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Isoelectric heterogeneity of human prorenin (inactive renin) in body fluids.

Using flat bed isoelectric focusing, inactive renins from various body fluids were compared. Normal plasma inactive renin demonstrated six consistent peaks at pH 4.98, 5.14, 5.29, 5.47, 5.63 and 5.91, the largest being 5.29, 5.47 and 5.63. The isoelectric pattern of ovarian follicular fluid inactive renin was similar, with slight shifts at major peaks to a higher pH. Pregnancy plasma had inactive renin patterns like normal human plasma as did pleural fluid, lymphocoele fluid and plasma from anephric humans. Amniotic fluid inactive renin showed a markedly different pattern with major peaks at pH 5.00, 5.16 and 5.30. Fetal plasma (cord blood) also showed differences with only four peaks at 5.24, 5.40, 5.58 and 5.83. We conclude that human prorenin isoelectric patterns are similar for plasma, ovarian and most tissue fluid prorenins, but different for fetal plasma and amniotic fluid suggesting that these forms of renin do not cross the placental barrier.

Amniotic Fluid

Detection of Haemophilus influenzae type b antigens in body fluids, using specific antibody-coated staphylococci.

Protein A-rich staphylococci coated with Haemophilus influenzae type b antiserum agglutinate specifically with homologous bacterial cells or with cell-free supernatant fluids of cultures of the organism. Antibody-coated staphylococci were used to detect soluble antigens in body fluids of patients infected with H. influenzae type b. Cerebrospinal fluid from 36 cases of meningitis caused by this orgainsm showed positive coagglutination tests in 86% of patients prior to initiation of therapy. Antigens could be detected in 46% of sterile cerebrospinal fluid specimens obtained from the same cases 1 to 10 days after therapy. Soluble antigens were also detectable in sera (58%) and urine specimens (67%) of patients with H. influenzae type b septicemia, when such specimens were tested within 10 days of onset of illness. No antigen could be detected in body fluids beyond 10 days. The coagglutination test was positive in 57% of all body fluids examined; contercurrent immunoelectrophoresis (CCIE) was positive in only 27%. All specimens positive by CCIE were also positive by coagglutination. No false-positive reactions were noted by either test in body fluids from controls. The coagglutination test is simple, specific, and more sensitive than the CCIE method and could be a valuable tool for detecting antigens in body fluids of patients with various infections.

Agglutination Tests

Effect of staphylococcal enterotoxin B on body fluid compartments in conscious rhesus monkeys.

Techniques for measuring changes in body fluid compartments after SEB injection i.v. in conscious rhesus monkeys are described and compared with base line values measured by identical techniques in normal monkeys. Although few changes were observed in body fluid volumes 3 h after i.v. staphylococcal enterotoxin B (SEB; 1 mg/kg), the F cell ratio and all fluid compartments except for RBC volume were decreased significantly by 5 h after i.v. SEB administration. Rapid intracellular dehydration and decreases in plasma and blood volumes appear to play a role in the development of circulatory shock during enterotoxemia in monkeys.

Animals

The rapid determination of the ABO group from body fluids (or stains) by dot enzyme-linked immunosorbent assay (dot-ELISA) using enzyme-labeled monoclonal antibodies.

Using ABH enzyme-labeled monoclonal antibodies, the authors could rapidly detect the ABO group from body fluids and body fluid stains by the dot enzyme-linked immunosorbent assay (dot-ELISA). In this test, the antigen was immobilized on nitrocellulose paper; the entire piece of paper was coated with an appropriate dilution of enzyme-labeled McAb directly against the antigen of interest; and, finally, 3,3'-diaminobenzidine (DAB) substrate solution was added. The site of a positive reaction is clearly visible as a brown spot. We analyzed 521 samples and got satisfactory results. We also analyzed 99 practical case samples by this method and achieved the same results as those obtained by other researchers using other methods. This method is accurate, simple, direct, rapid, and sensitive; it also produces easily observed results, requires no equipment, and can be completed in 30 min. The test proved to be clearly more sensitive for the detection of the ABO blood group in secretor saliva than the conventional hemagglutination inhibition test. Also saliva diluted 10(-4) to 10(-5) and the ABO group of nonsecretor saliva and urine could be easily detected by this method.

ABO Blood-Group System

[Cardiothoracic ratio and roentgenologic heart size as the indices of body fluid retention in uremics under hemodialysis].

