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Selected AbstractsEffects of metformin and oleic acid on adipocyte expression of resistinDIABETES OBESITY & METABOLISM, Issue 1 2006R Rea Aim:, The adipocyte-secreted hormone resistin has been implicated in obesity-induced insulin resistance and type 2 diabetes, but pharmacological and dietary factors that regulate resistin gene expression and the effects of resistin on cellular glucose uptake in muscle have not been clearly defined. Methods:, Expression of resistin mRNA was studied in differentiated 3T3-L1 adipocytes by using real-time semiquantitative reverse transcription-polymerase chain reaction. The effects of resistin on insulin-stimulated and insulin-independent 2-deoxyglucose uptake were evaluated in L6 muscle cells. Results:, Insulin 1 µm and rosiglitazone 10 µm markedly reduced resistin mRNA expression (relative to the control gene TF2D) by 4.7-fold (p < 0.05) and 5.3-fold (p < 0.02), respectively. Similar reductions in resistin mRNA were demonstrated with metformin 100 µm (6.2-fold reduction, p < 0.02) and oleic acid 100 µm (3.9-fold reduction, p < 0.03). Resistin 1 µm significantly reduced maximum insulin-stimulated 2-deoxyglucose uptake in L6 cells from 634 to 383% (relative to 100% for control, p < 0.001), and co-administration of rosiglitazone had no effect on resistin-induced insulin resistance. In the absence of insulin, however, resistin increased glucose uptake dose-dependently (e.g., 1.75-fold at 5 µm, p < 0.001) via a mitogen-activated protein kinase-dependent pathway. Conclusions:, These results demonstrate that various glucose-lowering therapies and oleic acid reduce resistin gene expression in isolated adipocytes, and that resistin impairs insulin-stimulated glucose uptake in skeletal muscle-derived cells. [source] Early intervention with second-generation antipsychotics in first-episode psychosis: results of an 8-week naturalistic studyEARLY INTERVENTION IN PSYCHIATRY, Issue 1 2010Richard C. Josiassen Abstract Objective: The objective was to compare short-term effectiveness of aripiprazole with three other second-generation antipsychotics (SGAs) in the treatment of first-episode psychosis. Method: In a naturalistic, ,single-blind' design, 60 subjects experiencing their first psychotic episode were treated for 8 weeks with aripiprazole (n = 19), risperidone (n = 16), olanzapine (n = 14) or quetiapine (n = 11). Medication and dosing decisions were made by treating psychiatrists, constrained to once-a-day dosing, low initial doses and no clozapine. Weekly ratings were obtained using the Positive and Negative Syndrome Scale (PANSS), Simpson-Angus Rating Scale and Barnes Akathasia Rating Scale. Weight and vital signs were also collected weekly. Results: The group presented with severe psychotic symptoms (mean baseline PANSS total score of 105.2), which were reduced rapidly (P < 0.0005). The between-group and group by time interaction terms were non-significant. Similar reductions were seen across all PANSS sub-scales. At Week 1 the mean PANSS Activation Scale score was reduced more with olanzapine than in the other groups (P < 0.002). Few instances of extrapyramidal symptoms occurred; all were sporadic and did not require treatment. Group body weight increased by 7.3% over the study. Vital signs remained unchanged. Conclusions: Early intervention with low doses of four SGAs led to rapid symptom reduction in first-episode psychotic patients with severe psychopathology. Although no clear medication advantages were observed in the short term, longer duration studies with larger samples will be required for determining efficacy, rates of compliance, relapse prevention and diminished incidence of extrapyramidal signs and symptoms. [source] Similar reductions in body-weight for placebo and green teaFOCUS ON ALTERNATIVE AND COMPLEMENTARY THERAPIES AN EVIDENCE-BASED APPROACH, Issue 2 2006Article first published online: 14 JUN 2010 [source] The effect of repeated professional supragingival plaque removal on the composition of the supra- and subgingival microbiotaJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 9 2000Laurie Ann Ximénez-Fyvie Abstract Background, aims: The purpose of the present investigation was to determine the effect of weekly professionally administered supragingival plaque removal on the composition of the supra and subgingival microbiota. Methods: 18 adult subjects with periodontitis who had been treated and were in a maintenance phase of therapy were clinically and microbiologically monitored at baseline, 3, 6 and 12 months. After the baseline visit, the subjects received scaling and root planing followed by professional supragingival plaque removal every week for 3 months. Clinical measures of plaque accumulation, bleeding on probing (BOP), gingival redness, suppuration, pocket depth and attachment level were made at 6 sites per tooth at each visit. Separate supra (N=1804) and subgingival (N=1804) plaque samples were taken from the mesial aspect of all teeth excluding third molars in each subject at each time point and evaluated for their content of 40 bacterial taxa using checkerboard DNA-DNA hybridization. Significance of changes in mean counts, prevalence and proportions of bacterial species over time in both supra and subgingival samples were determined using the Quade test and adjusted for multiple comparisons. Results: Mean % of sites exhibiting plaque, gingival redness and BOP were significantly reduced during the course of the study. Significant decreases in mean counts were observed in both supra and subgingival samples. Mean total DNA probe counts (×105, ±SEM) at baseline, 3, 6 and 12 months were: 133±19, 95±25, 66±6, 41±6 (p<0.001) for supragingival samples and 105±22, 40±10, 19±4, 13±3 (p<0.001) for subgingival samples. Mean counts of 22 of 40 and 34 of 40 species tested were significantly reduced in the supra and subgingival samples respectively over the monitoring period. For example, mean counts of Porphyromonas gingivalis×105 at baseline, 3, 6 and 12 months in the subgingival plaque samples were 2.0±0.4, 0.5±0.2, 0.6±0.3, 0.3±0.1 (p<0.001); Bacteroides forsythus 2.0±0.6, 0.4±0.1, 0.4±0.2, 0.1±0.2 (p<0.001); Treponema denticola 3.4±1.1, 0.8±0.3, 0.4±0.2, 0.3±0.3 (p<0.01). Similar reductions were seen in supragingival plaque samples. While counts were markedly reduced by professional plaque removal, the proportion and prevalence of the 40 test species were marginally affected. Conclusions: Weekly professional supragingival plaque removal profoundly diminished counts of both supra- and subgingival species creating a microbial profile comparable to that observed in periodontal health. This profile was maintained at the final monitoring visit, 9 months after completion of therapy. [source] Corneal hysteresis using the Reichert ocular response analyser: findings pre- and post-LASIK and LASEKACTA OPHTHALMOLOGICA, Issue 