In traditional Chinese herb medicine, Alismatis rhizoma has been used in treating edema, inflammation and increasing urine flow. Mechanism of Alismatis rhizoma¡¦s effect on these functions has not been elucidated. Since diuresis has been considered closely related to the reabsorption of sodium ion via the epithelium of tubule and collecting duct in kidney, we suspected that Alismatis rhizoma may influence transportation of salt and water. The measurement of short circuit current ( Isc ) has been used widely to estimate the ion transportation between mucosal and serosal side of epithelium. In the present experiment, we used the voltage ¡V clamp technique to demonstrate the effect of Alismatis rhizoma¡¦s extracts ( ARE ) on Isc and sodium ion conductance in frog skin. Our result showed that in control stage, the potential difference ( PD ) of frog skin is 64.81 ¡Ó 2.44 mV, the Isc is 59.82 ¡Ó 3.58 £gA / cm2 and the conductance is 1.09 ¡Ó 0.18 £gA / cm2 / mV. After ARE was applied to mucosal side of the frog skin, its Isc decrease from 62.63 ¡Ó 5.31 £gA / cm2 to 47.92 ¡Ó 5.41 £gA / cm2, which could further go down to 3.36 ¡Ó 1.06 £gA/cm2 by adding amiloride. Treating serosal side of frog skin with ARE decreased approximately 10% of its Isc. No apparent changes in conductance were observed by adding ARE to mucosal ( 0.98 ¡Ó 0.24 £gA / cm2 / mV ) or serosal side ( 0.96 ¡Ó 0.23 £gA / cm2 / mV ). Adding L-arginine ( the precursor of nitric oxide ) to the serosal side of the skin epithelium elevated the Isc for 17.10 ¡Ó 9.30 £gA/cm2. This effect can be inhibited by applying ARE or NG-L-nitro-arginine methyl ester ( L-NAME, NO synthase inhibitor ) before application of L-arginine. In summary, Alismatis rhizoma could affect Isc on both mucosal and serosal sides of the frog skin. Its effect on lowering Isc was more obvious when applied to the mucosal side than to the serosal side. The ARE may exert its effect on mucosal side by affecting amiloride¡Vsensitive sodium channel and on serosal side by affecting the NO signal transduction pathway.

Rhei Rhizoma, also named as rhubarb or Da Huang, has been used widely in oriental traditional medicine in treating constipation and edema. However, though much affection has been paid to the make of components on pharmaceutical mechanisms, few studies have been conducted to reveal chemical and physical mechanism of these effects. Studies have shown that diarrhea causes imbalance of chloride and sodium ion movements via epithelium, we wondered if similar mechanism may apply to Rhei Rhizoma, a herbal drug which has been used to treat constipation in oriental medicine for thousands of years. The measurement of short-circuit current (Isc) has been used widely to estimate the ion transportation between mucosal and serosal side of epithelium. In this study, we used Ussing chamber technique to examine the alternation in membrane potential and short-circuit currents. The result shows, at default, the Isc of frog skin we used was at 59.23¡Ó5.58£gA/cm², and the conductance was at 1.11¡Ó0.50£gA/cm²¡EmV. The lnjection of 1ml RRE to mucosal side of the frog skin leaded to a 90% elevation of the Isc. Followed by the application of Amiloride (sodium channel inhibitor) and Chlorothiazide (chloride channel inhibitor) to mucosal side of the epithelial skin, the observed Isc were then reduced 136% and 33% respectively. If RRE were applied after the adding of Amiloride or Chlorothiazide to the frog skin, then the Isc of the skin elevated only 24% and 70% respectively. These results show that Rhei Rhizoma Extract (RRE) significantly increases Isc upon application to the mucosal side of the skin epithelium. Amiloride and Chlorothiazide will both inhibit the Isc induced by RRE, indicating activation of chloride channel and Amiloride-sensitive sodium channel of the epithelial tissue by RRE. After the regular Ringer solution used in the preparation was replaced by Na-free and Cl-free Ringer solution, the inhibition of Isc by RRE application could still be observed although the inhibition was trivia. These results indicate that RRE acts dominantly on mucosa side of the epithelium and can be used to enhance sodium transport and to stimulate the secretion of Cl- in the epithelium.

Summary Clmatis Armandi has been used frequently in traditional Chinese medicine for the treatment of diuretic symptoms. The mechanism of its action is unclear. Possible action of this substance may involve alternation of electrolyte transport through the epithelia membranes. In this study¡Atransepithelial conductance of frog skin was measured in vitro in voltage-clamped Ussing chambers. Adding Clematis Armandi extracts to apical surface induced a conductance increment of 1.21 £gS and an apical to serosal Isc of 28.78 £gA/cm2. The Isc can not be completely blocked by apical application of amiloride. Nifedipine and TEA had no effect on Clematis Armandi induced Isc decrease. These data indicate that frog skin is highly responsive to the concentrated Clematis Armandi extracts. The increase in Isc reflects changes in transepithelial transport of Na+ ions modulated at apical membrane. The enormous increase in transepithelial conductance suggests that in additional to enhancement of amiloride-sensitive Na+ channels, Clematis Armandi may also modulate other pathways, such as Cl- ion channel modulation, which needs further investigation.

