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"Endogenous" benzodiazepine activity in body fluids of patients with hepatic encephalopathy.

Body fluids from patients with hepatic encephalopathy and from controls with no renal or hepatic disease were assayed for benzodiazepine immunoreactivity and benzodiazepine-receptor-binding activity. The subjects had taken no synthetic benzodiazepines for at least 3 months. Benzodiazepine receptor binding in cerebrospinal fluid was significantly higher in hepatic encephalopathy patients than in controls (210 [SE 50.2] vs 40.7 [7.3] oxazepam equivalents [ng/ml]). The severity of hepatic encephalopathy was directly and significantly correlated with the level of benzodiazepine activity by radioreceptor assay or radioimmunoassay in urine and in plasma. Benzodiazepine activity equivalent to levels of more than 900 ng/ml was found in patients with advanced encephalopathy. Although the chemical identity and source of this substance (or substances) are still unknown, its properties and the estimated levels of activity suggest it may have a role in the pathogenesis of the neural inhibition seen in hepatic encephalopathy.

Benzodiazepines

The measurement of endorphins in body fluids.

The measurement of endorphins in body fluids has been an important advance in clinical research attempting to link the endogenous opioid system to psychiatric illness and symptomatology. The consideration of methodologic differences in assay technique and in clinical methods is important in evaluating results of studies. Whereas findings in early clinical studies supported the notion of increased endorphin system function in patients with schizophrenia, cumulative data from the considerable number of studies carried out throughout world centers have been unable to demonstrate a consistent abnormality in levels of endorphins in CSF or plasma of patients with schizophrenia. Among the affective disorders, data suggest the possibility of relative changes in levels of opioids within individual manic-depressive patients when studied across state change from depression to mania. In studies of depressive illness there is accumulating evidence that the endogenous opioid system may relate or contribute to abnormality of the HPA axis. In our work measuring opioids in CSF we have observed relationships between anxiety and CSF opioids in normals and psychiatric patients and changes in CSF opioid activity in patients with anorexia nervosa accompanying weight change. These data are consistent with other evidence linking endorphins to CNS noradrenergic systems and to biologic response to stress.

Anorexia Nervosa

Specific capture of ABH blood group antigens of the red cell or body fluids by double antibody sandwich-ELISA.

A double antibody sandwich-ELISA method for the detection of the ABH blood group of each constituent of mixed stains is described. Extracts from mixed stains were applied to microtitration plates coated with rabbit polyclonal antisera to red cells or body fluids. ABH antigens in body fluid stains which were captured by the polyclonal antibodies were detected by monoclonal anti-A and -B and enzyme-conjugated anti-mouse immunoglobulin. By this procedure, ABH antigens of only saliva, semen or red cells could be detected from mixed stains, but no ABH antigen capture activity was observed using anti-sweat, -milk, -vaginal secretion, -erythrocyte membrane and -band 3 antibodies.

ABO Blood-Group System

Comparison of Papanicolaou's and Wright-Giemsa stains in the examination of body fluids for Hodgkin's disease.

We reviewed 36 body fluid specimens from 18 patients with Hodgkin's disease (HD) to characterize the cytologic features of HD as seen in Wright-Giemsa (WG)-stained cytocentrifuge preparations, and to compare diagnostic agreement between WG- and Papanicolaou-stained samples. Slides were examined independently by two pathologists without knowledge of the original diagnosis, and were classified as either positive, inconclusive, or negative for malignant cells. There was diagnostic agreement between both methods in 35 (97%) of 36 samples. Features in cytocentrifuged WG-stained specimens that were most helpful in recognizing HD included mirror image nuclei in typical Reed-Sternberg cells and an axis of symmetry in polylobate Reed-Sternberg variants, with even distribution of the nuclear material within the cytoplasm.

Adult

A possible physiological role of atrial natriuretic peptide in body fluid volume regulation.