2 2008Caitriona Kirwan Abstract. Purpose:, To evaluate and compare corneal hysteresis in patients prior to and following laser in situ keratomileusis (LASIK) and laser-assisted subepithelial keratectomy (LASEK) using the Reichert ocular response analyser (ORA). Methods:, Corneal hysteresis was recorded prior to and 3 months after corneal laser refractive surgery for myopia. Preoperative corneal hysteresis was correlated with age and preoperative central corneal thickness (CCT). Postoperative corneal hysteresis was correlated with postoperative CCT in both the LASIK and LASEK treatment groups. The correlations between postoperative change in hysteresis and stromal ablation depth, percentage of tissue ablated, optical zone and patient age were also examined. Results:, A total of 84 eyes of 84 patients were involved in the study. LASIK was performed in 63 eyes and LASEK in 21. Mean preoperative corneal hysteresis of all eyes was 10.8 ± 1.5 mmHg. Mean age, preoperative CCT, corneal hysteresis and ablation profile were similar in both groups. A statistically significant decrease in hysteresis occurred following LASIK (p < 0.01) and LASEK (p < 0.01) with similar decrements observed in both treatment groups. A moderate correlation was found between postoperative hysteresis and postoperative CCT in LASIK (r = 0.7) and LASEK (r = 0.7) treated eyes. A weak correlation was found between postoperative decrease in hysteresis and the parameters examined. Conclusion:, Corneal hysteresis decreased following LASIK and LASEK. Similar reductions occurred following both procedures, indicating that LASIK involving a thin 120-,m flap did not induce additional biomechanical change. Postoperative reduction in hysteresis did not correlate with the amount or percentage of corneal tissue removed, nor with optical zone or patient age. [source] Growth-induced changes in the proteome of Helicobacter pyloriELECTROPHORESIS, Issue 5-6 2006Christina Uwins Abstract Helicobacter pylori is a major human pathogen that is responsible for a number of gastrointestinal infections. We have used 2-DE to characterise protein synthesis in bacteria grown either on solid agar-based media or in each of two broth culture media (Brucella and brain heart infusion (BHI) broth). Significant differences were observed in the proteomes of bacteria grown either on agar-based or in broth media. Major changes in protein abundance were identified using principal component analysis (PCA), which delineated the profiles derived for the three key growth conditions (i.e. agar plates, Brucella and BHI broth). Proteins detected across the gel series were identified by peptide mass mapping and Edman sequencing. A number of proteins associated with protein synthesis in general as well as specific amino acid synthesis were depressed in broth-grown bacteria compared to plate-grown bacteria. A similar reduction was also observed in the abundance of proteins involved in detoxification. Two of the most abundant spots, identified as UreB and GroEL, in plate-grown bacteria showed a >140-fold drop in abundance in bacteria grown in Brucella broth compared to bacteria grown on agar plates. Two protein spots induced in bacteria grown in broth culture were both identified as glyceraldehyde 3-phosphate dehydrogenase based on their N -terminal amino acid sequences derived by Edman degradation. The underlying causes of the changes in the proteins abundance were not clear, but it was likely that a significant proportion of the changes were due to the alkaline pH of the broth culture media. [source] Chronic interleukin-6 alters the level of synaptic proteins in hippocampus in culture and in vivoEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 12 2007Elly J. F. Vereyken Abstract There is now considerable evidence that the level of expression of the proinflammatory cytokine, interleukin-6 (IL-6), is increased in the central nervous system (CNS) during neuroinflammatory conditions such as occurs in neurological disorders and in disease and injury. However, our understanding of the consequences of increased expression of IL-6 on the CNS is still limited, especially with respect to the developing nervous system, which is known to be particularly vulnerable to environmental factors. To address this issue, we investigated the properties of cultured hippocampal neurons exposed chronically to IL-6 during the main period of morphological and physiological development, which occurs during the first 2 weeks of culture. IL-6 was tested at 500 U/mL, considered to reflect a pathophysiologic concentration. The morphological features of neuronal development in the control and IL-6-treated cultures appeared similar. However, Western blot analysis showed a significant reduction in the level of Group-II metabotropic receptors (mGluR2/3) and L-type Ca2+ channels in the IL-6-treated cultures. A similar reduction in mGluR2/3 and L-type Ca2+ channel protein was observed in transgenic mice that over-express IL-6 in the CNS through astrocyte production starting early in development. Analysis of Ca2+ signals produced by spontaneous synaptic network activity in the hippocampal cultures and effects of a mGluR2/3 agonist and antagonist showed that the reduced levels of mGluR2/3 impact on the functional properties of hippocampal synaptic network activity. These results have important implications relative to the mechanisms responsible for altered CNS function during conditions associated with increased levels of IL-6 in the CNS. [source] SEXUAL SELECTION AND IMMUNE FUNCTION IN DROSOPHILA MELANOGASTEREVOLUTION, Issue 2 2008Kurt A. McKean The evolution of immune function depends not only on variation in genes contributing directly to the immune response, but also on genetic variation in other traits indirectly affecting immunocompetence. In particular, sexual selection is predicted to trade-off with immunocompetence because the extra investment of resources needed to increase sexual competitiveness reduces investment in immune function. Additional possible immunological consequences of intensifying sexual selection include an exaggeration of immunological sexual dimorphism, and the reduction of condition-dependent immunological costs due to selection of ,good genes' (the immunocompetence handicap hypothesis, ICHH). We tested for these evolutionary possibilities by increasing sexual selection in laboratory populations of Drosophila melanogaster for 58 generations by reestablishing a male-biased sex ratio at the start of each generation. Sexually selected flies were larger, took longer to develop, and the males were more sexually competitive than males from control (equal sex ratio) lines. We found support for the trade-off hypothesis: sexually selected males were found to have reduced immune function compared to control males. However, we found no evidence that sexual selection promoted immunological sexual dimorphism because females