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Plasma samples were thawed on ice for four hours then mixed by vortexing, prior to preparing for assays. Serum total cholesterol and triglyceride concentrations were determined enzymatically using kits from Roche Diagnostics (Sommerville, NJ). Serum HDL-cholesterol was determined by a direct method (Unimate HDL Direct ; Roche Diagnostics, Indianapolis, IN) that utilizes the combined action of polymers, polyanions, and detergent to solubilize cholesterol from HDL but not from VLDL, LDL, and chylomicrons as previously described . Analysis was performed on a Cobas-Fara II Clinical Analyzer (Montclair, NJ) using commercially available calibrators and quality control standards (Roche Diagnostics, Indianapolis, IN).
|
15369594_p6
|
15369594
|
Cholesterol Analysis
| 4.103221 |
biomedical
|
Study
|
[
0.9994953870773315,
0.0003499461163301021,
0.00015463147428818047
] |
[
0.9983298182487488,
0.0011768938275054097,
0.0003829405759461224,
0.00011034979979740456
] |
en
| 0.999997 |
Plasma from subjects was analyzed for plasma glutathione peroxidase using an ELISA kit (OXIS Internatl., Portland, OR). Two replicates per sample of 20 μl of plasma were diluted 1:25 with TRIS-HCl buffer then pippetted into pre-coated polyclonal antibodies microplate wells specific for human plasma glutathione peroxidase (GPX). The amount of enzyme present was determined by reaction with para-nitrophenyl-phosphate and was read using a microplate reader at 405 nm (Elx 808 Ultra Microplate Reader, Bio-Tek Instruments Inc., Winooski, VT). The concentration of plasma GPX was determined from a standard curve for each plate using five dilutions of GPX standard.
|
15369594_p7
|
15369594
|
Plasma Glutathione Peroxidase Assay
| 4.105367 |
biomedical
|
Study
|
[
0.9996048808097839,
0.00021617031597997993,
0.00017897339421324432
] |
[
0.9990252256393433,
0.0005673743435181677,
0.00034069447428919375,
0.00006664513784926385
] |
en
| 0.999997 |
Malondialdehyde compounds were determined colorimetrically using a commercial kit specific for measuring free and total malondialdehyde compounds (OXIS Internatl., Portland, OR). Two replicates per sample of 210 μl of plasma were added to each test tube with 11 μl of 500 mM butylated hydroxytoluene and 5.3 μl of concentrated hydrochloric acid. Tubes were capped, mixed then incubated at 60°C for 80 minutes, cooled to room temperature and 680 μl of N-methyl-2-phenylindole in acetonitrile was added. Then tubes were mixed, and centrifuged at 13,000 g for 5 minutes. New tubes were prepared and 660 μl of clear supernatant was added with 115 μl of concentrated HCl. Tubes were capped, mixed and incubated at 45°C for 60 minutes. Samples were centrifuged at 13,000 g for 5 min and the supernatant was read on a spectrophometer at 575 nm. Concentration of samples was determined using a five point standard curve.
|
15369594_p8
|
15369594
|
Plasma lipid peroxidation
| 4.110187 |
biomedical
|
Study
|
[
0.9994935989379883,
0.00028084873338229954,
0.00022556025942321867
] |
[
0.9974232912063599,
0.002107031876221299,
0.0003625962417572737,
0.0001070762809831649
] |
en
| 0.999998 |
This assay was conducted according to previously published methods . In brief, three reagents were used: 1) sodium acetate, acetic acid buffer (pH 3.6); 2)10 mmol/L solution of 2, 4,6-tripyridyl-s-triazine in a 40 mmol/L solution of hydrochloric acid (Sigma, St. Louis, MO); and 3) 20 mmol/L solution of ferric chloride hexahydrate prepared in double deionized water. The FRAP reagent was prepared daily with 25 ml of reagent one, 2.5 ml reagent two and three that were heated to 37°C before using . The assay was conducted with 10 uL of plasma that was diluted with 30 μl of ddi water. Sample was added to reagent in cuvettes with an autosampler and then read on a COBAS FARA II spectrofluorometric centrifugal analyzer (Roche, Montclair, NJ) at 593 nm at four minutes. FRAP values were determined from a five point curve using a trolox (vitamin E analog) standard. Standard curves were run after every 90 samples.
|
15369594_p9
|
15369594
|
Ferric reducing ability of plasma assay
| 4.107915 |
biomedical
|
Study
|
[
0.9995737671852112,
0.00021843025751877576,
0.00020782221690751612
] |
[
0.998963475227356,
0.0006373838405124843,
0.0003290293097961694,
0.00007006761006778106
] |
en
| 0.999998 |
Experimental procedures for the clinical trial were approved by the Institutional Review Board at the Johns Hopkins University Bloomberg School of Hygiene and Public Health; subjects gave their written informed consent to participate. The plasma cholesterol and antioxidant studies were approved by the Institutional Review Board at Oklahoma State University, Stillwater, OK. Data were analyzed using Proc Mixed Procedure and mean separation was performed using LSMEANS, correlation analysis was performed using Spearman's Correlation Coefficient Analysis (SAS Statistical Analysis Software, version 8.2, SAS Institute, Cary, NC).
|
15369594_p10
|
15369594
|
Ferric reducing ability of plasma assay
| 2.312385 |
biomedical
|
Study
|
[
0.9940213561058044,
0.0035790514666587114,
0.002399526070803404
] |
[
0.7558122873306274,
0.24163566529750824,
0.0009445901378057897,
0.0016073964070528746
] |
en
| 0.999997 |
Because there were significant four way interactions with gender × intervention period × treatment × weeks with MDA, FRAP, GPX and cholesterol analysis, trends by treatment, intervention period or week of treatment were not seen. Supplementing the diet with 20 mg/day of lycopene of either food did not change the plasma antioxidant status of the subjects and values ranged from 0.66–2.20, 540–1094, and 1296–2596 :mol/L for MDA, FRAP and GPX respectively. These levels are similar to levels reported for healthy subjects in other studie .
|
15369594_p11
|
15369594
|
Results
| 4.069653 |
biomedical
|
Study
|
[
0.9995212554931641,
0.000253973325015977,
0.0002247717638965696
] |
[
0.9993895292282104,
0.0002341218205401674,
0.00031865015625953674,
0.0000576733255002182
] |
en
| 0.999998 |
Intervention with 20 mg of lycopene to the diet of subjects did not alter their total cholesterol, HDL-C or triglyceride status. However, there were gender differences and the women had higher average levels of plasma triglycerides, total cholesterol and HDL-C than men . The higher cholesterol levels for women compared to men in this study were not unusual since women in this age range often have higher cholesterol levels than men, a phenomenon related to decreased estrogen production . In this study, menopausal information was not recorded.
|
15369594_p12
|
15369594
|
Results
| 3.378269 |
biomedical
|
Study
|
[
0.9991716146469116,
0.0004180776886641979,
0.0004103611863683909
] |
[
0.9985345602035522,
0.0011110714403912425,
0.000260831177001819,
0.00009356124064652249
] |
en
| 0.999998 |
There was a significant positive correlation between each pair of total cholesterol and MDA and MDA and FRAP and between HDL-C and MDA and HDL-C and GPX. A significant negative correlation was found between triglycerides and GPX (Table 2 ).
|
15369594_p13
|
15369594
|
Results
| 3.968662 |
biomedical
|
Study
|
[
0.9993844032287598,
0.00022014309070073068,
0.0003954308049287647
] |
[
0.9993935823440552,
0.00027072630473412573,
0.0002921986160799861,
0.000043458130676299334
] |
en
| 0.999997 |
Because of the correlation between cholesterol concentrations and antioxidant analysis, a preliminary analysis of data was conducted to determine if cholesterol levels impacted antioxidant results. Subjects were separated into two groups based upon baseline concentrations of plasma triglycerides, total cholesterol and LDL-C above or below 200, 180 and 160, respectively. Five subjects, two men and three women, fit the criteria of moderately hypercholesterolemic (Table 3 ).
|
15369594_p14
|
15369594
|
Results
| 3.96686 |
biomedical
|
Study
|
[
0.9993008375167847,
0.00040598318446427584,
0.0002931374474428594
] |
[
0.9992935657501221,
0.0004581484536174685,
0.00017818155174609274,
0.00007010527042439207
] |
en
| 0.999996 |
Analyses showed an interaction of cholesterol level × treatment period × treatment factor for MDA and FRAP analysis. Higher MDA and FRAP levels were found in the group having higher cholesterol levels compared to the other group (Table 4 ). No trend with cholesterol level and glutathione peroxidase was found.
|
15369594_p15
|
15369594
|
Results
| 3.986273 |
biomedical
|
Study
|
[
0.9992648959159851,
0.00029981418629176915,
0.0004352251999080181
] |
[
0.9994736313819885,
0.000238824199186638,
0.00023601345310453326,
0.000051508377509890124
] |
en
| 0.999997 |
We found no improvement in the antioxidant status of healthy middle-aged adults supplemented with two lycopene-containing foods. In previous antioxidant studies, reduced lipid peroxidation was reported in subjects supplemented from one to four weeks with 5 to 45 mg lycopene containing tomato products . However, in each of these studies, the diet was not controlled. When healthy elderly subjects in a diet controlled study were supplemented with 13.3 mg of tomato lycopene (LycoRed) for 12 weeks, lycopene intervention did not significantly change LDL oxidation, as measured by the rate of conjugated diene production .
|
15369594_p16
|
15369594
|
Discussion
| 4.048499 |
biomedical
|
Study
|
[
0.9995587468147278,
0.00022329376952257007,
0.0002179201110266149
] |
[
0.9991501569747925,
0.0002440701937302947,
0.000547737639863044,
0.000058002777223009616
] |
en
| 0.999995 |
The reports from lycopene intervention studies that measured FRAP activity are not in agreement. One study reported improvement in FRAP levels of plasma in subjects supplemented with tomato juice and olive oil , while two other tomato juice intervention studies reported no improvement in plasma antioxidant levels after lycopene supplementation as measured by Trolox equivalent antioxidant capacity (TEAC), radical trapping antioxidant parameter assay (TRAP), and FRAP . Researchers in one study found that the FRAP assay was more accurate when measuring the antioxidant activity of water-soluble antioxidants . They thought full expression of the antioxidant activity was not identified from lycopene in this assay since it is a lipophyllic compound. Curiously, both watermelon and tomato contain other water-soluble compounds that are reported to have antioxidant activity that reacts in vitro in the FRAP assay . In this study, contribution of these water-soluble compounds in changes in plasma FRAP activity with either food intervention compared to the control was not found.
|
15369594_p17
|
15369594
|
Discussion
| 4.066451 |
biomedical
|
Study
|
[
0.9995124340057373,
0.0002008750889217481,
0.0002866827417165041
] |
[
0.9880512952804565,
0.00031655110069550574,
0.011536642909049988,
0.00009549679816700518
] |
en
| 0.999997 |
Unlike a previous report by Fuhrman et al., neither lycopene intervention with watermelon nor tomato affected cholesterol levels . Differences in results may have been due to lycopene dosage level. In that study the subjects were supplemented with 60 mg/day for three months, however diet was not controlled.
