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19F-Nuclear magnetic resonance (NMR) spectroscopy facilitates the analysis of intracellular metabolism and membrane transport of 2-fluoroadenosine (F-AR) in human porcine aortic endothelial cells and erythrocytes. The extra- and intracellular fluorine signals can be differentiated from each other as they exhibit a distinct chemical shift both in endothelial cells and erythrocytes. Fluoroadenosine is quickly transformed into fluoro-ATP after its intake by endothelial cells and erythrocytes.

Impacts between urban and agricultural land cover on storm flow water quality are poorly understood for the Eastern Corn Belt Ecoregion in SW Ohio. Storm flow water samples were collected from May 2017 to October 2017 across seven SW Ohio watersheds which ranged in urban land cover from 6% to 92% and in agricultural land cover from 4% to 70%. Two watersheds contained water resource recovery facilities (WRRFs). Percent agricultural land cover in a watershed and storm magnitude were primary explanatory variables for total suspended solid and total phosphorus concentrations. Total nitrogen, nitrate, and phosphate concentrations were primarily explained by the presence of WRRFs and percent agricultural land cover. Increased dissolved nutrient concentrations in watersheds with WRRFs indicate that WRRFs in the study area are ineffectively removing nitrate and phosphate from effluent. Results suggest that to improve water quality during storm flows, additional management efforts need to be focused on agricultural watersheds and WRRFs. PRACTITIONER POINTS: Storm flow water quality in the study area is significantly affected by land cover, WRRF Q, and storm Q. TSS and TP concentrations are best explained by percent of agricultural land cover in a watershed and magnitude of storms. TN, NO 3 -N, and PO 4 3 -P concentrations are best explained by WRRF Q, followed by the percent agricultural land cover. This study shows that agricultural land cover and WRRFs play a significant role in water quality degradation in SW Ohio., (© 2019 Water Environment Federation.)

Motivating physicians to increase productivity and maximize patient satisfaction may result in conflicted behavior, raising questions about whether one must be sacrificed for the other. To determine if high satisfaction (measured by Press Ganey patient satisfaction survey) can be achieved while maintaining high productivity (measured in McGladrey relative value units, MRVU), longitudinal data collected from January 2002 to July 2004 were modeled using repeated measures regression. A total of 136 000 patient-completed satisfaction questionnaires evaluating 417 physicians were collected for analysis. Patient confidence (positively correlated; P = .001) and physician/patient time (inversely correlated; P = .001) were associated with higher physician productivity. Increases in MRVU were associated with decreases in patient perceptions of time with the physician (P = .003). The relationships between patient satisfaction and physician productivity were relatively small, suggesting that they are not necessarily incompatible and that both can be improved simultaneously.

Human term myometrium is poorly characterized as a source of proinflammatory mediators involved in parturition. We have investigated the basal expression of cytokines in myometrium, as well as the effects of CRH and lipopolysaccharide (LPS) on cytokine release. Explants from term myometrium were challenged with CRH or LPS (1 microg/mL each) in short-term tissue culture. Interleukin (IL)-1beta++, IL-6, IL-8, and tumor necrosis factor (TNF)alpha concentrations in the medium were quantified by enzyme immunoassay. The major cytokines released after 24 h were IL-6 and IL-8. All cytokines investigated were stimulated significantly by LPS (P: < 0. 05) but not by CRH. Messenger RNA levels of these cytokines were investigated by RT-PCR. IL-1beta+ and IL-6 messenger RNA were present in preterm and term myometrium before and during labor, whereas IL-8 and TNFalpha were expressed only by myometrium in active labor. Furthermore, myometrial CRH receptors and macrophages were characterized immunohistochemically. We conclude that human term myometrium is a site of production of proinflammatory cytokines and is involved in the inflammation-like reactions mediating the birth process. Cytokine release in term myometrium seems not to be under control of CRH.

