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Reducing blood and body fluid exposures among clinical laboratory workers. Meeting the OSHA standards.

This article examines risks and exposure patterns in clinical laboratories to help identify the most effective methods for implementing the OSHA standard for protection against bloodborne pathogens, especially hepatitis B virus and the human immunodeficiency virus. Using eye-opening figures, tables, and statistics, the authors identify areas where clinical laboratorians can improve their protection and conclude by offering three strategies for reducing blood and body fluid exposures in clinical laboratories.

Acquired Immunodeficiency Syndrome↗

Determination of triazolam by GC-MS in two autopsy cases: distribution in body fluids and organs.

A detailed procedure for analysis of triazolam by GC-MS was constructed in our laboratory. At the concentration of 100 ng/ml, recoveries of triazolam in plasma and urine were 84.9 and 91.0%, respectively. The coefficients of variation in terms of its recovery were 11.5 (plasma) and 10.2% (urine). The detection limit for quantitation by the method was approximately 5 ng/g. This method was applied to two autopsy cases, giving triazolam distribution in body fluids and organs. In one case (33-year-old woman), concentrations of triazolam in the heart blood, urine, brain, lung, liver, kidney, skeletal muscles and stomach contents were 83.9, 741, 106, 165, 507, 293, 125 and 343 ng/g, respectively. From these toxicological data together with autopsy findings, her cause of death was diagnosed as triazolam poisoning. In the other case (45-year-old man), triazolam concentrations in the urine and stomach contents were 7.81 and 41.1 ng/g, respectively, but it could not be detected in the pleural blood; his cause of death was judged to be a traumatic shock, based on autopsy findings.

Journal Article↗

The relationship between peritoneal glucose absorption and body fluid loss by ultrafiltration during continuous ambulatory peritoneal dialysis.

Eight continuous ambulatory peritoneal dialysis (CAPD) patients were studied during six consecutive days using dialysate with a different glucose concentration on each day. For all dialysate glucose concentrations ranging from 70 to 198 mmol/l, an inverse linear relationship was found between the percentage of absorbed glucose and the ultrafiltration rate. In each patient a linear correlation was demonstrated between the dialysate glucose concentration and the quantity of body fluid removed by ultrafiltration. Therefore the dialysate glucose concentration without net removal of fluid could be calculated for each patient: DGCUF = 0. This DGCUF = 0 is a parameter of the relationship between the transport of water and glucose over the peritoneal membrane and can thus be considered a marker of the individual ultrafiltration. The calculated DGCUF = 0 was compared with DGCUF = 0 predicted from the glucose absorption using the equation as derived from our data: DGCUF = 0 (mmol/l) = 2.1 X glucose absorption (%) -67 (mmol/l). As marked day to day variations were found, the results of these predicted DGCUF = 0 should be interpreted cautiously when used for long-term follow-up of peritoneal permeability in CAPD patients.

Adult↗

The effect of a 14-day flight on body fluids in primates.

Using isotopes, two Bion 11 monkeys were examined 36 hours after flight. Total body water and all fluid compartments, particularly extracellular and interstitial fluid, were found to diminish. Blood volume and red blood cell mass also decreased. Forty-two days after flight, fluid compartments returned to normal. After the simulated flight, fluid compartments also tended to decrease but in a lesser degree than after real flight.

Animals↗

Measurement of recombinant interferon levels by high performance immunoaffinity chromatography in body fluids of cancer patients on interferon therapy.

The technique of high performance immunoaffinity chromatography was used to measure the levels of recombinant interferon in chronic lymphocytic leukaemia patients enrolled in a phase II recombinant interferon clinical trial. The technique employed a short high pressure chromatography column packed with minute glass beads which had monoclonal antibody, directed against recombinant alpha interferon, immobilized to their surface. This system was used to measure interferon levels in a variety of different human body fluids. A good correlation was found when interferon levels, detected by chromatographic separation, were compared to levels obtained by a conventional radioimmunoassay.

