Taraxacum officinale extract shows antitumor effects on pediatric cancer cells and enhance mistletoe therapy.

OBJECTIVES: While Taraxacum officinale (dandelion) extracts showed antitumor effects on adult cancer cells, effects on pediatric tumor cells as a single agent or in combination with mistletoe extracts are hitherto unknown. MATERIAL AND METHODS: The anti-proliferative effects of an aqueous fermented Taraxacum officinale extract (Taraxacum) on a pediatric cancer cell line panel were assessed by cell viability assays (MTT). In two neuroblastoma cell lines, SH-SY5Y and Kelly, the effects on cell cycle distribution (PI staining), mitochondrial integrity (MitoTracker staining), invasion (Boyden chamber assay) and migration (Scratch-assay) as well as the synergistic effects of the co-treatment of Taraxacum and mistletoe preparations (Iscucin® Tiliae or Iscucin® Pini) were investigated. RESULTS: All tested cancer cell lines were more susceptible to Taraxacum than the normal human fibroblast cell line, NHDF-C. In neuroblastoma cell lines Taraxacum caused apoptosis and loss of mitochondrial integrity as well as an inhibition of invasion and migration. The simultaneous therapy of Taraxacum and the mistletoe extracts revealed synergistic effects. CONCLUSION: This preclinical data support the use of Taraxacum as a potential adjuvant application in pediatric oncology.

An industrial engineering approach to laboratory automation for high throughput screening.

Across the pharmaceutical industry, there are a variety of approaches to laboratory automation for high throughput screening. At Sphinx Pharmaceuticals, the principles of industrial engineering have been applied to systematically identify and develop those automated solutions that provide the greatest value to the scientists engaged in lead generation.

Increased concentrations of interleukin 1-beta in whole blood cultures supernatants after 12 weeks of moderate endurance exercise.

The aim of the study was to investigate whether a regular moderate endurance exercise programme influenced the in vitro cytokine synthesis by stimulated whole blood cultures. To this end, eight healthy subjects exercised moderately by running for 3-5 h a week over a period of 12 weeks, whilst seven other healthy subjects served as the control group. The intensity of the exercise was determined by lactic acid concentrations in the blood which were maintained between 1.8 and 2.5 mmol x l(-1). Over the period of training the running velocity producing the 4 mmol x l(-1) lactic acid threshold increased from 2.86 (SD 0.83) m x s(-1) to 3.06+/-0.79 m x s(-1) (P < or = 0.008). Blood samples were taken at rest before and after the training programme. The following blood parameters were determined: leucocyte count, differential leucocyte count, lymphocyte subpopulations [CD14 positive (+)/CD45+, CD4+/ CD25+, CD8+, CD16+/CD122+]. Whole blood cultures were stimulated with lipopolysaccarides [interleukin (IL)-1 beta and IL-6] and staphylococcal enterotoxin B [IL-2, soluble interleukin 2 receptor (sIL2-R) and interferon (IFN)-gamma]. Cytokine concentrations in the supernatants were measured using an enzyme-linked immunosorbent assay. The white blood cell count, differential leucocyte count, lymphocyte subset distribution and the expression of the CD25 and CD122 antigen on lymphocytes were unchanged by training. After the training programme the IL-1 beta production changed significantly [1496 (SD 264) pg ml(-1) before, compared to 2127 (SD 672) pg ml(-1) after training, P < or = 0.008]. In the control group these parameters remained unchanged. With respect to changes in the values in both groups the syntheses of IL-1 beta (P < or = 0.023) and IL-6 (P < or = 0.021) were significantly higher after regular training. The syntheses of IL-2, sIL-2 and INF-gamma were not significantly influenced. Regular endurance exercise influenced the in vitro production of monocyte derived cytokines, while the effect of exercise on the cytokines synthesized by T-cells appeared to be of lesser importance.

Developing patient and family education programs for a transplant center.