Popularly used impedance-based methods need voltage and current waveform as well as line impedance per unit length to estimate distance to fault location. For a non-homogenous system with different line configuration, these methods assume that the system is homogenous and use the line impedance of the most frequently occurring line configuration. Load present in the system before fault is an important parameter which affects fault location accuracy. Impedance-based methods like Takagi and positive-sequence method assume that the load is lumped beyond the fault point which may not be true for a typical distribution system. As a result, accuracy of the impedance-based methods in estimating distance to fault is affected. Another short-coming of impedance-based methods are that they are unable to identify the branch in which the fault may be located. To minimize these errors, this thesis proposes a short-circuit fault current profile approach to complement impedance-based algorithms. In the short-circuit fault current profile approach, circuit model of the distribution feeder is used to place faults at every bus and the corresponding short-circuit fault current is plotted against reactance or distance to fault. When a fault occurs in the distribution feeder, fault current recorded by relay is extrapolated on the current profile to get location estimates. Since the circuit model is directly used in building the current profile, this approach takes into account load and non-uniform line impedance. Using the estimates from short-circuit fault current profile approach and impedance-based methods, the path on which the fault is located is identified. Next to improve fault location estimates, a median value of the estimates is computed. The median is a more robust estimate since it is not affected by outliers. The strategy developed above is tested using modified IEEE 34 Node Test Feeder and validated against field data provided by utilities. For the IEEE 34 Node Test Feeder, it is observed that the median estimate computed from impedance-based methods and the short-circuit fault current profile approach is very close to the actual fault location. Error in estimation is within 0.58 miles. It was also observed that if a 0.6 mile radius is built around the median estimate, the fault will lie within that range. Now the IEEE 34 Node Test Feeder represents a typical distribution feeder and has also been modeled to represent the worst case scenario, i.e. load current is around 51% of the fault current for the farthest bus. Hence the 0.6 mile radius around the median estimate will hold true for most distribution feeders and will be used when computing the fault range for field case events. For the field events, it was seen that the actual faults indeed lie within the 0.6 mile radius built around the median estimate and the path of the fault location has also been accurately estimated. For certain events, voltage waveform was not useful for analysis. In such situations, short-circuit fault current profile alone could be used to estimate fault location. Error in estimation is within 0.1 miles, provided the circuit model closely represents the distribution feeder. / text

The thesis deals with the problematics of short-circuit current of synchronous machine. The first section focuses on the theoretical process of short-circuit current of synchronous machine and describes how this transient process changes throughout it's duration and with what reactances and time constants it is defined. Subsequently, this section deals with the methodics of tests of the snychronous machine,which may be used to set it's reactances. The second part of the work focuses on measurement of the short-circuit current in a specific synchronous machine. Further, there are measurements of processes of the short-circuit in time and their analysis. The last section of the thesis deals with the simulation of short-circuit current process. The simulation is ran by the Matlab/Simulink software. Simulated processes of short-circuit are then compared with the results of measurements from the previous section of the thesis.

Doctor of Philosophy / Department of Anatomy and Physiology / Bruce D. Schultz / The first aim of this project is to define the cellular mechanisms that account for the low Na[superscript]+ concentration in human milk. MCF10A cells, which were derived from human mammary epithelium and grown on permeable supports, exhibit amiloride- and benzamil-sensitive short circuit current (I[subscript]sc), suggesting activity of the epithelial Na[superscript]+ channel, ENaC. When cultured in the presence of cholera toxin (Ctx), MCF10A cells exhibit greater amiloride sensitive I[subscript]sc at all time points tested, an effect that is not reduced with Ctx washout for 12 hours or by cytosolic pathways inhibitors. Ctx increases the abundance of both beta and gamma-ENaC in the apical membrane and increases its monoubiquitination but without changing total protein and mRNA levels. Additionally, Ctx increases the levels of both the phosphorylated and the nonphosphorylated forms of Nedd4-2, a ubiquitin-protein ligase that regulates ENaC degradation. The results reveal a novel mechanism in human mammary gland epithelia by which Ctx regulates ENaC-mediated Na[superscript]+ transport. The second project aim is to develop a protocol to isolate mammary gland epithelia for subsequent in vitro culture. Caprine (1[superscript]0CME) and bovine mammary epithelia (1[superscript]0BME) were isolated and cultured on permeable supports to study hormone- and neurotransmitter-sensitive ion transport. Both 1[superscript]0CME and 1[superscript]0BME cells were passed for multiple subcultures and all passages formed electrically tight barriers. 1[superscript]0CME were cultured in the presence of hydrocortisone and exhibited high electrical resistance and amiloride-sensitive I[subscript]sc, suggesting the presence of ENaC-mediated Na[superscript]+ transport. 1[superscript]0BME were grown in a complex media in the presence or absence of dexamethasone. In contrast to 1[superscript]0CME, 1[superscript]0BME exhibited no detectable amiloride-sensitive I[subscript]sc in either culture condition. However, 1[superscript]0BME monolayers responded to an adrenergic agonist, norepinephrine, and a cholinergic agonist, carbamylcholine, with rapid increases in I[subscript]sc. Thus, this protocol for isolation and primary cell culture can be used for future studies that focus on mammary epithelial cell regulation and functions. In conclusion, the results from these projects demonstrate that mammary epithelial cells form electrically tight monolayers and can exhibit neurotransmitter- and/or hormone-induced net ion transport. The mechanisms that regulate Na[superscript]+ transport across mammary gland may provide clues to prevent or treat mastitis.