To study whether or not atrial natriuretic peptide (ANP) is physiologically involved in body fluid volume regulation, we examined the relationship between plasma ANP level and renal function during NaCl loading and ANP infusion. In study I, six normotensives (NTs) and seven hypertensives (HTs) were placed on 7-day low (3 g/day) and then 7-day high NaCl diets (20 g/day). The plasma ANP level increased by 60% (p less than 0.01) on the high NaCl diet. Although the plasma ANP level was higher in HTs than in NTs, the changes in plasma ANP due to NaCl loading were similar between the two groups. Furthermore, increases in urinary Na excretion due to ANP infusion at 25 ng/kg/min were greater for the high NaCl diet than for the low NaCl diet (p less than 0.02). In study II, graded doses of ANP were infusion into 16 other HTs and Nts on an 8 g/day NaCl diet. ANP infusion at 2.5 ng/kg/min increased the plasma levels of ANP by 80% (p less than 0.001). Such increments were associated with an increase in urinary Na excretion by 25% (p less than 0.02) in both HTs and NTs. This rise in plasma ANP was comparable to that induced by high NaCl intake. Thus, a slight increase in plasma ANP induced by dietary Na loading seems to augment renal Na excretion, suggesting that ANP may play a physiological role in body fluid volume regulation.

Adult

Elevation of the panting threshold of the desert iguana, Dipsosaurus dorsalis, during dehydration: potential roles of changes in plasma osmolality and body fluid volume.

Dehydration of the desert iguana, Dipsosaurus dorsalis, resulted in a progressive elevation in the magnitude of the skin temperature necessary to elicit thermal panting (i.e., the panting threshold). Panting threshold increased from 43.4 +/- 0.8 degrees C at 100% initial body weight (IBW) to 45.4 +/- 1.2 degrees C at 90% IBW to 45.7 +/- 0.9 degrees C at 80% IBW. Plasma osmolality showed no significant change with dehydration to 80% IBW. Changes in plasma osmolality, whether induced by NaCl or non-ionic sucrose loading, had a significant impact on panting threshold. Increasing plasma osmolality resulted in an elevation of panting threshold while decreasing plasma osmolality resulted in lower panting thresholds. Decreasing body fluid volume by exsanguination of 1 ml whole blood/100 g body weight resulted in a mean increase in panting threshold by 0.7 +/- 0.2 degrees C. Volume loading with 160 mM NaCl (approximately isosmotic) had no significant effect on panting threshold. These data suggest that plasma osmolality and decreases in body fluid volume may be potent modulators of panting threshold during periods of water deprivation. However, at least in desert iguanas, increases in plasma osmolality would not appear to be an important factor in the elevation of panting threshold during dehydration to 80% IBW.

Animals

ABO and Lewis typing of semen, saliva and other body fluids.

It is well known that ABH group specific substances are usually present in high concentrations in body fluids of secretors. In normal circumstances these substances can withstand drying and retain their antigenic activity over a prolonged period. This enables the forensic serologist to assist in the investigation of various crimes by grouping stains of body fluids such as semen and saliva. It is possible, for instance, to group saliva and lip mucosa stains on cigarette ends, gags, masks, postage stamps and envelope flaps etc. but it is the grouping of seminal stains in the investigation of sexual crimes which predominates. The results of such tests can be extremely valuable in either including or excluding suspects.

ABO Blood-Group System

Body fluid compartments in rabbits on exposure to acute hypobaric hypoxia.

Body fluid compartments were studied in rabbits divided into three groups--control, exposed to acute hypoxia, and exposed to hypoxia after treatment with 2 mg frusemide intramuscularly. Total body water, extracellular body water, and plasma space were determined using the triple radiotracer technique. Total body water decreased insignificantly with no change in extracellular body space on exposure to hypoxia. Plasma volume and blood volume showed a significant decrease with a significant increase in haematocrit. In rabbits pretreated with frusemide, total body water, extracellular body water, plasma volume, blood volume, and interstitial fluid space decreased significantly on hypoxic exposure. This study suggested hypohydration on acute hypoxic exposure with a loss of intracellular water, while pretreatment with frusemide resulted in further hypohydration with a loss from both intracellular and extracellular compartments. The results have been discussed in relation to suggested use of diuretics on induction to high altitude.

