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Tong Wang

Publications and source records attributed to Tong Wang.

At least 73 records · Page 4Linked to original sources

Renal and intestinal transport defects in Slc26a6-null mice.

SLC26A6 (PAT1, CFEX) is an anion exchanger that is expressed on the apical membrane of the kidney proximal tubule and the small intestine. Modes of transport mediated by SLC26A6 include Cl-/formate exchange, Cl-/HCO3- exchange, and Cl-/oxalate exchange. To study its role in kidney and intestinal physiology, gene targeting was used to prepare mice lacking Slc26a6. Homozygous mutant Slc26a6-/- mice appeared healthy and exhibited a normal blood pressure, kidney function, and plasma electrolyte profile. In proximal tubules microperfused with a low-HCO3-/high-Cl- solution, the baseline rate of fluid absorption (Jv), an index of NaCl transport under these conditions, was the same in wild-type and null mice. However, the stimulation of Jv by oxalate observed in wild-type mice was completely abolished in Slc26a6-null mice (P<0.05). Formate stimulation of Jv was partially reduced in null mice, but the difference from the response in wild-type mice did not reach statistical significance. Apical membrane Cl-/base exchange activity, assayed with the pH-sensitive dye BCPCF in microperfused proximal tubules, was decreased by 58% in Slc26a6-/- animals (P<0.001 vs. wild types). In the duodenum, the baseline rate of HCO3- secretion measured in mucosal tissue mounted in Ussing chambers was decreased by approximately 30% (P<0.03), whereas the forskolin-stimulated component of HCO3- secretion was the same in wild-type and Slc26a6-/- mice. We conclude that Slc26a6 mediates oxalate-stimulated NaCl absorption, contributes to apical membrane Cl-/base exchange in the kidney proximal tubule, and also plays an important role in HCO3- secretion in the duodenum.

Animals↗

NHE2-mediated bicarbonate reabsorption in the distal tubule of NHE3 null mice.

NHE3(-/-) mice display a profound defect in proximal tubule bicarbonate reabsorption but are only mildly acidotic owing to reduced glomerular filtration rate and enhanced H(+) secretion in distal nephron segments. In vivo microperfusion of rat distal tubules suggests that a significant fraction of bicarbonate reabsorption in this nephron segment is mediated by NHE2. Two approaches were used to evaluate the role of distal tubule NHE2 in compensating for the proximal defect of H(+) secretion in NHE3(-/-) mice. First, renal clearance experiments were used to assess the impact of HOE694, an inhibitor with significant affinity for NHE2, on excretion of bicarbonate in NHE3(-/-) and NHE2(-/-) mice. Second, in vivo micropuncture and microperfusion were employed to measure the concentration of bicarbonate in early distal tubule fluid and to measure distal bicarbonate reabsorption during a constant bicarbonate load. Our data show that HOE694 had no effect on urinary bicarbonate excretion in NHE3(+/+) mice, whereas bicarbonate excretion was higher in NHE3(-/-) mice receiving HOE694. HOE694 induced a significant increase in bicarbonate excretion in mice given an acute bicarbonate load, but there was no effect during metabolic acidosis. Bicarbonate excretion was not affected by HOE694 in bicarbonate-loaded NHE2(-/-) mice. In vivo micropuncture revealed that early distal bicarbonate concentration was elevated in both bicarbonate-loaded and NHE3(-/-) mice. Further, microperfusion experiments showed that HOE694-sensitive bicarbonate reabsorption capacity was higher in acidotic and NHE3 null animals. We conclude that NHE2 contributes importantly to acidification in the distal tubule, and that it plays a major role in limiting urinary bicarbonate losses in states in which a high luminal bicarbonate load is presented to the distal tubule, such as in NHE3 null mice.

Acidosis↗

Mechanosensory function of microvilli of the kidney proximal tubule.