Adult↗

Effects of a 60-day confinement on the blood pressure, hormonal responses and body fluids of a mixed crew.

During the EXEMSI experiment, an international crew of 4 subjects (1 woman and 3 men) was confined for 60 days in a normobaric diving chamber (with 1060 mbar atmospheric pressure) to simulate life in a space station and to assess the effects of confinement on psychological and physiological factors. Blood pressure and blood volume regulating hormones (atrial natriuretic peptide, renin, aldosterone) and urine data (24-h urine outputs, ionogram) were measured before (BDC: baseline data collection), during (D: day) and after (R: recovery) confinement. We also measured energy expenditure and total body water, 14 days before, and after 27 days of confinement, by the double-labeled water method. We found a marked increase in 24-h urine output during most of the confinement in the men and the woman. Body weight (-1.8 +/- 0.9 kg) and energy expenditure (-1064 +/- 143 kcal/d, p<0.01) decreased in the 3 men. The total body water (TBW) decreased by 1.5 +/- 1.2 l in the men. Stress was not indicated by plasma and urine catecholamines but plasma growth hormone was elevated on D2 (p<0.01 vs. BDC) in the men. This study shows that confinement conditions can modify body fluid (increases in 24-h urine outputs and TBW changes) and energetic metabolisms.

Adaptation, Physiological↗

Determination of alpha-aminoadipic acid in brain, peripheral tissues, and body fluids using GC/MS with negative chemical ionization.

alpha-Aminoadipic acid (alphaAA) is a structural homolog of the excitatory amino acid glutamate and a natural product of lysine metabolism in mammalian cells. Under experimental conditions, alphaAA can influence various elements of glutamatergic neurotransmission. Moreover, as a selective inhibitor of kynurenine aminotransferase II, alphaAA is capable of decreasing the levels of the neuroinhibitory metabolite kynurenic acid in the brain. We now describe the identification of this potential endogenous neuromodulator in tissues and body fluids by gas chromatography/mass spectrometry (GC/MS) analysis of its pentafluorobenzyl (PFB) derivative. alphaAA was recovered from the GC column with a retention time of approximately 7 min. Subsequent MS analysis using electron capture with negative ionization revealed two separate ions for alphaAA (m/z 520, approximately 45% and m/z 322, approximately 55%). Both of these ions were positively identified with two different GC methodologies. In the rat, alphaAA levels ranged from 5 to 30 microM in various brain areas and from 8 to 40 microM in peripheral organs, whereas serum and urine contained only 1-2 microM alphaAA. Levels in the human brain were 18.7+/-2.4 microM (cortex) and 18.0+/-1.7 microM (striatum) alphaAA (n=9 each), and the mouse forebrain contained 8.3+/-1.9 microM alphaAA (n=6). Neuronal depletion, caused in rats by an intrastriatal injection of NMDA (300 nmol/2.5 microl), did not alter the striatal content of alphaAA, indicating that brain alphaAA resides at least in part in glial cells. alphaAA may therefore function as a glia-derived modulator of excitatory neurotransmission.

2-Aminoadipic Acid↗

A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater, shellfish, and mammalian body fluid.

We developed a competitive enzyme-linked immunosorbent assay (ELISA) to analyze brevetoxins, using goat anti-brevetoxin antibodies obtained after immunization with keyhole limpet hemocyanin-brevetoxin conjugates, in combination with a three-step signal amplification process. The procedure, which used secondary biotinylated antibodies, streptavidine-horseradish peroxidase conjugate, and chromogenic enzyme substrate, was useful in reducing nonspecific background signals commonly observed with complex matrices. This competitive ELISA detected brevetoxins in seawater, shellfish extract and homogenate, and mammalian body fluid such as urine and serum without pretreatment, dilution, or purification. We investigated the application of this technique for shellfish monitoring by spiking shellfish meat with brevetoxins and by analyzing oysters from two commercial shellfish beds in Florida that were exposed to a bloom of Karenia brevis (formerly Gymnodinium breve). We performed brevetoxin analysis of shellfish extracts and homogenates by ELISA and compared it with the mouse bioassay and receptor binding assay. The detection limit for brevetoxins in spiked oysters was 2.5 microg/100 g shellfish meat. This assay appears to be a useful tool for neurotoxic shellfish poisoning monitoring in shellfish and seawater, and for mammalian exposure diagnostics, and significantly reduces the time required for analyses.