Altitude Sickness

Maternal ingested methadone, body fluid methadone, and the neonatal withdrawal syndrome.

The relationship between the quantity of methadone ingested by the pregnant mother, the quantity of methadone in maternal and neonatal body fluids, and the subsequent neonatal withdrawal course was studied. The severity of the neonatal withdrawal syndrome was found to be related to the total dose of methadone ingested by the mother during the last 12 weeks of pregnancy (p less than 0.02), the maternal dose of methadone at delivery (p less than 0.01), and the intrapartum serum methadone levels (p less than 0.01). The cord blood levels of methadone were consistently lower than the maternal serum levels. Amniotic fluid methadone levels were not constantly related to maternal or neonatal serum methadone levels. Concentrations of methadone 10 to 60 times greater than that of cord blood were found in neonatal urine.

Amniotic Fluid

Body fluid variations and endogenous digitalis-like compounds during chronic NaCl loading in Wistar rats.

Circulating digitalis-like compounds have been proposed to be raised in volume expanded hypertension and to participate in Na+ homeostasis. We have investigated the temporal relationships between the activity of these circulating digitalis-like compounds, blood pressure and body fluid volume variations during a chronic NaCl load in the Wistar rat. Characteristics of salt-loaded rats were compared to those of weight-matched controls. At one week, when extracellular fluid volume (ECFV) was elevated, the capacity of plasma extracts to inhibit the Na+K+ATPase activity begun to rise. At two weeks, ECFV remained elevated, and plasma volume, blood pressure and the activity of plasma digitalis-like compounds increased. After 13 weeks, the continuous rise in plasma digitalis-like activity and in blood pressure was accompanied by the return of body fluid volumes towards control values. These changes in plasma digitalis-like activity, body fluid volumes, and systolic blood pressure during a high NaCl diet are compatible with the proposed role of circulating digitalis-like compounds as natriuretic and hypertensive factors.

Animals

Body fluid analysis of 1,3-diphenylguanidine for mutagenicity as detected by Salmonella strains.

The toxicity and mutagenicity of 1,3-diphenylguanidine (DPG) were monitored in Salmonella bacteria using the direct incorporation protocol. The test consisted of either direct incorporation of DPG or analysis of body fluid and faecal material derived from animals exposed to DPG. The data from direct incorporation of DPG suggested that the compound is a direct acting mutagen and that in the presence of metabolic activation system there was a reduction in the number of histidine revertants. The data further showed that while higher dosage levels of DPG without metabolic activation were extremely toxic and thus generated significantly fewer revertants, similar concentrations proved to be moderately mutagenic in the presence of S-9 mix. From the data on mutagenic activity of body fluids and faeces, it is suggested that (1) the rate of excretion of i.p.-administered DPG into extracellular fluids, urinary and gastrointestinal tracts is concentration dependent, and (2) a greater proportion of the compound is eliminated through the urinary tract.

Animals

Vascular responsiveness and body fluid status in patients on chronic hemodialysis.

The vascular responsiveness and the body fluid distribution were studied in 5 hypotensive and 11 normotensive patients on long-term hemodialysis. A diminished vascular responsiveness to intravenously infused norepinephrine or angiotensin II was found in uremic patients, especially in the hypotensive group. A slightly larger, but non-significant circulating blood volume (CBV) and a less extracellular fluid volume (ECF) was found in the hypotensive group as compared with those of the normotensive group. These results suggest that with autonomic nervous dysfunction, such a diminution of vascular responsiveness may be an another factor responsible for the development of hypotension.

Adult

p53 immunostaining as a marker of malignancy in cytologic preparations of body fluids.

