the principal means for disposal of nitrogen derived from amino acid metabolism. Specialized phloretin-inhibitable urea transporters are expressed in kidney medulla and play a central role in urea excretion and water balance. These transporters allow accumulation of urea in the medulla and enable the kidney to concentrate urine to an osmolality greater than systemic plasma. Recently, expression cloning with Xenopus oocytes has led to the isolation of a novel phloretin-inhibitable urea transporter (UT2) from rabbit, and subsequently from rat kidney. UT2 from both species has the characteristics of the phloretin-sensitive urea transporter previously defined in kidney by in vitro perfused tubule studies. Based on these advances, Ripoche and colleagues cloned a homologous urea transporter (HUT11) from erythrocytes. UT2 and HUT11 predict 43 kDa polypeptides and exhibit 64% amino acid sequence identity. Since regulation of urea transport in the kidney plays an important role in the orchestration of the antidiuretic response, we have studied the regulation of urea transporter in rat kidney at the mRNA level. On Northern blots probed at high stringency, rat UT2 hybridized to two transcripts of 2.9 kb and 4.0 kb, which have spatially distinct distributions within the kidney. Northern analysis and in situ hybridization of kidneys from rats maintained at different physiologic states revealed that the 2.9 and 4.0 kb transcripts are regulated by separate mechanisms. The 4 kb transcript was primarily responsive to changes in the dietary protein content, whereas the 2.9 kb transcript was highly responsive to changes in the hydration state of the animal. We propose that the two UT2 transcripts are regulated by distinct mechanisms to allow optimal fluid balance and urea excretion.

Animals↗

PubMed1996

Functional differentiation of the human red blood cell and kidney urea transporters.

The recent cloning of two urea transporters will allow to better understand their role in the urinary concentrating mechanism. This physiological approach needs to be sustained by a knowledge of their functional characteristics. We compared the pharmacological properties of the human red blood cell and kidney urea transporters (HUT11 and HUT2) in the Xenopus oocyte expression system. Both proteins allow the rapid transfer of urea but not of water. Both are inhibited by phloretin, although with different half-maximal inhibitory concentrations (IC50; 75 microM, for HUT11 and 230 microM for HUT2). Whereas para-chloromercuribenzene sulfonate inhibits HUT11 with an IC50 of 150 microM, it does not inhibit HUT2, whatever the concentration used. We demonstrate that thiourea diffuses through HUT11 with a Michaelis constant (Km) of 40 mM, but not through HUT2. In contrast, it inhibits urea transport through both proteins. This identification of a substrate binding site independent from the transport activity is the first step in the understanding of the molecular events underlying urea transport.

4-Chloromercuribenzenesulfonate↗

PubMed1995

Molecular characteristics of Na(+)-coupled glucose transporters in adult and embryonic rat kidney.

Two distinct Na(+)-coupled glucose transporters (SGLTs) with either a high or a low affinity for glucose were shown to provide reabsorption of filtered glucose in the kidney. We have previously reported the characteristics of the high affinity Na+/glucose cotransporter SGLT1 from rabbit, rat, and human kidney and the low affinity Na+/glucose cotransporter SGLT2 from human kidney. Because the molecular identity of SGLT2 as the kidney cortical low affinity Na+/glucose cotransporter has been recently challenged based on studies of the porcine low affinity Na+/glucoe cotransporter SAAT-pSGLT2 (Mackenzie, B., Panayotova-Heiermann, M., Loo, D. D. F., Lever, J.E., and Wright, E. M. (1994) J. Biol. Chem. 269, 22488-22491), we have reevaluated the properties of SGLT2 in greater detail. We furthermore report new data on the regulation of SGLT1 and SGLT2 during kidney development. To analyze and compare SGLT1 and SGLT2 in adult and embryonic kidney, we have cloned and characterized SGLT2 from rat kidney and determined its tissue distribution based on Northern analysis and in situ hybridization. When expressed in Xenopus oocytes, rat SGLT2 stimulated transport of alpha-methyl-D-glucopyranoside (2 mM) in oocytes up to 4.5-fold over controls with an apparent Km of 3.0 mM. The transport properties (i.e. a Na+ to glucose coupling of 1:1 and lack of galactose transport) generally matched those of the kidney cortical low affinity system. We show that expression of rat SGLT2 mRNA is kidney specific and that it is strongly and exclusively expressed in proximal tubule S1 segments. Hybrid-depletion studies were performed to conclusively determine whether SGLT2 corresponds to the kidney cortical low affinity system. Injection of rat kidney superficial cortex mRNA into oocytes stimulated the uptake of alpha-methyl-D-glucopyranoside (2 mM) 2-3-fold. We show that hybrid depletion of this kidney RNA using an SGLT2 antisense oligonucleotide completely suppresses the uptake. These data strongly indicate that SGLT2 is the major kidney cortical low affinity glucose transporter. We therefore propose that SAAT-pSGLT2 be renamed SGLT3. Experiments addressing the expression of SGLT1 and SGLT2 mRNAs in embryonic rat kidneys reveal that the two Na+/glucose cotransporters are developmentally regulated and that there may be a different splice variant for SGLT2 in embryonic kidney compared to the adult.