Normal variations in glomerular filtration induce proportional changes in proximal tubule Na+ reabsorption. This "glomerulotubular balance" derives from flow dependence of Na+ uptake across luminal cell membranes; however, the underlying physical mechanism is unknown. Our hypothesis is that flow-dependent reabsorption is an autoregulatory mechanism that is independent of neural and hormonal systems. It is signaled by the hydrodynamic torque (bending moment) on epithelial microvilli. Such signals need to be transmitted to the terminal web to modulate Na+-H+-exchange activity. To investigate this hypothesis, we examined Na+ transport and tubular diameter in response to different flow rates during the microperfusion of isolated S2 proximal tubules from mouse kidneys. The data were analyzed by using a mathematical model to estimate the microvillous torque as function of flow. In this model, increases in luminal diameter have the effect of blunting the impact of flow velocity on microvillous shear stress and, thus, microvillous torque. We found that variations in microvillous torque produce nearly identical fractional changes in Na+ reabsorption. Furthermore, the flow-dependent Na+ transport is increased by increasing luminal fluid viscosity, diminished in Na+-H+ exchanger isoform 3 knockout mice, and abolished by nontoxic disruption of the actin cytoskeleton. These data support our hypothesis that the "brush-border" microvilli serve a mechanosensory function in which fluid dynamic torque is transmitted to the actin cytoskeleton and modulates Na+ absorption in kidney proximal tubules.

Animals↗

Paranemic crossover DNA: a generalized Holliday structure with applications in nanotechnology.

Paranemic crossover (PX) DNA is a four-stranded coaxial DNA complex containing a central dyad axis that relates two flanking parallel double helices. The strands are held together exclusively by Watson-Crick base pairing. The key feature of the structure is that the two adjacent parallel DNA double helices form crossovers at every point possible. Hence, reciprocal crossover points flank the central dyad axis at every major or minor groove separation. This motif has been modeled and characterized in an oligonucleotide system; a minor groove separation of five nucleotide pairs and major groove separations of six, seven, or eight nucleotide pairs produce stable PX DNA molecules; a major groove separation of 9 nucleotide pairs is possible at low concentrations. Every strand undergoes a crossover every helical repeat (11, 12, 13, or 14 nucleotides), but the structural period of each strand corresponds to two helical repeats (22, 24, 26, or 28 nucleotides). Nondenaturing gel electrophoresis shows that the molecules are stable, forming well-behaved complexes. PX DNA can be produced from closed dumbbells, demonstrating that the molecule is paranemic. Ferguson analysis indicates that the molecules are similar in shape to DNA double crossover molecules. Circular dichroism spectra are consistent with B-form DNA. Thermal transition profiles suggest a premelting transition in each of the molecules. Hydroxyl radical autofootprinting analysis confirms that there is a crossover point at each of the positions expected in the secondary structure. These molecules are generalized Holliday junctions.

Circular Dichroism↗

Mechanical stimuli induce cleavage and nuclear translocation of the polycystin-1 C terminus.

Polycystin-1, which is encoded by a gene that is mutated in autosomal dominant polycystic kidney disease (ADPKD), is involved in cell-matrix interactions as well as in ciliary signaling. The precise mechanisms by which it functions, however, remain unclear. Here we find that polycystin-1 undergoes a proteolytic cleavage that releases its C-terminal tail (CTT), which enters the nucleus and initiates signaling processes. The cleavage occurs in vivo in association with alterations in mechanical stimuli. Polycystin-2, the product of the second gene mutated in ADPKD, modulates the signaling properties of the polycystin-1 CTT. These data reveal a novel pathway by which polycystin-1 transmits messages directly to the nucleus.

Amino Acid Sequence↗

Expression of ENaC and serum- and glucocorticoid-induced kinase 1 in the rat intestinal epithelium.

Increase in epithelium sodium channel (ENaC) activity induced by aldosterone in the distal tubule of the kidney has been attributed to serum- and glucocorticoid-induced kinase 1 (sgk1). The distal colon constitutes another classical aldosterone-responsive epithelium that expresses both ENaC and sgk1 in an aldosterone-dependent manner. However, the site of expression and the temporal relationship of the aldosterone induction of these two proteins have not been investigated. Here, we examined the distribution and abundance of sgk1 in the rat intestine under basal conditions and after changes in the concentration of aldosterone and glucocorticoids. Results indicate that sgk1 is expressed in the distal colon and also in the ileum and jejunum. Abundance of sgk1 was high in control animals, and it did not change significantly after sodium depletion or after a single dose of aldosterone; however, it decreased after adrenalectomy. In contrast, the three subunits of ENaC were markedly induced in the distal colon by acute and chronic increases in aldosterone levels. Results indicate differential regulation of sgk and ENaC subunits by aldosterone in the distal colon. Distribution of sgk1 in the intestine beyond the aldosterone-responsive segments suggests that sgk1 may additionally regulate other sodium transporters in the intestinal epithelium.