The accurate identification of suspicious cells in cytologic preparations is a common problem in diagnostic cytopathology. Recent studies have shown that mutation of the p53 gene may be the most common genetic event in human malignancy. Mutation leads to altered conformation and increased half-life of the p53 protein, resulting in detectability by immunocytochemistry. The usefulness of p53 immunocytochemical staining as a marker of malignancy in the cytologic analysis of body fluids was investigated in the present study. One hundred fifty-four serial samples of body fluids submitted for cytologic diagnosis were also examined for p53 immunoreactivity. Of 121 cases reported as cytologically benign, 3 (2.5%) stained positively for p53; 16 samples were cytologically malignant, and 7 (43.7%) of these were positive for p53 (P < .001). Of those reported as suspicious but not conclusively malignant, 4 of 17 (23.5%) showed p53 immunoreactivity. On review, two of the three patients whose samples were benign cytologically yet showed positive p53 staining had histologic evidence of malignancy. The third patient died without a postmortem examination. Of the 17 cytologically suspicious cases, 16 (94.1%) were later proven to be malignant, and p53 was positive in 4 (25%). These results suggest that p53 immunostaining could be of value as a marker of malignancy in the cytologic examination of body fluids. The presence of p53 immunoreactivity in cytologic samples is strongly suggestive of malignancy, though its absence does not exclude neoplasia.

Body Fluids

An improved method for quantification of extra domain A-containing cellular fibronectin (EDAcFN) in different body fluids.

A quantitative direct enzyme immunoassay for the extra domain A-containing isoform of cellular fibronectin (EDAcFN) was established for screening of large series of blood samples and various body fluids of different pH and viscosity. The method is based on the monoclonal antibody DH1 recognizing the extra domain A in cellular fibronectin (EDAcFN). Studies on the effect of dilution of plasma and serum samples in this direct assay indicated that the measured concentration of cFN in the samples greatly depend on the ratio of sample dilution. The linearity of the assay was improved with sample dilution and the optimal dilution was 1:5. Stored diluted samples retained their cFN content at +4 degrees C, and -20 degrees C and -70 degrees C for months in contrast to samples stored undiluted. With this direct EIA the detection limit was 0.05 micrograms/ml and the linear portion of the standard curve could be extended above 30 micrograms/ml. Thus, the cFN concentration of blood samples could be measured reliably without inhibition also in samples with very high concentration of cFN. This is particularly important when measuring blood samples from cancer patients, since these samples may contain more than 20 micrograms/ml EDAcFN. The assay was standardized for blood samples but, due to the possibility of sample dilution, it also enabled reliable quantification of EDAcFN in various other body fluids. Undiluted some of the samples with non-neutral pH (urine, bile) or with high viscosity (seminal plasma) interfered with the assay. In addition to blood samples, the EDAcFN concentration was determined in samples of urine, bile, amniotic fluid, cervicovaginal secretions, seminal fluid, cerebrospinal fluid, bronchoalveolar lavage fluid, pleural fluid and saliva. Thereby, this modified method was shown to be applicable to various body fluids.

Amniotic Fluid

The periventricular anteroventral third ventricle (AV3V): its relationship with the subfornical organ and neural systems involved in maintaining body fluid homeostasis.

The periventricular tissue surrounding the anteroventral third ventricle (AV3V) is critically involved in the maintenance of normal body fluid balance and distribution. The present review examines the anatomical, neurochemical, and functional relationship of the AV3V with neural systems subserving body fluid homeostasis. In particular, the nature of AV3V afferents from the subfornical organ (SFO) and from brainstem noradrenergic cell groups is discussed. A model is presented proposing that specific structures within the AV3V, particularly along the ventral lamina terminalis, function to integrate information derived from blood-borne angiotensin II (via the SFO) with input arising from vascular pressure/volume receptors. The resultant of this integration is important for the generation of a normal component of thirst (i.e., drinking) associated with extracellular dehydration.

Afferent Pathways

Body fluid composition in chronic renal failure.

