A preliminary investigation into the design of pressure cushions and their potential applications for forearm robotic orthoses.

BACKGROUND: Load cells are often used in rehabilitation robotics to monitor human-robot interaction. While load cells are accurate and suitable for the stationary end-point robots used in rehabilitation hospitals, their cost and inability to conform to the shape of the body hinder their application in developing affordable and wearable robotic orthoses for assisting individuals in the activities of daily living. This exploratory work investigates the possibility of using an alternative technology, namely compliant polymeric air cushions, to measure interaction forces between the user and a wearable rigid structure. METHODS: A polymeric air cushion was designed, analyzed using a finite element model (FEM), and tested using a bench-top characterization system. The cushions underwent repeatability testing, and signal delay testing from a step response while increasing the length of the cushion's tubes. Subsequently, a 3D printed wrist brace prototype was integrated with six polymeric air cushions and tested in static conditions where a volunteer exerted isometric pronation/supination torque and forces in vertical and horizontal directions. The load measured by integrating data recorded by the six sensors was compared with force data measured by a high quality load cell and torque sensor. RESULTS: The FEM and experimental data comparison was within the error bounds of the external differential pressure sensor used to monitor the pressure inside the cushion. The ratio obtained experimentally between the pressure inside the pressure cushion and the 8 N applied load deviated by only 1.28% from the FEM. A drift smaller than 1% was observed over 10 cycles. The rise times of the cushion under an 8 N step response for a 0.46, 1.03, and 2.02 m length tube was 0.45, 0.39, and 0.37 s. Tests with the wrist brace showed a moderate root mean square error (RMSE) between the force estimated by the pressure cushions and the external load cells. Specifically, the RMSE was 13 mNm, 500 mN, and 1.24 N for forearm pronation/supination torque, vertical force, and horizontal force, respectively. CONCLUSIONS: The use of compliant pressure cushions was shown to be promising for monitoring interaction forces between the forearm and a rigid brace. This work lays the foundation for the future design of an array of pressure cushions for robotic orthoses. Future research should also investigate the compatibility of these polymeric cushions for data acquisition during functional magnetic resonance imaging in shielded rooms.

Characterization of primary rabbit kidney cultures that express proximal tubule functions in a hormonally defined medium.

Primary cultures of rabbit-kidney epithelial cells derived from purified proximal tubules were maintained without fibroblast overgrowth in a hormone-supplemented serum-free medium (Medium RK-1). A hormone-deletion study indicated that the primary cultures derived from purified rabbit proximal tubules required all of the three supplements in Medium RK-1 (insulin, transferrin, and hydrocortisone) for optimal growth but did not grow in response to EGF and T3. In contrast, the epithelial cells in primary cultures derived from an unpurified preparation of rabbit kidney tubules and glomeruli grew in response to EGF and T3, as well as insulin, transferrin, and hydrocortisone. These observations suggest that kidney epithelial cells derived from different segments of the nephron grow differently in response to hormones and growth factors. Differentiated functions of the primary cultures derived from proximal tubules were examined. Multicellular domes were observed, indicative of transepithelial solute transport by the monolayers. The proximal tubule cultures also accumulated alpha-methylglucoside (alpha-MG) against a concentration gradient. However, little or no alpha-MG accumulation was observed in the absence of Na+. Metabolic inhibitor studies also indicated that alpha-MG uptake by the primaries is an energy-dependent process, and depends upon the activity of the Na+/K+ ATPase. Phlorizin at 0.1 mM significantly inhibited 1 mM alpha-MG uptake whereas 0.1 mM phloretin did not have a significant inhibitory effect. Similar observations have been made concerning the Na+-dependent sugar-transport system located on the lumenal side of the proximal tubule, whereas the Na+-independent sugar transporter on the peritubular side is more sensitive to inhibition by phloretin than phlorizin. The cultures also exhibited PTH-sensitive cyclic AMP synthesis and brush-border enzymes typical of proximal cells. However, the activities of the enzymes leucine aminopeptidase, alkaline phosphatase, and gamma-glutamyl-transpeptidase were lower in the cultures than in purified proximal-tubule preparations from which they are derived.