Age Factors↗

PubMed1995

Ultrastructural changes following overhydration with irrigating fluids.

We studied ultrastructural alterations caused by intravenous infusion of 150 ml/kg of irrigating fluid containing mannitol and/or sorbitol in 15 rabbits. The osmolality of the solutions varied between 165 and 350 mosmol/kg. Specimens for electron microscopic examination were obtained from the kidney, brain and lung. Regardless of the choice of solute, the hypo-osmotic solutions caused damage to kidney and brain tissue by inducing marked interstitial and cellular oedema. No such changes were observed after administration of isosmotic irrigating fluid. We conclude that ultrastructural changes following overhydration with irrigating fluid vary with the osmolality of the solution but not with the choice of solute. Moreover, the consistent absence of tissue damage in the lungs suggests that the lung is not a primary target organ for the "TUR syndrome".

Animals↗

PubMed1995

Mammalian ion-coupled solute transporters.

Active transport of solutes into and out of cells proceeds via specialized transporters that utilize diverse energy-coupling mechanisms. Ion-coupled transporters link uphill solute transport to downhill electrochemical ion gradients. In mammals, these transporters are coupled to the co-transport of H+, Na+, Cl- and/or to the countertransport of K+ or OH-. By contrast, ATP-dependent transporters are directly energized by the hydrolysis of ATP. The development of expression cloning approaches to select cDNA clones solely based on their capacity to induce transport function in Xenopus oocytes has led to the cloning of several ion-coupled transporter cDNAs and revealed new insights into structural designs, energy-coupling mechanisms and physiological relevance of the transporter proteins. Different types of mammalian ion-coupled transporters are illustrated by discussing transporters isolated in our own laboratory such as the Na+/glucose co-transporters SGLT1 and SGLT2, the H(+)-coupled oligopeptide transporters PepT1 and PepT2, and the Na(+)- and K(+)-dependent neuronal and epithelial high affinity glutamate transporter EAAC1. Most mammalian ion-coupled organic solute transporters studied so far can be grouped into the following transporter families: (1) the predominantly Na(+)-coupled transporter family which includes the Na+/glucose co-transporters SGLT1, SGLT2, SGLT3 (SAAT-pSGLT2) and the inositol transporter SMIT, (2) the Na(+)- and Cl(-)-coupled transporter family which includes the neurotransmitter transporters of gamma-amino-butyric acid (GABA), serotonin, dopamine, norepinephrine, glycine and proline as well as transporters of beta-amino acids, (3) the Na(+)- and K(+)-dependent glutamate/neurotransmitter family which includes the high affinity glutamate transporters EAAC1, GLT-1, GLAST, EAAT4 and the neutral amino acid transporters ASCT1 and SATT1 reminiscent of system ASC and (4) the H(+)-coupled oligopeptide transporter family which includes the intestinal H(+)-dependent oligopeptide transporter PepT1.

Amino Acid Sequence↗

PubMed1995

Cloning and regulation of expression of the rat kidney urea transporter (rUT2).