|
15369594_p18
|
15369594
|
Discussion
| 2.799562 |
biomedical
|
Study
|
[
0.9985358715057373,
0.00035747792571783066,
0.001106712268665433
] |
[
0.9918081164360046,
0.0050484114326536655,
0.0029114047065377235,
0.00023204725584946573
] |
en
| 0.999997 |
Fruits and vegetables are excellent sources of antioxidant compounds and the average American consumes only 1.5 and 3.1 servings per day . In many of the studies where antioxidant protection with lycopene containing foods was reported, subjects consumed their normal diet that may or may not have met the recommended servings of fruits and vegetables . Increasing fruit and vegetable consumption to 12 servings per day compared to 5.8 servings, without the addition of other diet interventions, reduced a biomarker of DNA oxidative damage (8-hydroxydeoxyguanosine) by 32% . In a controlled trial where subjects were supplemented with tomato juice but restricted in total fruit and vegetable consumption and exposed to low levels of ozone, researchers found reduced DNA strand breaks compared to placebo controls . Because this study controlled for other phytochemical containing fruits and vegetables, the DNA protection was attributed to tomato juice phytochemicals .
|
15369594_p19
|
15369594
|
Discussion
| 4.046551 |
biomedical
|
Study
|
[
0.9996305704116821,
0.00015575243742205203,
0.00021360743266995996
] |
[
0.9966313242912292,
0.00039179096347652376,
0.002909225644543767,
0.00006757978553650901
] |
en
| 0.999997 |
The positive correlation between total cholesterol and MDA antioxidant analysis has been reported in studies with hypercholesterolemic subjects compared to normocholesterolemic subjects . The MDA assay measures lipid peroxidation products, and a higher level of lipids available to react with peroxidizing agents results in higher MDA values .
|
15369594_p20
|
15369594
|
Discussion
| 3.976813 |
biomedical
|
Study
|
[
0.9997434020042419,
0.00009133932326221839,
0.00016527494881302118
] |
[
0.9939520955085754,
0.0035611819475889206,
0.002383890561759472,
0.00010280239075655118
] |
en
| 0.999996 |
The trend correlating higher FRAP with higher cholesterol levels has not been previously reported. The significance of this trend is speculative, since the FRAP assay measures the oxidation and reduction potential of compounds based on the reduction of the ferric to ferrous iron , lipid peroxidation products may have contributed to the oxidation/reduction potential of the reaction.
|
15369594_p21
|
15369594
|
Discussion
| 3.481783 |
biomedical
|
Study
|
[
0.9993414282798767,
0.00016310556384269148,
0.0004954249016009271
] |
[
0.9873091578483582,
0.01167045347392559,
0.0008148679626174271,
0.00020559004042297602
] |
en
| 0.999995 |
Long-term supplementation studies where diet is controlled will probably be necessary to identify the benefits provided by lycopene. There may be real health benefits associated with lycopene especially since it is stored in various tissues and exhibits strong antioxidant activity in vitro . Also the body of epidemiological evidence points to the protection provided against cardiovascular disease and some cancers with lycopene containing foods . Recent cancer intervention studies have reported beneficial effects on prostate cancer from lycopene food supplementation . The health benefits associated with diets providing lycopene are most likely long-term. Therefore, the findings of the present study should not be interpreted as a lack of health benefits from regular consumption of lycopene-rich foods.
|
15369594_p22
|
15369594
|
Conclusions
| 3.960757 |
biomedical
|
Study
|
[
0.9996795654296875,
0.00012489059008657932,
0.00019561743829399347
] |
[
0.9822850227355957,
0.0014676116406917572,
0.0160916056483984,
0.00015573347627650946
] |
en
| 0.999996 |
The interaction between cholesterol levels and antioxidant values needs more research. Contradictory findings of this study with other ex vivo antioxidant studies may be due to the cholesterol levels of subjects thus warranting further research.
|
15369594_p23
|
15369594
|
Conclusions
| 2.358308 |
biomedical
|
Study
|
[
0.996099591255188,
0.00046938416198827326,
0.003430989570915699
] |
[
0.854636549949646,
0.1358306109905243,
0.008528931997716427,
0.0010039618937298656
] |
en
| 0.999998 |
None declared.
|
15369594_p24
|
15369594
|
Competing interests
| 0.828596 |
other
|
Other
|
[
0.19056588411331177,
0.00526782963424921,
0.804166316986084
] |
[
0.017649328336119652,
0.97890704870224,
0.0020668187644332647,
0.0013767849886789918
] |
it
| 0.999996 |
Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. All programs and services of the U.S. Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap. The article cited was prepared by a USDA employee as part of his/her official duties. Copyright protection under U.S. copyright law is not available for such works. Accordingly, there is no copyright to transfer. The fact that the private publication in which the article appears is itself copyrighted does not affect the material of the U.S. Government, which can be freely reproduced by the public.
|
15369594_p25
|
15369594
|
Note
| 1.076356 |
other
|
Other
|
[
0.00470685726031661,
0.0008088318281807005,
0.9944842457771301
] |
[
0.0007642676937393844,
0.9984732270240784,
0.00039327304693870246,
0.0003692721656989306
] |
en
| 0.999996 |
JKC: conception and design of the study, drafted the manuscript, BHA: design of study, editing, PLC:design of the study, statistical analysis, PPV: conception of study, editing, RAB: editing, technical assistance, BAC:conception of study, editing. All authors read and approved the final manuscript.
|
15369594_p26
|
15369594
|
Authors' contributions
| 0.934445 |
other
|
Other
|
[
0.05491210147738457,
0.0016817302675917745,
0.9434061646461487
] |
[
0.0025823067408055067,
0.9964101910591125,
0.0006221096264198422,
0.0003854396636597812
] |
en
| 0.999998 |
Atherogenesis involves the cellular infiltration of several cell types, including monocytes, T lymphocytes, and mast cells. Cytokine secretion by these cells and endothelial cells are contributing factors in the growth and propagation of atherosclerotic plaques as well as the stability and degradation of fibrous caps. Cytokines implicated in atherogenesis include Interleukin (IL)-1β, IL-6, IL-8, IL-13, and Tumor Necrosis Factor (TNF) .
|
15383152_p0
|
15383152
|
Background
| 4.086964 |
biomedical
|
Study
|
[
0.999377429485321,
0.00031301312264986336,
0.00030960285221226513
] |
[
0.5573585033416748,
0.014798927120864391,
0.42709654569625854,
0.0007460314081981778
] |
en
| 0.999996 |
IL-1β is secreted mainly by macrophages and virtually by every cell type in the body. IL-1β is produced in response to various stimulants, such as cytokines, bacteria, and viruses, but most interestingly to epinephrine . IL-1β has a broad range of functions which includes activation of neutrophils, endothelial cells, monocytes, T-cells, and mast cells. It may also induce procoagulant changes in endothelial tissue. IL-6 induces an acute phase response consisting of increased fibrinogen synthesis and thrombocytosis with increased vascular permeability. The detection of IL-6 in the blood of patients suffering from unstable angina suggests that nuclear factor-kappa B (NF-κB) activation may be occurring at the vascular level in patients with heart disease . IL-8 is in the CXC family of chemokines and functions to recruit neutrophils to the site of inflammation. IL-13 exerts multiple effects on cell differentiation and function of monocytes/macrophages. It can also suppress the cytotoxic function of monocytes/macrophages and the production of proinflammatory cytokines by these cells .
|
15383152_p1
|
15383152
|
Background
| 4.632305 |
biomedical
|
Study
|
[
0.9990457892417908,
0.00039231355185620487,
0.0005618512514047325
] |
[
0.7542337775230408,
0.007952239364385605,
0.23697200417518616,
0.000841895816847682
] |
en
| 0.999997 |
Mast cells are found preferentially around blood vessels and beneath the epithelium of the skin and mucus membranes . Traditionally, mast cells are responsible for allergy and asthma pathogenesis. Typically, mast cell activation occurs in response to cross-linkage of the high affinity IgE receptor (FcεRI) by antigen and IgE . Activation may also occur in response to a range of agents, such as pathogens, cytokines, and even oxidized low density lipoprotein (ox-LDL). After activation, key mediators secreted by mast cells include preformed mediators like histamine, proteoglycans, proteases, and several cytokines and growth factors . Mast cells have been observed in both aortic atherosclerotic lesions and in coronary arteries. The large numbers of mast cells found in the adventitia of arteries and in the intima are in proportion to the severity of heart disease . The study of the distribution, activation, and phenotype of mast cells in lesions of 250 specimens of human carotid arteries by Jeziorski, et al. further supports the role of mast cells in atherogenesis . They demonstrated significant numbers and focal accumulations of mast cells in association with macrophages and extensive activation/degranulation at all developmental stages of atherosclerotic lesion development. It now appears likely that inflammatory events and mast cells play an important role in atherogenesis as recently reviewed by us .
|
15383152_p2
|
15383152
|
Background
| 4.461092 |
biomedical
|
Review
|
[
0.9974652528762817,
0.0012501373421400785,
0.0012844983721151948
] |
[
0.14268463850021362,
0.0016550793079659343,
0.8549514412879944,
0.0007088897400535643
] |
en
| 0.999996 |
Stress is known to influence immune function . An immunoregulatory effect of the sympathetic nervous system in stress has been indicated for some time . Catecholamines, such as epinephrine, norepinephrine, and dopamine, are elevated in stress responses, and mediate vasoconstriction and an increase in blood pressure as a result of increased peripheral vascular resistance. In disorders such as sepsis, cardiovascular disease, or cocaine abuse, catecholamines are elaborated in excess. Sustained increases in circulating catecholamines by infusion of epinephrine or norepinephrine have been shown to cause moderate cardiovascular and metabolic effects . Catecholamines induce aggravation of aortic and coronary atherosclerosis in monkeys and play a direct role in atherogenesis and cardiovascular disease .
|
15383152_p3
|
15383152
|
Background
| 4.148834 |
biomedical
|
Study
|
[
0.9992783665657043,
0.0003847925108857453,
0.00033684642403386533
] |
[
0.5323867201805115,
0.004330415744334459,
0.46255043148994446,
0.0007324189064092934
] |
en
| 0.999996 |
Epinephrine and norepinephrine increase the uptake of low density lipoprotein in atheroscelotic plaques in rabbits and rats as well as enhance proliferation of rat endothelial and smooth muscle cells . It has been reported that norepinephrine increases adherence and chemotaxis of macrophages . Epinephrine also upregulates the surface expression of L-selectin on monocytes in vitro . Most recently, we have reported that nitric oxide production from macrophages induced by LPS is enhanced by catecholamines . Both epinephrine and IL-1 are involved in acute phase responses seen in stress and in coronary artery disease. Studies have shown that norepinephrine can induce IL-1β mRNA in mycocardial tissue and that infusion of IL-1β in animal models can induce expression of catecholamines . These data suggest that, in some conditions, both IL-1β and catecholamines can be delivered to tissues that can then mediate additive or modulatory effects. Moreover, as reviewed by Gidron Y et al., stress in conjunction with the release of catecholamines and proinflammatory cytokines, can potentiate atherogenesis. Hence, studies of the interactions between catecholamines, monokines and inflammatory cell activation are especially relevant. The aim of the study was to determine whether epinephrine affects IL-1β induced proatherogenic cytokine production in mast cells, a phenomenon previously not described. Our results indicated that epinephrine synergized with IL-1β in the production of proatherogenic cytokines, suggesting a potential role for this interaction in inflammatory and atherogenic states.