Patient and family education is an important component of the organ transplant programs at the University of Nebraska Medical Center. The Medical Center is in the process of planning a new transplant center which will employ the family-centered, educationally-intensive cooperative care concept. This approach was chosen as the model for the delivery of care at the Lied Transplant Center because it emphasizes efficient, effective clinical care by requiring active participation by the family or essential other, thereby better preparing both the patient and the family for the transition to home and to the community. This article presents the evolution of patient education in our transplant programs, discusses the educational needs of transplant patients across the continuum of care, provides insight into the process of planning educational programs for the new center and provides a sample module for teaching which is based on the Cooperative Care concept.

A novel approach to the placement of Palmaz-Schatz biliary stents in saphenous vein grafts.

The placement of Palmaz-Schatz biliary stents has become a successful method to treat stenoses in large saphenous vein grafts. Unlike coronary stents, the biliary stents are not routinely delivered with a selective delivery sheath and may be more difficult to deliver and prone to detachment from the balloon and even embolization during delivery. In order to enhance the ability to deliver these stents, a guide-within-a-guide system was developed. A 7F guiding catheter was used as a selective delivery sheath within a standard guiding catheter. Nineteen biliary stents were successfully placed in vein grafts in 15 patients using this system. One procedure was complicated by an embolic event documented angiographically following intragraft delivery of urokinase but prior to stent implantation.

Politics and the nurse manager.

Sometimes, the "games being played" in our job settings cripple our ability to be powerful, effective participants in the world of work. Power, which is the ability to obtain, retain and move resources, requires two sets of attributes: competence and political savoir-faire. Much attention is given to improving competence but little time is spent in learning the intricacies of political behavior. The most important strategy is to learn to "read the environment" through observation, listening, reading, detachment and analysis.

Adaptor device for shortening guide catheters to access distal lesions in coronary angioplasty.

One of the challenges in interventional cardiology is accessing distal stenoses in both native coronary arteries and bypass grafts. This article reports the fabrication and use of an adaptor device that can be connected to the proximal segment of a shortened guide catheter for easier and safer access to these distal lesions.

Balloon angioplasty of chronic total coronary artery occlusions: what does it cost in radiation exposure, time, and materials?

Coronary angioplasty of chronic total occlusions is known to have a lower success rate and higher frequency of restenosis than angioplasty of subtotal stenoses, but there are no data describing the additional time, resources, and radiation exposure associated with this procedure. The purpose of this study was to compare these features in 90 consecutive patients who underwent angioplasty of a total occlusion (group 1) to those of 100 consecutive patients who underwent angioplasty of a subtotal stenosis (group 2). Angioplasty was successful in 60% of group 1 and in 94% of group 2 patients. Procedures were longer in group 1 and significantly more guide catheters, angioplasty balloon catheters, and guide wires were required per patient compared to group 2. The volume of contrast media used in each group was similar. Estimates of radiation entry exposure, calculated from fluoroscopy exposure times and using data from phantom studies, were 53 R (roentgen) per patient in group 1 vs. 34 R in group 2. These procedures were performed using pulsed progressive fluoroscopy; radiation exposure would be considerably higher using conventional fluoroscopic systems. Cineangiographic radiation exposures were similar for each group and accounted for an average additional exposure of 14-22 R for each procedure. Total radiation exposure was reduced in both groups when the angioplasty procedure was combined with the diagnostic angiogram. These observations should be considered when dilation of a total occlusion is planned. Efforts to reduce the amount of radiation exposure should remain a priority in interventional catheterization laboratories.

Relation of stenosis morphology and clinical presentation to the procedural results of directional coronary atherectomy.