Aldosterone↗

ROMK is required for expression of the 70-pS K channel in the thick ascending limb.

Apical potassium recycling is crucial for salt transport by the thick ascending limb (TAL). Loss-of-function mutations in the K channel, ROMK (Kir1.1; KCNJ1), cause Bartter syndrome, a genetically heterogeneous disorder characterized by severe reduction in salt absorption by the TAL, Na wasting, polyuria, and hypokalemic alkalosis. ROMK(-/-) null mice exhibit a Bartter phenotype and lack the small-conductance (30-pS) apical K channel (SK) in the TAL. However, a distinct 70-pS K channel can also significantly contribute to the apical conductance of TAL. We now examine the effect of ROMK deletion on the functional expression of the 70-pS K channel in the TAL. Functional expression of the 70-pS K channel was low [average channel activity (NP(o)) = 0.02] in ROMK(+/+) mice on a control K diet but increased to 0.27 by high-K intake for 2 wk. In contrast, the high-K diet decreased NP(o) of SK by approximately 30%, from 2.04 to 1.44. In ROMK heterozygous (+/-) mice on a control K diet, SK activity was about one-half of that observed in ROMK(+/+) mice (0.95 vs. 2.04). The high-K diet also reduced SK activity in ROMK(+/-) mice by approximately 40% (from 0.95 to 0.55) but increased NP(o) of the 70-pS K channel from 0 to 0.09 in ROMK(+/-) mice. This corresponds to approximately 30% of channel activity (NP(o) = 0.27) observed in ROMK(+/+) mice. Neither the 70-pS nor the 30-pS K channels were observed in TAL cells from ROMK(-/-) mice on either the normal or high-K diets. Thus functional expression of the 70-pS K channel is enhanced by increasing dietary K and requires expression of ROMK. It is likely that ROMK forms a critical subunit of the 70-pS K channel, accounting for the loss of apical K secretory channel activity in ROMK Bartter syndrome.

Administration, Oral↗

Intracellular Na concentration and Rb uptake in proximal convoluted tubule cells and abundance of Na/K-ATPase alpha1-subunit in NHE3-/- mice.

Proximal solute and fluid absorption is greatly reduced in mice in which the gene encoding the Na/H exchanger isoform 3 has been ablated (NHE3-/-). To obtain information on the intracellular functional consequences of such selective NHE3 deficiency, Na, Cl and K concentrations and cell Rb uptake were measured using electron microprobe analysis after a 30-s infusion of Rb (an index of basolateral Na/K-ATPase activity) in proximal convoluted tubule (PCT) cells of NHE3-/- and wild-type (NHE3+/+) mice. In addition, the relative abundance of the alpha1-subunit of the Na/K-ATPase in the outer cortex was determined by Western blot analysis. PCT cell Na concentration in NHE3-/- mice was slightly but significantly lower than in NHE3+/+ [13.1+/-0.6 ( n=64) vs. 14.9+/-0.6 ( n=62) mmol/kg wet wt.; means +/-SEM]. The lower intracellular Na concentration was associated with significantly reduced Rb uptake rates [9.7+/-0.6 ( n=59) vs. 14.8+/-0.8 ( n=50) mmol/kg wet wt./30 s], but the abundance of the alpha1-subunit of the Na/K-ATPase was not different between NHE3-/- and NHE3+/+ mice. Intracellular Cl concentration was higher (14.2+/-0.4 vs. 12.8+/-0.4 mmol/kg wet wt.) and K concentration unchanged (122.7+/-2.7 vs. 121.6+/-2.5 mmol/kg wet wt.) in PCT cells in NHE3-/- compared with NHE3+/+ mice. These findings suggest that the elimination of apical NHE3 in PCT cells of NHE3-/- mice reduces apical Na entry and, due to lower cell Na concentrations, Na/K-ATPase activity. The observed changes in intracellular Na concentration did not affect the expression of Na/K-ATPase in the renal cortex of NHE3-/- mice. There were no significant changes of cell Na concentration and Rb uptake in distal convoluted tubule, connecting tubule, principal and intercalated cells.