Developing a culturally valid and reliable quality of life questionnaire for diabetes mellitus.

The aim of this study was to design a culturally adapted questionnaire for studying quality of life (QOL) among type 1 and 2 adult diabetes patients in the Islamic Republic of Iran. The 41 items on the questionnaire were based on qualitative research and covered general and health-related QOL. In a descriptive survey, 104 patients completed the questionnaire; 68 (65.4%) were female. Mean age was 50.5 years (standard deviation 12.8). Most patients (86.5%) had type 2 diabetes. Cronbach's alpha coefficient for the questionnaire was 0.98. The questionnaire successfully distinguished the lower QOL of patients suffering from pain in the limbs, loss of appetite, fatigue, constipation and itching. The questionnaire could determine both general and health-related QOL.

Effect of polycations on prostaglandin synthesis in cultured glomerular mesangial cells.

The presence of uniform negative charges on the surface of cultured rat glomerular mesangial cells was demonstrated by an ultrastructural marker, cationized ferritin. Interaction between cell surface negative charges and protamine sulfate, stimulated the synthesis of prostaglandins E2, F2 alpha, 6-keto-PGF1 alpha and thromboxane B2 (TXB2) in a dose-dependent manner, reaching a maximum response at protamine concentration of 50 micrograms/ml. The effect of protamine sulfate was reversed by 25 units/ml heparin. The polyanions, L-glutamic and L-aspartic acids, reversed the protamine effect in a dose-dependent manner. Excess substrate, arachidonic acid, masked the protamine sulfate-stimulated PGE2 synthesis by mesangial cells. The effect of protamine sulfate on PGE2 synthesis was rapid, peaked in 5 min and was independent of extracellular Ca2+. A synthetic cation, poly(L-lysine) hydrobromide, exerted a similar effect on cellular PGE2 synthesis in mesangial cells. The effect of poly(L-lysine) was dependent on the molecular mass of the cationic species employed and was maximum at 17 to 90 kDa. The use of large molecular mass polymers of L-lysine (175 and 565 kDa) resulted in a decline in PGE2 synthesis. These observation indicate that, in mesangial cells, changes in cell membrane electrical charge are linked to enhanced biosynthetic activity and eicosanoid synthesis.

Sodium-dependent glucose transport by cultured proximal tubule cells.

The cotransport of sodium ion and alpha-methyl glucose, a non-metabolized hexose, was studied in rabbit proximal tubule cells cultured in defined medium. The rate of uptake of alpha-methyl glucose shows saturation kinetics, in which Km, but not Vmax, is dependent upon the Na+ concentration in the medium. The transport system was found to be of the high-affinity type, characteristic of the straight portion of the proximal tubule. Analysis of the rates of initial uptake within the context of a generalized cotransport model, suggests that two Na+ ions are bound in the activation of the hexose transport. The steady-state level of accumulation of alpha-methyl glucose also depends upon sodium concentration, consistent with the initial rate findings. The uptake of alpha-methyl glucose is inhibited by other sugars with the relative potencies of D-glucose greater than alpha-methyl glucose greater than D-galactose = 3-O methylglucose. L-Glucose, D-fructose, and D-mannose show no inhibition. Phlorizin inhibits the alpha-methyl glucose uptake with a Ki of 9 X 10(-6) M. Ouabain (10(-3) M) decreases the steady-state alpha-methyl glucose accumulation by 60%. In the absence of sodium, the accumulation of alpha-methyl glucose is 7-fold less than at 142 mM Na+, reaching a level comparable to the sodium-independent accumulation of 3-O-methyl-D-glucose. These findings are similar to those observed in the proximal tubule of the intact kidney.

Decreased degradative enzymes in mesangial cells cultured in high glucose media.