Studies were performed to assess body fluid composition alterations occurring in 10 patients undergoing chronic hemodialysis. Red cell mass (RCM), plasma volume (PV), extracellular fluid volume (ECF), and total body water (TBW) were assessed 24 hours following hemodialysis to dry weight, following infusion of normal saline to increase body weight by approximately 2 kg, and following hemodialysis to decrease body weight by approximately 2 kg. Results demonstrate that TBW constituted from 48.5 to 51.0% of body weight. The ECF constituted from 41.8 to 46.3% of the total body water. The ratio of PV to interstitial fluid (ISF) volume (ISF = ECF - PV) approximated 1.1 to 1.2:3. Salt and water alterations (isotonic saline infusion and ultrafiltration) were restricted to the ECF component of the TBW. In the ECF, it was principally the ISF that buffered salt and water expansion or depletion. There was no evidence for volume alterations in the intracellular fluid (ICF = TBW - ECF). We conclude that the ISF is the buffer zone which maintains the proper balance and relationship between vascular capacity and volume.

Adult

Effects of nonsteroidal anti-inflammatory drugs (NSAID) on renal excretion of sodium and water, and on body fluid volume in rats.

Effects of nonsteroidal anti-inflammatory drugs (NSAID) on urine volume and urinary sodium excretion, and on plasma volume and extracellular fluid volume were examined in conscious rats. The basal urine volume and urinary sodium excretion were decreased and the increased urine volume and urinary sodium excretion elicited by saline load (25 ml/kg) and by hydrochlorothiazide (10 mg/kg) were inhibited after oral administration of NSAID in doses which inhibited the rat carrageenin-induced hind paw edema (indomethacin, 1--10 mg/kg; tolmetin, 3--30 mg/kg; phenylbutazone, 3--30 mg/kg; aspirin, 30--300 mg/kg), but aminopyrine (30--300 mg/kg) did not show such an effect. The inhibitory activity on renal function was diminished gradually with repeated administration of NSAID. NSAID (indomethacin, 3 mg/kg; tolmetin, 10 mg/kg; phenoxybenzamin, 10 mg/kg; aspirin, 100 mg/kg) increased plasma volume and extracellular fluid volume of rats after repeated medication for 3 or 5 days, but the body fluid volume expansion disappeared with further repeated administration of NSAID. These results suggest that NSAID may inhibit the intrarenal role of prostaglandins and decrease sodium and water excretion in urine with resulting increased body fluid volume. Tolerance to these actions of NSAID developed after repeated administration.

Animals

Concentration of placental protein 19 in body fluid and placental tissues.

Placental protein 19 (PP19) is one of the new placental tissue proteins identified in extracts from human term placenta by Bohn and Winkler. We measured the PP19 concentration in body fluids and placental tissue by radioimmunoassay; the minimum detectable dose of standard was 1.5 ng/ml. Although ethylene diamine tetraacetic acid (EDTA-2K) inhibited the immunoreaction between PP19 (225/242) and anti-PP19 antibody (632 ZA), the PP19 concentration did not differ between serum and heparin and sodium citrate plasmas. The serum PP19 concentration was increased by hemolysis. In blood cell fractions separated by the Ficoll-Paque/Macrodex method, polymorphonuclear leukocyte fraction contained the highest PP19 concentration. The circulating serum PP19 concentration was 4.5 +/- 1.1 ng/ml (mean +/- standard deviation) in the proliferative phase (n = 8) and 5.1 +/- 1.6 ng/ml in the secretory phase (n = 7) for nonpregnant women, and 4.6 +/- 2.2 ng/ml from men (n = 12). Seminal plasma (n = 8) contained 212.2 +/- 99.7 ng/ml. The maternal serum PP19 concentration in 291 normal pregnancies increased from 6.2 ng/ml (median) at 6-7 weeks of gestation to 34.1 ng/ml at 38-39 weeks. The mean PP19 concentration was higher in amniotic fluid and retroplacental blood, but lower in umbilical cord blood than that in circulating maternal serum. In hydatidiform mole, vesicular fluid contained high PP19 concentration (1154.6 +/- 659.5 ng/ml), although these maternal serum concentration was not statistically higher than normal range. The chorionic villous trophoblast contained more PP19 than decidua, chorion, and amnion. These results suggest that PP19 has an extraplacental source, even though the chorionic villous trophoblast may be the main source throughout pregnancy.

Adult