Animals↗

Role of PKC and calcium in modulation of effects of angiotensin II on sodium transport in proximal tubule.

It has been well documented that low concentrations of ANG II (10(-11) to 10(-10) M) stimulate, whereas high concentrations of ANG II (10(-8) to 10(-5) M) inhibit Na(+) transport in proximal tubules of rat and rabbit kidneys. Measured ANG II concentration in proximal tubular fluid is in the nanomolar range. In the present study, we investigated the role of PKC, intracellular Ca(2+), and cAMP in modulating the effects of luminal ANG II on Na(+) absorption by microperfusion techniques in rabbit superficial segment of proximal tubules in vitro. We confirmed that ANG II (10(-9) M) had no change on fluid absorption (J(v)); however, fluid absorption increased significantly when 10(-9) M ANG II and 3,4,5-trimethoxybenzoic acid-8-(diethylamino)octyl ester (TMB-8), a blocker of intracellular calcium mobilization, were added together. In contrast, ANG II significantly decreased J(v) when PKC was inhibited. When 10(-9) M ANG II was present together with 1-(5-isoquinolinesulfonyl)-2-mehtylpiperazine and TMB-8, no significant change of J(v) occurred. Inhibition of endogenous cAMP activity by a PKA inhibitor did not change either basal or ANG II-stimulated fluid absorption. Our results indicate that ANG II regulates Na(+) absorption by a cAMP-independent mechanism and that PKC and intracellular calcium both play a critical role in modulating the effects of physiological concentration of ANG II on proximal tubule transport. Balance between these two cytosolic messengers modulates the effects of ANG II on fluid absorption in the proximal tubule.

Angiotensin II↗

Metabolic partitioning of endogenous fatty acid in adipocytes.

OBJECTIVE: To develop an accurate new method to measure the partitioning of adipocyte endogenous fatty acids among different metabolic pathways, a critical step toward understanding the regulatory mechanism by which fat disposition is modulated. RESEARCH METHODS AND PROCEDURES: Isolated primary rat adipocytes were pre-incubated with isotope-labeled fatty acids. This allows determination of the specific activity of labeled fatty acids in the endogenous lipid pool. After the removal of exogenous fatty acids, the disposition of endogenous fatty acids into the three major metabolic pathways, namely, oxidation, re-esterification, and release into the medium, was measured independently. This was compared with the total lipolytic release of endogenous fatty acids, as measured by glycerol release. Adipocytes from normal fed and fasted animals were used to determine the effects of physiological variations on the metabolic fate of endogenous fatty acids. RESULTS: In normal fed animals, 0.2% of endogenous fatty acids were oxidized, 50.1% were released, and 49.7% were re-esterified. Fasting doubled the partitioning of fatty acids toward oxidation (p < 0.05) in association with increased lipolysis (1.4-fold increase) (p < 0.05). This effect was completely abolished by the addition of insulin to the cells (61% reduction) (p < 0.05). DISCUSSION: The endogenous fatty acids in adipocytes are actively oxidized. This process can be regulated by altered physiological conditions or by insulin. Over time, it is possible that a small shift of fatty acids toward oxidation could have a significant impact on body fuel economy. This hypothesis needs to be tested.

Adipocytes↗

Octanoate inhibits triglyceride synthesis in 3T3-L1 and human adipocytes.

To understand how medium-chain fatty acids (FA) influence lipid metabolism in adipocytes, we studied the effects of octanoate on the oxidation of glucose and endogenous palmitate, cellular O(2) consumption, mitochondrial membrane potential, lipid synthesis from long-chain FA, glucose and lactate. We found that octanoate significantly suppressed the esterification of oleate into triglycerides (TG) in both 3T3-L1 and human adipocytes. Octanoate also significantly suppressed de novo FA synthesis. These effects were associated with octanoate-mediated reductions in the activities of acyl CoA:1,2-diacylglycerol acyltransferase (DGAT) and acetyl CoA carboxylase (ACC). Cells pretreated with octanoate had reduced mRNA levels for a number of lipid metabolism genes, including of DGAT, ACC and stearoyl CoA desaturase-1. On the other hand, octanoate did not acutely perturb cellular O(2) consumption or mitochondrial membrane potential. Together, these results suggest that octanoate affected adipocyte function by reducing TG synthesis but not by enhancing oxidation.