Abnormalities in extracellular matrix degradation may play a pathogenetic role in diabetic nephropathy. Cultured renal mesangial cells are known to synthesize increased amounts of matrix proteins when incubated in high glucose media (e.g., 30 mmol/l). However, the effect of glucose loading on degradative enzymes is unknown. Primary cultures of rat mesangial cells were grown until confluent in the presence of fetal calf serum (FCS) and insulin (0.67 U/ml). Cells were then cultured for 7 days in plastic wells in either 10 or 30 mmol/l glucose media containing neither FCS nor insulin. Collagenase activity in media were determined by zymography and quantitative spectrofluorometry. Cathepsin B and D activities in cell extracts were measured by spectrofluorometry (using the fluorescent substrate Z-Arg-Arg-7-amido-4-methylcoumarin) and 125I-labeled hemoglobin digestion, respectively. Gelatin-degrading activity of live mesangial cells was also determined. mRNA levels for collagenase IV, cathepsin B, and cathepsin D were determined by Northern analysis. A major band of collagenase activity with a molecular size of 72 kDa was observed in all mesangial cell media. Exposure of cells to high glucose media resulted in significant reductions in collagenase and cathepsin B activities as well as impairment in gelatin-degrading activity. Collagenase IV and cathepsin B and D mRNA levels were also decreased by glucose loading. To exclude the possibility that glucose loading was injurious to cells, 3H-leucine uptake (as a measure of protein synthesis) and membrane alkaline phosphatase activity (as a biochemical marker of viability) were not affected by the high glucose condition.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of angiotensin II in glucose-induced inhibition of mesangial matrix degradation.

Accumulation of mesangial matrix in diabetic nephropathy is caused by increased synthesis and decreased degradation. We have previously demonstrated that incubation in high-glucose medium decreases mesangial cell collagenase activity (Diabetes 44:929-935, 1995). Because angiotensin II (AII) is involved in the pathogenesis of diabetic nephropathy, the present studies were performed to determine if AII mediates glucose-induced 1) inhibition of mesangial collagenase activity, 2) mesangial matrix accumulation, and 3) in-crease in transforming growth factor (TGF)-beta1 secretion in mesangial cells. The direct effect of high glucose on AII generation in mesangial cells was also determined. Primary mesangial cells from normal Sprague-Dawley rats were used in all studies. Collagenase activity in cell medium was determined using three methods: 1) zymography; 2) quantitative assay using fluoresceinated gelatin as substrate; and 3) a new enzyme-linked immunosorbent assay (ELISA) that specifically measures 72-kDa collagenase (MMP-2), the principal collagenase synthesized by mesangial cells. Matrix accumulation was estimated by immunoperoxidase assay on cell layers using anti-glomerular basement membrane (GBM) antibodies. TGF-beta1 and AII levels were determined by ELISA. Exposure of mesangial cells to 30 mmol/l glucose (high glucose) vs. 5 mmol/glucose (normal glucose) for 5 days resulted in a significant decrease in collagenase activity (25%) that was normalized by 10(-4) mol/l losartan, a type 1 angiotensin II (AT1) receptor antagonist. High glucose increased anti-GBM binding compared with normal glucose; this effect of glucose was reversed by losartan. Incubation of cells with 30 mmol/l glucose increased total TGF-beta1 secretion, which was also normalized by losartan. Addition of AII (10(-6) mol/l) for 24 h to the culture medium inhibited collagenase activity by 33%; losartan (10(-4) mol/l) blocked this inhibition of enzyme activity. Also, AII decreased collagenase (MMP-2) levels but stimulated TGF-beta1 secretion in mesangial cells. Finally, glucose increased mesangial AII generation in a concentration-dependent manner, with incubation in 30 mmol/l glucose increasing AII by 25% compared with 5 mmol/l glucose. We conclude that glucose increases AII production by mesangial cells, which results in stimulation of TGF-beta1 secretion, decreased matrix degradation, and increased matrix accumulation. These effects of AII are mediated by the AT1 receptor.

Bimanual elbow exoskeleton: Force based protocol and rehabilitation quantification.