|
15383152_p4
|
15383152
|
Background
| 4.359574 |
biomedical
|
Study
|
[
0.9994524121284485,
0.00033922752481885254,
0.00020837686315644532
] |
[
0.9987683892250061,
0.00032641744473949075,
0.0007933590095490217,
0.00011179722787346691
] |
en
| 0.999996 |
Human mast cell line, HMC-1, was incubated with IL-1β at various concentrations for 24 hours. The cell free supernatants of the cultures were harvested and subjected to IL-6 assay. HMC-1 cultured in medium alone produced trace amounts of IL-6. The IL-6 production from HMC-1 cultures treated with IL-1β was significantly increased in a dose-dependent manner . Since there was no significant difference in the IL-6 production induced by IL-1β at concentrations of 10 and 50 ng/ml, 10 ng/ml of IL-1β has been used to induce cytokine production in HMC-1 for the rest of the experiments. Epinephrine (Epi) alone at a concentration of 10 -3 M did not induce production of IL-6 in HMC-1 . When epinephrine at 10 -3 to 10 -7 M concentration was added simultaneously with IL-1β into the cultures, the production of IL-6 was enhanced significantly (p < 0.05) compared with that induced by IL-1β alone . Since the physiological concentration of epinephrine in plasma is 0.11 – 0.27 × 10 -6 M , we decided to use epinephrine at a supramaximal concentration of 1 × 10 -5 M for the rest of the experiments. In addition to IL-6, the enhancing effect of epinephrine was also observed in the production of IL-8 and IL-13 from IL-1β-induced HMC-1 cells .
|
15383152_p5
|
15383152
|
Epinephrine enhances IL-1β-induced IL-6, IL-8, and IL-13 production in mast cells
| 4.137032 |
biomedical
|
Study
|
[
0.9995404481887817,
0.00026128694298677146,
0.00019824126502498984
] |
[
0.9994219541549683,
0.00020630363724194467,
0.000314286386128515,
0.00005741556378779933
] |
en
| 0.999999 |
To measure proatherogenic cytokine gene expression, HMC-1 were treated with IL-1β, epinephrine, and IL-1β plus epinephrine for 6 hours and harvested for transcriptional analysis via RT-PCR. IL-1β-treated HMC-1 showed increased IL-6 mRNA transcription as seen with densitometry, while epinephrine alone appeared to have no effect. When IL-1β and epinephrine were added together to HMC-1, IL-6 mRNA expression increased over IL-1β treatment alone . The intensities of the cytokine and house keeping gene (HPRT) bands were measured by densitometry, and the ratio of the cytokine to the house keeping gene was calculated and assigned as the intensity index. The intensity indices for IL-6 were 0.36 for the control, 0.39 for IL-1β alone, 0.33 for epinephrine alone, and 0.54 for IL-1β plus epinephrine. IL-1β activated IL-8 mRNA production but epinephrine had no effect on IL-8 transcripts. IL-1β and epinephrine treatment together further increased IL-8 mRNA production . The intensity indices for IL-8 were 0.17 for the control, 0.52 for IL-1β alone, 0.20 for epinephrine alone, and 0.64 for IL-1β plus epinephrine. The results with IL-13 expression showed the same pattern. IL-1β was a good inducer of IL-13 transcription while epinephrine alone only minimally induced IL-13 mRNA. The combined stimulus of IL-1β and epinephrine significantly increased IL-13 mRNA production over that seen with each stimulus alone . Intensity indices for IL-13 were 0.22 for the control, 0.57 for IL-1β alone, 0.20 for epinephrine alone, and 0.64 for IL-1β plus epinephrine. To evaluate further the ability of epinephrine to induce IL-13 transcription at a molecular level, we transiently transfected HMC-1 cells with minimal promoter sequences as described in the materials and methods. IL-1β at 10 ng/ml significantly increased IL-13 promotor activity as detected by luciferase expression (data not shown). Epinephrine did not enhance IL-13 promoter activity suggesting that post-transcriptional mechanisms may be involved in the IL-13 induction. It is likely that epinephrine either prolongs IL-13 mRNA half life and/or enhances IL-13 secretory processes from the mast cell in response to IL-1-stimulation.
|
15383152_p6
|
15383152
|
Epinephrine enhances IL-1β-induced IL-6, IL-8, and IL-13 production in mast cells
| 4.337569 |
biomedical
|
Study
|
[
0.9993795156478882,
0.0004172671469859779,
0.00020330294501036406
] |
[
0.9991437196731567,
0.00030898101977072656,
0.000431226595537737,
0.00011616582196438685
] |
en
| 0.999995 |
Since our previous study has shown that the effect of epinephrine on nitric oxide synthesis is mediated by β-adrenoceptors , β-adrenergic receptor antagonists (propranolol and atenolol) were used to block the enhancing effect of epinephrine on proatherogenic cytokine production in HMC-1. Propranolol (Pro) and atenolol (Ate) at a concentration of 1 × 10 -4 M did not affect the cell viability in the cultures (88 and 90%, respectively, while that of the medium control was 85%), nor induced production of IL-6, IL-8 or IL-13 . When propranolol at 1 × 10 -4 and 1 × 10 -5 M was used in the culture, it significantly reduced the enhancing effect of epinephrine on IL-6 production . Propranolol at 1 × 10 -4 M also significantly reduced the enhancing effect of epinephrine on IL-8 production , and at 1 × 10 -4 and 1 × 10 -5 M significantly reduced the enhancing effect of epinephrine on IL-13 production . However, atenolol only significantly reduced the enhancing effect of epinephrine on IL-13 production , but not on IL-6 or IL-8 production .
|
15383152_p7
|
15383152
|
Enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1 is down regulated by adrenoceptor antagonists
| 4.148464 |
biomedical
|
Study
|
[
0.9995142221450806,
0.00028323358856141567,
0.00020260526798665524
] |
[
0.9992961883544922,
0.00021582591580227017,
0.0004223102005198598,
0.0000656351403449662
] |
en
| 0.999996 |
In order to further identify whether the enhancing effect of epinephrine on proatherogenic cytokine production is through the β-adrenoceptor, HMC-1 cells were incubated with rabbit polyclonal antibodies against β 1 and β 2 adrenergic receptors followed by a FITC-labeled second antibody. By flow cytometry analysis, β 1 and β 2 adrenergic receptors were found in small amounts on HMC-1 (18.6 and 11.7% respectively) .
|
15383152_p8
|
15383152
|
Expression of β 1 and β 2 adrenergic receptors on mast cells
| 4.115514 |
biomedical
|
Study
|
[
0.9995618462562561,
0.00021548506629187614,
0.00022264826111495495
] |
[
0.9993394017219543,
0.0003788590547628701,
0.0002257593732792884,
0.00005594527465291321
] |
en
| 0.999996 |
Since glucocorticoids are very effective treatment strategies for inflammatory disease, dexamethasone was used to determine its effect on atherogenic cytokine production in HMC-1. Dexamethasone (Dex, 1 × 10 -7 M) alone did not induce proatherogenic cytokine production . However, Dex significantly inhibited the enhancing effect of epinephrine on IL-6 production . The cell viability of the cultures was not different between the medium control (70%) and Dex (64%) groups. Dex also significantly inhibited the enhancing effect of epinephrine on IL-8 and IL-13 production . When Dex was included in the IL-1β-treatment, it slightly decreased the cytokine production when compared to the IL-1β alone, but the decrease was not significant .
|
15383152_p9
|
15383152
|
Enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1 is down regulated by immunosuppressants
| 4.095145 |
biomedical
|
Study
|
[
0.9995487332344055,
0.00025736287352629006,
0.00019382171740289778
] |
[
0.9993211030960083,
0.0002198209404014051,
0.0003984833601862192,
0.000060651487729046494
] |
en
| 0.999997 |
NF-κB is an important transcription factor that mediates the transcription of many proinflammatory cytokine genes. To study the role NF-κB plays in the enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1, NF-κB activation was analyzed in HMC-1 cultures. NF-κB translocation, as seen by a shift in oligonucleotide binding in EMSA gels, was not seen in control or epinephrine treated cells . A marked increase of NF-κB nuclear binding activity was observed in samples stimulated with IL-1β and IL-1β plus epinephrine for one hour but started to diminish after two hours . Not only did IL-1β plus epinephrine have no further effects on NF-κB translocation over IL-1β treatment alone, it seemed to decrease after one and two hours of stimulation.
|
15383152_p10
|
15383152
|
Role of NF-κB activation in the enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1
| 4.141466 |
biomedical
|
Study
|
[
0.9995508790016174,
0.000261124805547297,
0.00018803862622007728
] |
[
0.9993991851806641,
0.00024021344142965972,
0.0002942003193311393,
0.00006644880340900272
] |
en
| 0.999996 |
Because of its importance in cytokine signaling, phosphorylated p38 MAPK was also assayed. After 30 minutes of activation, the HMC-1 were lysed to be analyzed for p38 activation by Western blot. The presence of phosphorylated p38 was greatly increased in the epinephrine and IL-1β plus epinephrine samples . IL-1β alone had small effects on p38 activation at this time point while control levels were virtually nonexistent.
|
15383152_p11
|
15383152
|
Role of p38 MAPK activation in the enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1
| 4.058121 |
biomedical
|
Study
|
[
0.9994827508926392,
0.00018997423467226326,
0.00032735569402575493
] |
[
0.9994210004806519,
0.00035268449573777616,
0.00018118448497261852,
0.00004513343083090149
] |
en
| 0.999998 |
To confirm the role of NF-κB and p38 MAPK in the enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1, Bay 11, an NF-κB inhibitor , and SB203580, a specific inhibitor of p38 MAPK , were added to the cultures. By themselves, neither Bay 11 (1 × 10 -5 M) nor SB203580 (1 × 10 -5 M) affected the cell viability of the cultures (92 and 87%, respectively, while that of the medium control was 92%), nor did they induce proatherogenic cytokine production . However, Bay 11 and SB203580 significantly inhibited the enhancing effect of epinephrine on IL-6 production . Bay 11 decreased the IL-1β-epinephrine induced IL-8 production but not significantly, however SB203580 did significantly inhibit the enhancing effect of epinephrine on IL-8 production . Bay 11 and SB203580 significantly inhibited the enhancing effect of epinephrine on IL-13 production .
|
15383152_p12
|
15383152
|
Enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced HMC-1 is down regulated by NF-κB and p38 MAPK inhibitors
| 4.123474 |
biomedical
|
Study
|
[
0.9995051622390747,
0.0002720117336139083,
0.0002228673838544637
] |
[
0.9993483424186707,
0.0002129630302079022,
0.00037931703263893723,
0.00005933519423706457
] |
en
| 0.999996 |
Inflammatory cytokines play an important role in atherogenesis. Acute phase response (APR) proteins have been demonstrated as risk factors for atherosclerotic heart disease . Recent studies also suggest a prominent role for the APR in cerebrovascular disease and brain ischemia . The APR culminates in the secretion of inflammatory cytokines such as IL-6, TNF-α", and IL-1 resulting in the synthesis of several proteins including C-reactive protein, fibrinogen, serum amyloid A protein, and ceruloplasmin . These cytokines are intimately involved with the stress response . These cytokines can also induce transcriptional regulation of complement genes that have been shown to play a role in cardiovascular disease . Catecholamines are elaborated in stress responses which mediate vasoconstriction and elevate systemic vascular resistance and blood pressure. Catecholamines induce aggravation of aortic and coronary atherosclerosis in monkeys and play a direct role in atherogenesis and cardiovascular disease . Thus, it is important to understand the interaction between epinephrine and IL-1β with respect to atherogenic cytokine production.