3T3 Cells↗

The effects of the potassium channel opener minoxidil on renal electrolytes transport in the loop of henle.

ATP-sensitive potassium channels (K(ATP)) in the thick ascending limb of the loop of Henle play an important role in apical K(+) recycling, a mechanism essential for maintaining the activity of the Na/2Cl/K-cotransporter. We have previously demonstrated that inhibition of K(ATP) decreases Na(+) and K(+) absorption in the loop of Henle and induces diuretic and natriuretic effects. In the present study, we used renal clearance and in vivo microperfusion techniques to evaluate the effects of the K(ATP) opener minoxidil on the urinary excretion and absorption in the loop of Henle of Na(+), K(+), Ca(2+), and Mg(2+). Intravenous injection of minoxidil (1.5 mg/kg) significantly decreased fractional Na(+) (FENa) and Mg(2+) (FEMg) excretion and urine volume with a moderate decrease in blood pressure (12%) and glomerular filtration rate (15%). Urine volume decreased 63%, and FENa and FEMg decreased 58 and 37%, respectively. In contrast, K(+) and Ca(2+) excretion did not change significantly. In the microperfusion of the loop of Henle, addition of minoxidil to the perfusion fluid significantly increased fluid (J(v)), Na(+) (J(Na)), Cl(-) (J(Cl)), and K(+) (J(K)) absorption. J(v) increased 44% (from 8.32 to 11.95 nl/min), J(Na) increased 14% (from 1.96 to 2.34 nmol/min), J(Cl) increased 21% (from 1.72 to 2.08 nmol/min), and J(K) increased 57% (from 35.8 to 56.4 pmol/min). We conclude that the activation of K(ATP) leads to stimulation of Na/2Cl/K-cotransporter activity and increases the rates of Na(+), Cl(-), and K(+) absorption in the loop of Henle, an effect contributing to the antidiuretic and antinatriuretic action of this K channel opener.

Animals↗

Inhibition of heme oxygenase decreases sodium and fluid absorption in the loop of Henle.

We previously demonstrated that carbon monoxide (CO) stimulates the apical 70-pS K+ channel in the thick ascending limb (TAL) of the rat kidney (Liu HJ, Mount DB, Nasjletti A, and Wang WH. J Clin Invest 103: 963-970, 1999). Because the apical K+ channel plays a key role in K+ recycling, we tested the hypothesis that heme oxygenase (HO)-dependent metabolites of heme may affect Na+ transport in the TAL. We used in vivo microperfusion to study the effect of chromium mesoporphyrin (CrMP), an inhibitor of HO, on fluid absorption (Jv) and Na+ absorption (JNa) in the loop of Henle and renal clearance methods to examine the effect of CrMP on renal sodium excretion. Microperfusion experiments demonstrated that addition of CrMP to the loop of Henle decreased Jv by 13% and JNa by 20% in animals on normal rat chow and caused a decrease in Jv (39%) and JNa (40%) in rats on a high-K+ (HK) diet. The effect of CrMP is the result of inhibition of HO because addition of MgPP, an analog of CrMP that does not inhibit HO, had no effect on Jv. Western blot analysis showed that HO-2 is expressed in the kidney and that the level of HO-2 was significantly elevated in animals on a HK diet. Renal clearance studies demonstrated that the infusion of CrMP increased the excretion of urinary Na+ (ENa) and volume (UV) without changes in glomerular filtration rate. The effect of CrMP on ENa and UV was larger in HK rats than those kept on normal chow. We conclude that HK intake increases HO-2 expression in the kidney and that HO-dependent metabolites of heme, presumably CO, play a significant role in the regulation of Na+ transport in the loop of Henle.

Absorption↗

Effect of lidocaine on ovalbumin and egg albumin foam stability.

Foam fractionation is a simple separation process that can remove and concentrate hydrophobic molecules such as proteins, surfactants, and organic wastes from an aqueous solution. Bovine serum albumin and ovalbumin have been widely used as model proteins due to their strong foaming potential and low price. Here, we study the effect of lidocaine on albumin foam, since drugs like lidocaine are known to bind with albumin. We observed that lidocaine not only enhances the amount of foam produced but also the stability of that foam as well. The foam stability was evaluated as the decay rate constant of the foam, determined from a change in height (or volume) of the foam over a given time period.