Animals↗

Mannose in body fluids as an indicator of invasive candidiasis.

Using gas-liquid chromatography, we measured five mannose in the serum of six nondiabetic patients with autopsy-proven invasive candidiasis. In all patients serum mannose concentrations were higher than mannose levels found in serum from normal adults and children or from patients with catheter-associated candidemia, mucosal candidiasis, and other mycoses. Spinal fluid from two patients with Candida meningitis showed increased free mannose as compared to seven non-inflammatory spinal fluid samples. However, free mannose in the serum of poorly controlled diabetics (blood glucose of greater than or equal to 300 mg/dl) did overlap concentrations in patients with invasive candidiasis. In vitro culture of Candida albicans demonstrated increasing concentrations of mannose associated with growth of the organism. We conclude that physical and chemical assay for mannose in body fluids may be a useful technique to assist in the diagnosis in invasive candidiasis.

Adolescent↗

[Medical results from STS 1-4: analysis of body fluids].

Venous blood was drawn from the eight crewmembers of Space Shuttle flights STS-1 through STS-4 three times before lift-off and twice after landing, and the characteristics of biomedical blood components were evaluated. Twenty-four-hour urine pools were collected 30 days before flight and on landing day or day 4 after landing, and electrolytes, selected hormones and other components were measured. The results indicated that although fluid and electrolyte losses occur during space flight, conservation of these substances is begun almost immediately upon cessation of weightlessness. Enzyme and hormone measurements indicated that landing may have caused some stress on crewmembers.

Blood Chemical Analysis↗

In vitro differences of hydroxyapatite from different resources in simulated body fluid.

Hydroxyapatite (HA; Ca10(PO4)6(OH)2), is one of the significant implant materials used in Orthopaedics and Dental applications. However, synthetically produced HA may not be stable under ionic environment, which it will unavoidably encounter during its applications. In this paper, the in vitro effects of three HA materials derived from different resources, i.e. commercial HA (HAC), synthesised HA from pure chemicals (HAS) and synthesised HA from kapur sireh; derived traditionally from natural limestone (HAK), were studied. The HA disc samples were prepared and immersed in simulated body fluid (SBF) for 31-day period. The evaluation conducted focuses on the changes of the pH and the Calcium ion (Ca-ion) and Phosphate ion (P-ion) concentrations in the SBF solution, as well as the XRD and SEM data representing the reactions on the HA materials. From the XRD, it was found that HAK has the smallest crystallite sizes, which in turn affect the pH of the SBF during immersion. The Ca and P-ion concentrations generally decrease over time at different rates for different HA. Upon 1-day immersion in SBF, apatite growth was observed onto all three surfaces, which became more pronounced after 3-day immersion. However, the appetites formed were observed to be different in shapes and sizes. The reasons for the difference in the apatite-crystals and their subsequent effects on cells are still being investigated.

Bone Substitutes↗

Body fluid homeostasis in congestive heart failure and cirrhosis with ascites.

The urinary excretion of salt and water in man is regulated by a variety of renal and extrarenal mechanisms that respond to changes in dietary sodium intake as well as to alterations in the holding capacity of the vascular and interstitial compartments. Changes in extracellular fluid volume are detected by volume sensors located in the intrathoracic vascular bed, the kidney and other organs. These sensing mechanisms gauge the adequacy of intravascular volume relative to capacitance at various sites within the circulation. Congestive heart failure and cirrhosis with ascites are two disease states of man in which a hemodynamic disturbance within a given circulatory subcompartment is perceived by these sensing mechanisms and results in renal sodium retention. While the primary disturbance in both of these conditions originates outside the kidney, a variety of renal effector mechanisms respond to the perceived circulatory disturbance and result in enhanced tubule reabsorption of salt and water. These effector mechanisms involve physical adjustments in renal microvascular hemodynamics, tubule fluid composition and flow rate and transtubular ion gradients. These in turn are partially regulated by a variety of neural and humoral pathways including the renin-angiotensin-aldosterone axis, prostaglandins, and kinins.