|
15383152_p13
|
15383152
|
Discussion
| 4.387598 |
biomedical
|
Study
|
[
0.9993457198143005,
0.0003636903129518032,
0.000290579569991678
] |
[
0.9033831357955933,
0.00151452689897269,
0.09472205489873886,
0.00038029710412956774
] |
en
| 0.999996 |
In this study, IL-1β, an acute phase cytokine, activated mast cells to produce proatherogenic cytokines, IL-6, IL-8, and IL-13, in a dose-dependent manner . These results confirm our previous report that IL-1β regulates mast cell function . These results also show that epinephrine significantly up regulated the IL-1β induction of proatherogenic cytokines in mast cells giving new insight into neuronal regulation of the immune system. The gene expression of these proatherogenic cytokines was also increased in IL-1β-induced HMC-1 cells by addition of epinephrine, suggesting that the enhancing effect of proatherogenic cytokine production is a result of increased cytokine gene transcription . These data provide a novel role for epinephrine in inflammation and atherogenesis.
|
15383152_p14
|
15383152
|
Discussion
| 4.179298 |
biomedical
|
Study
|
[
0.9995648264884949,
0.00029259503935463727,
0.00014253717381507158
] |
[
0.999131977558136,
0.0002481892879586667,
0.0005246829823590815,
0.00009523132030153647
] |
en
| 0.999997 |
IL-1β signaling probably synergizes with β 2 -adrenoreceptor-mediated signaling pathways in inducing proatherogenic cytokine production. Several reports have shown that the effect of catecholamines on immune function is due to β-adrenoceptors . Flow cytometry data indicated that HMC-1 cells express both β 1 and β 2 adrenoceptors in small amounts . The result showed that the enhancing effect of proatherogenic cytokine production by epinephrine is down regulated by β 1 and β 2 adrenoceptor antagonist, propranolol, but not by β 1 specific adrenoceptor antagonist, atenolol, further suggesting the enhancing effect involves β 2 adrenoceptors . The down regulation by propranolol does not appear to be due to cytotoxicity of the antagonist since there is no difference in viabilities between the propranolol-treated and untreated cell cultures. It was interesting to see that propranolol caused a reduction of production of IL-13 to an amount that was much lower than that treated with IL-1β only . It may be that epinephrine-induced enhancement of IL-13 production is more sensitive to the propranolol blocking.
|
15383152_p15
|
15383152
|
Discussion
| 4.234129 |
biomedical
|
Study
|
[
0.9995929598808289,
0.0002292675490025431,
0.00017773572471924126
] |
[
0.9993239641189575,
0.0002751992142293602,
0.0003261113306507468,
0.00007482184446416795
] |
en
| 0.999997 |
Activated NF-κB has been demonstrated in atheromatous plaques and has been shown to play a role in atherogenesis . To study the mechanism of the enhancing effect of epinephrine on proatherogenic cytokine production from IL-1β-induced mast cells, NF-κB and p38 MAPK activations were investigated. Control samples and epinephrine alone samples did not induce NF-κB activation. However, a marked increase in NF-κB activation was observed in samples stimulated with IL-1β and IL-1β plus epinephrine . NF-κB activation was seen early at one hour and began to fade by two hours. NF-κB also was not increased by the addition of epinephrine to IL-1β and even seemed to decrease it at both time points suggesting that NF-κB is needed for cytokine induction but not for the enhancing effect. The presence of phosphorylated p38 MAPK was greatly increased in the epinephrine and IL-1β plus epinephrine samples but only minimally activated with IL-1β alone at a 30 minute incubation time point . SB203580 blocked the IL-1β and IL-1β plus epinephrine effect on IL-6, IL-8, and IL-13 expression suggesting that p38 plays an important role in signaling from both IL-1β and epinephrine. The double stimulation of p38, early by IL-1β and later by epinephrine, may explain the enhancing effect on the production of IL-6, IL-8, and IL-13 in mast cells.
|
15383152_p16
|
15383152
|
Discussion
| 4.30144 |
biomedical
|
Study
|
[
0.999478280544281,
0.00036071176873520017,
0.00016110582510009408
] |
[
0.999007523059845,
0.000303871463984251,
0.0005723772337660193,
0.00011624074977589771
] |
en
| 0.999996 |
The enhancing effect of epinephrine on proatherogenic cytokine production was also down regulated by immunosupressants, such as Dex. Dex at the concentration used in this study did not affect the cell viability of the culture, suggesting the down regulation effect of the drugs is not due to toxic effect. Dex also slightly, but not significantly, decreased IL-1β-induced cytokine production in mast cells . Taken all together, these results indicate that β 2 -adrenoceptor antagonists and glucocorticoids may have clinical potential in stress-mediated disease and atherogenesis.
|
15383152_p17
|
15383152
|
Discussion
| 4.08513 |
biomedical
|
Study
|
[
0.99964439868927,
0.00021513037791009992,
0.00014049612218514085
] |
[
0.9976224303245544,
0.0002913846692536026,
0.0019999491050839424,
0.0000861886001075618
] |
en
| 0.999997 |
All the signaling pathways induced by IL-1β and epinephrine in mast cells are complex and beyond the scope of this manuscript. However, two important inflammatory pathways, NF-κB and p38 MAPK, have been shown. IL-1β release from immune challenge and epinephrine elevated from stress response can jointly stimulate mast cells to increase IL-6, -8, and -13 production above that which is seen with either stimulus alone. The exact mechanisms are unclear, but we have shown that IL-1β is a strong inducer of NF-κB while epinephrine is a strong inducer of p38 MAPK. Neither NF-κB nor p38 MAPK was activated further by IL-1β plus epinephrine compared to either stimulus alone nor was the promotor activity of IL-13 increased by the double stimulus as seen by luciferase activity of a IL-13 reporter gene construct. These data would suggest that IL-1β is activating IL-6, IL-8, and IL-13 by NF-κB while p38 MAPK activation is enhancing protein production by inducing other transcription factors, stabilizing the gene mRNA, or other forms of post-translational modification. These mechanisms are summarized in Fig. 10 .
|
15383152_p18
|
15383152
|
Discussion
| 4.245473 |
biomedical
|
Study
|
[
0.9995458722114563,
0.00027108972426503897,
0.00018302974058315158
] |
[
0.9991211295127869,
0.00023434303875546902,
0.000565302383620292,
0.00007922865188447759
] |
en
| 0.999999 |
In conclusion, stress related catecholamines, such as epinephrine, synergized with IL-1β in gene expression and production of proatherogenic cytokines, IL-6, IL-8, and IL-13 in mast cells. The enhancing effect of proatherogenic cytokine production by epinephrine on IL-1β-induced mast cells was down regulated by β-adrenoceptor antagonist, propranolol, and the immunosuppressant Dex. These data support a novel role for catecholamines in disorders such as inflammation and atherogenesis. These data also indicate that β-adrenoceptor antagonists and immunosuppressants may be used preventively and therapeutically for modulation of the catecholamine – proatherogenic cytokine axis in disease states.
|
15383152_p19
|
15383152
|
Conclusions
| 4.166011 |
biomedical
|
Study
|
[
0.9996216297149658,
0.00025489498511888087,
0.00012341277033556253
] |
[
0.997222900390625,
0.0004609854076988995,
0.002199246548116207,
0.00011686199286486953
] |
en
| 0.999997 |
HMC-1 cell line, established from a patient with mast cell leukemia, were graciously provided by Dr. Butterfield (Mayo Clinic, Rochester, MN). These cells were maintained in RPMI 1640 media (GibcoBRL, Rockville, MD), supplemented with 5 × 10 -5 M 2-mercaptoethanol (Sigma Chemical Company, St. Louis, MO), 10 mM HEPES (GibcoBRL), Gentamycin 50 μg/ml, 5 μg/ml insulin transferrin, 2 mM L-glutamine, and 5% heat inactivated fetal bovine serum (Atlanta Biologicals, Atlanta, GA), at 37°C and in 5% CO 2 mixture . HMC-1 cells were cultured and maintained in 25 cm 2 flasks. To each well of a 6 well culture plate, two ml of HMC-1 mast cells at 0.75 × 10 6 cells/ml concentration were cultured with epinephrine at 1 × 10 -5 M concentration in the presence and absence of IL-1β (10 ng/ml) for 24 hrs. The cultures were carried out in triplicate. Supernatants were harvested for measuring IL-6, IL-8, and IL-13 by ELISA and cell viability and numbers of the culture were analyzed.
|
15383152_p20
|
15383152
|
Mast cell culture and the induction of cytokine production in HMC-1 cells
| 4.121021 |
biomedical
|
Study
|
[
0.9995806813240051,
0.0002290477423230186,
0.0001903477677842602
] |
[
0.9990144968032837,
0.0006564536597579718,
0.0002575581893324852,
0.00007143850234569982
] |
en
| 0.999996 |
Cytokine ELISA was performed for the following cytokines: IL-6, IL-8, and IL-13. ELISA was carried out on cell-free culture supernatants using commercially available ELISA kits, according to manufacturers instructions as earlier described (R&D Systems, Minneapolis, MN; Immunotech, Westbrook, ME; Genzyme, Cambridge, MA). Results were analyzed on an ELISA plate reader .
|
15383152_p21
|
15383152
|
ELISA for cytokine proteins
| 3.966443 |
biomedical
|
Study
|
[
0.9996100068092346,
0.00015314806660171598,
0.00023689810768701136
] |
[
0.9981277585029602,
0.0013850746909156442,
0.0004154692869633436,
0.00007169449963839725
] |
en
| 0.999996 |
At the end of incubation, the cells were subjected to the viability count by trypan blue (TB) dye exclusion technique. Two tenths ml of cell cultures were mixed with 0.05 ml of TB, applied to hemocytometer, and counted under a microscope. The cell viability is calculated by dividing the number of live cells (unstained cells) by the total number of all cells (TB-stained and unstained cells) and expressed as a percent.