BACKGROUND: Directional coronary atherectomy has recently become available to treat coronary stenoses. This study was performed to determine the relation of patient characteristics and stenosis morphology to procedural outcome with directional coronary atherectomy to gain insight into which patients might be best treated with this device. METHODS AND RESULTS: Four hundred stenoses from 378 patients consecutively treated at six major referral institutions were analyzed. Angiographic data were assessed at a central angiographic laboratory using standardized morphological criteria and computer-assisted quantitative dimensional analyses. Procedural success was achieved in 87.8% of stenoses, and major ischemic complications (death, myocardial infarction, and emergency bypass surgery) occurred in 6.3% of patients. Lesion success and complications were closely correlated with recognized modified American College of Cardiology/American Heart Association Task Force lesion morphological criteria. Observed for type A stenoses were 93% success and 3% complication rates; for type B1 stenoses, 88% success and 6% complication rates; and for type B2 stenoses, 75% success and 13% complication rates, respectively. There were too few type C stenoses treated to analyze. Furthermore, multivariate testing demonstrated stenosis angulation (multivariate p less than 0.001), proximal tortuosity (p less than 0.001), decreased preatherectomy minimum lumen dimension (p = 0.032), and calcification (p = 0.041) to correlate independently with adverse outcome and complex, probably thrombus-associated stenoses to have a favorable outcome (p = 0.055). Operator experience (p = 0.020) and a history of restenosis (p = 0.022) also favorably influenced outcome. CONCLUSIONS: The procedural outcome of directional coronary atherectomy is highly associated with coronary stenosis morphology. Furthermore, after appropriate stratification for morphology and clinical presentation, overall atherectomy procedural outcome may be similar to that achieved with coronary angioplasty. However, specific subsets of patients may have relatively better outcome with either atherectomy or balloon angioplasty.

Preliminary observations on the relationship between gas production and microbial protein synthesis in vitro.

The relationship between gas production and microbial protein synthesis was studied in vitro using the method of MENKE et al. (1979). 150 mg starch or cellulose or a mixture of 10% glucose, 40% starch and 50% cellulose was used as the carbohydrate source. The microbial protein synthesis and gas production occurring during 2 hrs after the 5th, 10th, and 23rd hr of incubation were studied. Total and net microbial synthesis were estimated using 32P as a microbial marker and by the net disappearance of NH3-N respectively. The data indicate that the type of carbohydrate and the rate of carbohydrate fermentation influence microbial protein synthesis per unit volume of gas produced. However, the relationship between total synthesis and cumulative gas production (up to 8 hrs incubation) with carbohydrate mixture as the substrate was linear. With reference to these observations, the possibilities and difficulties in using cumulative gas production as an index of microbial growth potential of the feedstuffs are discussed.

A randomized comparison of the nephrotoxicity of iopamidol and diatrizoate in high risk patients undergoing cardiac angiography.

Three hundred seven high risk patients with renal impairment (serum creatinine greater than or equal to 1.5 mg/dl) were randomized in a double-blind manner to either iopamidol (a nonionic, low osmolar radiocontrast agent) or diatrizoate (a conventional radiocontrast agent) at cardiac angiography with subsequent follow-up study of renal function. Baseline clinical and angiographic variables were similar in the iopamidol (n = 155) and diatrizoate (n = 152) groups. Change in renal function after angiography was less pronounced with iopamidol compared with diatrizoate as measured by mean ( +/- SD) increase in 24 h serum creatinine (0.11 +/- 0.2 versus 0.22 +/- 0.26 mg/dl, p less than 0.001), mean maximal increase in serum creatinine (0.2 +/- 0.44 versus 0.38 +/- 0.73 mg/dl, p less than 0.0001) and percent of patients with a maximal increase in serum creatinine greater than 0.5 mg/dl (8% versus 19%, p less than 0.01). Such differences could not be documented in diabetic patients using insulin. There was no significant difference between agents in the number of patients developing clinically severe acute renal dysfunction. It is concluded that iopamidol is less nephrotoxic than diatrizoate in high risk patients at cardiac angiography. However, the difference in nephrotoxicity is small, of no major clinical significance in the majority of high risk patients and could not be documented in insulin-using diabetic patients. Iopamidol may be the preferred agent in certain patients with advanced renal impairment, but further study is warranted.

Interaction of progesterone derivatives with the digitalis target enzyme: impact of glycosidation on inhibitory potency.

As a function of the structural modification of the steroid nucleus, the inhibitory interaction of 11 progesterone derivatives with human Na/K-ATPase (Na+/K(+)-transporting ATPase, EC 3.6.1.37), through C3-O-rhamnosylation, is either much decreased or weakly up to strongly increased, so that the rhamnosyl residue contributes to the complementary Gibbs energy of interaction, at the most, the same Gibbs energy increment as realized in ouabain. After C3 beta-O-rhamnosylation, the activity of some progesterone derivatives considerably surpasses that of 3 beta-O-rhamnosyl-chlormadinolacetate, which has been known to elicit positive inotropy in cats. The progesterone derivatives (aglycons and glycosides), that have been analysed more closely, produce their effects by the same molecular mechanism of interaction with Na/K-ATPase as characteristic for digitalis aglycons and glycosides. The results promise to pave the way for the identification of the chemical nature of endogenous digitalis and for the design of novel inotropic drugs.