We evaluated the cardiothoracic ratio (CTR) and the roentgenologic heart size (RHS) as the indices of body fluid retention in 31 of uremic patients under hemodialysis. The maximum changes of the roentgenologic thoracic diameter, measured monthly for one year in 31 patients, varied from 10 to 68 mm. The average differences of RHS and CTR between maximum inspiration and forced inspiration in 18 healthy people were 1 mm (RHS) and 2.5% (CTR), respectively, and the change of roentgenologic thoracic diameter was 12 mm. The error of repeated CTR measurements in a period of one year could be estimated more than 2%. In 22 patients the body weight reduction of 1 kg corresponded to a 4 mm decrease of RHS (p less than 0.005) and 1.5% of CTR (p less than 0.05), respectively. In relation to the change of body fluid balance assessed by the change of body weight, RHS showed a higher correlation than CTR. These results could be attributed that RHS was hardly influenced by the respiration, whereas CTR was surprisingly affected. It is concluded that the measurement of RHS is more useful as an index of control of body fluid balance than CTR.

Adult

Heartworm migration toward right atrium following artificial pulmonary arterial embolism or injection of heartworm body fluid.

Heartworms harboring in the pulmonary arteries migrated toward the right atrium following insertion of dead heartworms or heartworm-like silicone tubes, or intravenous injection of body fluid extract of a female heartworm. The migration occurred within 3 hr (early group) or 1 to 7 days (late group) after insertion of dead worms, 1 to 11 days after insertion of silicone tubes, and immediately after infusion of heartworm-body fluid. The cardiac output decreased to an unmeasurably low level, and the pulmonary arterial pressure was also reduced in the early group. Although the output decreased, the pulmonary arterial pressure and the total pulmonary resistance increased gradually in the late group. In dogs with heartworm migration, in which silicone tubes had been inserted, the changes in cardiopulmonary values were the same as those in dogs of the late group. In dogs to which the body fluid had been administered intravenously, the changes in cardiopulmonary values were well accord with those in the early group. The systemic blood pressure also fell immediately after the administration with the shock-like state. These results suggest that the death of a part of the heartworms may be closely associated with the migration of heartworms toward the right atrium through the pulmonary arterial embolism and/or shock-like reaction by heartworm body fluid.

Animals

Inactivation of human immunodeficiency virus (HIV) by ionizing radiation in body fluids and serological evidence.

A method to use ionizing radiation to inactivate HIV (Human Immunodeficiency Virus) in human body fluids was studied in an effort to reduce the risk of accidental infection to forensic science laboratory workers. Experiments conducted indicate that an X-ray absorbed dose of 25 krad was required to completely inactivate HIV. This does not alter forensically important constituents such as enzymes and proteins in body fluids. This method of inactivation of HIV cannot be used on body fluids which will be subjected to deoxyribonucleic acid (DNA) typing.

Acquired Immunodeficiency Syndrome

The use of mass fragmentography for the detection of tetrodotoxin in human body fluids.

The authors report on a convenient electron impact ionization-mass fragmentographic method for determining the amount of tetrodotoxin (TTX) present in body fluids. To explain this method briefly, the TTX present in body fluids was purified with activated charcoal and transformed into 2-amino-6-hydroxymethyl-8-quinazoline (C9-base) with alcoholic KOH (1%). Then the C9-base was injected into a gas chromatograph-mass spectrometer in the form of trimethylsilyl derivative. In this manner the minimum detectable amount of TTX in body fluids was found to be about 0.01 microgram/g. When this method was applied to sera, urine, and the stomach contents of 2 decedents suspected of puffer fish poisoning, 0.053 microgram/g of TTX was detected in stomach contents of one victim. This new analytic method is simpler and more sensitive than other assay methods and thus is considered useful in forensic work.

Body Fluids

Body fluids and plasma atrial peptide after its chronic infusion in hypertensive rats.

To determine the role of body fluid volume in the chronic hypotensive effect of atrial natriuretic factor (ANF), spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats were infused with the peptide (Arg 101-Tyr 126) at a rate of 100 ng/h/rat for 5 days. Blood pressure (BP) was decreased from 176 +/- 4 to 133 +/- 3 mmHg in the SHR group 4 days after ANF infusion was initiated, whereas no changes were observed in ANF-infused WKY animals. Starting 5 days after the infusion began, body fluid measurements revealed no differences in plasma, blood and extracellular fluid volumes or in interstitial spaces. BP and plasma ANF concentrations were determined in another set of experiments before, during and after chronic ANF infusion. BP declined from 169 +/- 3 to 133 +/- 5 mmHg in SHR 5 days after the infusion commenced, but returned to basal values by day 10 or 11. Plasma ANF was significantly higher in SHR than in WKY rats throughout the observation period. However, there were no discernible changes in this parameter in ANF-infused SHR compared to non-infused SHR. A 3-fold rise in plasma ANF was noted in infused WKY rats at day 3 only. It is concluded that the chronic hypotensive effect of ANF in hypertensive animals is not related to changes in either body fluid volume or distribution. Moreover, the finding that chronic ANF infusion reduces BP in SHR without altering its plasma levels suggests a rapid ANF turnover.