Aldosterone↗

Evidence of gestational heroin exposure by comparative analysis of fetal and maternal body fluids, tissues, and hair in a heroin-related death.

In order to determine the fetal-maternal distribution of heroin and its main metabolites (6-monoacetylmorphine and morphine) the drug concentrations were measured in autopsy material. The heroin-related death of a pregnant drug abuser (approximately the 32nd week of gestation) provided the fetal and maternal material. Fetal and maternal hair was analyzed in order to obtain long-term information on the transplacental opiate transfer. Morphine and 6-monoacetylmorphine were detected in toxic concentrations in maternal as well as in fetal tissues and body fluids. The drug concentrations in the fetal blood were significantly lower than in the maternal blood. The ratio of fetal-to-maternal (F/M) blood morphine concentration was found to be 0.39, whereas the F/M ratio of 6-monoacetylmorphine in blood was 0.15. In fetal hair analysis, morphine, heroin, and, for the first time, 6-monoacetylmorphine were measured. The resulting F/M ratios were 0.49, 0.36, and 0.6, respectively.

Adult↗

HIV excretion patterns and specific antibody responses in body fluids.

HIV excretion patterns and specific antibody responses were evaluated in blood, semen, female genital secretions, saliva, and crevicular fluid. Samples were examined for infectious virus, viral antigens, viral nucleic acid, HIV specific IgG, IgA, anti-nef, and anti-p24. Viral load in peripheral blood appeared to increase with disease progression. The proportion of patients with antibody responses specific for nef and p24 was also lower among patients with more advanced disease. Infectious virus and viral antigens were detected infrequently and at lower levels in body fluids than in blood, which may reflect the presence of local antibodies. HIV nucleic acid was detected in some semen and saliva samples in the absence of infectious virus.

Female↗

Effects of lower body negative pressure (LBNP) on the distribution of body fluids.

Reduction of pressure on the lower part of the human body is believed to be capable of altering the normal fluid distribution. This could serve as a therapeutic method for emergency relief of the effects of hypertension, or it could be used as counterbalance for zero gravity and assist in preparing astronauts for reentry into the gravitational field. The present work describes a technique of assessment of the fluid shift caused by such pressure reduction. Thirty experiments were performed on four healthy subjects. The shift of fluid was assessed by measuring changes in body weight distribution of the tested subjects. The shift of fluid was assessed by measuring changes in body weight distribution of the tested subjects. The experimental results show that two processes may have taken place within the human body: shift of blood within the vascular system and diffusion of fluids from the circulatory system to the tissues. The total fluid shift ranges between 1 and 2 kg. It was proportional to the pressure gradient and exponential with time. The systolic blood pressure was reduced with the pressure reduction while the pulse rate increased.

Body Fluids↗

Analysis of LSD in human body fluids by high-performance liquid chromatography, fluorescence spectroscopy and radioimmunoassay.

A scheme of analysis is described in which the particular advantages of high-performance liquid chromatography (HPLC), fluorescence spectroscopy and radioimmunoassay (RIA) are exploited to the greatest effect. RIA affords a rapid and sensitive preliminary screening method, while the subsequent HPLC analysis using fluorimetric detection yields quantitative chromatographic evidence together with characteristic fluorescence spectra. Fractionation of samples by HPLC followed by RIA of the fractions gives further confirmation of the presence of LSD and its metabolites. The combined methodology has been applied to the analysis of LSD in body fluids for forensic and clinical purposes. Levels down to 0.5 ng of LSD per ml can be detected using the minimum of sample.

Chromatography, High Pressure Liquid↗