Transport and phosphorylation of 2-fluoroadenosine (F-AR) were studied in human erythrocytes and porcine aortic endothelial cells by 19F-nuclear magnetic resonance (NMR) spectroscopy. F-AR (590 microM) added to a human erythrocyte suspension (15% hematocrit) was rapidly incorporated into adenine nucleotides at a rate of 38 nmol.min-1.ml red blood cells-1. Intracellular F-AR could be distinguished from extracellular F-AR due to a chemical shift difference of 0.43 +/- 0.03 ppm (n = 5 experiments). Compared with F-AR, fluoro-ATP purified by high-performance liquid chromatography (HPLC) exhibited a chemical shift of -0.052 ppm, which was too small to differentiate intracellular F-AR and fluoro-ATP in vivo. F-AR uptake was decreased by inhibition of membrane transport with dipyridamole (25 microM) or blockade of adenosine kinase by iodotubercidin (10 microM). The time course of F-AR uptake suggested that the rate-limiting step was not membrane transport but the intracellular phosphorylation by adenosine kinase. In porcine aortic endothelial cells grown on microcarrier beads and perfused within the magnet, there was a linear relation between the F-AR concentration applied (2, 4, 8, or 32 microM) and net uptake measured (27-827 pmol.min-1.mg-1). Intra- and extracellular fluoroadenine compounds were separated by 0.12 ppm, and HPLC analysis confirmed F-AR conversion to fluoroadenine nucleotides. Our findings demonstrate that cellular transport and metabolism of F-AR can be noninvasively studied and analyzed by 19F-NMR.

The dependence of macromolecule permeability (MP; indicator fluorescein isothiocyanate-labeled albumin) of endothelial cells on their energetic state was investigated using confluent monolayers of rat coronary microvascular endothelial cells and porcine aortic macrovascular endothelial cells. When oxidative and glycolytic energy productions were inhibited (5 mM KCN plus 5 mM 2-deoxy-D-glucose) 90% of the endothelial ATP contents were lost within 15 min, followed by a progressive increase of MP, disintegration of the actin cytoskeleton, and the opening of intercellular gaps. Elution of the blocker and a subsequent 3-h incubation in complete culture medium reversed the effects of 2-h metabolic blockade, completely for MP and partially for ATP levels. Coronary microvascular and aortic macrovascular endothelial cells responded similarly to energy depletion and repletion, the microvascular cells being more sensitive. The results demonstrate that 1) energetic inhibition augments endothelial macromolecule permeability when both oxidative and glycolytic energy production are inhibited, 2) increased macromolecule permeability in energy-depleted endothelial monolayers is caused by the opening of intercellular gaps, and 3) endothelial cells reversibly tolerate up to 2 h of almost complete ATP depletion.

The ability of coronary endothelial cells in 14 day confluent cultures to metabolize glucose, palmitate, lactate and various amino acids was investigated. Under aerobic conditions, 99% of glucose, (5 mM) was degraded to lactate and only 0.04% was oxidized in the Krebs cycle. One percent of the glucose catabolized was directed into the hexose monophosphate pathway, but this fraction could be increased by 81% by 0.4 mM methylene blue. Glucose oxidation in the Krebs cycle was increased at glucose concentrations lower than 1 mM, or by the uncoupler 2,4-dinitrophenol. Oxidation to CO2 of palmitate (300 microM), lactate (1 mM), and glutamine (0.5 mM) was diminished in the presence of glucose (5 mM) by 80, 66, and 48%, respectively. These results demonstrate that coronary endothelial cells utilize exogenous glucose, at physiological concentration, predominantly for glycolytic energy production. The metabolic pattern is characteristic of the Crabtree effect. In these cells, glucose not only effectively suppresses the oxidation of the substrates lactate and palmitate, i.e. of substrates preferred by the whole heart, but also of glutamine, which is a major oxidative substrate for coronary endothelial cells. Absolute rates of substrate catabolism are low as compared to those of the beating heart indicating a low energy demand of coronary endothelial cells.