An aging population, along with the increase in cardiovascular disease incidence that accompanies this demographic shift, is likely to increase both the economic and medical burden associated with stroke in western societies. Rehabilitation, the standard treatment for stroke, can be expanded and augmented with state of the art technologies, such as robotic therapy. This paper expands upon a recent work involving a force-feedback master-slave bimanual exoskeleton for elbow rehabilitation, named a Bimanual Wearable Robotic Device (BWRD). Elbow force data acquired during the execution of custom tasks is analyzed to demonstrate the feasibility of tracking patient progress. Two training tasks that focus on applied forces are examined. The first is called "slave arm follow", which uses the absolute angular impulse as a metric; the second is called "conditional arm static", which uses the rise time to target as a metric, both presented here. The outcomes of these metrics are observed over three days.

Role of angiotensin II in diabetic nephropathy.

Considerable evidence suggests that the intrarenal renin-angiotensin system plays an important role in diabetic nephropathy. Angiotensin-converting enzyme (ACE) inhibitors and angiotensin II (Ang II) receptor blockers (ARBs) can attenuate progressive glomerulosclerosis in disease models and can slow disease progression in humans. Because agents that interfere with Ang II action may decrease glomerular injury without altering glomerular pressures, it has been suggested that Ang II has direct effects on glomerular cells to induce sclerosis independent of its hemodynamic actions. To study nonhemodynamic effects of Ang II on matrix metabolism, many investigators have used cell culture systems. Glucose and Ang II have been shown to produce similar effects on renal cells in culture. For instance, incubation of mesangial cells in high-glucose media or in the presence of Ang II stimulates matrix protein synthesis and inhibits degradative enzyme (e.g., collagenase, plasmin) activity. Glucose and Ang II also can inhibit proximal tubule proteinases. Glucose increases expression of the angiotensinogen gene in proximal tubule cells and Ang II production in primary mesangial cell culture, which indicates that high glucose itself can activate the renin-angiotensin system. The effects of glucose and Ang II on mesangial matrix metabolism may be mediated by transforming growth factor-beta (TGF-beta). Exposure of mesangial cells to glucose or Ang II increases TGF-beta expression and secretion. Their effects on matrix metabolism can be blocked by anti-TGF-beta antibody or ARBs such as losartan, which also prevents the glucose-induced increment in TGF-beta secretion. Taken together, these findings support the hypothesis that the high-glucose milieu of diabetes increases Ang II production by renal, and especially, mesangial cells, which results in stimulation of TGF-beta secretion, leading to increased synthesis and decreased degradation of matrix proteins, thus producing matrix accumulation. This may be an important mechanism linking hyperglycemia and Ang II in the pathogenesis of diabetic nephropathy.

Comparison of national gestational weight gain guidelines and energy intake recommendations.

Although data showing adverse effects with high and low gestational weight gain (GWG) come from a large number of countries, a variety of guidelines about the GWG exist. Our objectives were to compare existing GWG and energy recommendations across various countries, as well as the rationale or evidence on which they were based. We used the United Nations' Human Developmental Index to determine the ranking of the country to ensure broad sampling and then searched for guidelines. We first searched the national government websites, and if necessary searched Medline and EMBASE, Global Health databases, and bibliographies of published articles for both guidelines and the studies on which they were based. We found guidelines for 31% of the countries, and 59% of these had a GWG recommendation, 68% had an energy intake recommendation (EIR), and 36% had both. About half of the GWG guidelines are similar to the 2009 American Institutes of Medicine (IOM) and 73% of the EIRs are similar to the 2006 IOM. Despite the documented relationship between both high GWG and adverse outcomes for women and infants and low GWG and adverse outcomes in infants, there are a wide variety of guidelines for GWG and energy recommendations by different countries around the world.

Preferred patterns of diastolic timed vibrations for pre-hospitalization treatment of acute coronary ischemia.

This paper presents and evaluates preferred patterns of vibrations and active breaking techniques for the Diastolic Timed Vibrator (DTV). DTV uses low frequency mechanical vibrations applied to the chest to help in clot dissolution in pre-hospitalization treatment of acute coronary ischemia. In this work, we argue that random and ramp type vibration patterns increase the performance of the DTV method. Furthermore, we present results for various methods of vibration stopping aiming at reduction of vibration overspill into the systole of heart cycle of the patient.

Depression, anxiety, activities of daily living, and quality of life scores in patients undergoing renal replacement therapies.