showed a similar reduction in immune function. We found no evidence of evolutionary changes in the condition-dependent expression of immunocompetence contrary to the expectations of the ICHH. Lastly, we compared males from the unselected base population that were either successful (IS) or unsuccessful (IU) in a competitive mating experiment. IS males showed reduced immune function relative to IU males, suggesting that patterns of phenotypic correlation largely mirror patterns of genetic correlation revealed by the selection experiment. Our results suggest increased disease susceptibility could be an important cost limiting increases in sexual competitiveness in populations experiencing intense sexual selection. Such costs may be particularly important given the high intersex correlation, because this represents an apparent genetic conflict, preventing males from reaching their sexually selected optimum. [source] The effect of contact load reduction on the fatigue life of pearlitic rail steel in lubricated rolling,sliding contactFATIGUE & FRACTURE OF ENGINEERING MATERIALS AND STRUCTURES, Issue 8 2000D. I. Fletcher Twin-disc contact simulation tests were carried out to investigate the influence of contact pressure variation on rail steel fatigue life. Both a colloidal suspension of molybdenum disulphide in an oil carrier fluid (similar to a commercial flange lubrication product) and water were used as lubricants. It was found that the reduction from 1500 to 900 MPa of the maximum Hertzian contact pressure (at which a molybdenum,disulphide-lubricated and previously worn rail sample was tested) extended the fatigue life of the rail steel by over five times. For water lubrication a similar reduction in contact pressure produced only a marginal increase in fatigue life. The results were found to be in qualitative agreement with the predictions of the newly developed Three Mechanism (TM) model of rolling contact fatigue, which is introduced here. This model combines the mechanisms of ratcheting and the fracture mechanics-based mechanisms of both shear stress- and tensile stress-driven, fluid-assisted, crack growth. [source] Severe alterations of endothelial and glial cells in the blood-brain barrier of dystrophic mdx miceGLIA, Issue 3 2003Beatrice Nico Abstract In this study, we investigated the involvement of the blood-brain barrier (BBB) in the brain of the dystrophin-deficient mdx mouse, an experimental model of Duchenne muscular dystrophy (DMD). To this purpose, we used two tight junction markers, the Zonula occludens (ZO-1) and claudin-1 proteins, and a glial marker, the aquaporin-4 (AQP4) protein, whose expression is correlated with BBB differentiation and integrity. Results showed that most of the brain microvessels in mdx mice were lined by altered endothelial cells that showed open tight junctions and were surrounded by swollen glial processes. Moreover, 18% of the perivascular glial endfeet contained electron-dense cellular debris and were enveloped by degenerating microvessels. Western blot showed a 60% reduction in the ZO-1 protein content in mdx mice and a similar reduction in AQP4 content compared with the control brain. ZO-1 immunocytochemistry and claudin-1 immunofluorescence in mdx mice revealed a diffuse staining of microvessels as compared with the control ones, which displayed a banded staining pattern. ZO-1 immunogold electron microscopy showed unlabeled tight junctions and the presence of gold particles scattered in the endothelial cytoplasm in the mdx mice, whereas ZO-1 gold particles were exclusively located at the endothelial tight junctions in the controls. Dual immunofluorescence staining of ,-actin and ZO-1 revealed colocalization of these proteins. As in ZO-1 staining, the pattern of immunolabeling with anti,,-actin antibody was diffuse in the mdx vessels and pointed or banded in the controls. ,-actin immunogold electron microscopy showed gold particles in the cytoplasms of endothelial cells and pericytes in the mdx mice, whereas ,-actin gold particles were revealed on the endothelial tight junctions and the cytoskeletal microfilaments of pericytes in the controls. Perivascular glial processes of the mdx mice appeared faintly stained by anti-AQP4 antibody, while in the controls a strong AQP4 labeling of glial processes was detected at light and electron microscope level. The vascular permeability of the mdx brain microvessels was investigated by means of the horseradish peroxidase (HRP). After HRP injection, extensive perivascular areas of marker escape were observed in mdx mice, whereas HRP was exclusively intravascularly localized in the controls. Inflammatory cells, CD4-, CD8-, CD20-, and CD68-positive cells, were not revealed in the perivascular stroma of the mdx brain. These findings indicate that dystrophin deficiency in the mdx brain leads to severe injury of the endothelial and glial cells with disturbance in ,-actin cytoskeleton, ZO-1, claudin-1, and AQP4 assembly, as well as BBB breakdown. The BBB alterations suggest that changes in vascular permeability are involved in the pathogenesis of the neurological dysfunction associated with DMD. GLIA 42:235,251, 2003. © 2003 Wiley-Liss, Inc. [source] Inactivation of root canal medicaments by dentine, hydroxylapatite and bovine serum albuminINTERNATIONAL ENDODONTIC JOURNAL, Issue 3 2001I. Portenier Abstract Aim This study examined and compared the inhibition of the antibacterial effect of saturated calcium hydroxide solution, chlorhexidine acetate and iodine potassium iodide by dentine, hydroxylapatite and bovine serum albumin. MethodologyEnterococcus faecalis strain A197A prepared to a suspension of 3 × 108 cells per ml in 0.5% peptone water was used. Fifty µL of saturated calcium hydroxide solution, 0.05% chlorhexidine acetate or 0.2/0.4% iodine potassium iodide were incubated at 37 °C with 28 mg dentine powder (DP), hydroxylapatite (HA) or bovine serum albumin (BSA) in 50 µL water for 1 h before adding 50 µL of the bacterial suspension. Samples for bacterial culturing were taken from the suspension 1 and 24 h after adding the bacteria. In further experiments, the amount of dentine was stepwise reduced from 28 mg 150 µL,1 to 2.8 mg 150 µL,1. Results Calcium hydroxide was totally inactivated by the presence of 28 mg of DP, HA or BSA. Chlorhexidine (0.05%) was strongly inhibited by BSA and slowed down by dentine. However, HA had little or no inhibitory effect on chlorhexidine. The antibacterial effect of 0.2/0.4% iodine potassium iodide on E. faecalis was totally inhibited by dentine (28 mg), but was practically unaffected by HA or BSA. A stepwise reduction of dentine from 28 mg 150 µL,1 to 2.8 mg 150 µL,1 was followed by a similar reduction of the inhibition of the antibacterial activity of chlorhexidine. Iodine potassium iodide was not inhibited at all with dentine amounts less than 28 mg. However, the effect of saturated calcium hydroxide