Animals

Nucleotide pyrophosphatase and phosphodiesterase. I. Organ distribution and activities in body fluids.

We estimated nucleotide pyrophosphatase and phosphodiesterase I activities in human and rat organs and in body fluids from man and dog. The highest organ activities were found in epididymis, kidney, liver, and intestine. In body fluids, the activity was highest in seminal plasma, followed by intestinal lymph, serum, heart lymph, cerebrospinal fluid, milk, and urine. The ratio nucleotide pyrophosphatase/phosphodiesterase I and the urea resistance of phosphodiesterase I differed among human organs, body fluids, and blood cells. Different isoenzymes probably exist. The activities in serum share several properties with those in several organs--e.g. pH-optimum 9.6-9.8, dependency on Zn2+, and the effects of inhibitors. Phosphodiesterase I in erythrocytes, which has not been described previously, differs from enzyme from other sources by lower pH optimum (8.5), dependency on Mg2+, inhibition by Zn2+, and stimulation by dithiothreitol.

Adult

Effects of insulin on plasma renin activity, plasma atrial natriuretic peptide and body fluid volume in diabetes mellitus.

The effects of insulin treatment on plasma renin activity (PRA), plasma atrial natriuretic peptide (ANP) and body fluid volume were studied in 16 hospitalized patients with insulin-independent diabetes mellitus. Parameters were recorded for 2 days during treatment by diet alone and for 3 weeks after starting insulin. Blood samples were obtained weekly from 9 patients for the measurement of fasting plasma glucose, hematocrit, PRA and plasma ANP. A 24-hr urine sample was collected to determine the urinary excretion of glucose and sodium. In a separate group of 7 patients, plasma volume and extracellular fluid volume were determined by the Evans blue and sodium thiocyanate dilution tests, respectively. In the group of 9 diabetic patients, significant (p less than 0.05) reductions in fasting plasma glucose, hematocrit and the urinary excretion of sodium and glucose were seen with insulin treatment. PRA fell significantly (p less than 0.05) from 5.2 +/- 1.2 ng/ml/hr (mean +/- SEM) on the control days to 2.3 +/- 0.5 on the 21st day after starting treatment. Plasma levels of ANP averaged 35 +/- 5 pg/ml on the control days and these did not change significantly. In the other group of 7 patients, both plasma volume and extracellular fluid volume increased significantly (p less than 0.05) with insulin treatment. A sodium-retaining effect of insulin and a decrease in osmotic diuresis may have increased the body fluid volume and caused the fall in PRA. Thus, a vasodilatory action of insulin may assist in compensation for the increase in body fluid volume, preventing a rise in plasma ANP levels.

Atrial Natriuretic Factor

Increased monoamine turnover in the subfornical organ area following body fluid depletion.

To clarify whether monoaminergic inputs to the subfornical organ (SFO) area participate in fluid regulatory systems, we examined the effects of body fluid depletion on monoamine turnover in the region of the SFO using microdialysis techniques in rats. An iso-osmotic reduction of fluid volume following subcutaneous treatment with polyethylene glycol (PEG) significantly increased dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIAA) concentrations in the SFO area. 5-hydroxytryptamine (5-HT) in the SFO area could be detected after the PEG treatment, while 5-HT was undetectable before the treatment. The data imply that both dopaminergic and serotonergic systems in the SFO area may be involved in controlling body fluid balance.

Animals

Pleural effusion disease in rabbits. Interferon in body fluids and tissues after experimental infection.

The distribution of interferon in body fluids and tissues was studied in 18 rabbits infected experimentally with the agent of pleural effusion disease (PED). Circulating interferon of the classical type was demonstrable 12 h after inoculation, and a maximum response was attained 2-3 days later. Circulating interferon disappeared between 6 and 8 days after inoculation. Interferon titres of serum were closely correlated with the early phase of febrile response and probably also with the initial growth phase of the PED agent. The interferon titres of pleural fluid exceeded by far the titres of other body fluids and tissues. No interferon could be demonstrated in brain, liver and urine.

Animals

Mass fragmentographic determination of vanilmandelic acid, homovanillic acid and isohomovanillic acid in human body fluids.

Vanilmandelic, homovanillic and isohomovanillic acids in body fluids were efficiently isolated by liquid chromatography on an Amberlite XAD-4 column and by organic extraction in a special apparatus. The purified metabolites were converted into their trifluoroacetylhexafluroisopropanol esters and analyzed by mass fragmentography. The working curves of the metabolites were linear from 0.5 to 5 ng injected. The minimum detectable concentrations of all the metabolites were 2 ng/ml for plasma and cerebrospinal fluid, and 120 ng/ml for urine. The metabolite concentrations in plasma, cerebrospinal fluid and urine of normal persons and patients were determined.

