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Quantitative estimation of Holarrhena antidysenterica bark total alkaloids in crude drugs and in the body fluids of man and rat.

A turbidimetric method was developed for the quantitative estimation of the total alkaloids of kutaj bark (Holarrhena antidysenterica) in crude medicinal preparations and in the body fluids of man and rat. The alkaloids were colloidally precipitated with Dragendorff's reagent as complex salts of potassium iodobismuthate in extremely dilute solutions. The finely subdivided orange-brown precipitate gave a coloured, clear homogeneous suspension in the presence of gum arabic. Optical density of such suspensions changed linearly with the change in alkaloid concentration, when prepared within the standardized experimental conditions that included control of ion concentration and temperature of the reaction mixture. Observations revealed the reversible nature of the alkaloid-reagent reaction. Crude medicinal preparations from three different pharmaceutical sources contained varying concentrations of the alkaloids. Complete recovery of the alkaloids was possible from plasma and urine, while significant amounts of the alkaloids were lost to blood cells and faecal contents in man and rat.

Alkaloids↗

Anxiety and perception of risk of HIV and hepatitis B infection among health-care workers reporting accidental exposures to blood and other body fluids.

We noticed considerable variation in anxiety among staff reporting blood exposure incidents and therefore undertook a study to investigate this. We studied 100 consecutive staff reporting blood or other body fluid exposures to the Occupational Health Unit. The nurse seeing the staff member administered a questionnaire about worries related to the incident, knowledge of HIV and hepatitis B transmission risks, perception of risk from the particular incident and predicted reaction of others that would be told. Level of anxiety was recorded on a visual analogue scale. Staff were then given information and counselling as usual, and asked to re-attend after a week, when the questionnaire was repeated. We found that the initial level of anxiety was not related to knowledge of HIV or hepatitis B transmission risks, but was related to perception of risk from the incident and to predicted reaction of others that would be told. The eight staff involved in exposures to known HIV-infected blood were not more anxious than the remainder. There was a reduction in anxiety between visits, which was significantly greater in women, in those who had a non-parental exposure and in those where the source patient was known. Knowledge of transmission risks also improved significantly between visits. This study underlines the importance of adequate counselling of staff who have suffered blood exposures.

Adult↗

Body fluids, circadian blood pressure and plasma renin during growth hormone administration: a placebo-controlled study with two growth hormone doses in healthy adults.

Side effects that can be related to fluid retention are common during the initial phases of growth hormone (GH) administration. The aim of this study was to examine the changes in body fluid compartments, diurnal blood pressure and plasma renin concentration during GH administration with two different dosages in healthy adults. Eight healthy male subjects aged 24-32 years were examined during three 2-week study periods in a double-blind placebo controlled study. They received, in random order, GH (3 or 6 IU m-2 daily) or placebo during 2 weeks. Bio-impedance was measured every 2nd day, and extracellular volume (ECV) and plasma volume (PV) were isotopically determined at day 6. Blood samples were obtained regularly. Diurnal blood pressure was recorded and 24-h urinary samples were collected at days 0, 6 and 14. ECV (l) was increased by GH (placebo, 19.58 +/- 0.82; 3 IU m-2, 20.77 +/- 1.22; 6 IU m-2, 20.65 +/- 0.94; p<0.01), whereas PV (l) was unaffected (placebo, 3.91+/- 0.20; 3 IU m-2, 4.04 +/- 0.22; 6 IU m-2, 3.90 +/- 0.27). Total body water (l) increased significantly during GH administration (placebo, 50.8 +/- 2.6; 3 IU m-2, 52.6 +/- 2.3; 6 IU m-2, 53.9 +/- 1.8, p<0.05). After 6 days of treatment a significant increase in renin (p = 0.03) was observed. Mean diurnal blood pressure levels remained unchanged, whereas mean diurnal heart rate (min-1) increased significantly (placebo, 75 +/- 3.6; 3 IU m-2, 79 +/- 3.2; 6 IU m-2, 79 +/- 3.7; p<0.01). In conclusion, GH administration induces an elevation in total body water which may involve a stimulation of plasma renin and an increased ECV without any changes in PV or diurnal blood pressure.