The aim of this study was to investigate anxiety, depression, quality of life (QOL), and activities of daily living (ADL) among our hemodialysis and renal transplant subjects. Psychological disorders were prevalent among patients with end-stage renal disease (ESRD), adversely affecting QOL and ADL. All patients > or =18 years under renal replacement therapy were asked to participate in the study. Patients completed 4 questionnaires including the Symptom Checklist-90 subscales of depression and anxiety, the Nottingham Extended ADL scale, and the Duke Health Profile questionnaire. We examined a total of 100 transplant and 63 hemodialysis patients. Transplant patients were significantly younger, better educated, and with lower morbidity than the hemodialysis patients. Transplant patients also showed less depression and anxiety as well as better ADL and physical health. Of the variance in anxiety scores, 12.2% was explained by gender and the presence of cardiac disease (P = .022). For depression, replacement therapy and gender explained 16.4% of the variance (P = .004). Replacement therapy accounted for 35.3% of the variance of the ADL score (P < .001), while together with the additional factors of age, gender, and education these 52.8% (P = .007) was accounted for. For the general health score, gender, and cardiac disease explained 11.6% of the variance (P < .001). This study demonstrated that depression and anxiety are more prevalent among hemodialysis patients compared with kidney transplant subjects; the method of treatment was a major contributor to the variance in ADL and depression scores among ESRD patients.

Events that stimulate release of thromboxane B2 in passive heymann nephritis.

The cause of proteinuria in passive Heymann nephritis has been attributed to the activation of the C5b-9 membrane attack complex following the antibody binding on the glomerular epithelial cell. Previous studies have shown an association between release of prostaglandin thromboxane B2 (TxB2) and proteinuria. Whether this release is dependent on antibody binding per se, or on secondary actions subsequent to antibody binding has not been clarified. The present study was designed to address this issue. Antibody binding event was experimentally separated from the proteinuria by employing a rabbit antibody which produces equivalent glomerular binding equal to that produced by a sheep antibody but without causing proteinuria. Comparisons were made with animals injected with the sheep antibody which produces all the hallmarks of the disease, including proteinuria. Animals injected with the rabbit antibody showed glomerular immunofluorescent deposits which were identical to the deposits produced by the control sheep antibody. However, rabbit antibody failed to produce the typical electron-dense subepithelial deposits, complement binding and proteinuria. Comparison of prostaglandin profile in isolated glomeruli revealed that TxB2 was unchanged in rabbit antibody-injected glomeruli (compared with its nonimmune antibody control). On the other hand, glomeruli from sheep antibody-injected animals released 45% higher TxB2 compared with their respective nonimmune antibody control. These data suggest that the binding of antibody per se may not be a sufficient stimulus for TxB2 release. Subsequent events of subepithelial electron dense deposit formation, complement activation, and proteinuria are associated with TxB2 release.

Prostaglandin and thromboxane synthesis by highly enriched rabbit proximal tubular cells in culture.

Prostaglandin synthetic profiles were studied in monolayers of highly enriched rabbit renal proximal tubular cells cultured in serum-free, hormone-supplemented, defined media. The cultures were initiated from glomeruli-free cortical suspensions. Cells in culture demonstrated morphologic and functional characteristics highly suggestive of proximal tubular cells. The basal and stimulated synthesis of immunoassayable prostaglandin (PG) E2, PGF2 alpha, 6-keto-PGF1 alpha, and thromboxane (Tx) B2 in response to various agonists, as well as the effect of two cyclooxygenase inhibitors, was assessed. Under both basal and stimulated conditions, PGE2 was the major product synthesized. PGF2 alpha and 6-keto-PGF1 alpha were synthesized to a lesser extent, and TxB2 was undetectable. The basal synthesis of PGE2 and PGF2 alpha in cultured cells was found to be higher than in isolated proximal tubular fragments by sevenfold and fivefold, respectively. Exogenous arachidonate, angiotensin II, and the divalent cation ionophore A23187 stimulated all three immunoassayable prostaglandins in a dose-dependent manner. Arginine vasopressin (10(-5) mol/L) had no stimulatory effect. In Ca++-free media or in the presence of 10(-5) mol/L Ca++ channel blocker, verapamil, the stimulatory effects of angiotensin II and A23187 were ameliorated. The stimulatory effect of angiotensin II was inhibited by saralasin (10(-5) mol/L), indicating that receptor binding could mediate PGE2 synthesis. Both indomethacin and sulindac sulfide (10(-5) mol/L) reversibly inhibited PGE2 synthesis.