Test validity is determined by the proportion of results that are diagnostically confirmed and predicted on the measures used to identify the disease process. This article summarizes the results of a series of 224 stable high-risk infants who were screened by automated (ALGO-1) and conventional (Bio-logics LT) ABR instrumentation. Failure criteria was defined as the absence or prolongation of a replicable wave V response (conventional) or Refer by the automated system. The overall failure rates at a 35 dB screening level were comparable between devices. Sensitivity and specificity measures for the ALGO-1 unit were 100 and 96 percent, respectively. Permanent hearing loss was demonstrated in 5 percent of the newborns screened in this study. Advantages of the automated system include a dual artifact rejection system, attenuating ear couplers, and a battery operated design. These findings suggest that the automated ABR screener is a viable alternative to conventional ABR instrumentation for the limited purpose of neonatal auditory screening.

We report the use of hyperbaric oxygen in four neonates with delayed wound healing. Three presented with cyanotic congenital heart disease and had wounds associated with surgical procedures; the fourth had a nonhealing wound as a result of a complication of an umbilical-artery catheter. All were treated in a hyperbaric chamber with 100% oxygen at 2 atmospheres absolute. All wounds healed after institution of hyperbaric therapy. There was no evidence of serious side effects in any patient. These observations suggest, but do not prove, the efficacy of hyperbaric oxygen therapy for neonates with delayed wound healing.

Using light and electron-microscopic immunolocalization techniques, and gel electrophoresis combined with immunoblotting, we have examined the expression of cytoskeletal proteins in normal human fetal, child and adult lenses, in human anterior capsular cataract and in bovine lens cells in vivo and in vitro. In this report, we focus our observations on the pattern of actin-isoform expression during normal and pathological situations in vivo and culture conditions. We have noted that cells of developing and mature human lenses as well as bovine lens cells in situ contain only beta- and gamma-actins. In contrast, alpha-smooth muscle (alpha-sm) actin, an isoform typical of smooth muscle differentiation, was demonstrated in bovine lens cells at different times of culture. Moreover, the multilayered cells observed in the subcapsular zone of human anterior capsular cataract were characterized by the presence of alpha-sm actin. Thus, extensive changes in actin-isoform expression take place in lens cells growing in culture and may also occur during cataractogenesis. The biological meaning of the appearance of a marker of myoid differentiation in the ectodermally derived lens-forming cells is discussed.

Endothelial cells (EC) contribute to the control of local vascular diameter by formation of an endothelium derived relaxant factor (EDRF) (1). Whether nitric oxide (NO) is identical with (EDRF) or might represent only one species of several EDRFs has not been decided as yet (2-5). Therefore, we have directly compared in cultured EC the kinetics of NO formation determined in a photometric assay with the vasodilatory effect of EDRF and NO in a bioassay. Basal release of NO was 16, 4 pmol/min/ml packed EC column. After stimulation with bradykinin (BK) and ATP onset of endothelial NO release and maximal response preceded the EDRF-mediated relaxation. Concentrations of NO formed by stimulated EC were quantitatively sufficient to fully explain the smooth muscle relaxation determined in the bioassay. Our data provide convincing evidence that under basal, BK and ATP-stimulated conditions 1. endothelial cells release nitric oxide as free radical, 2. nitric oxide is solely responsible for the vasodilatory properties of EDRF.

"This paper provides a one-period normative model that may be used as a guide or benchmark by which the economic planner may develop policies and plans for regional economic development. The model can accomodate a region that could be as large as a small country or as small as a city, provided sufficient relevant data are available." The authors outline "a procedure for modeling and solving economic planning and analysis problems with 'intergoal trade-offs' while retaining the deviational variable as a measure of satisfaction....[They present] a generalized regional polynomial goal programming model with some suggested objectives and constraints [and illustrate] the approach with a hypothetical example.", (excerpt)

The practice of injecting pregnant women with methylene blue to detect membrane rupture and differentiate twin sacs is discussed. The authors recommend caution in the use of substances with potential adverse fetal effects.