solution was totally eliminated by dentine, in all four concentrations. Conclusion Inhibition by dentine of the antibacterial activity of calcium hydroxide, chlorhexidine and iodine potassium iodide occurs by different mechanisms. Different components of dentine may be responsible for the inhibition of these three medicaments. Calcium hydroxide was particularly sensitive to inhibition by both inorganic and organic compounds. [source] Addition of nimesulide to small intestinal submucosa biomaterial inhibits postsurgical adhesiogenesis in ratsJOURNAL OF BIOMEDICAL MATERIALS RESEARCH, Issue 1 2010Mark A. Suckow Abstract Adhesion formation is a common complication in abdominal surgery with incidence as high as 93% and small bowel obstruction a common complication. Because the extracellular matrix material, small intestinal submucosa (SIS), is commonly used in various surgical procedures, methods to inhibit adhesiogenesis are of great interest. This study was undertaken to determine if incorporation of nimesulide (NM), a selective cyclooxygenase (COX)-2 inhibitor, could reduce the extent and tenacity of intraabdominal adhesion formation associated with SIS implantation. Female Sprague,Dawley rats underwent a cecal abrasion surgical procedure to induce adhesiogenesis. Rats were either left untreated or treated by direct application over the injured cecum with polypropylene mesh (PPM); SIS; SIS containing a low dose of NM; or SIS containing a high dose of NM. Rats were euthanized 21 days later, and adhesion extent and tenacity were evaluated using standard scales (0 = minimal adhesiogenesis; 4 = severe adhesiogenesis). Addition of NM to SIS resulted in a significant (p < 0.05) reduction in adhesion extent and in a similar reduction in adhesion tenacity for SIS containing a low dose of NM. Adhesions typically extended from the abraded cecal surface to the body wall and were characterized histologically by fibrous tissue adherent to the cecal wall. In conclusion, addition of the nonsteroidal anti-inflammatory, COX-2 selective drug, NM, to SIS attenuates adhesion extent and tenacity when compared with surgical placement of SIS or PPM alone. © 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2010 [source] Targeted Cytotoxic Analogue of Luteinizing Hormone-Releasing Hormone (LH-RH) Only Transiently Decreases the Gene Expression of Pituitary Receptors for LH-RHJOURNAL OF NEUROENDOCRINOLOGY, Issue 1 2002M. Kovacs Abstract A cytotoxic analogue of LH-RH, AN-207, consisting of 2-pyrrolinodoxorubicin (AN-201) linked to carrier [D-Lys6]LH-RH, was developed for targeted therapy of cancers expressing LH-RH-receptors. To determine its possible side-effects on the pituitary gland, we investigated the gene expression of pituitary LH-RH-receptors and LH secretion in ovariectomized female and normal male rats after treatment with the maximum tolerated dose of AN-207. The effect of AN-207 on the gene expression of the pituitary GH-RH-receptors and GH secretion was also assessed in male rats. Five hours after a single i.v. injection of AN-207 at 175 nmol/kg, there was a 39,51% decrease in mRNA expression for the pituitary LH-RH-receptors in male and female rats. The carrier, at an equimolar dose, caused a similar reduction (37,39%), whereas the cytotoxic radical AN-201, at an equitoxic dose (110 nmol/kg), produced only a 12,24% decrease (NS) in the mRNA expression of LH-RH-receptors. AN-207 and the carrier analogue induced a comparable 90,100-fold increase in serum LH concentrations in male rats, and the same 12-fold elevation in OVX rats at 5 h. Seven days after treatment with AN-207, the mRNA levels for the LH-RH receptors and the serum LH concentration were back to normal in both sexes. AN-207, the carrier, and AN-201 had no significant effect on the expression of mRNA for GH-RH-receptors in the pituitary. In vitro, a continuous perfusion of pituitary cells with 10 nM AN-207 did not affect the hormone-releasing function of the targeted LH cells or the nontargeted GH cells. Our results demonstrate that cytotoxic LH-RH analogue AN-207, at the maximum tolerated dose causes only a transient decrease in the gene expression of the pituitary LH-RH receptors, and the levels of mRNA for LH-RH receptor fully recover within 7 days. Moreover, the carrier hormone moiety, and not the cytotoxic radical in AN-207 is responsible for this transient suppression. Our findings suggest that the therapy with cytotoxic LH-RH analogues will not inflict permanent damage to pituitary function. [source] Modulation of ERK and JNK activity by transient forebrain ischemia in ratsJOURNAL OF NEUROSCIENCE RESEARCH, Issue 3 2006Deborah A. Shackelford Abstract The mitogen-activated protein (MAP) kinase families of ERK and JNK participate in numerous intracellular signaling pathways and are abundantly expressed in the CNS. Activation of ERK and JNK during reperfusion of ischemic tissue is implicated in promoting cell death, insofar as inhibition of either pathway reduces neuronal cell death. However, ERK or JNK activation provides protection in other neuronal injury models. In this study, we monitored the concurrent modulation of ERK and JNK activity in the hippocampus, neocortex, and striatum during ischemia and immediately upon reperfusion in a rat model of transient global ischemia. All three regions incur a similar reduction in blood flow during occlusion but show different extents and temporal patterns of injury following reperfusion. ERK and JNK were active in the normal rat forebrain, and phosphorylation was reduced by ischemia. Upon reperfusion, ERK was rapidly activated in the hippocampus, neocortex, and striatum, whereas JNK phosphorylation increased in the hippocampus and striatum but not in the neocortex. The response of JNK vs. ERK more closely reflects the susceptibility of these regions. JNK1 was the predominant phosphorylated isoform. A minor pool of phosphorylated JNK3 increased above the control level after reperfusion in hippocampal but not in neocortical particulate fractions. In addition, a novel 32,35-kDa c-Jun kinase activity was detected in the hippocampus, neocortex, and striatum. The results show that ERK and JNK activities are rapidly, but not identically, modulated by ischemia and reperfusion and indicate that the MAP kinase pathways contribute to regulating the response to acute CNS injury. © 2006 Wiley-Liss, Inc. [source] The effect of adding cadmium and lead alone or in combination to the diet of pigs on their growth, carcase composition and reproductionJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 13 2003Clive Phillips Abstract Limits for cadmium and lead concentrations in food animal products have been established independently, whereas these two toxic metals often co-exist in polluted regions. Weaned pigs (60) were allocated to ten treatments: control; low (0.5 mg kg,1), medium (1 mg kg,1) or high cadmium (2.5 mg kg,1) in feed; low (5 mg