Effect of protamine on the permeability and structure of rat peritoneum.

The effect of protamine, a polycationic protein, on the mesothelial permeability and ultrastructure was evaluated in a rat model of peritoneal dialysis. The peritoneal permeability to urea and inulin were measured after the intraperitoneal instillation of protamine sulfate in varying concentrations. A functional correlation was made with the ultrastructure of omentum. Protamine concentrations between 5 and 30 micrograms/ml decreased the permeability to inulin without significantly altering that of urea. At concentrations from 30 to 75 micrograms/ml, protamine increased permeability to urea by 50% and to inulin by 20%. Mesothelial cells revealed a loss of microvilli and minor degrees of disorganization of submembranous, cytoplasmic microfilaments without significant changes in the intramembranous structure of occluding junctions. Effects were partially reversible. At a concentration of 100 micrograms/ml, there was an irreversible doubling in permeability to inulin without a comparable effect on permeability to urea associated with the disruption of occluding junctions in focal areas. This complex response to protamine suggests at least two transperitoneal diffusion pathways, transcellular and paracellular. The mesothelial cell occluding junctions may be a diffusion barrier in the latter pathway.

Plasma lipids and acyltransferase activities in experimental nephrotic syndrome.

Lecithin:cholesterol acyltransferase (LCAT) and lysolecithin acyltransferase (LAT) are two activities carried out by the same plasma enzyme, but require different apoprotein activators. The LCAT reaction takes place primarily on high density lipoproteins (HDL) and is activated by serum albumin, whereas LAT takes place on low density lipoproteins (LDL) and is inhibited by albumin. In nephrotic syndrome (NS), the levels of serum albumin are reduced, whereas the LDL levels are increased, and therefore, the ratio of LAT/LCAT activities should be increased. To test this hypothesis, we estimated the lipid levels and the two enzyme activities in experimental NS induced in rats by the injection of anti-Fx1A antibody (passive Heymann nephritis). As found in other nephrotic conditions, the plasma lipid levels rose progressively as the proteinuria increased and the serum albumin concentration declined. In addition, the ratio of LAT/LCAT activities increased by about fourfold after nine days of induction of nephritis. The LCAT activity correlated positively and the LAT activity negatively with serum albumin levels. The esterified cholesterol correlated positively with LCAT activity in normal rats but negatively in nephrotic animals, indicating that most of the cholesteryl esters in NS may be non-LCAT derived. The free cholesterol/lecithin ratio, a known risk factor for atherosclerosis, increased significantly in nephrotic rats. Furthermore, since the increase in the LAT activity produces more disaturated lecithins, another putative risk factor, the cumulative risk of coronary heart disease may be increased in long-term NS.

Electrophysiologic study of rabbit proximal tubular cell monolayers in primary culture.

Primary cultures of renal cortical cells prepared by selective sieves have been found to display some characteristics of renal proximal tubular epithelium but their site of origin has not been confirmed by electrophysiologic studies. Cells were cultured in a defined medium on collagen gels. Confluency was approached after 7-10 days but gels were found to have zero transepithelial resistance unless they were allowed to contract spontaneously. With the appearance of a nonzero resistance, there was a change in morphology to a more columnar cell with better developed microvilli. These structural features were particularly prominent in clusters of proliferating cells observed on and around remnants of original tubules embedded in the gel. In noncontracted cultures there was no focal cell clustering and cells were squamous-like with rudimentary microvilli, similar in appearance to cells grown on plastic culture dishes. Measurements made in contracted monolayers yielded an average transepithelial resistance of 6.5 omega cm2, a spontaneous transepithelial potential difference of +0.9 mV, measured with respect to the serosa, and an apical membrane potential of -75 mV when cells were bathed in 0.4 mM K and -49 mV when cells were bathed in 4 mM K media. Mucosal protamine (50 micrograms/ml) increased transepithelial resistance by 22%, suggesting that the epithelial cell tight junctions were responsive to external stimuli. Monolayers were anion selective, giving a dilution potential (lumen-directed NaCl gradient) of -2.6 mV with respect to the serosa. These experiments show that primary culture of rabbit renal cortical cells separated by differential sieves displays electrophysiologic and morphologic characteristics of a proximal renal tubular epithelium. Confluency and attainment of differentiated morphology and function are promoted when monolayer cells are not bound to an unyielding substrate.