|
15383152_p22
|
15383152
|
Measurement of cell viability of the cultures
| 4.100532 |
biomedical
|
Study
|
[
0.9995684027671814,
0.00020429264986887574,
0.0002274025318911299
] |
[
0.9959264993667603,
0.003496340475976467,
0.00047658346011303365,
0.00010067093535326421
] |
en
| 0.999996 |
HMC-1 were treated with the appropriate reagents and allowed to incubate at 37°C before being harvested for RNA. RNA was extracted from HMC-1 (3 × 10 6 cells) by the addition of 1 ml of RNAzol B (Tel-Test, Inc., Friendswood, Texas) . After shaking for 1 minute the samples were centrifuged at 12,000 × g for 15 minutes at 4°C. The aqueous layer was washed twice with 0.8 ml phenol : chloroform (1:1, v/v), centrifuged at 12,000 × g for 15 minutes at 4°C, washed once with 0.8 ml of chloroform and centrifuged at 12,000 × g for 15 minutes at 4°C again. Isopropanol was added to the aqueous phase, and the preparation was frozen at -20°C overnight. The following day, the samples were centrifuged at 12,000 × g for 30 minutes at 4°C. The RNA pellet was washed with 1 ml 75% ethanol and allowed to air dry until all moisture was gone. The pellet was resuspended in DEPC water and quantitated by optical density readings at 260 nm. cDNA was synthesized with murine leukemia virus reverse transcriptase (2.5 U/μl), 10 × PCR buffer (500 mM KCl, 100 mM Tris-HCl, pH 8.3), 1 mM each of the nucleotides dATP, dCTP, dGTP and dTTP; RNase inhibitor (1 U/μl), MgCl 2 (5 mM), and oligo(dT) 16 (2.5 μM) as a primer. The samples were incubated at 42°C for 20 minutes, 99°C for 20 minutes, and 5°C for 5 minutes in a DNA thermocycler (Perkin-Elmer Corp., Norwalk, CT) for reverse transcription. PCR of cDNA was done with MgCl 2 (1.8 mM), each of the dNTPs (0.2 mM), AmpliTaq polymerase (1 U/50 μl), and paired cytokine-specific primers (0.2 nM of each primer) to a total volume of 50 μl. Cycles consisted of 1 cycle of 95°C for 2 min, 35 cycles of 95°C for 45 sec, 60°C for 45 sec, and 72°C for 1 min 30 sec, and lastly, 1 cycle of 72°C for 10 min. Ten microliters of the sample were electrophoresed on a 2% agarose gel and stained with ethidium bromide for viewing. Primer sequences used are as follows: HPRT: 5' CGA GAT GTG ATG AAG GAG ATG G 3' and 5' GGA TTA TAC TGC CTG ACC AAG G 3'; IL-6: 5' ATG AAC TCC TTC TCC ACA AGC GC 3' and 5' GAA GAG CCC TCA GGC TGG ACT G 3'; IL-8: 5' ATG ACT TCC AAG CTG GCC GTG GCT 3' and 5' TCT CAG CCC TCT TCA AAA ACT TCT C 3'; and IL-13: 5' GGA AGC TTC TCC TCA ATC CTC TCC TGT T 3' and 5' GCG GAT TCG TTG AAC CGT CCC TCG CGA AA 3'. Densitometry was done by normalizing target genes to house keepers using Un-Scan-It Version 5.1 software (Orem, UT). The PCR experiment was repeated twice.
|
15383152_p23
|
15383152
|
Analysis of cytokine gene expression by RT-PCR
| 4.284801 |
biomedical
|
Study
|
[
0.9992659687995911,
0.0005041830008849502,
0.0002297613536939025
] |
[
0.9969570636749268,
0.002331559546291828,
0.00054015131900087,
0.00017123470024671406
] |
en
| 0.999996 |
HMC-1 were stimulated with PMA, IL-1β and/or epinephrine and then harvested for EMSA analysis . Cells were washed with PBS and mixed with one hundred microliters of hypotonic buffer which contains: 10 mM HEPES pH 7.9, 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM dithiothreitol (DTT), 0.5 mM phenylmethylsulfonyl fluoride (PMSF), 1 μM aprotinin, 1 μM pepstatin, 14 μM leupeptin, 50 mM NaF, 30 mM β-glycerophosphate, 1 mM Na 3 VO 4 , and 20 mM p-nitrophenyl phosphate. Cells were incubated over ice for 30 minutes and then vortexed after the addition of 6.25 μl of 10% of Nonidet P-40. After 2 minutes of centrifugation at 30,000 × g, supernatants were kept at -80°C while the pellets were collected and vortexed every 20 minutes for 3 hours in 60 ml of a hypertonic salt solution: 20 mM HEPES pH 7.9, 0.4 M NaCl, 1 mM EDTA, 1 mM EGTA, 12 mM DTT, 1 mM PMSF, 1 μM aprotinin, 1 μM pepstatin, 14 μM leupeptin, 50 mM NaF, 30 mM β-glycerophosphate, 1 mM Na 3 VO 4 , and 20 mM p-nitrophenyl phosphate. Nuclear translocation of NF-κB was analyzed by the Electrophoretic Mobility Shift Assay (EMSA) using the nuclear fraction. Seven micrograms of nuclear protein were added to 2 ml of binding buffer (Promega, Madison, WI), and 35 fmol of double stranded NF-κB consensus oligonucleotide (5' AGT TGA GGG GAC TTT CCC AGG C 3') (Promega, Madison, WI) end labeled with γ-P 32 ATP (Amersham Biosciences, Piscataway, NJ). The samples were incubated at room temperature for 20 minutes and run on a 5% nondenaturing polyacrylamide gel for 2 hours. The gel was then dried on a Gel-Drier (Bio-Rad Laboratories, Hercules CA) and exposed to Kodak X-ray film at -80°C.
|
15383152_p24
|
15383152
|
NF-κB assay in HMC-1
| 4.178737 |
biomedical
|
Study
|
[
0.9994407296180725,
0.0003269432345405221,
0.00023237778805196285
] |
[
0.9985761642456055,
0.0010085977846756577,
0.00032681183074600995,
0.00008845172123983502
] |
en
| 0.999998 |
Cells were treated and lysed in lysis buffer (50 mM Tris HCL, 150 mM NaCl, 1 mM EDTA, 1% Triton × 100, and 0.1% SDS) to be analyzed for p38 MAPK activation by Western blot . Briefly, 50 μg of sample protein was diluted 1:2 with Laemmli buffer (Bio-Rad laboratories, Hercules, CA) and boiled for 10 minutes in a sand bath. The resultant sample was then run in a Bio-Rad Modular Mini Electrophoresis System (Hercules, CA) on a 10% polyacrylamide gel for 1 hour and then transferred to a 0.2 μm nitrocellulose membrane (Bio-Rad laboratories, Hercules, CA) for 1 hour. The blot was then incubated in blocking buffer (1% BSA and 0.1% Tween in PBS) for 1 hour at room temperature with gentle agitation. Rabbit anti-human Phospho-p38 MAPK (Thr180/Tyr182) polyclonal antibody (Calbiochem, San Diego, CA) was diluted 1:1000 in blocking buffer and incubated on the blot overnight at 4°C with gentle agitation. After the primary antibody was removed the blot was washed three times for 10 minutes each with agitation in the wash buffer (0.1% Tween in PBS). The blot was then incubated in horse radish peroxidase conjugated mouse anti-rabbit Ig's antibody (human adsorbed, Santa Cruz Biotechnology, Santa Cruz, CA) diluted 1:5000 in blocking buffer. The blot remained in the secondary antibody for 1 hour at room temperature. The blot was then washed again and covered with Super Signal West Pico Chemiluminescent Substrate (Pierce, Rockford, IL) for 5 minutes. The blot was then exposed to acetate transparency film (Kodak, Rochester, NY) and developed. The same protocol was repeated for total p38 MAPK analysis.
|
15383152_p25
|
15383152
|
Detection of p38 MAPK by Western blot
| 4.227527 |
biomedical
|
Study
|
[
0.9992522597312927,
0.000479584326967597,
0.00026808350230567157
] |
[
0.9921579360961914,
0.006875597406178713,
0.0007575341733172536,
0.00020898204820696265
] |
en
| 0.999995 |
Resting HMC-1 were centrifuged, washed in PBS at room temperature, and resuspended in 100 μl of PBS. The cells were incubated for 20 minutes with rabbit polyclonal anti β 1 or β 2 adrenergic receptor antibodies (Santa Cruz, Santa Cruz, CA) using normal rabbit serum as a control. The samples were washed with PBS with 0.01% sodium azide and resuspended in 100 ml PBS. FITC labeled goat anti-rabbit Ig's antibody was added to the samples and allowed to bind for 20 minutes. The samples were once again washed with PBS with 0.01% sodium azide and resuspended in 100 μl of PBS. In addition, HMC-1 were pretreated with normal rabbit serum and incubated with FITC labeled goat anti-rabbit Ig's antibody as a control for nonspecific binding . Cell suspensions were then gated and analyzed based on fluorescence using a Becton Dickinson FACSCalibur 4-color flow cytometer (San Diego, CA) and histograms generated on WinMDI 2.8 software (kindly provided by Joseph Trotter over the internet).
|
15383152_p26
|
15383152
|
Analysis of β-adrenoceptor by flow cytometry
| 4.094995 |
biomedical
|
Study
|
[
0.999517560005188,
0.00026681163581088185,
0.00021566712530329823
] |
[
0.9975583553314209,
0.001998855033889413,
0.0003544004575815052,
0.00008844047988532111
] |
en
| 0.999998 |
HMC-1 were treated with IL-1β (10 ng/ml), epinephrine (10 -5 M), and IL-1β plus epinephrine to investigate IL-13 promotor activity. Untreated cells were used as a control. Transient transfection assays were performed using a reporter gene construct containing the minimal promoter sequence of IL-13. The promoter sequence (-233 to + 50, relative to the transcription initiation site) of the IL-13 gene was fused to the luciferase coding sequence. Plasmid DNA was obtained with double-cesium chloride purification (BioServe Biotechnologies, Laurel, MD), while SuperFect reagent (Qiagen) was used for transient transfections of HMC-1 cells. Two micrograms of plasmid DNA and 8 μl SuperFect reagent were used for transfection of 1 × 10 6 HMC-1 cells. Luciferase expression was monitored by chemiluminescence of cell lysates 24 hrs after transfections using the Enhanced Luciferase Assay Kit (Analytical Luminescence Laboratory, Ann Arbor, MI).
|
15383152_p27
|
15383152
|
IL-13 promotor analysis
| 4.112407 |
biomedical
|
Study
|
[
0.9995660185813904,
0.00023709946253802627,
0.00019682291895151138
] |
[
0.9993388056755066,
0.0003394650120753795,
0.00026443161186762154,
0.00005726069139200263
] |
en
| 0.999996 |
All experiments were done in triplicate. The data were analyzed by Student's two-tailed t -test using Statistica software (StatSoft, Inc., Tulsa, OK). All data were reported as means ± SE. A p -value of less than 0.05 was considered significant.
|
15383152_p28
|
15383152
|
Statistical analysis of the data
| 2.806252 |
biomedical
|
Study
|
[
0.9984896183013916,
0.00033774826442822814,
0.0011726062512025237
] |
[
0.9387653470039368,
0.05862291529774666,
0.0020999128464609385,
0.0005118457484059036
] |
en
| 0.999998 |
HMC-1, human mast cell – 1
|
15383152_p29
|
15383152
|
List of abbreviations used
| 2.286855 |
biomedical
|
Other
|
[
0.993644118309021,
0.0017199463909491897,
0.004635963123291731
] |
[
0.03937317430973053,
0.9569436311721802,
0.0017125861486420035,
0.001970697194337845
] |
en
| 0.999995 |
Epi, epinephrine
|
15383152_p30
|
15383152
|
List of abbreviations used
| 2.019962 |
biomedical
|
Other
|
[
0.9515482783317566,
0.004050311166793108,
0.04440135136246681
] |
[
0.02616344951093197,
0.9713367819786072,
0.0014648468932136893,
0.0010350050870329142
] |
ro
| 0.428571 |
Pro, propranolol
|
15383152_p31
|
15383152
|
List of abbreviations used
| 1.419508 |
biomedical
|
Other
|
[
0.8505718111991882,
0.012861893512308598,
0.13656634092330933
] |
[
0.047124918550252914,
0.9440017342567444,
0.0054204752668738365,
0.0034528695978224277
] |
it
| 0.999994 |
Ate, atenolol
|
15383152_p32
|
15383152
|
List of abbreviations used
| 1.785231 |
biomedical
|
Other
|
[
0.9321759939193726,
0.011140943504869938,
0.05668307840824127
] |
[
0.013209075666964054,
0.9834203720092773,
0.0018315912457183003,
0.0015390294138342142
] |
it
| 0.999993 |
Dex, dexamethasone
|
15383152_p33
|
15383152
|
List of abbreviations used
| 2.055018 |
biomedical
|
Other
|
[
0.9678703546524048,
0.002688413253054023,
0.029441174119710922
] |
[
0.03341039642691612,
0.9637342691421509,
0.001599860843271017,
0.0012554398272186518
] |
en
| 0.857138 |
MAPK, mitogen-activated protein kinase
|
15383152_p34
|
15383152
|
List of abbreviations used
| 2.539467 |
biomedical
|
Other
|
[
0.9888755083084106,
0.002337801270186901,
0.00878670159727335
] |
[
0.012277449481189251,
0.9862273335456848,
0.0008199642179533839,
0.000675269344355911
] |
en
| 0.999996 |
DSC designed experiments, oversaw research, and wrote paper. SMF designed and conducted experiments and wrote paper. SP helped with experiments. KC conducted experiments. EK conducted experiments. SAL conducted experiments. SKH conducted experiments. GK oversaw research.