In mammals, urea is the predominant end-product of nitrogen metabolism and plays a central role in the urinary-concentrating mechanism. Urea accumulation in the renal medulla is critical to the ability of the kidney to concentrate urine to an osmolality greater than systemic plasma. Regulation of urea excretion and accumulation in the renal medulla depends on the functional state of specialized phloretin-sensitive urea transporters. To study these transporters and their regulation of expression we isolated a cDNA which encodes the rat homologue (rUT2) of rabbit UT2 (You, G., C.P. Smith, Y. Kanai, W.-S. Lee, M. Stelzner, and M.A. Hediger, et al. Nature (Lond.). 1993. 365:844-847). Rat UT2 has 88% amino acid sequence identity to rabbit UT2 and 64% identity to the recently cloned human erythrocyte urea transporter, HUT11 (Olives, B., P. Neav, P. Bailly, M.A. Hediger, G. Rousselet, J.P. Cartron, and P. Ripoch J. Biol. Chem. 1994. 269:31649-31652). Analysis of rat kidney mRNA revealed two transcripts of size 2.9 and 4.0 kb which had spatially distinct distributions. Northern analysis and in situ hybridization showed that the 4.0-kb transcript was primarily responsive to changes in the protein content of the diet whereas the 2.9-kb transcript was responsive to changes in the hydration state of the animal. These studies reveal that the expression levels of the two rUT2 transcripts are modulated by different pathways to allow fluid and nitrogen balance to be regulated independently. Our data provide important insights into the regulation of the renal urea transporter UT2 and provide a basis on which to refine our understanding of the urinary concentrating mechanism and its regulation.

Amino Acid Sequence↗

PubMed1994

The human kidney low affinity Na+/glucose cotransporter SGLT2. Delineation of the major renal reabsorptive mechanism for D-glucose.

The major reabsorptive mechanism for D-glucose in the kidney is known to involve a low affinity high capacity Na+/glucose cotransporter, which is located in the early proximal convoluted tubule segment S1, and which has a Na+ to glucose coupling ratio of 1:1. Here we provide the first molecular evidence for this renal D-glucose reabsorptive mechanism. We report the characterization of a previously cloned human kidney cDNA that codes for a protein with 59% identity to the high affinity Na+/glucose cotransporter (SGLT1). Using expression studies with Xenopus laevis oocytes we demonstrate that this protein (termed SGLT2) mediates saturable Na(+)-dependent and phlorizin-sensitive transport of D-glucose and alpha-methyl-D-glucopyranoside (alpha MeGlc) with Km values of 1.6 mM for alpha MeGlc and approximately 250 to 300 mM for Na+, consistent with low affinity Na+/glucose cotransport. In contrast to SGLT1, SGLT2 does not transport D-galactose. By comparing the initial rate of [14C]-alpha MeGlc uptake with the Na(+)-influx calculated from alpha MeGlc-evoked inward currents, we show that the Na+ to glucose coupling ratio of SGLT2 is 1:1. Using combined in situ hybridization and immunocytochemistry with tubule segment specific marker antibodies, we demonstrate an extremely high level of SGLT2 message in proximal tubule S1 segments. This level of expression was also evident on Northern blots and likely confers the high capacity of this glucose transport system. We conclude that SGLT2 has properties characteristic of the renal low affinity high capacity Na+/glucose cotransporter as previously reported for perfused tubule preparations and brush border membrane vesicles. Knowledge of the structural and functional properties of this major renal Na+/glucose reabsorptive mechanism will advance our understanding of the pathophysiology of renal diseases such as familial renal glycosuria and diabetic renal disorders.

Animals↗

PubMed1993

Cloning and characterization of the vasopressin-regulated urea transporter.

Urea is the principal end product of nitrogen metabolism in mammals. Movement of urea across cell membranes was originally thought to occur by lipid-phase permeation, but recent studies have revealed the existence of specialized transporters with a low affinity for urea (Km > 200 mM)2. Here we report the isolation of a complementary DNA from rabbit renal medulla that encodes a 397-amino-acid membrane glycoprotein, UT2, with the functional characteristics of the vasopressin-sensitive urea transporter previously described in in vitro-perfused inner medullary collecting ducts. UT2 is not homologous to any known protein and displays a unique pattern of hydrophobicity. Because of the central role of this transporter in fluid balance and nitrogen metabolism, the study of this protein will provide important insights into the urinary concentrating mechanism and nitrogen balance.

Amino Acid Sequence↗