kg,1) medium (10 mg kg,1) or high (25 mg kg,1) lead in feed; and low, medium and high cadmium plus lead in feed. Growth rates and concentrations of cadmium and lead in body tissues (kidney, liver, spleen, lungs, heart, testicle, ribs, hair and teeth) were measured after 137 days. There was a similar reduction in weight gain for pigs in the cadmium and lead treatments, compared with the control, and a greater reduction for the pigs in the cadmium plus lead treatments. The reduction increased with the level of metal included. There was an increase in cadmium concentration of all tissues and blood with increasing feed cadmium concentration, which was usually less when lead was also included in the feed. There was also an increase in tissue lead concentration with increasing dietary lead, and this was in most cases increased when cadmium was also included in the feed. The most sensitive tissues for cadmium and lead exposure were the kidney, liver, hair and teeth, and regression equations were developed for the accumulation rates in these tissues. Tissue and blood cadmium concentrations increased gradually with increasing dietary lead, whereas tissue lead concentration was not sensitive to dietary cadmium, except in the ribs and heart. In a second experiment, 10 sows were allocated to a control diet or the same diet but with a supplement of cadmium and lead. The birth weight of piglets was decreased by the supplement and their mortality increased. Lead accumulated most in the ovary and oviduct of the sows, and there were increases in the lead and, to a lesser extent, cadmium concentrations of tissues of the piglets from these sows. Copyright © 2003 Society of Chemical Industry [source] Bioactivity of falcarinol and the influenceof processing and storage on its content in carrots (Daucus carota L)JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 10 2003Susanne L Hansen Abstract The concentration-dependent activity of the polyacetylene falcarinol ((9Z)-heptadeca-1,9-dien-4,6-diyn-3-ol), isolated from carrots, was investigated in a bioassay with primary mammary epithelial cells in collagen gels and compared with that of ,-carotene, the orange pigment in carrots. Falcarinol showed biphasic activity, having stimulatory effects between 0.01 and 0.05 µg ml,1 and inhibitory effects between 1 and 10 µg ml,1, whereas ,-carotene showed no effect in the concentration range 0.001,100 µg ml,1. The results are discussed in relation to the health-promoting effects of carrots and related vegetables. Falcarinol was quantified in the carrot cultivars Bolero, Rodelika and Fancy by analytical reverse phase HPLC, subjected to various processing and storage conditions in order to study how long-term storage, blanching, freezing and boiling influence the content of falcarinol. Long-term storage of raw carrot cubes (1 cm3) reduced the falcarinol content by almost 35%. A similar reduction was found in steam-blanched carrot cubes (1 cm3). Long-term storage at ,24 °C of steam blanched carrot cubes did not reduce the falcarinol content further. A reduction of almost 70% in the falcarinol content was found in carrot pieces boiled in water for 12 min compared with raw carrots. Copyright © 2003 Society of Chemical Industry [source] Masitinib, a c-kit/PDGF receptor tyrosine kinase inhibitor, improves disease control in severe corticosteroid-dependent asthmaticsALLERGY, Issue 8 2009M. Humbert Background:, Masitinib is a tyrosine kinase inhibitor targeting stem cell factor receptor (c-kit) and platelet-derived growth factor (PDGF) receptor, which are expressed on several cell types including mast cells and bronchial structural cells, respectively. We hypothesized that c-kit and PDGF receptor inhibition may decrease bronchial inflammation and interfere with airway remodeling, which are crucial features of severe asthma. Objectives:, The primary endpoint was the percent change from baseline in oral corticosteroids after 16 weeks of treatment. Change in asthma control (asthma control questionnaire), exacerbation rate, pulmonary function tests, rescue medication requirement and safety were secondary endpoints. Methods:, A 16-week randomized, dose-ranging (3, 4.5, and 6 mg/kg/day), placebo-controlled study was undertaken in 44 patients with severe corticosteroid-dependent asthma who remained poorly controlled despite optimal asthma management. Results:, At 16 weeks of treatment, a comparable reduction in oral corticosteroids was achieved with masitinib and placebo (median reduction of 78% and 57% in the masitinib and placebo arms, respectively). Despite this similar reduction, the Asthma Control Questionnaire score was significantly better in the masitinib arm as compared to placebo with a reduction by 0.99 unit at week 16 (P < 0.001) vs 0.43 unit in the placebo arm. Masitinib therapy was associated with more transient skin rash and edema. Conclusions:, Masitinib, a c-kit and PDGF-receptor tyrosine kinase inhibitor, may represent an innovative avenue of treatment in corticosteroid-dependent asthma. These preliminary results warrant further long-term clinical studies in severe asthma (ClinicalTrials.gov Identifier: NCT00842270). [source] The detection of small simulated field defects using multifocal VEPsOPHTHALMIC AND PHYSIOLOGICAL OPTICS, Issue 3 2003H. L. Chan Abstract Introduction: The multifocal visual-evoked potential (mfVEP) has been widely investigated in the study of diseases of the visual system. However, the sensitivity of the mfVEP in objective detection of field defects has not been determined. This study investigates the variation of the mfVEP responses whilst simulating field defects by using different sizes of mask on the stimulus pattern. Methods: Simulated field defects of four different sizes (2, 3, 5, and 7 degrees) at two different eccentricities (10 and 16 degrees) were generated on a standard mfVEP dartboard stimulus using opaque masks. These masks were placed at the centre of each dartboard sector and the modified stimuli were used to elicit mfVEPs from 10 normal subjects. The response densities and latencies of N1, P1 of the mfVEP were compared, without and with small simulated field defects. Results: The minimum size of simulated field defect causing significant response density reduction in P1 and N1 was 5 degrees at both retinal eccentricities. N1 showed similar reduction in response density at both retinal eccentricities, but P1 showed larger reduction at the 10-degree location than at the 16-degree location. There was no change in latencies with simulated field defect at either location. Conclusions: The mfVEP is only sensitive to a simulated field defect equal to or larger than 5 degrees in diameter, and mfVEP has greater sensitivity at 10-degree eccentricity than at 16-degree eccentricity. [source] The development of the RTS,S malaria vaccine candidate: challenges and lessonsPARASITE IMMUNOLOGY, Issue 9 2009W. R. BALLOU Summary