|
15383152_p35
|
15383152
|
Author's contributions
| 0.830318 |
other
|
Other
|
[
0.11950717121362686,
0.0019447332015261054,
0.8785480856895447
] |
[
0.0038794169668108225,
0.9949570298194885,
0.0006913308752700686,
0.00047218482359312475
] |
en
| 0.999997 |
Many patients do not eat and drink sufficiently during hospitalisation. Thus, 30–50% of the elderly patients are undernourished and most of these patients' protein and energy requirements are not met . Their muscular tissue, including their heart and respiratory muscles, is adversely affected by this situation and their immune function is suppressed . The clinical consequences include lassitude, difficulty in mobilising, prolonged convalescence and an increased risk of pressure wounds , phlebitis and infections . Patients often have reduced appetite, nausea or aversion towards certain types of food, which may partly explain the inadequacy of their food and liquid intake. Intervention studies have shown that by offering food or in-between meals rich in energy and protein, it is possible to increase the patient's protein and energy intake . However, most of these intervention studies only use quantitative data. The present intervention study offers data, both quantitative on patients' food intake and qualitative on the occupational groups' attitudes and experiences in relation to the intervention, the food service and the nutritional care in general. These data can contribute to raise our knowledge of nutritional care in general and to identify issues crucial to an improvement of hospitalised patients' food intake in particular.
|
15363101_p0
|
15363101
|
Background
| 4.088354 |
biomedical
|
Study
|
[
0.9984489679336548,
0.0012344681890681386,
0.00031645086710341275
] |
[
0.9977893829345703,
0.0004872657300438732,
0.0015473744133487344,
0.0001759977894835174
] |
en
| 0.999998 |
The first aim of this research was to examine to which extent standard hospital food met hospitalised medical patients' protein and energy requirements. The second aim was to introduce nursing procedures focusing on the nutritional care based on the Danish nutritional recommendations for inpatients to investigate the effect of this intervention on the patients' intake of protein and energy. The third aim was to explore the involved occupational groups' attitudes towards nutritional intervention and nutritional care in general. Particular attention was paid to the identification of problems possibly related to insufficient patient nutrition.
|
15363101_p1
|
15363101
|
Background
| 3.981482 |
biomedical
|
Study
|
[
0.9966742992401123,
0.002171215135604143,
0.0011545432498678565
] |
[
0.9990620017051697,
0.0005997731350362301,
0.00022082468785811216,
0.00011748788529075682
] |
en
| 0.999996 |
The setting was an endocrinology ward with 49 beds and 3481 patients discharged during 2002 (divided into bed sections IA and IB) and a cardiology ward with 53 beds and 4542 patients discharged during 2002 (divided into bed sections IIA and IIB) . All hospital food was produced in a central hospital kitchen and transported in heated containers to the bed sections where it was portioned out and served to the patients.
|
15363101_p2
|
15363101
|
Setting
| 1.903013 |
biomedical
|
Other
|
[
0.807397186756134,
0.17150263488292694,
0.021100280806422234
] |
[
0.2784005403518677,
0.7041912078857422,
0.0012078846339136362,
0.016200345009565353
] |
en
| 0.999997 |
Medical patients' pre-intervention dietary protein and energy intakes were assessed by 72-hour weighed food records at four bed sections (two wards) to include the appropriate number of patients. Before the intervention the bed sections at each ward was randomised to intervention or control. After a five-month intervention period, patients' dietary protein and energy intakes were assessed to evaluate the effect of intervention. After intervention the occupational groups involved in the nutritional care and the food service at the two intervention sections were interviewed in focus groups or by individual interview.
|
15363101_p3
|
15363101
|
Design of the study
| 3.938376 |
biomedical
|
Study
|
[
0.9606466889381409,
0.038210444152355194,
0.00114289834164083
] |
[
0.9824665784835815,
0.015553056262433529,
0.0007980451919138432,
0.0011823662789538503
] |
en
| 0.999996 |
Both acute and referred medical patients at all ages participated. The inclusion criteria were defined as: 1) the patient was not placed on a prescribed diet, 2) the patient had no contact with, or had not previously received dietary advice from a clinical dietician, 3) the patient did not belong to an ethnic minority, and 4) the patient was hospitalised for at least five days. Patients with dementia and patients who were severely mentally or physically impaired were excluded.
|
15363101_p4
|
15363101
|
Participants
| 2.807259 |
biomedical
|
Study
|
[
0.810509204864502,
0.18623407185077667,
0.0032568229362368584
] |
[
0.968268632888794,
0.02533753216266632,
0.0012290423037484288,
0.005164734087884426
] |
en
| 0.999997 |
Typical patient diagnoses included acute or chronic lung disease (e.g. chronic obstructive lung disease, asthma, bronchitis), acute or chronic cardiovascular disease (e.g. hypertension, angina, thrombosis, apoplexy), metabolic disorders (e.g. thyrotoxicosis, osteoporosis) or infectious disease (e.g. pneumonia, cystitis). The nursing staff selected the patients meeting all the criteria. The patients received oral and written information about the investigation underlining the voluntary nature of their participation. Three or four patients from a bed section participated at the same time, providing data for the food records.
|
15363101_p5
|
15363101
|
Participants
| 2.30541 |
biomedical
|
Study
|
[
0.9574646353721619,
0.04023241624236107,
0.0023029805161058903
] |
[
0.9506514072418213,
0.04416614770889282,
0.0009753710473887622,
0.004207023419439793
] |
en
| 0.999998 |
The occupational groups participating in the focus group interviews were: nurses, health care support staff and nurse aides on day or evening duty from one of the two intervention sections IB and IIB (four interviews), two nurses in charge from the two intervention sections (one interview), three maids from the two intervention sections (one interview), two clinical dieticians from the two intervention sections (one interview), and one catering officer from the kitchen (one individual interview). In total 26 informants participated in eight interviews.
|
15363101_p6
|
15363101
|
Participants
| 1.846439 |
biomedical
|
Study
|
[
0.7645626664161682,
0.018769310787320137,
0.2166679948568344
] |
[
0.852945864200592,
0.14461639523506165,
0.0009820683626458049,
0.0014556600945070386
] |
en
| 0.999997 |
Data on patient age, date of and diagnosis on hospitalisation, second diagnosis, oedema, dehydration, body weight on hospitalisation (if measured) were collected by the investigators from hospital records. Body temperature (if fever) was collected from the hospitals records during the 72-hour food recording. Patients' body weights were recorded twice: at hospitalisation (or when they were included in the study) and on discharge. This weighing was standardised according to time of the day, the patients dress and the scales. The changes in body weight during hospitalisation were recorded for the patients not having oedema or dehydration. Patient height was measured and body mass index (BMI, kg/m 2 ) calculated. The patient was asked about ability to chew and swallow and recorded as 'effortless', 'slight difficulty' or 'with difficulty'. On discharge they ascribed to the meals during hospitalisation was recorded as 'very important', 'of some importance', 'almost no importance' and 'no importance'.
|
15363101_p7
|
15363101
|
Patients' characteristics
| 3.89149 |
biomedical
|
Study
|
[
0.8723084330558777,
0.12643428146839142,
0.0012573057319968939
] |
[
0.987869918346405,
0.007986113429069519,
0.0009719619411043823,
0.0031719037797302008
] |
en
| 0.999995 |
The patients had their food and drink weighed for 72 hours at breakfast, lunch, afternoon coffee and supper by the investigators. The patients, relatives or staff recorded the last-meal-of-the-day and individual between-meals as estimated records. The investigators contacted the patients three to five times a day to follow up on these estimated records. The weight of the between meals provided by visitors was estimated by weighing similar food items.
|
15363101_p8
|
15363101
|
Food records
| 2.28533 |
biomedical
|
Study
|
[
0.8329859972000122,
0.16260920464992523,
0.004404854495078325
] |
[
0.8780621290206909,
0.1119496151804924,
0.0014756984310224652,
0.008512523956596851
] |
en
| 0.999997 |
The food items were weighed in the form received from the kitchen. Potatoes, mashed potatoes, sauce, meat, etc. were weighed separately. Standardized menus such as stew, open sandwiches, sandwiches, etc. were weighed in full. The total weight of the food items each patient was served was weighed before serving as was plate waste after the meal. Drinks were estimated and recorded when poured into a glass or feeding cup one centimetre below the rim.
|
15363101_p9
|
15363101
|
Food records
| 1.991823 |
biomedical
|
Other
|
[
0.9499989151954651,
0.016547847539186478,
0.03345317021012306
] |
[
0.24660731852054596,
0.7490906119346619,
0.0009453290258534253,
0.003356770146638155
] |
en
| 0.999995 |
24-hour food records were checked and coded to calculate protein (gram) and energy (kJ) intake by the investigators. The calculations were based on the data from the recipes used in the hospital kitchen and the Danish Database 'Dankost 2000', which contains data from the Danish food tables .
|
15363101_p10
|
15363101
|
Dietary intake of protein and energy
| 2.634221 |
biomedical
|
Study
|
[
0.9974545836448669,
0.0009665913530625403,
0.0015788179589435458
] |
[
0.9842578172683716,
0.014889249578118324,
0.000500933441799134,
0.000352041854057461
] |
en
| 0.999997 |
Each patient's physical activity was recorded every hour during three days and nights (72 hours) in the period of food recording. It was recorded whether the patient was 'lying asleep', 'lying awake', 'sitting', 'walking' or 'training'. For walking or training the approximate duration of activity was recorded as a fraction of an hour, and a factor of physical activity was estimated for each 24-hour period . The investigators contacted the patients three to five times a day to follow up on the recording of the physical activity.
