Na transport in sheep rumen is modulated by voltage-dependent cation conductance in apical membrane.
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The effects of clamping the transepithelial potential difference (PDt; mucosa reference) have been studied in sheep rumen epithelium. Pieces of ruminal epithelium were examined in Ussing chambers, in a part of the experiments combined with conventional intracellular recordings. After equilibration, the tissue conductance (Gt) was 2.50 +/- 0.09 mS/cm(2), the potential difference of the apical membrane (PD(a)) was -47 +/- 2 mV, and the fractional resistance of the apical membrane (fRa) was 68 +/- 2% under short-circuit conditions. Hyperpolarization of the tissue (bloodside positive) depolarized PDa, decreased fRa, and increased Gt significantly. Clamping PDt at negative values caused converse effects on PDa and fRa. All changes were completely reversible. The determination of individual conductances revealed that the conductance of the apical membrane increased almost linearly with depolarization of PDa. The PD-dependent changes were significantly reduced by total replacement of Na. These observations support the assumption of a PD-dependent conductance in the apical membrane that permits enhanced apical uptake of Na even at depolarized PDa. This mechanism appears to be important for the regulation of osmotic pressure in forestomach fluid. (+info)
Single- and multifrequency models for bioelectrical impedance analysis of body water compartments.
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The 1994 National Institutes of Health Technology Conference on bioelectrical impedance analysis (BIA) did not support the use of BIA under conditions that alter the normal relationship between the extracellular (ECW) and intracellular water (ICW) compartments. To extend applications of BIA to these populations, we investigated the accuracy and precision of seven previously published BIA models for the measurement of change in body water compartmentalization among individuals infused with lactated Ringer solution or administered a diuretic agent. Results were compared with dilution by using deuterium oxide and bromide combined with short-term changes of body weight. BIA, with use of proximal, tetrapolar electrodes, was measured from 5 to 500 kHz, including 50 kHz. Single-frequency, 50-kHz models did not accurately predict change in total body water, but the 50-kHz parallel model did accurately measure changes in ICW. The only model that accurately predicted change in ECW, ICW, and total body water was the 0/infinity-kHz parallel (Cole-Cole) multifrequency model. Use of the Hanai correction for mixing was less accurate. We conclude that the multifrequency Cole-Cole model is superior under conditions in which body water compartmentalization is altered from the normal state. (+info)
Reliability and validity of body composition measures in female athletes.
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The purpose of this investigation was to determine the reliability and validity of bioelectrical impedance (BIA) and near-infrared interactance (NIR) for estimating body composition in female athletes. Dual-energy X-ray absorptiometry was used as the criterion measure for fat-free mass (FFM). Studies were performed in 132 athletes [age = 20.4 +/- 1.5 (SD) yr]. Intraclass reliabilities (repeat and single trial) were 0.987-0.997 for BIA (resistance and reactance) and 0.957-0.980 for NIR (optical densities). Validity of BIA and NIR was assessed by double cross-validation. Because correlations were high (r = 0.969-0.983) and prediction errors low, a single equation was developed by using all 132 subjects for both BIA and NIR. Also, an equation was developed for all subjects by using height and weight only. Results from dual-energy X-ray absorptiometry analysis showed FFM = 49.5 +/- 6.0 kg, which corresponded to %body fat (%BF) of 20.4 +/- 3.1%. BIA predicted FFM at 49.4 +/- 5.9 kg (r = 0.981, SEE = 1.1), and NIR prediction was 49. 5 +/- 5.8 kg (r = 0.975, SEE = 1.2). Height and weight alone predicted FFM at 49.4 +/- 5.7 kg (r = 0.961, SEE = 1.6). When converted to %BF, prediction errors were approximately 1.8% for BIA and NIR and 2.9% for height and weight. Results showed BIA and NIR to be extremely reliable and valid techniques for estimating body composition in college-age female athletes. (+info)
Bioelectrical impedance: fat content of beef and pork from different size grinds.