Cultured heart cells from adult rats were exposed to anoxia in a substrate-free Tyrode's solution at constant pH. In this system the metabolic and the morphologic pattern can be investigated simultaneously. Anoxic changes develop gradually above 2 mumol adenosine triphosphate (ATP)/ gww . Morphometry reveals that the morphologic changes are closely related to the energetic state: creatine phosphate (CP) decay is accompanied by the loss of small mitochondrial matrix granules (r = 0.97). The fall of ATP is coincident with sarcomere shortening (r = 0.95) and, below 4 mumol/ gww , with mitochondrial swelling (r = -0.88). The number of lipid droplets correlates with the ATP level during anoxia and reoxygenation (r = -0.92). The early energetic depletion is accompanied by a moderate release of cytosolic enzymes and morphologic changes: the appearance of sarcolemmal microblebs and an increase in subsarcolemmal vesicles. Below an average ATP level of 2 mumol/ gww an increasing number of individual cells fail to recover when reoxygenated . However, that failure is accompanied neither by massive enzyme release nor by ultrastructural damage regarded as typical for the "oxygen paradox."

In substrate-free anoxia, activities of released cytosolic enzymes (LDH, MDH) correlate inversely with the actual ATP level (for both: r = -0.98). At the same time there is a close correlation between lactate production from glycogen and the ATP content (r = 0.98). With external glucose present enzyme release is greatly delayed, but this could be due to the stimulation of glycolysis as well as to the maintenance of high ATP levels. When glycolysis is blocked by iodoacetate under aerobic conditions, the cells also become depleted of high-energy phosphates. This depletion is delayed in the presence of pyruvate. Cytosolic enzyme release again is correlated with total ATP contents, by the same relation in the presence or absence of pyruvate. Glycolytic energy production is negligible in both cases and does not seem to determine enzyme release directly.

To achieve early cellular attachment of isolated adult cardiac myocytes in primary culture, various procedures were tested. A selective attachment of rod-shaped cells can be obtained either on serum-preincubated tissue-culture plastic dishes, or on modified plastic materials to which serum proteins are bound covalently, or on laminin. For the pretreatment of tissues culture dishes fetal calf serum is better suited than other sera.

Ca2+-tolerant isolated adult heart cells can be exposed to 1 mM EGTA and then again to 1 mM CaCl2 without developing irreversible hypercontracture. Thus, they are not subject to the calcium paradox, even though they apparently become more permeable to Na+ during Ca2+-free incubation. When these cells are incubated anoxically without substrate they slowly lose their energetic reserves. The process resembles that of the arrested anoxic myocardium. The appearance of irreversibly hypercontracted cells is neither accompanied by a parallel increase in enzyme release nor by an aggravation of the anoxia-induced damage by reoxygenation. Thus these cells are not subject to the oxygen paradox. It is suggested that the differences between the myocytes' behaviour in tissue and in the isolated state are due to the fact that the isolated cells are free from mechanical interactions with adjacent cells.

Catherization of the posterior tibial artery provides an alternative to cannulation of other arteries. We successfully catheterized this artery 17 times in 15 neonates whose weights ranged from 800 to 3,000 g. The catheters were placed percutaneously 13 times and by cutdown four times, and they remained in place for as long as 12 days (mean, 96 hours). Compromise of distal circulation was found in only two babies, both of whom had had umbilical artery catheters removed because of cyanosis of the toes that had resolved itself prior to posterior tibial artery cannulation. Our patients experienced no short-term complications from this procedure.

Cultured adult cardiocytes were exposed to anoxia. The initial decrease of high-energy phosphates was accompanied by a moderate release of cytosolic enzymes and morphological changes: the appearance of sarcolemmal 'microblebs' (approximately 1 micron in diameter) and an increase of subsarcolemmal vesicles. At ATP levels above 2 mumol/gww, metabolic and morphological alterations were reversible. Probably the sarcolemmal changes are causally related to the loss of macromolecules from reversibly injured cells. At ATP levels below 2 mumol/gww, an increasing number of cells become irreversibly hypercontracted. In these cells cytoplasmic masses are protruded into large 'macroblebs' (10-30 micron in diameter), however sarcolemmal continuity is preserved. Thus, enzyme release, irreversible contracture and cytolysis do not occur simultaneously in anoxic isolated cardiocytes.