|
15363101_p11
|
15363101
|
Physical activity
| 2.996074 |
biomedical
|
Study
|
[
0.9308311939239502,
0.06730660051107407,
0.0018621399067342281
] |
[
0.9785634279251099,
0.018370188772678375,
0.0007377677829936147,
0.0023286903742700815
] |
en
| 0.999997 |
Official Danish food recommendations for institutions propose that patients with chronic diseases have 1.0–1.5 gram of protein per kilogram body weight depending on the degree of stress metabolism . A factor of 1.2 gram was used as an estimate of moderate metabolic stress . However, they did not allow for the underestimation of underweight and overestimation of overweight patients' requirements. The calculations were adjusted accordingly in the following way: If the BMI was below 20, the recommended requirement was calculated as 1.5 gram per kilogram bodyweight per 24 hours. If the BMI was above 30, the recommended requirement was calculated as 1.0 gram per kilogram bodyweight per 24 hours .
|
15363101_p12
|
15363101
|
Estimation of protein and energy requirements
| 3.779575 |
biomedical
|
Other
|
[
0.9945687055587769,
0.004266366362571716,
0.0011650433298200369
] |
[
0.09911291301250458,
0.863227128982544,
0.03590809926390648,
0.0017517979722470045
] |
en
| 0.999997 |
The estimated energy requirement was calculated as 'basal metabolic rate' (Harris Benedict equation ) x 'the factor of physical activity' x 'the factor required to increase the body weight (if the BMI was below 20)' or a 'factor of stress (if the BMI was above or equal to 20)' . If the BMI was below 20, the factor 1.3 was used instead of the stress factor. The factor required to increase the body weight was an estimate of the amount of energy the patient was able to consume . The stress factor was applied in patients judged to have metabolic stress because of their pathological condition. The stress factor range was 1.1–1.4 for patients with chronic lung disease, chronic heart disease and apoplexy: severe infections were given a factor of 1.3. The stress factor was determined by the temperature and was set at 1.2 at a temperature of 38°C, 1.3 at 39°C and 1.4 at 40°C. Only one factor of stress was used, and the temperature stress factor had the highest priority .
|
15363101_p13
|
15363101
|
Estimation of protein and energy requirements
| 4.106159 |
biomedical
|
Study
|
[
0.9981186985969543,
0.0016239729011431336,
0.0002572855446487665
] |
[
0.9908347129821777,
0.008075929246842861,
0.0007483303779736161,
0.00034091953421011567
] |
en
| 0.999997 |
The mean recorded protein (g) and energy (kJ) intake was compared with the estimated protein (g) and energy requirements (kJ), and the degree (in percent) to which the patient's 24-hour requirements were met.
|
15363101_p14
|
15363101
|
Estimation of protein and energy requirements
| 3.233977 |
biomedical
|
Study
|
[
0.9903675317764282,
0.008949409238994122,
0.0006830603815615177
] |
[
0.9917603135108948,
0.006874214857816696,
0.000610831193625927,
0.0007545910775661469
] |
en
| 0.999999 |
The nurses in charge from the two intervention bed sections IB and IIB received information a) specifying to which degree the patients' protein and energy requirements were being met before intervention and b) detailing the Danish Recommendations for Hospitalised Patients . In order to introduce and facilitate continuous staff monitoration of the patients' nutritional status during their hospitalisation the following intervention procedures were formulated in collaboration with the two nurses in charge. Such monitoration would allow the staff to identify patients at risk of under nourishment and would secure continuous registration, which was seen as a precondition for optimising the patients' uptake of nutrients. The procedures were formulated as one standard applying to be used to all non-diet patients admitted to bed sections IB and IIB: The patient's nutritional status is assessed on admission and during hospitalisation.
|
15363101_p15
|
15363101
|
Intervention
| 3.399446 |
biomedical
|
Other
|
[
0.5064963102340698,
0.48722344636917114,
0.00628021452575922
] |
[
0.401949942111969,
0.574975848197937,
0.005001167766749859,
0.018073076382279396
] |
en
| 0.999996 |
As recommended by the nurses in charge three forms (A, B and C) with different purpose were made to support the staff in relation to the nutritional care.
|
15363101_p16
|
15363101
|
Intervention
| 1.391155 |
biomedical
|
Other
|
[
0.5227901935577393,
0.36724433302879333,
0.10996546596288681
] |
[
0.01449489500373602,
0.9765834808349609,
0.0017230012454092503,
0.007198547013103962
] |
en
| 0.999996 |
In Form A patient data related to the nutritional care were collected upon admission: height, body weight, BMI, usual body weight and changes in body weight for a defined period (if possible), oedema or dehydration, the date and by whom the patient had been informed about the food service, the result of the first assessment of the nutritional status (result from form B), a short description of 1) the patient's ability to eat and drink, and 2) of the action taken by the staff 3) the date of the next assessment of the nutritional status. The form also allowed room for the results of the next five assessments of the patient's nutritional status.
|
15363101_p17
|
15363101
|
Intervention
| 2.605767 |
clinical
|
Other
|
[
0.30469024181365967,
0.6899882555007935,
0.005321478005498648
] |
[
0.3841959238052368,
0.5500597357749939,
0.0034960403572767973,
0.06224822252988815
] |
en
| 0.999998 |
The purpose of Form B was to assess the patient's nutritional status/risk score and suggesting the action the staff could take. Actions were performed according to detailed English Standards . The assessment parameters were body weight for height (BMI), appetite and ability to eat. The patient was assessed at 'low risk' when BMI was normal, the appetite good and the patient fully independent. The patient was assessed at 'moderate risk' when underweight but stable, the appetite poor, and the patient needed help with feeding or had some swallowing difficulties. The patient was assessed at 'high risk' when severely under weight or actively lost weight, ate very little or have had no food for the last four meals, and was dependent on others for feeding or had severe swallowing difficulties. Form B allowed the patient's nutritional status to be recorded six times to ensure continuity of the assessment of nutritional status. A short guide to action was provided to the staff for each of the assessment categories; For the 'low risk' patient 'no action necessary, but check weight weekly'. For the patient at 'moderate risk' the action could be 'check weight weekly, encourage with eating and drinking, replace missed meals with supplements and repeat score after one week and ask medical staff to refer patient to clinical dietician if no improvement'. For the patient at 'high risk' the action was to 'focus on encouraging with eating and drinking, replace missed meals with supplements and repeat score after three to four days and ask medical staff to refer patient to clinical dietician'.
|
15363101_p18
|
15363101
|
Intervention
| 3.349289 |
clinical
|
Other
|
[
0.06524080038070679,
0.9309984445571899,
0.003760737832635641
] |
[
0.13429823517799377,
0.6835837960243225,
0.007944860495626926,
0.17417317628860474
] |
en
| 0.999995 |
In Form C the estimated record of the patient's protein and energy intake could be calculated and compared to data in the nutritional handbook describing what the food items contained of protein and energy. This handbook contained a standardised description of all meals delivered from the kitchen in household measurements (spoons, pieces, decilitre, etc.) and the estimated protein (g) and energy (kJ) content.
|
15363101_p19
|
15363101
|
Intervention
| 2.791748 |
biomedical
|
Other
|
[
0.9294190406799316,
0.06541543453931808,
0.00516549265012145
] |
[
0.11400573700666428,
0.8802207112312317,
0.0018118065781891346,
0.003961803391575813
] |
en
| 0.999997 |
The investigators convened meetings with the nursing staff and the domestic helpers at the two intervention sections IB and IIB. The rationale of the standard was explained detailed and both oral and written instructions about the use of the forms were given. Four meetings were held in bed section IB and six in bed section IIB. At these meetings problems, ideas, etc. related to the standard and the forms were discussed and adjusted according to these discussions. The investigators contacted the staff in bed sections IB and IIB once or twice a week during the five-month investigation period to give support if wanted.
|
15363101_p20
|
15363101
|
Introducing the standard
| 1.874039 |
biomedical
|
Study
|
[
0.750356137752533,
0.15082579851150513,
0.09881812334060669
] |
[
0.5448908805847168,
0.44689688086509705,
0.0017861009109765291,
0.006426215171813965
] |
en
| 0.999999 |
The intervention at the two bed sections had no influence on the food production in the hospital. But before the intervention the kitchen produced a 'unrestricted diet' to all patients not placed on a prescribed diet, which contain about 8250 kJ and 70–80 gram of protein with about 15, 41 and 43% of energy from protein, fat and carbohydrates . During the intervention period the kitchen changed the production to two different diets to meet the Danish Nutritional Recommendations for Diseased People ; From the kitchen the diets were introduced in the following way; one diet for the elderly and people with little appetite – the so-called 'hospital diet' – and one diet for all patients with ischemic heart disease and diabetes mellitus – the so-called 'normal diet'. The 'hospital diet' contained about 10000 kJ and 90 gram of protein with 18, 40 and 42% of energy from protein, fat and carbohydrates. The 'normal diet' contained about 9000 kJ and 80 gram of protein with 10–15, 30 and 55–60% of energy from protein, fat and carbohydrates . The changes in the diets were introduced to the staff by the clinical dieticians. Besides these diets different commercial and no-commercial protein- and energy supplements, stewed fruit, soup etc. were available from the kitchen.
|
15363101_p21
|
15363101
|
Introducing the standard
| 3.713127 |
biomedical
|
Study
|
[
0.938947319984436,
0.059057269245386124,
0.0019954112358391285
] |
[
0.888506293296814,
0.10438022762537003,
0.0023733649868518114,
0.0047400533221662045
] |
en
| 0.999999 |
The number of patients required was calculated in the following way:The clinically relevant difference between the average extent to which the patient's protein and energy requirement was met before and after the intervention was estimated to 15% . Patients' dietary protein and energy intakes were estimated to lie 0–50% below their requirements (standard deviation (SD) 12.5–15.0%). A 5% significance level was chosen and the power was chosen to lie at 90%. The t-test was used to calculate an appropriate sample size for the control and intervention groups, viz. a minimum of 21 patients.
|
15363101_p22
|
15363101
|
Statistical methods
| 4.093786 |
biomedical
|
Study
|
[
0.9888206720352173,
0.01063739974051714,
0.0005418473738245666
] |
[
0.9741315245628357,
0.024331986904144287,
0.0006722923717461526,
0.0008642775937914848
] |
en
| 0.999996 |
The dietary protein and energy intake was calculated as a 24-hour mean (SD) for each patient and for each group (SD) of patients at each bed section. The outcome measure was the percentage degree to which the patient's actual protein and energy requirement was covered compared with his/her estimated requirement. Confidence intervals for the outcome measures were estimated.
|
15363101_p23
|
15363101
|
Statistical methods
| 3.891556 |
biomedical
|
Study
|
[
0.9961975812911987,
0.0034718604292720556,
0.0003305971040390432
] |
[
0.9972335696220398,
0.0020829590503126383,
0.00044355422141961753,
0.0002399902732577175
] |
en
| 0.999998 |
The effect of intervention for independent groups of patients were tested by one-way analysis of variance (ANOVA) using the SPSS version 9.0. The assumptions of independence, normality and identical variances were fulfilled. Analyses of covariance were described for non-comparable variables for the four patient groups after intervention.
|
15363101_p24
|
15363101
|
Statistical methods
| 3.750183 |
biomedical
|
Study
|
[
0.9993270635604858,
0.0003313679189886898,
0.0003415649989619851
] |
[
0.9982109069824219,
0.0014216949930414557,
0.00028881101752631366,
0.00007859218749217689
] |
en
| 0.999998 |
An interview guide was designed for each of five occupational groups: 1) nurses, health care support staff and nurse aides (four interviews), 2) charge nurses, 3) maids, 4) clinical dieticians and 5) one catering officer from the kitchen . In the interview the investigator focused on the informants' actions, attitudes, experiences and reflections in relation to the intervention and nutritional care. Focus group interviews were considered the most appropriate form of data collection given the intent of the study . All the 26 informants shared experience from the intervention study and from the situations where patients' meals were served. Eight focus group interviews were carried out at the hospital during the working hours in rooms familiar to the voluntary informants. The focus group interviews were tape-recorded with the permission of the informants, who were informed that they could read the transcribed interview, should they wish so. The qualitative data were analysed as a text.