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Multiple linear regression equations were developed for predicting the percentage of fat content of beef and pork. The predictor variables were bioelectrical resistance, temperature, and weight of product. Equations were developed for trim and product ground through a .95- or a .32-cm plate. The trim, .95-cm, and .32-cm grinds had 64, 108, and 96 observations, respectively, for beef product and 56, 101, and 92 observations, respectively, for pork product. Each of these observations was the average of bioelectrical impedance measurements taken in triplicate. The fat percentage ranges were 4 to 50% for beef and 7.5 to 50% for pork. The prediction equation applied to beef trim provided the following values: R2 = .80, Mallows's C(P) = 5.1, and root mean square error = 6.64. The R2 for equations predicting fat percentage in .95- and .32-cm ground beef were .84 and .95, respectively. The prediction equation applied to pork trim provided the following values: R2 = .77, Mallow's C(P) = 5.0, and root mean square error = 6.2. The R2 for equations predicting fat percentage in .95- and .32-cm ground pork were .87 and .96, respectively. The analyses were repeated with data sets of observations with less than 35% fat. The sample sizes and R2 for the trim, .95-, and .32-cm ground beef were 48, .36; 76, .60; and 65, .86; respectively. The sample sizes and R2 for the trim, .95-, and .32-cm ground pork were 42, .64; 62, .66; and 58, .92; respectively. Resistance, temperature, and weight remained as predictor variables for ground product with less than 35% fat. The smaller the grind, the more accurate the prediction. These results are positive for developing inexpensive, on-line systems for efficiently mixing ground product to a specific fat percentage. (+info)
A new experimental approach in endothelium-dependent pharmacological investigations on isolated porcine coronary arteries mounted for impedance planimetry.
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1. The aim of this study was to investigate whether the balloon-based impedance planimetry technique could be a useful tool in endothelium-dependent investigations. 2. Porcine large coronary arteries contracted with prostaglandin F2alpha (PGF2alpha, 10 microM) did not relax to bradykinin (0.1 nM - 0.1 microM), but did relax to sodium nitroprusside (SNP, 10 microM). However, after eversion of the segments, bradykinin induced relaxations with pD2 values and maximal responses of 8.78+/-0.09 and 75+/-2% (n=6), respectively. 3. Incubation with captopril (1 microM) did not reveal a relaxation to bradykinin in the normal vessel configuration and had no influence on the concentration-relaxation relationship in everted segments. 4. Lowering the luminal pressure in contracted segments from 131+/-5 mmHg (isometric, n=5) to 60 mmHg (isobaric, n=5) did not facilitate the action of bradykinin. 5. Eversion of segments did not influence the concentration-response relationship for K+ (4.7 - 125 mM), PGF2alpha (0.3 - 30 microM), and SNP (30 nM - 30 microM), although the time-courses of responses were faster when the agents were added from the intimal compared to the adventitial side of the preparation. 6. In the same everted segment contracted with PGF2alpha, the concentration-response relationship for bradykinin was not different under isometric and isobaric conditions. 7. These results indicate that, (1) reduced endothelium-dependent relaxations to adventitially administered substances can be ascribed to a diffusion barrier in the vessel wall, while enzymatic degradation, luminal pressure and precontractile responses seem not to play a role, (2) impedance planimetry applied to everted cylindrical segments could be a useful experimental approach in pharmacological studies of endothelium-dependent responses under isobaric and isometric conditions. (+info)
Measurement of fluid shift in CAPD patients using segmental bioelectrical impedance analysis.