RTS,S is the world's most advanced malaria vaccine candidate and is intended to protect infants and young children living in malaria endemic areas of sub-Saharan Africa against clinical disease caused by Plasmodium falciparum. Recently, a pivotal Phase III efficacy trial of RTS,S began in Africa. The goal of the programme has been to develop a vaccine that will be safe and effective when administered via the Expanded Program for Immunization (EPI) and significantly reduce the risk of clinically important malaria disease during the first years of life. If a similar reduction in the risk of severe malaria and other important co-morbidities associated with malaria infection can be achieved, then the vaccine could become a major new tool for reducing the burden of malaria in sub-Saharan Africa. Encouraging data from the ongoing phase II programme suggest that these goals may indeed be achievable. This review discusses some of the unique challenges that were faced during the development of this vaccine, highlights the complexity of developing new vaccine technologies and illustrates the power of partnerships in the ongoing fight against this killer disease. [source] The hydrodynamic and conformational properties of denatured proteins in dilute solutionsPROTEIN SCIENCE, Issue 1 2010Guy C. Berry Abstract Published data on the characterization of unfolded proteins in dilute solutions in aqueous guanidine hydrochloride are analyzed to show that the data are not fit by either the random flight or wormlike chain models for linear chains. The analysis includes data on the intrinsic viscosity, root-mean-square radius of gyration, from small-angle X-ray scattering, and hydrodynamic radius, from the translational diffusion coefficient. It is concluded that residual structure consistent with that deduced from nuclear magnetic resonance on these solutions can explain the dilute solution results in a consistent manner through the presence of ring structures, which otherwise have an essentially flexible coil conformation. The ring structures could be in a state of continual flux and rearrangement. Calculation of the radius of gyration for the random-flight model gives a similar reduction of this measure for chains joined at their endpoints, or those containing loop with two dangling ends, each one-fourth the total length of the chain. This relative insensitivity to the details of the ring structure is taken to support the behavior observed across a range of proteins. [source] The contribution of intracellular calcium stores to mEPSCs recorded in layer II neurones of rat barrel cortexTHE JOURNAL OF PHYSIOLOGY, Issue 2 2002Christopher R. L. Simkus Loading slices of rat barrel cortex with 50 ,m BAPTA-AM while recording from pyramidal cells in layer II induces a marked reduction in both the frequency and amplitudes of mEPSCs. These changes are due to a presynaptic action. Blocking the refilling of Ca2+ stores with 20 ,m cyclopiazonic acid (CPA), a SERCA pump inhibitor, in conjunction with neuronal depolarisation to activate Ca2+ stores, results in a similar reduction of mEPSCs to that observed with BAPTA-AM, indicating that the source for intracellular Ca2+ is the endoplasmic reticulum. Block or activation of ryanodine receptors by 20 ,m ryanodine or 10 mm caffeine, respectively, shows that a significant proportion of mEPSCs are caused by Ca2+ release from ryanodine stores. Blocking IP3 receptors with 14 ,m 2-aminoethoxydiphenylborane (2APB) also reduces the frequency and amplitude of mEPSCs, indicating the involvement of IP3 stores in the generation of mEPSCs. Activation of group I metabotropic receptors with 20 ,m (RS) -3,5-dihydroxyphenylglycine (DHPG) results in a significant increase in the frequency of mEPSCs, further supporting the role of IP3 receptors and indicating a role of group I metabotropic receptors in causing transmitter release. Statistical evidence is presented for Ca2+ -induced Ca2+ release (CICR) from ryanodine stores after the spontaneous opening of IP3 stores. [source] Salinity-induced changes in essential oil, pigments and salts accumulation in sweet basil (Ocimum basilicum) in relation to alterations of morphological developmentANNALS OF APPLIED BIOLOGY, Issue 2 2010N. Bernstein The objective of the project was to study salinity-induced effects on essential oil, pigments and salts accumulation in sweet basil (Ocimum basilicum, the cultivar Perrie) in relation to the alteration of plant morphological development and yield production. Hydroponically grown plants were exposed to one of six NaCl concentrations (1, 25, 50, 75, 100 and 130 mM NaCl). Inhibitory effects of salinity on biomass production of the shoot and the root, and area of individual leaves were apparent already under cultivation with 25 mM NaCl. Elevation of salinity from 1 to 100 mM NaCl induced 63% and 61% reductions in fresh and dry herb biomass production, respectively. The stress-induced reduction of foliage biomass sourced mainly from inhibition of leaf area development rather than reduction of internode and leaf number. Cl and Na concentrations in the leaves, stems and roots increased with elevation of NaCl concentration in the cultivation solution. While the extent of Cl accumulation was leaves>stems>roots, Na was largely excluded from the leaves and was preferentially accumulated in roots and the stems, potentially accounting for the moderate sensitivity of the leaf tissue to salinity. Salt stress increased the contents of essential oil and carotenoids in the leaves that may further account for the moderate sensitivity of sweet basil to salinity and suggest a potential for agro-industrial production. A twofold increase in both carotenoid concentration and the percent of essential oil in the fresh tissue was observed by elevation of the salinity from 1 to 130 mM NaCl. Overall, the stress induced increase of the percent of essential oil in the tissue in the salinity range 1,75 mM NaCl was about 50%, and thereby compensated for the similar reduction of biomass production in this salinity range, so that oil production on per plant basis was not reduced by salinity. [source] Does stand structure influence susceptibility of eucalypt floodplain forests to dieback?AUSTRAL ECOLOGY, Issue 3 2010SHAUN C. CUNNINGHAM Abstract Forest dieback is a worldwide problem that is likely to increase with climate change and increasing human demands for resources. Eucalyptus camaldulensis forests are an acute example of forest dieback, with 70% of the Victorian Murray River floodplain in some state of dieback. If we are to halt dieback in these floodplain forests, we need to understand what makes stands susceptible to dieback. Forest diebacks are often related to stand structure, with dieback more severe in senescent or high-density stands. We determined whether certain stand structures make these forests more susceptible to dieback. We undertook an extensive survey of 176 