|
15363101_p25
|
15363101
|
The interview in the occupational groups
| 2.866555 |
biomedical
|
Study
|
[
0.9752691388130188,
0.006935709156095982,
0.017795201390981674
] |
[
0.9965454936027527,
0.0030407689046114683,
0.00018867224571295083,
0.000225060575758107
] |
en
| 0.999996 |
The study fulfilled the declaration of Helsinki II and was approved by the Local Scientific Ethics Committee.
|
15363101_p26
|
15363101
|
Ethical approval
| 1.045085 |
biomedical
|
Other
|
[
0.9283490777015686,
0.006603012792766094,
0.06504786759614944
] |
[
0.0336640290915966,
0.9623197913169861,
0.001982150599360466,
0.0020340606570243835
] |
en
| 0.999995 |
Food records were completed for 48 patients before and 60 patients after intervention. Table 1 summarises the baseline characteristics of the participating patients. The patient groups were comparable with regard to BMI, stress factor and ability to chew and swallow. The average age of the medical patients was 72 ± 11 years. Before the intervention 17 patients out of 22 lost body weight. After the intervention 20 patients out of 37 lost body weight (table 1 ). Before the intervention, 56% of the patients participating in the study were weighed on admission (defined as within 48 hours from their arrival to the bed section). After intervention 52% of the patients in the control sections and 45% in the intervention sections were weighed on admission by the staff.
|
15363101_p27
|
15363101
|
Patient characteristics
| 3.773773 |
biomedical
|
Study
|
[
0.9739965796470642,
0.025144381448626518,
0.0008589404751546681
] |
[
0.9957242012023926,
0.0028409126680344343,
0.00040819740388542414,
0.0010266995523124933
] |
en
| 0.999998 |
In table 2 the average degree to which protein requirements were met before and after intervention are summarized. In table 3 the corresponding figures for energy requirements. There were no significant pre-intervention differences between the groups concerning the average degree to which their estimated protein (p = 0.918) and energy (p = 0.367) requirements were met.
|
15363101_p28
|
15363101
|
Patient requirement and protein and energy intake
| 3.191835 |
biomedical
|
Study
|
[
0.9973371624946594,
0.0010395003482699394,
0.0016233731294050813
] |
[
0.9989312291145325,
0.0006824817974120378,
0.00029655866092070937,
0.00008973098738351837
] |
en
| 0.999996 |
The results of the intervention was different at bed section IB and IIB; The intervention significantly improved the degree to which the energy and protein requirements were met among patients in intervention section IB compared with patients in the control sections IA and IIA (protein p = 0.009 and energy p = 0.010). On average, the former had an intake of 85% of their calculated protein requirement and 103% of their energy requirement. In intervention section IIB, the patients only had an intake reaching 56% of their protein and 76% of their energy requirement. These values were on average much lower than for patients in section IB and they were comparable to those obtained in the control sections IA and IIA. Analysis of co-variance for the non-comparable variables age, patient mobility, BMI, type of diet and number of bed-days showed no significant effect on the outcome measure for the degree of meeting the patients' requirement of protein and energy. The patients ability to chew and swallow, and the importance of the meals to the patients during hospitalisation were comparable in the four groups of patients before and after the intervention.
|
15363101_p29
|
15363101
|
Patient requirement and protein and energy intake
| 4.140936 |
biomedical
|
Study
|
[
0.9961689114570618,
0.003392799524590373,
0.0004383159684948623
] |
[
0.9985325336456299,
0.0007466422975994647,
0.000529601180460304,
0.0001912756124511361
] |
en
| 0.999998 |
In the control sections the diet met 61% of the patients' protein and 75% of their energy requirements after intervention. These levels were not significantly different from those recorded before the intervention, but 11% and 14% lower than before the kitchen changed the diets.
|
15363101_p30
|
15363101
|
Patient requirement and protein and energy intake
| 2.502019 |
biomedical
|
Study
|
[
0.995194137096405,
0.0032073291949927807,
0.0015985562931746244
] |
[
0.987743616104126,
0.011050241068005562,
0.0006338324747048318,
0.0005723267677240074
] |
en
| 0.999998 |
During the intervention period, the nursing staff in bed section IB used the forms for assessing the nutritional care of three patients. In intervention section IIB the forms was used assessing the nutritional care of 17 patients. The patients nutritional status/risk score were not determined otherwise.
|
15363101_p31
|
15363101
|
The intervention and the occupational groups
| 1.819032 |
biomedical
|
Study
|
[
0.8573452234268188,
0.12991152703762054,
0.01274329237639904
] |
[
0.6998759508132935,
0.28133252263069153,
0.0025723727885633707,
0.01621917448937893
] |
en
| 0.999998 |
Analysis of the qualitative data from the eight interviews extracted five templates with questions relevant to an increased risk of insufficient nutritional care:
|
15363101_p32
|
15363101
|
The intervention and the occupational groups
| 1.896506 |
biomedical
|
Study
|
[
0.9660962820053101,
0.00545076048001647,
0.028453027829527855
] |
[
0.953559935092926,
0.04315296560525894,
0.0023483065888285637,
0.0009388116886839271
] |
en
| 0.999997 |
1. Divergent attitudes towards intervention.
|
15363101_p33
|
15363101
|
The intervention and the occupational groups
| 1.419236 |
other
|
Other
|
[
0.4701954126358032,
0.00901678018271923,
0.5207878351211548
] |
[
0.051359839737415314,
0.9237440824508667,
0.02261386625468731,
0.0022822157479822636
] |
en
| 0.999995 |
2. Lack of flexibility during meals.
|
15363101_p34
|
15363101
|
The intervention and the occupational groups
| 1.376902 |
biomedical
|
Other
|
[
0.7169497013092041,
0.06026745215058327,
0.22278288006782532
] |
[
0.005901368334889412,
0.9886558055877686,
0.0023754476569592953,
0.0030673311557620764
] |
en
| 0.999997 |
3. Lack of knowledge about nutritional care for patients.
|
15363101_p35
|
15363101
|
The intervention and the occupational groups
| 1.537139 |
biomedical
|
Other
|
[
0.8733041882514954,
0.0452362522482872,
0.08145962655544281
] |
[
0.023654915392398834,
0.9684092402458191,
0.004759702365845442,
0.0031760532874614
] |
en
| 0.999997 |
4. Nutrition – a subordinate part of the care.
|
15363101_p36
|
15363101
|
The intervention and the occupational groups
| 1.408339 |
biomedical
|
Other
|
[
0.6550657153129578,
0.01669107750058174,
0.328243225812912
] |
[
0.008310102857649326,
0.9850992560386658,
0.005176971200853586,
0.0014136560494080186
] |
en
| 0.999996 |
5. Lack of recognition of responsibility for nutritional care.
|
15363101_p37
|
15363101
|
The intervention and the occupational groups
| 1.548132 |
biomedical
|
Other
|
[
0.7032615542411804,
0.06153688579797745,
0.23520158231258392
] |
[
0.012254726141691208,
0.9821261763572693,
0.0033931953366845846,
0.002225912408903241
] |
en
| 0.999996 |
Analysis showed that the staff in the intervention sections had not been using the nutritional records systematically. Several nurses thought that the records were too comprehensive and overwhelming. Many mentioned that they had not had the time to learn how to use the records and they were clearly perceived as an extra workload. The nurses in charge mentioned that it was not unproblematic to burden staff with material they did not have the time or resources to read. However, a few staff members, among them two nurse students from bed section IIB, had learned how to use the records. They found that they were utilizable and easy to use.
|
15363101_p38
|
15363101
|
Divergent attitudes towards intervention
| 1.978168 |
biomedical
|
Study
|
[
0.7290788292884827,
0.18896792829036713,
0.08195324242115021
] |
[
0.8773181438446045,
0.114722840487957,
0.002231465419754386,
0.0057276589795947075
] |
en
| 0.999997 |
The two nurses in charge had divergent views on the usability of the intervention study. The charge nurse in bed section IIB thought that the intervention had improved their work with the patients' nutrition. The staff had previously accepted that patients would lie without eating for seven to ten days. Intervention caused the staff to use a feeding tube on threatened patients earlier than before the intervention. However, the charge nurse from bed section IB declared that the staff in her section had not shown much commitment to the intervention. The staff had not taken 'ownership' of the intervention study because the decision to participate in the project had not been a staff decision but one taken by the central management. She emphasized that the staff's attitude was rooted in the fact they had to take in new ideas and instructions all the time.
|
15363101_p39
|
15363101
|
Divergent attitudes towards intervention
| 1.787821 |
clinical
|
Other
|
[
0.15876546502113342,
0.7960954904556274,
0.04513898119330406
] |
[
0.252034455537796,
0.6469166278839111,
0.003897454822435975,
0.09715142846107483
] |
en
| 0.999996 |
Several care providers in bed section IB thought that it was a sizeable extra workload to use the records for recording patients' nutritional statuses and that this had constituted a barrier to their active participation in the process. Other nurses in bed section IB declared that they did not think that it was necessary to continuously register a patient's nutritional status. It sufficed for some nurses to use their 'clinical judgement' and on this basis monitor the patient's weight status. These nurses were not interested in any new initiatives and in tools for nutritional care.
|
15363101_p40
|
15363101
|
Divergent attitudes towards intervention
| 1.835289 |
clinical
|
Other
|
[
0.33443012833595276,
0.6067350506782532,
0.05883480980992317
] |
[
0.08007892966270447,
0.8984399437904358,
0.0025378605350852013,
0.018943315371870995
] |
en
| 0.999996 |
The records were not – and are still not – an integral part of the nutritional care in the intervention sections. This impacted on care continuity. The few staff members who had actually been using the records and had been able to identify patients at risk of insufficient nutrition reported that their observations had not been translated into action.
|
15363101_p41
|
15363101
|
Divergent attitudes towards intervention
| 1.618136 |
clinical
|
Other
|
[
0.4567655920982361,
0.48526662588119507,
0.05796774849295616
] |
[
0.19709022343158722,
0.775296151638031,
0.004390242975205183,
0.023223353549838066
] |
en
| 0.999998 |
Although the food records were only used to a minor extent, the staff generally agreed to the relevance of focusing on the patients' nutrition. Several nurses had not previously paid much attention to the patient's nutrition, but the intervention had made them more conscious of this issue:
|
15363101_p42
|
15363101
|
Divergent attitudes towards intervention
| 1.56597 |
biomedical
|
Other
|
[
0.6810450553894043,
0.25182873010635376,
0.06712615489959717
] |
[
0.1322881579399109,
0.8473143577575684,
0.0045030745677649975,
0.015894325450062752
] |
en
| 0.999995 |
Subsets and Splits
SQL Console for rntc/test-pp-aa
The query retrieves a sample of documents that are clinical cases with an educational score above 3, providing limited analytical value.
Clinical Cases Sample
Returns a sample of 100 clinical case documents, providing a basic overview of the document type's content.