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OBJECTIVE: To determine, using segmental bioelectrical impedance analysis (BIA), the characteristics of fluid shift of each body segment in continuous ambulatory peritoneal dialysis (CAPD) patients during and after peritoneal dialysis (PD) solution exchange. DESIGN: Observational study with repeated measurements of bioelectrical impedance, which is inversely related to tissue fluid content. PATIENTS: Thirteen clinically stable CAPD patients. METHODS: Bioelectrical impedance was measured at frequencies of 5, 50, 250, and 500 kHz in each body segment four times: (1) before and (2) after drainage of dialysate, and (3) at 1 hour and (4) at 2 hours after exchange of new 1.5% dextrose PD solution. RESULTS: Impedance of both arms was significantly increased at 1 hour post exchange at all frequencies. In the trunk, impedance at all frequencies increased significantly after drainage, decreased significantly at 1 hour post exchange, and then increased again for the next hour without significance. Impedance of both legs showed a decreasing tendency at all frequencies during and after exchange. Net calculated water volume changes between the time before drainage and 2 hours post exchange were -0.5 L in the trunk, -0.25 L in both arms, +0.47 L in both legs, -0.28 L in total. The change in body weight between the time before drainage and 2 hours post exchange was -0.21 kg, on average, and significantly correlated with total net calculated water volume change (p = 0.009). CONCLUSIONS: Each body segment of the CAPD patient has its own characteristic pattern of fluid shift in response to PD solution exchange or dwell. Segmental BIA may be a useful tool for understanding the physiological changes in fluid shift in CAPD patients. (+info)
Clostridium perfringens enterotoxin fragment removes specific claudins from tight junction strands: Evidence for direct involvement of claudins in tight junction barrier.
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Claudins, comprising a multigene family, constitute tight junction (TJ) strands. Clostridium perfringens enterotoxin (CPE), a single approximately 35-kD polypeptide, was reported to specifically bind to claudin-3/RVP1 and claudin-4/CPE-R at its COOH-terminal half. We examined the effects of the COOH-terminal half fragment of CPE (C-CPE) on TJs in L transfectants expressing claudin-1 to -4 (C1L to C4L, respectively), and in MDCK I cells expressing claudin-1 and -4. C-CPE bound to claudin-3 and -4 with high affinity, but not to claudin-1 or -2. In the presence of C-CPE, reconstituted TJ strands in C3L cells gradually disintegrated and disappeared from their cell surface. In MDCK I cells incubated with C-CPE, claudin-4 was selectively removed from TJs with its concomitant degradation. At 4 h after incubation with C-CPE, TJ strands were disintegrated, and the number of TJ strands and the complexity of their network were markedly decreased. In good agreement with the time course of these morphological changes, the TJ barrier (TER and paracellular flux) of MDCK I cells was downregulated by C-CPE in a dose-dependent manner. These findings provided evidence for the direct involvement of claudins in the barrier functions of TJs. (+info)
A role for intracellular calcium in tight junction reassembly after ATP depletion-repletion.
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The integrity of the tight junction (TJ), which is responsible for the permeability barrier of the polarized epithelium, is disrupted during ischemic injury and must be reestablished for recovery. Recently, with the use of an ATP depletion-repletion model for ischemia and reperfusion injury in Madin-Darby canine kidney cells, TJ proteins such as zonula occludens-1 (ZO-1) were shown to reversibly form large complexes and associate with cytoskeletal proteins (T. Tsukamoto and S. K. Nigam, J. Biol. Chem. 272: 16133-16139, 1997). In this study, we examined the role of intracellular calcium in TJ reassembly after ATP depletion-repletion by employing the cell-permeant calcium chelator 1, 2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-AM (BAPTA-AM). Lowering intracellular calcium during ATP depletion is associated with significant inhibition of the reestablishment of the permeability barrier following ATP repletion as measured by transepithelial electrical resistance and mannitol flux, marked alterations in the subcellular localization of occludin by immunofluorescent analysis, and decreased solubility of ZO-1 and other TJ proteins by Triton X-100 extraction assay, suggesting that lowering intracellular calcium potentiates the interaction of TJ proteins with the cytoskeleton. Coimmunoprecipitation studies indicated that decreased solubility may partly result from the stabilization of large TJ protein-containing complexes with fodrin. Although ionic detergents (SDS and deoxycholate) appeared to cause a dissociation of ZO-1-containing complexes from the cytoskeleton, sucrose gradient analyses of the solubilized proteins suggested that calcium chelation leads to self-association of these complexes. Together, these results raise the possibility that intracellular calcium plays an important facilitatory role in the reassembly of the TJ damaged by ischemic insults. Calcium appears to be necessary for the dissociation of TJ-cytoskeletal complexes, thus permitting functional TJ reassembly and paracellular permeability barrier recovery. (+info)