Adult↗

Modified nucleosides in body fluids of tumor-bearing patients.

The catabolism of nucleic acids, particularly tRNA, produces a variety of modified nucleosides which are not reutilized by mammalian cells. Investigation of these compounds in body fluids, mainly urine, has recently provided evidence of altered metabolic situations in tumor-bearing patients. The factors involved in the alterations of modified nucleosides formation are connected with altered tRNA-modifying enzymes and/or altered turnover of subpopulations of tRNA. A common pattern in tumor cells or tissues is the presence of isoaccepting tRNA species containing aberrant nucleoside modifications. Several modified nucleosides have been detected and quantitated by HPLC analysis of the urine of normal subjects and cancer patients. Results obtained, in the authors' laboratory, among others, indicate a possible correlation between urinary excretion of these compounds and the course of the disease, with implications for the follow-up of therapeutic treatment. Particular reference should be made to psi, which appears to be a suitable marker for monitoring these subjects. The data from the authors' laboratory also show that the analysis of modified nucleosides in blood may be considered a useful tool in the search for proper markers associated with the cancer status. In this respect psi is suggested as a biochemical indicator for cancer patients.

Chromatography, High Pressure Liquid↗

[Occupational exposure to blood and body fluids in haemodialysis health care workers: epidemiological data and prevention in France].

Health care workers (HCW) in haemodialysis units are confronted with a significant risk of occupational exposure to blood and body fluids. The prevalence of bloodborne viruses is high among haemodialysis patients. The data presented here relates to the extraction of 121 occupational exposures notified to occupational health departments by haemodialysis HCW, documented between January 1995 and December 1999 in a network of 54 volunteer hospitals in Northern France. The exposures notified in haemodialysis wards were needlestick injuries in 85 cases (70.2%), splashes to the eyes or non-intact skin in 30 cases (24.8%) and cuts in 6 cases (5.0%). Connection and disconnection of dialysis catheters to fistulae, blood sampling procedures and injections alone were involved in approximately 3 notified exposures out of 4 (46.3, 14.9 and 11.6% respectively for a total of 72.8%). The principal mechanisms for exposure were the handling of blood-soiled needles and instruments or involved the handling of sharps containers. Haemodialysis fistula needles were involved in only 12 (13.2%) of notified percutaneous injuries. Nearly 2/3 (63%) of 91 notified percutaneous injuries could have been avoided by the observance of universal/standard precautions alone and the use of safety devices which were available at the time. The collection and analysis of occupational exposures can serve as basis for an assessment of practices, devices and safety equipment to increase HCW safety in haemodialysis wards.

Blood-Borne Pathogens↗

A radioimmunoassay for measurement of human pancreatic secretory trypsin inhibitor in different body fluids.

A radioimmunoassay for measurement of human pancreatic secretory trypsin inhibitor in nanogram quantities has been developed. The sensitivity of the assay now permits examination of the inhibitor content of various body fluids, wherein other methods exhibit serious short-comings. In healthy blood donors the serum level was 8.1 microgram/l. In patients with acute pancreatitis levels as high as 320 microgram/l have been measured, and patients who underwent endoscopic retrograde cholangiopancreatography showed an elevated inhibitor level in serum immediately after the examination without any clinical signs of disease, the highest registered value being 128 microgram/l. In peritoneal lavage fluid from patients with severe acute pancreatitis levels of 5-304 microgram/l have been measured. In urine the inhibitor level is about 14 microgram/l in healthy persons. The urine from one patient with proteinuria of glomerulo-tubular type contained 380 microgram/l.

Acute Disease↗

Pneumocystis carinii peritonitis. Antemortem confirmation of disseminated pneumocystosis by cytologic examination of body fluids.