Adolescent

[Ganglioside content and profile in the ovarian tumor tissues and in the biological body fluids of patients].

Lipid-bound sialic acid (LSA) content and ganglioside composition in resected tumours and body fluids of patients with benign ovarian tumours, borderline ovarian tumours, ovarian cancer and also in unaltered ovaries of patients with uterine cancer were examined. LSA levels in tissues and relation of the main gangliosides GD3/GM3 progressively decreased from unaltered ovaries to ovarian cancer. Distribution of gangliosides GD3 and GM3 in the borderline tumours was not uniform. Absolute content of GD3 increased more than twice in profuse growth from the inner surface and decreased almost three times in the cyst capsule as compared to the intact tumour. Ganglioside GD3 content decreased in malignant ovarian tumours but increased in ascitic fluid of cancer patients as compared to GM3. These results suggest that ganglioside GD3 is shed more intensively in the borderline ovarian tumours.

Ascitic Fluid

Changes in haemodynamics and body fluid volume due to enalapril in patients with essential hypertension on chronic diuretic therapy.

In 12 patients with essential hypertension who remained hypertensive despite chronic chlorthalidone treatment, the effect of 2 weeks of additional therapy with the converting enzyme inhibitor (CEI) enalapril on blood pressure and body fluid volumes has been evaluated. The objective was to examine the influence of a diuretic-stimulated renin-angiotensin-aldosterone system (RAAS) on haemodynamics and body fluid volume. Mean arterial pressure (MAP -21%), total peripheral resistance index (TPRI -22%) and plasma aldosterone concentration (PAC -39%) were decreased, and plasma renin activity (PRA 660%) was increased. The average heart rate (HR), cardiac index (CI), plasma volume (PV), blood volume (BV), extracellular fluid volume (ECFV) and body weight (BW) remained unchanged. A negative correlation was found between the per cent changes in ECFV and PAC. Thus, body fluid volumes during chronic diuretic treatment are well preserved even when the RAAS with its sodium retaining properties is suppressed by CEI. Possible mechanisms are a volume (not angiotensin II) - dependent stimulation of aldosterone and a fall in blood pressure.

Body Fluids

[Body fluids during 120-day anti-orthostatic hypokinesia].

Body fluid variations were examined during 120-day antiorthostatic (-5 degrees) hypokinesia in 21 test subjects, 9 of which comprised a control group (Group 1). The remaining 12 subjects formed three groups (four subjects each) who received drugs to normalize mineral and lipid metabolism (Group 2), or performed specially developed exercises (Group 3), or were on the combined treatment (Group 4). Total body water (TBW), intracellular fluid volume (IFV), extracellular fluid volume (EFV), and EFV composition were measured by nuclear physical methods. Measurements were taken prior to exposure, on head-down tilt days 1, 60 and 120, and on recovery day 15. Body composition and K40 content were investigated before and after head-down tilt. The controls showed body dehydration that began on tilt day 1 at the expense of vascular fluid and continued by day 60 at the expense of interstitial fluid and by day 120 at the expense of IFV. Group 2 subjects exhibited variations that were similar to those of controls. Group 3 subjects were in better shape due to the beneficial effect of exercise which diminished as head-down tilt continued. In the recovery period the above changes regressed.

Blood Volume

AUR Memorial Award 1992. Quantification of amino acids in human body fluids by 1H magnetic resonance spectroscopy. A specific test for the identification of abscess.

RATIONALE AND OBJECTIVES: When polymorphonucleocytes are incubated in proteinaceous fluid, they cause extensive protein degradation, which leads to accumulation of free amino acids. The authors tested whether these free amino acids, particularly valine and leucine, also accumulate in human abscess fluids, but not in other body fluids, and thus could be a specific and distinguishing marker for the presence of an abscess. METHODS: Thirty fluids, obtained by percutaneous drainage from 28 patients, were lyophilized and reconstituted in 2H2O before in vitro 1H magnetic resonance (MR) spectroscopy. Concentrations of valine and leucine were determined by comparison of spectra before and after addition of known amounts of valine and leucine. Two chart reviewers, blinded to the spectroscopic results, categorized cases as abscess (n = 14), non-abscess (n = 15), or infection but not abscess (n = 1). RESULTS: The concentration of valine and leucine was significantly higher in the abscess fluids, 2.57 +/- 1.90 mM than in the non-abscess fluids, 0.25 +/- 0.33 mM (P < .001). The one infected fluid which was not an abscess had no amino acids. Using 0.8 mM as the threshold concentration of valine and leucine necessary for the diagnosis of abscess resulted in a sensitivity rate of 86% and a specificity rate of 94%. CONCLUSION: The authors conclude that identification of high concentrations of valine and leucine by 1H MR spectroscopy may be a specific test for the diagnosis of abscess. This technique merits further investigation in vivo.

Abscess