Chemical models for the chemical nature of endogenous digitalis.

The inability or the capacity to promote the phosphorylation of Na+/K(+)-transporting ATPase (Na/K-ATPase) from [32P]Pi is shown to differentiate between mechanistically digitalis-unlike and digitalis-like inhibitors of this enzyme known to be the receptor for all digitalis actions. A negative or positive response in the phosphorylation promotion assay introduced here appears thus to be suitable to diagnose the chemical species in the isolates of animal origin related to the putative endogenous digitalis. Various digitalis-congeneric C/D-cis steroids, progesterone-congeneric C/D-trans steroids and the Erythrophleum alkaloid cassaine promote the enzyme phosphorylation and show a similar pattern of discrimination between three Na/K-ATPase variants. Thus, their cyclopentanoperhydrophenanthrene or perhydrophenanthrene nuclei appear to serve as the minimal pharmacophoric lead structures for bimolecular recognition and to represent chemical models for the chemical nature of endogenous digitalis. Specifically, the hormonal C/D-trans steroids could provide the basic skeleton in endogenous digitalis biosynthesis.

Coronary atherectomy: first 50 patients at the Mayo Clinic.

The safety and efficacy of a novel coronary atherectomy device were evaluated in the first 50 patients who underwent 53 such procedures at the Mayo Clinic. This treatment was used for atheromatous lesions in 27 patients and for restenosis after percutaneous transluminal coronary angioplasty in 23; in 3 patients, it was repeated for recurrent stenosis (2) or a persistent intimal flap (1). Successful results, defined as a reduction of the stenosis by 40% or more in association with removal of tissue, absence of Q-wave infarction, and no need for a bypass surgical procedure during in-hospital follow-up, were achieved in 47 of the 53 procedures (89%). Atherectomy reduced the stenoses from a mean of 87% to 15%. Failures occurred in 6 of the 53 procedures (11%). Three of these patients subsequently underwent successful conventional balloon angioplasty, and three required surgical treatment. Six patients had minor complications--embolization of atheromatous material in two, intimal dissection in two, and side-branch occlusion and transient thrombosis in one each. Coronary atherectomy with use of this device promises to be a relatively safe and efficient treatment of coronary artery stenosis.

Studies on in vitro degradation and in vivo digestion of a slow ammonia releasing urea product.

a slow ammonia releasing urea product (Uromalt) has been produced by autoclaving and drying of germinated barley and urea. In vitro degradability of nitrogen (IVDN, Raab et al., 1983) was in average 62 +/- 9% in 4 hours, compared to 76 +/- 8% with an autoclaved and dried mixture of urea and not-germinated barley. Mixtures of urea with Leucaena l. or Azadarichta indica (Neem cake) showed degradabilities of about 80 and 93%, respectively, when dried at 39 degrees C, but of 65% when dried at 100 degrees C. For comparison, degradabilities of proteins of ground nut, guar, sunflower, mustard, rape and sesame cake/meal have been determined. When growing lambs were fed a ration containing 15% Uromalt, faecal N-excretion was significantly higher on the expense of urinary N-excretion, compared with groups fed isonitrogenous rations containing soya bean meal with or without urea and molasses. N-retention was not significantly affected. It is concluded that this slow ammonia releasing urea product can be used in cases where molasses or other feedingstuffs rich in rapidly fermentable carbohydrates are not available.

5 Beta,14 beta-androstane-3 beta,14-diol binds to the digitalis receptor site on Na/K-ATPase.

5 beta,14 beta-Androstane-3 beta,-14-diol, the lead (minimum) structure in digitalis compounds, shows the same characteristics of interaction with Na/K-ATPase as ordinary digitalis compounds judged by the following six criteria: (I) shape of the concentration-inhibition curves, (II) species differences in affinity for the enzyme, (III) apparent competition with K+, (IV) competition with digitoxigenin for binding to the enzyme, (V) stabilization of phosphoenzyme formed from ATP, and (VI) enhancement of phosphorylation from orthophosphate.