Antifoaming Agents↗

[Research report of experimental database establishment of digitized virtual Chinese No.1 female].

OBJECTIVE: To establish digitized virtual Chinese No.1 female (VCH-F1) image database. METHODS: A 19 years old female cadaver was scanned by CT, MRI, and perfused with red filling material through formal artery before freezing and em- bedding. The whole body was cut by JZ1500A vertical milling machine with a 0.2 mm inter-spacing. All the images was produced by Fuji FinePix S2 Pro camera. RESULTS: The body index of VCH-F1 was 94%. We cut 8 556 sections of the whole body, and each image was 17.5 MB in size and the whole database reached 149.7 GB. We have totally 6 versions of the database for different applications. CONCLUSIONS: Compared with other databases, VCH-F1 has good representation of the Chinese body shape, colorful filling material in blood vessels providing enough information for future registration and segmentation. Vertical embedding and cutting helped to retain normal human physiological posture, and the image quality and operation efficiency were improved by using various techniques such as one-time freezing and fixation, double-temperature icehouse, large-diameter milling disc and whole body cutting.

Anatomy, Cross-Sectional↗

[Effect of particle size distribution of dental poly (methyl methacrylate) beads on the microstructure of denture].

OBJECTIVES: To investigate the effect of particle size distribution (PSD) of dental poly (methyl methacrylate) (PMMA) beads on the microstructure of denture. METHODS: It was compared and analyzed that the porosity of three groups of spherical specimens which were made of PMMA beads with different PSD at proper polymer/monomer ratios. And the final microstructure of each group was investigated using Digital High Definition Microscope. RESULTS: With proper polymer/monomer ratio increasing, the occurrence of porosity tended to decrease. The final microstructure of denture varied with PSD of PMMA beads. CONCLUSION: By means of optimizing PSD of beads, the microstructure of denture could be improved, and the occurrence of porosity could be reduced.

Dental Materials↗

Control effects of p(epsilon) and pH on the generation and stability of chlorine dioxide.

A new method, without assistance of activity ratio diagram, was applied to construct the p(epsilon)-pH diagrams for chlorine system. The optimal pH range for generation of C1O2 by contacting Cl2 (g) directly with ClO2- solution is within pH 1.35-1.94, particularly within pH 1.35-4.00 only if minimizing the formation of Cl2. It is unachievable to synthesize pure ClO2 from the reaction of Cl2 and ClO2- . Conversely, ClO2 may be present a variation of stability in different waters owing to the changed p(epsilon) and pH. ClO2 could be relatively stable if not disproportionate into ClO3-, coexisting with ClO2- (p(epsilon) 17.63 and pH > 9.68), Cl2(pH < or = 0.92) or Cl- (pH 0.92-9.68). When chlorine system has already reached the ultimate equilibria, ClO2 is a stable species in strongly acid media. As the acidity decreases, ClO2 disproportionates into ClO3- and Cl2. Aqueous ClO2 is unstable within the normal pH range. This work initially, theoretically elucidates the generation and stability of ClO2 by way of the p(epsilon)-pH diagrams.

Chlorine Compounds↗

[Rheological properties of photo-polymerized composite resin reinforced with retentive filler].

OBJECTIVE: To determine the rheological properties of the photo-polymerized composite resin reinforced with retentive filler (RF) and its rheological difference with normal filler (NF) composite resin. METHODS: Rheological properties of the composite resins, such as viscosity, shear stress and creep compliance, were measured with dynamic stress rheometer at room temperature (25 degrees C). RESULTS: The viscosity of the composite resin reinforced with RF is higher than that of NF composite resin (P < 0.01); at the beginning, the viscosity of the former has little change with the rising of shear stress and the latter decreases, then the viscosities of the two composite resins increase with the rising of shear stress as soon as over 203.18 Pa of shear stress; the creep compliance of the composite resin reinforced with RF is significant smaller than that of NF composite resin (P < 0.01). CONCLUSIONS: The rheological properties of the two composite resins have significant difference, so they have different clinical using properties.

Composite Resins↗