Histologic confirmation of extrapulmonary Pneumocystis carinii infection in the acquired immunodeficiency syndrome has usually required organ biopsy when the diagnosis was made antemortem. Three cases of Pneumocystis peritonitis were studied in which confirmation of extrapulmonary dissemination was achieved by cytologic examination of ascitic fluid. Patients presented with characteristic choroidal lesions, transudative ascites, profound hypoalbuminemia, and hepatic dysfunction. Cytologic examination of ascitic fluid confirmed extrapulmonary dissemination of pneumocystis. All three patients died despite a minimum of 2 weeks of standard therapy. Cytologic examination of body fluids to confirm dissemination of Pneumocystis may obviate the need for organ biopsy. Disseminated pneumocystosis should be included in the differential diagnosis of ascites or peritonitis in a patient at risk for human immunodeficiency virus--associated opportunistic infections. The presence of transudative ascites may be characteristic of this syndrome.

Acquired Immunodeficiency Syndrome↗

Measurement of bactericidal/permeability-increasing protein in human body fluids by sandwich ELISA.

A sensitive sandwich ELISA has been developed to measure levels of native bactericidal/permeability-increasing protein (BPI) as well as two recombinant forms of BPI (rBPI and rBPI23) in human body fluids. The linear range for the rBPI and rBPI23 standard curves were 100-6000 pg/ml and 25-800 pg/ml respectively. Recovery of different concentrations of rBPI spiked into pooled human plasma samples averaged 83% and ranged from 65% at 300 ng/ml to 97% at 3 ng/ml. Recovery of rBPI23 averaged 56% and ranged from 30% at 0.5 ng/ml to 90% at 50,000 ng/ml. Because LBP is present in normal human plasma and shares sequence homology with BPI, the effects of rLBP on the BPI ELISA were also evaluated. Under standard assay conditions, rLBP caused minimal interference with BPI detection. At 100 micrograms/ml, rLBP generated a signal equivalent to 3 ng/ml of rBPI and 0.6 ng/ml of rBPI23. Matched serum and plasma samples were collected from 20 healthy adults to measure endogenous levels of BPI. The range of BPI concentrations was < 0.2-2.1 ng/ml in plasma and 4.9-72.1 ng/ml in serum. Western blot analysis indicated that the BPI ELISA immunoreactivity in plasma and serum correlated with the presence of a protein doublet (M(r) approximately 60,000), which comigrated with native BPI extracted from human neutrophils. These data demonstrate that low levels of holo-BPI are present in plasma, and suggest that additional quantities of BPI were released from neutrophils during the process of coagulation.

Acute-Phase Proteins↗

Rapid determination of low pg/mg amounts of N-nitrosodiethylamine in rodent body fluid and tissue samples by isotope dilution high resolution mass spectrometry.

A rapid, sensitive, specific, and cost-effective method for the determination of N-nitrosodiethylamine (NDEA) in rodent tissue and body fluid samples is reported. The method involves isolation of the NDEA from the samples with subsequent detection and quantitation of observed NDEA against mass-labeled N-(15N)-nitrosodiethyl-(15N)-amine (15N2DEA) by high resolution gas chromatography/high resolution mass spectrometry selected ion monitoring. Validation of the method included linearity studies of NDEA response, calculation of the relative response factor (RRF) for NDEA versus 15N2DEA, and absolute recovery studies. Estimates of precision, detection limit, and minimum quantifiable limit are also reported.

Animals↗

[Simultaneous determination of catecholamines, serotonin, and their precursors and metabolites in body fluid by an HPLC system with multi-electrode electrochemical detector].

We have developed two HPLC systems for the simultaneous determination of CA, 5-HT and their precursors and metabolites in human body fluid. One system consisted of two reversed-phase columns, a column switching device, a pair of electrodes for elimination of interferents and two sets of new electrochemical detectors with four electrodes. Using this system, adequate separation of peaks for 17 kinds of CA, 5-HT and their related metabolites in a standard solution required only 17 min. NE, MHPG, DOPAC, 5-HIAA, HVA and TRP levels in cerebrospinal fluid (CSF) were determined without any pretreatment of the CSF samples. The concentrations of DOPAC, 5-HIAA and HVA in CSF were significantly lower in Alzheimer's disease, vascular dementia and Parkinson's disease than in the controls. The other system was developed for determination of CA acidic metabolites and 5-HIAA in human urine. This system consisted of a mixed-mode column (C18/anion) and 8-channel electrochemical detector with isocratic elution of citrate buffer. Detection limits, precision and analytical recoveries of this method were satisfactory for clinical use.

Aged↗