stands across 100 000 ha of forest, covering the range of stand condition on this floodplain. Large and small trees (20-, 40-, 80- and 120-cm diameter) showed a similar reduction in the probability of being alive with decreasing stand condition. A slight improvement in stand condition was found at higher densities and basal areas, which may reflect the higher productivity or younger age of these stands. Stand condition was moderately, positively correlated with longitude, with stand condition being higher in the east of the Murray River floodplain where flooding frequencies are currently higher. This suggests that dieback of these floodplain forests would be more effectively mitigated by increased water availability through flooding than by altering stand structure. [source] Role of Humoral Mediators in, and Influence of a Liposomal Formulation on, Acute Amphotericin B NephrotoxicityBASIC AND CLINICAL PHARMACOLOGY & TOXICOLOGY, Issue 4 2001Ramzi Sabra Both direct effects of amphotericin B on contractile vascular cells, and indirect effects, due to humoural mediators, have been proposed. This study examines the role of nitric oxide, endothelin and angiotensin II in the acute nephrotoxic effects of amphotericin B in rats, and compares the antifungal and nephrotoxic effects of liposomal amphotericin B and amphotericin B-deoxycholate. Anaesthetized rats were given infusions of amphotericin B-deoxycholate in the presence or absence of N-nitro-L-arginine, PD 145065, a non-specific endothelin receptor antagonist, and L-158809, an angiotensin II type I receptor antagonist, or increasing doses of liposomal amphotericin B. Amphotericin B-deoxycholate (0.03 mg/kg/min intravenously) caused a significant 44% reduction in glomerular filtration rate and 65% maximal fall in renal blood flow. N-Nitro-L-arginine-treated rats had a lower renal blood flow and glomerular filtration rate at baseline, but sustained similar reduction of 53% and 75% in these parameters, respectively. PD145065 and L-158809 did not modify these effects either. Increasing doses of liposomal amphotericin B (from 0.01 up to 0.50 mg/kg/min.) induced no change in either glomerular filtration rate or renal blood flow. In vitro susceptibility tests revealed similar potency for liposomal amphotericin B and amphotericin B-deoxycholate in their fungistatic effects and slightly higher potency for amphotericin B-deoxycholate in their fungicidal effect. These results suggest that endogenous endothelin, angiotensin II or nitric oxide systems are not involved in the nephrotoxic effects of amphotericin B. The liposomal amphotericin B results suggest that amphotericin B nephrotoxicity is due to a direct interaction of amphotericin B with renal cells that is prevented by its encapsulation in liposomes. [source] Effect of montelukast pretreatment on inducible nitric oxide synthase mRNA expression in the lungs of antigen-challenged allergic miceCLINICAL & EXPERIMENTAL ALLERGY, Issue 12 2003K. Sade Summary Background Growing evidence suggests that inducible nitric oxide synthase (iNOS) is the main source of the high output of exhaled nitric oxide (NO) in asthma. Treatment of asthmatic patients with glucocorticoids reduces high levels of exhaled NO mainly by inhibiting the transcription of iNOS. A similar reduction in exhaled NO was recently observed in patients treated with the leukotriene receptor antagonists, but the exact interaction between these drugs and iNOS remains obscure. Objective The purpose of this study was to evaluate the effect of a leukotriene receptor antagonist, montelukast, on the expression and activity of iNOS in a murine model of allergic asthma. Methods Twenty-four BALB/c mice were sensitized to OVA and were equally divided into 3 groups (Groups 1,3). Eight additional mice were sham sensitized and served as a negative control group (Group 4). Group 1 received montelukast 1 mg/kg/day in their drinking water, Group 2 received dexamethasone 1 mg/kg/day in their drinking water and Groups 3 and 4 received plain tap water. After 1 week, the animals were challenged by inhalation of OVA and, 3 h later, they were killed and their lung cells were isolated by enzymatic tissue digestion. NO generation was measured by a Griess assay, and iNOS mRNA was studied by RT-PCR. Results A significant increase in iNOS mRNA expression and in NO generation was evident after allergen challenge compared with the controls. Pretreatment with montelukast mildly decreased NO production without producing a concomitant significant decrease in iNOS mRNA expression. Conclusion: Unlike pretreatment with glucocorticoids, we failed to find compelling evidence for a major role for montelukast treatment in the modulation of iNOS mRNA in a murine model of acute asthma. [source] INVOLVEMENT OF PROLYLCARBOXYPEPTIDASE IN THE EFFECT OF RUTAECARPINE ON THE REGRESSION OF MESENTERIC ARTERY HYPERTROPHY IN RENOVASCULAR HYPERTENSIVE RATSCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 3 2009Xu-Ping Qin SUMMARY 1Previous studies indicate that rutaecarpine blocks increases in blood pressure and inhibits vascular hypertrophy in experimentally hypertensive rats. The aim of the present study was to determine whether the effects of rutaecarpine are related to activation of prolylcarboxypeptidase (PRCP). 2Renovascular hypertensive rats (Goldblatt two-kidney, one-clip (2K1C)) were developed using male Sprague-Dawley rats. Chronic treatment with rutaecarpine (10 or 40 mg/kg per day) or losartan (20 mg/kg per day) for 4 weeks to the hypertensive rats caused a sustained dose-dependent attenuation of increases in blood pressure, increased lumen diameter and decreased media thickness, which was accompanied by a similar reduction in the media cross-sectional area : lumen area ratio in mesenteric arteries compared with untreated hypertensive rats. 3Angiotensin (Ang) II expression was significantly increased in mesenteric arteries of hypertensive rats compared with sham-operated rats. No significant differences in plasma AngII levels were observed between untreated hypertensive and sham-operated rats. Hypertensive rats treated with high-dose rutaecarpine had significantly decreased Ang II levels in both the plasma and mesenteric arteries. 4Expression of PRCP protein or kallikrein mRNA was significantly inhibited in the right kidneys and mesenteric arteries of hypertensive rats. However, expression of PRCP protein and kallikrein mRNA was significantly increased after treatment with rutaecarpine or losartan (20 mg/kg per day). 