Peritoneal permeability in the rat: modulation by microfilament-active agents.

A model of peritoneal dialysis in the rat was used to determine the effects of cytochalasins on ultrastructure and peritoneal permeability to molecules of varying molecular weight. The permeability to urea, inulin, and plasma albumin were determined after intraperitoneal administration of cytochalasin B (2 to 10 X 10(-6) M) and cytochalasins D and E (2 X 10(-6) M). Cytochalasin B (20 X 10(-6) M) increased the permeability to inulin, urea, and albumin by 30, 60, and 150%, respectively. These effects were, to a large degree, reversible. Cytochalasins D and E produced greater increments in permeability for all molecules; this increase was only partially reversible. Ultrastructure analysis by scanning electron microscopy revealed extensive development of membrane protuberances (zeiotic knobs) on mesothelial cells exposed to cytochalasin B. A return to a normal apical cell surface was apparent although incomplete at 24 hr. Tight junctions were not grossly altered and major changes in intramembranous junctional strands were not observed. The major effect of cytochalasins on the cell surface may be responsible for the increased permeability to urea, predominately a transcellular probe. Inulin, which follows a paracellular route, was less affected. Altered protein permeability may be due to the action of cytochalasin on the exposed capillary endothelium in subdiaphragmatic areas where the mesothelium is discontinuous.

Angiotensin-induced sodium excretion patterns in cirrhosis: role of renal prostaglandins.

Changes in renal hemodynamics and sodium excretion induced by an angiotensin II (AII) infusion were correlated with urinary prostaglandin E2 (PGE2) excretion in 15 patients with cirrhosis and ascites. All induced natriuretic responses in 47% and antinatriuretic responses in 53% of the patients. Natriuresis was accompanied by an increase; antinatriuresis by a decrease in PGE2 excretion. Although there was no change in GFR (CIn), renal blood flow (CPAH) decreased. Patients were clinically indistinguishable. Antinatriuretic responders tended, however, to have higher baseline PGE2 excretion rates, were more sensitive to effects of prostaglandin blockade, and were less sensitive to the pressor effect of AII. Following partial inhibition of renal prostaglandin synthesis by indomethacin, AII-induced natriuretic responses were accentuated. GFR, RBF, and urine flow rate markedly decreased in both groups. There was no difference in pressor sensitivity to AII following prostaglandin synthesis blockade. We conclude that in patients with hepatic cirrhosis, the sodium excretion pattern induced by an exogenous AII challenge may depend on the prior state of intrarenal prostaglandin activity. Our findings also support the hypothesis that renal hemodynamic parameters in patients with cirrhosis and ascites are crucially dependent on renal prostaglandins.

Induction of epithelial tight junctions by a light chain protein isolated from a patient with Fanconi's syndrome.

Tight junctions have not been described in the adult human glomerulus. In a 22-year-old patient with kappa light chain proteinuria, tight junctions were observed between glomerular epithelial cell foot processes (kidney biopsy). The light chain isolated from urine proved to have a relatively high isoelectric point. When the light chain was exposed in vitro to the mucosal surface of a 'leaky' epithelium, the Necturus gallbladder, transepithelial resistance and potential difference increased in a concentration-dependent, reversible manner and NaCl dilution potentials decreased, consistent with a reduction in tight-junctional ionic permeability. Gallbladders fixed in situ and freeze-fractured during peak electrophysiological responses revealed an increase in tight-junctional depth. This report indicates that certain light chains, possibly by virtue of a positive charge, may induce changes in epithelial tight junction structure and/or permeability.