Origin of differences of inhibitory potency of cardiac glycosides in Na+/K+-transporting ATPase from human cardiac muscle, human brain cortex and guinea-pig cardiac muscle.

The inhibitory potency of altogether 95 steroidal compounds (including cardenolides, bufadienolides and their glycosides) on the Na/K-ATPases (Na+/K+-transporting ATPases, EC 3.6.1.37) from human cardiac muscle, human brain cortex and guinea-pig cardiac muscle was compared to probe the complementary chemotopology of the inhibitor binding site areas on the three enzyme variants. The changes of potency, resulting from systematic variations of the geometry of steroid skeleton and the character as well as the structure of side chains at C3 or/and C17 of steroid backbone, allowed the following major conclusions. With the human cardiac and cerebral enzyme forms, the paired K0.5 (K'D) values for 77 steroid derivatives, covering seven orders of ten, were highly correlated. On an average, the total of compounds showed a 1.5-fold higher affinity to the cardiac enzyme. This tiny differentiation did not appear to be connected with an important difference in the chemotopology of the complementary subsites for steroid nucleus binding on the two enzyme forms. With the human and guinea-pig cardiac enzyme variants, the K0.5 values for 69 steroid derivatives, covering six orders of ten, were determined. For 41 5 beta, 14 beta-androstane derivatives only, the paired K0.5 values showed a close correlation. Here, the human enzyme variant exhibited 27-fold higher affinity. However, the paired K0.5 values determined on both enzymes for 28 steroid derivatives of differing structural features were but poorly correlated. Essentially, the geometries of the steroid nucleus determined the differential contributions of the side chains at C3 and C17 to the integral inhibitory potency on the two enzyme variants. Thus, the species differences in the potency of cardiac glycosides were traced to species differences in the complementarity of the steroid binding subsites. Hence, estimates of the potency of new steroidal compounds obtained on the guinea-pig cardiac enzyme can be neither quantitatively nor qualitatively easily extrapolated to the human cardiac enzyme. The extrathermodynamic analysis of the data opened major new insights in the structure-activity relationships concerning the role of C14 beta-OH, the character of the lead structure in cardioactive steroid lactones, and the significance of the configuration of A/B ring junction.

Evidence against parallel operation of sodium/calcium antiport and ATP-driven calcium transport in plasma membrane vesicles from kidney tubule cells.

The present study aimed to clarify the existence of a Na+/Ca2+ antiport device in kidney tubular epithelial cells discussed in the literature to represent the predominant mechanistic device for Ca2+ reabsorption in the kidney. Inside-out oriented plasma membrane vesicles from tubular epithelial cells of guinea-pig kidney showed an ATP-driven Ca2+ transport machinery similar to that known to reside in the plasma membrane of numerous cell types. It was not affected by digitalis compounds which otherwise are well-documented inhibitors of Ca2+ reabsorption. The vesicle preparation contained high, digitalis-sensitive (Na+ + K+)-ATPase activities indicating its origin from the basolateral portion of plasma membrane. The operation of a Na+/Ca2+ antiport device was excluded by the findings that steep Ca2+ gradients formed by ATP-dependent Ca2+ accumulation in the vesicles were not discharged by extravesicular Na+, and did not drive 45Ca2+ uptake into the vesicles via a Ca2+-45Ca2+ exchange. The ATP-dependent Ca2+ uptake into the vesicles became increasingly depressed with time by extravesicular Na+. This was not due to an impairment of the Ca2+ pump itself, but caused by Na+/Ca2+ competition for binding sites on the intravesicular membrane surface shown to be important for high Ca2+ accumulation in the vesicles. Earlier observations on Na+-induced release of Ca2+ from vesicles pre-equilibrated with Ca2+, seemingly favoring the existence of a Na+/Ca2+ antiporter in the basolateral plasma membrane, were likewise explained by the occurrence of Na+/Ca2+ competition for binding sites. The weight of our findings disfavors the transcellular pathway of Ca2+ reabsorption through tubule epithelium essentially depending on the operation of a Na+/Ca2+ antiport device.