5The data suggest that the repression of increases in systolic blood pressure and reversal of mesenteric artery remodelling by rutaecarpine may be related to increased expression of PRCP in the circulation and small arteries in 2K1C hypertensive rats. [source] Randomized-controlled study comparing post-operative pain between coblation palatoplasty and laser palatoplasty,CLINICAL OTOLARYNGOLOGY, Issue 2 2006Belloso A. Objectives:, This study aimed to evaluate differences in post-operative pain comparing KTP laser-assisted uvulopalatoplasty without tonsillectomy (LAUP) with a new described surgical method: coblation uvulopalatoplasty with tonsillectomy (CP). We also evaluate the impact of each surgical technique in reduction of snoring loudness. Material and methods:, Single blind randomized-controlled trial. From a population of 41 consecutive patients on the waiting list for uvulopalatoplasty for simple snoring, the study group was reduced to 17 CP and 13 LAUP. Post-operative pain and reduction of snoring loudness were recorded using visual analogue scales (VAS) during the first 15 post-operative days. Post-operative snoring loudness was documented for 1-year period. Results:, Both groups had similar post-operative pain during the first seven post-operative days. A statistically significant reduction in post-operative pain was observed in the CP group after day 8, and maintained until the end of the study. Reduction of snoring loudness was significant in both groups, but no differences were observed between them. Discussion:, Coblation uvulopalatoplasty compared with LAUP demonstrates a reduction in post-operative pain, significant after the first post-operative week. The collateral thermal injury caused by laser is responsible for the slow-healing rate and maintained post-operative pain. Coblation dissociates tissue at lower temperatures with minimal collateral thermal injury and consequently faster and less painful recovery. Both surgical procedures have significant and similar reduction in snoring loudness. Conclusions:, Both methods are adequate treatment options for snoring. The less painful recovery in CP promotes this surgical technique as our preferred choice for palate surgery. [source] Nerve growth factor expression in parasympathetic neurons: regulation by sympathetic innervationEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 12 2000Wohaib Hasan Abstract Interactions between sympathetic and parasympathetic nerves are important in regulating visceral target function. Sympathetic nerves are closely apposed to, and form functional synapses with, parasympathetic axons in many effector organs. The molecular mechanisms responsible for these structural and functional interactions are unknown. We explored the possibility that Nerve Growth Factor (NGF) synthesis by parasympathetic neurons provides a mechanism by which sympathetic,parasympathetic interactions are established. Parasympathetic pterygopalatine ganglia NGF-gene expression was examined by in situ hybridization and protein content assessed by immunohistochemistry. Under control conditions, NGF mRNA was present in ,,60% and NGF protein was in 40% of pterygopalatine parasympathetic neurons. Peripheral parasympathetic axons identified by vesicular acetylcholine transporter-immunoreactivity also displayed NGF immunoreactivity. To determine if sympathetic innervation regulates parasympathetic NGF expression, the ipsilateral superior cervical ganglion was excised. Thirty days postsympathectomy, the numbers of NGF mRNA-positive neurons were decreased to 38% and NGF immunoreactive neurons to 15%. This reduction was due to a loss of sympathetic nerve impulse activity, as similar reductions were achieved when superior cervical ganglia were deprived of preganglionic afferent input for 40 days. These findings provide evidence that normally NGF is synthesized by parasympathetic neurons and transported anterogradely to fibre terminals, where it may be available to sympathetic axons. Parasympathetic NGF expression, in turn, is augmented by impulse activity within (and presumably transmitter release from) sympathetic axons. It is suggested that parasympathetic NGF synthesis and its modulation by sympathetic innervation provides a molecular basis for establishment and maintenance of autonomic axo-axonal synaptic interactions. [source] Straightforward Strategy for the Stereoselective Synthesis of Spiro-Fused (C-5)Isoxazolino- or (C-3)Pyrazolino-(C-3)quinolin-2-ones from Baylis,Hillman Adducts by 1,3-Dipolar Cycloaddition and Reductive Cyclization,EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 32 2008Virender Singh Abstract A straightforward and general approach for the stereoselective synthesis of spiro-fused (C-5)isoxazolino- or (C-3)pyrazolino-(C-3)quinolin-2-ones from the adducts offorded from the Baylis,Hillman reaction of 2-nitrobenzaldehyde and ethyl acrylate by sequential 1,3-dipolar cycloaddition and reductive cyclization is presented. It was found that the reductive cyclization of the isoxazoline derivatives proceeded efficiently in the presence of In/HCl, whereas similar reductions of pyrazolines gave better yields when carried out in the presence of an Fe/AcOH mixture. However, similar attempts employing the Baylis,Hillman adduct of 2-nitrobenzaldehyde and methyl vinyl ketone did not yield the desired compounds.(© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008) [source] Hypertonic Saline Treatment of Severe Hyperkalemia in Nonnephrectomized DogsACADEMIC EMERGENCY MEDICINE, Issue 9 2000Justin L. Kaplan MD Abstract. Objectives: To determine whether a hypertonic saline bolus improves cardiac conduction or plasma potassium levels more than normal saline infusion within 15 minutes of treatment for severe hyperkalemia. Previously with this model, 8.4% sodium chloride (NaCl) and 8.4% sodium bicarbonate (NaHCO3) lowered plasma potassium equally effectively. Methods: This was a crossover study using ten conditioned dogs (14-20 kg) that received, in random order, each of three intravenous (IV) treatments in separate experiments at least one week apart: 1) 2 mmol/kg of 8.4% NaCl over 5 minutes (bolus); 2) 2 mmol/kg of 0.9% NaCl over one hour (infusion); or 3) no treatment (control). Using isoflurane anesthesia and ventilation (pCO2= 35-40 torr), 2 mmol/kg/hr of IV potassium chloride (KCl) was infused until conduction delays (both absent p-waves and ,20% decrease in ventricular rate in ,5 minutes) were sustained for 15 minutes. The KCl was then decreased to 1 mmol/kg/hr (maintenance) for 2 hours and 45 minutes. Treatment (0 minutes) began after 45 minutes of maintenance KCl. Results: From 0 to 15 minutes, mean heart rate increased 29.6 (95% CI = 12.2 to 46; p < 0.005) beats/min more with bolus than infusion and 23.4 (95% CI = 2.6 to 43.5; p < 0.03) beats/min more with bolus than control. No clinically or statistically significant difference was seen in heart rate changes from 0 to 30 minutes. Decreases in potassium from 0 to 15 minutes were similar with bolus, infusion, and control. Conclusions: In this model, 8.4% NaCl bolus reversed cardiac conduction abnormalities within the first 15 minutes after treatment, more rapidly than did the 0.9% NaCl infusion or control. This reversal occurred despite similar reductions in potassium levels. [source] |