On serum precoated tissue culture dishes, isolated ventricular myocytes attach firmly during 4 hours of incubation. Since in this monoculture cells do not divide and show only minimal signs of cytoplasmic spreading, individual cells mostly lie isolated from others. However, when occasionally two cells attach in close vicinity, new cell-cell contact structures are formed already during the first hours in culture. The de-novo formation of these communications is demonstrated by the finding that they often connect cells in an end-to-side manner that does not occur in vivo.

In Ca-tolerant adult cardiomyocytes the contribution of endogenous substrates (glycogen, tri- and diacylglycerol) to oxidative substrate metabolism was investigated. After 4 h in culture medium (M 199 plus 4% fetal calf serum) the cellular triacylglycerol content is 3.6-fold higher than in fresh myocardium and reflects the free fatty acid composition of the medium. When triacylglycerol is degraded, all long-chain fatty acids are hydrolysed at equal rates. In these quiescent cells, the activity of pyruvate dehydrogenase is low (10% of full activity, in Tyrode solution with 5 mM glucose). Up to 30% of full pyruvate dehydrogenase activity, the contribution of non-lipid substrates (glycogen, glucose, lactate and pyruvate) to oxidative energy production is correlated to pyruvate dehydrogenase activity. At 5 mM medium concentration, glucose, lactate and pyruvate share in energy production the proportions of 15, 36 and 50%, whereas endogenous lipolysis accounts for 78, 61 and 46%. It is concluded that these quiescent cardiomyocytes represent cardiac metabolism in a basal state in which the preference for fatty acids, especially from endogenous lipids, is very pronounced. The utilization of endogenous substrates therefore has to be considered in all studies investigating the oxidative metabolism of these isolated cells.

In medium 199 plus 20% fetal calf serum adult rat cardiomyocytes establish a long-term culture (25 days). During the first 10 days they change their gross morphology from the typical elongated in vivo shape (day 1), to a smooth spherical intermediate form (days 2 to 5), to a spread cell type beating spontaneously (days 10 to 15). During the first 10 days in culture, protein content per cell increases and the cell population decreases. By the tenth day, protein content has doubled, and about half of the cells originally plated remain. Thereafter both the protein content and the number of cells are essentially constant for the remainder of the 25-day period investigated. On days 1, 15 and 25 adenine nucleotide contents (213, 216 and 225 nmol/10(6) cells) and values of adenylate energy charge (0.91, 0.87 and 0.88) were similar. At all times in culture, palmitate (0.1 mM) is oxidized at higher rates than lactate (1 mM) and glucose (5 mM). At all times in culture glycolytic flux is sensitive to insulin with half maximal effect seen around 10(-9) M. Oxidation rates for all exogenous substrates are maximal at 15 days in culture, indicating maximal energy demand at this time. The conversion of glucose to lactate, however, progressively increases, so that at 25 days in culture, 70% of ATP derived from degradation of exogenous glucose is glycolytic. The results of this study demonstrate that oxidative metabolism of cardiomyocytes in long-term culture resembles, in its basic characteristics, that of the intact heart. In their increased glycolytic activity, however, they are clearly different.

Nineteen cases of nonimmunologic hydrops fetalis occurring during a nine-year period were reviewed. The pregnancies were complicated by hydramnios (78%) and preterm delivery (84%). Hydramnios appears to be the most useful indicator of the pregnancy at risk; its occurrence should prompt ultrasonographic investigation for evidence of hydrops. Modalities available for antenatal diagnosis of underlying fetal abnormalities include amniocentesis, serologic tests, fetal cardiac monitoring, radiography, hemoglobin electrophoresis and glucose tolerance testing. A specific cause for the hydrops may not be detectable (42% of our cases were idiopathic). Management of affected pregnancies is influenced by the frequent occurrence of fetal asphyxia and premature delivery. Outcome is poor: only 32% of the babies survived beyond the neonatal period. Symptomatic treatment for the neonate includes fluid restriction, maintenance of blood sugar, support of ventilation and attention to the complications of asphyxia.