In this study, we add an additional layer above and under the CIGS absorber layer as a secondary absorption layer respectively. We made the conventional structure of ZnO/CdS/CIGS/Mo becomes the structure of ZnO/CdS/CIGS/InGaP/Mo and ZnO/CdS/InGaP/CIGS/Mo which can improve the conversion efficiency. And we translate the thickness proportion of Ga and the doping concentration to find out the best parameter. According to the simulation, the wavelength of EQE in 600 nm ~ 1200 nm for our proposed CIGS solar cell which the additional layer under CIGS layer has been improved when compared to the conventional CIGS solar cell. The short-circuit current density has been increased about 9 %. And the conversion efficiency has also been increased about 9 %.When the additional layer above the CIGS absorber layer, according to the simulation, the wavelength of EQE in 300 nm ~ 600 nm for our proposed CIGS solar cell is improved when compared with the conventional CIGS solar cell. The short-circuit current density has been improved about 7.7 %, the open-circuit voltage about 7.1 %, and the conversion efficiency about 20.6 %. The main reason is that when the InGaP absorption layer under the CIGS layer which can catch the light which can¡¦t be absorbed by CIGS layer. The InGaP absorption layer above the CIGS layer which can catch the light immediately.

PCWE has been used widely in oriental traditional medicine in treating edema and diarrhea. Recent studies have shown that PCWE may also have anti-tumor and anti-inflammation acts. However, few studies have been conducted to reveal the mechanisms of these effects. In the present study, we tried to elucidate the mechanisms by investigating the effects on PCWE on the regulation of ion transport across the epithelial membranes of colon, which is also useful in explaining the anti-diarrhea and anti-edema effects. Alternation in membrane potential and short-circuit current (Isc) were examined using the Ussing chamber technique. Our results showed that PCWE decreased Isc upon application to the apical side. Amiloride inhibited this Isc induced by PCWE indicating that PCWE acted on amiloride-sensitive sodium channel of the epithelium. However, when PCWE was applied to the serosal side, the Isc was not changed, indicating a minimal influence of this substance on ATP-driving Na+/K+ counter transporters. Our data also showed that the Isc decreased by PCWE could be inhibited by bumetanide and chlorothiazide (Cl¡Â channel inhibitors). We therefore concluded that PCWE could both enhance sodium transport and stimulate the secretion of Cl¡Â in colon epithelium.

Wortmannin is a potent inhibitor of phosphatidylinositol 3-kinase (PI3K) and membrane trafficking in many cells. To test the hypothesis that cystic fibrosis transmembrane conductance regulator (CFTR) traffics into and out of the plasma membrane during cAMP-stimulated epithelial Cl- secretion, we have studied the effects of wortmannin on forskolin-stimulated Cl- secretion by the human colonic cell line T84. At the PI3K inhibitory concentration of 100 nM, wortmannin did not affect significantly forskolin-stimulated Cl- secretion measured as short-circuit current (I(SC)). However, 500 nM wortmannin significantly inhibited forskolin-stimulated I(SC). cAMP activation of apical membrane CFTR Cl- channels in α-toxin-permeabilized monolayers was not reduced by 500 nM wortmannin, suggesting that inhibition of other transporters accounts for the observed reduction in T84 Cl- secretion. Forskolin inhibits apical endocytosis of horseradish peroxidase (HRP), but wortmannin did not alter forskolin inhibition of apical HRP endocytosis. In the absence of forskolin, wortmannin stimulated HRP endocytosis significantly. We conclude that, in T84 cells, apical fluid phase endocytosis is not dependent on PI3K activity and that CFTR does not recycle through a PI3K-dependent and wortmannin-sensitive membrane compartment.

No description available.