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PubMed1975

Removal by dialysis of methylguanidine from body fluids.

The dialysis clearance of methylguanidine (MG) was found to be lower 'in vivo' than 'in vitro' and to decrease during a haemodialysis. Its protein binding, which rises as the plasma pH rises during haemodialysis, accounts for its dialysis behaviour. The low dialysis clearance of MG 'in vivo' explains why the post dialysis percentage decrease in its plasma levels is lower than that of urea (U) and of creatinine (CR). The slow transfer of MG from tissue during dialysis, shown by direct measurements on plasma and muscle tissue of anuric dogs, accounts for its high plasma rebound level after haemodialysis. Rebounds after peritoneal dialysis were lower and the plasma MG levels 12 hr after the termination of the two dialysis procedures were not different. The dialytic behaviour of MG is different from that of U and of CR which have similar molecular weights and the conclusions drawn from the behaviour of the latter two metabolites cannot be applied to MG. On the contrary, the removal of MG from body fluids of uraemic patients by dialysis is equal with peritoneal dialysis and with the various haemodialysis schedules that ensure good clinical results, while leaving plasma U and CR concentrations in a high range.

Animals↗
PubMed1979

Post-proline cleaving enzyme. Synthesis of a new fluorogenic substrate and distribution of the endopeptidase in rat tissues and body fluids of man.

Synthesis and application of the first fluorogenic substrate, N-carbobenzoxyglycylprolyl-4-methylcoumarinyl amide (Z-Gly-Pro-MeCouNH) for the determination of the post-proline cleaving enzyme (EC 3.4.21.-) were reported. Maximal activity of the enzyme purified from lamb kidney for the new substrate was observed at pH 7.0. This substrate showed a higher affinity (Km = 0.02 mM) for the enzyme than the proline containing substrates studied previously and allowed the detection of 10-50 ng post-proline cleaving enzyme activity per ml sample after a 1 min incubation period. Distribution of post-proline cleaving enzyme and other proline specific peptidases in rat tissues was studied using Z-Gly-Pro-MeCouNH and other proline-containing substrates. High post-proline cleaving enzyme activity was observed in testis, liver and skeletal muscle. Inhibition experiments indicated that post-proline cleaving enzyme activity was completely inactivated by 0.1 mM diisopropylphosphofluoridate and Z-Gly-Pro-chloromethylketone, as had been found in the case of the enzyme isolated from lamb kidney. Activity in human body fluids was also tested for levels of post-proline cleaving enzyme activity using Z-Gly-Pro-MeCouNH and semen was found to show the highest cleaving activity.

Aminohydrolases↗
PubMed1976

Acclimatization in a hot, humid environment: body fluid adjustments.

Four trained men worked 4 h/day at 40-50% of their maximum aerobic capacity first for 3 days at 25 degrees C db, 18 degrees C wb and then for 10 consecutive days at 45 degrees C db, 32 degrees C wb. Between days 1 and 2 of heat exposure mean total circulating protein (TCP) and plasma volume (PV) increased 11.6% and 9%, respectively. Preexposure TCP and PV increased until day 6 of heat exposure. Of the protein fractions beta-globulins underwent the largest relative increase. During work movement of protein into and out of the vascular compartment was similar in control and acclimatizing subjects but the latter generally maintained a greater amount of protein and fluid within the vascular volume. There was no evidence of salt and water retention. The increase in vascualr volume was ascribed to transfer of interstitial protein and water to the vascular volume. Regression coefficients indicated significant correlations for changes in plasma volume versus heart rate, stroke volume, and cardiac output during acclimatization. It was concluded that the most critical event in heat acclimatization is the expansion of the plasma volume.

Acclimatization↗
PubMed1979

Immunoassay of the adenosine deaminase complexing proteins of human tissues and body fluids.

A sensitive immunoassay for the adenosine deaminase binding protein (complexing protein) of human kidney has been developed. Impetus for the development of the assay was provided by the observations that (a) antibody to complexing protein does not react with the catalytically active adenosine deaminase monomer, and (b) binding of antibody to complexing protein does not affect the binding or catalytic activity of the enzyme monomer. Preformed immune precipitate prepared from rabbit anti-kidney complexing protein serum and goat anti-rabbit gamma-globulin serum is used to selectively insolubilize complexing protein. Quantitation is accomplished by measuring the intrinsic adenosine deaminating activity or adenosine deaminase binding capacity of the protein held in the immune precipitate. As little as 1 ng of kidney complexing protein can be accurately quantitated with the assay. The assay was used to demonstrate that complexing proteins from liver, lung, spleen, fibroblasts, plasma, and urine react with antibody to kidney complexing protein. The shared capacity to bind adenosine deaminase coupled with their antigenic similarity suggests that the complexing proteins of a number of human tissues and body fluids may be products of the same gene.

Adenosine Deaminase↗
PubMed1992

Nursing sickness in lactating mink (Mustela vison). II. Pathophysiology and changes in body fluid composition.

An investigation of the pathophysiological characteristics of nursing sickness in mink was carried out as a follow-up study of a previous epidemiological survey at a Danish fur research farm during the 1989 breeding season. In a total of 48 nursing females of the Standard Black and Pastel type, concentrations of several pertinent biochemical constituents of whole blood, plasma, urine and skeletal muscle were determined in order to identify nutritional and metabolic factors involved in the origin and development of the disease. Compared to the reference data obtained in 17 apparently normal lactating dams the findings in 31 females suffering from nursing sickness presented varying clinical and biochemical signs of progressive dehydration and emaciation: aldosteronism, hypovolemia, hyponatremia, hyperkalemia (in the face of muscle potassium depletion), hyperglycemia and azotemic acidemia. Neither ketosis nor severe lactacidemia was observed. The urine was almost devoid of sodium and chloride, and urinary potassium concentration diminished by approximately 50%. The concentrating ability of the kidneys was reduced to less than one third of the maximum value. The results were consistent with severe dehydration and emaciation due to heavy losses of energy, water and body mass along with increasing milk production. The progressive nature of the disease supported the hypothesis that nursing sickness is due to the combined effects of heavy milk production and excessive tissue catabolism along with reduced or ceased dietary intake, and maybe increasing environmental stress. In the advanced stage of the disease coma and death appear to be the inevitable outcome of the metabolic strains for continuing milk production.

Acid-Base Equilibrium↗
PubMed1991

[A HPLC method for determining piroxicam in body fluids].

Minimal detectable concentration of serum piroxicam by using HPLC reported in literature was mostly around 50 ng.ml-1 serum. Though the sensitivity was enough for pharmacokinetics study, it could not meet the needs of drug formulation screen study in developing precision drug delivery system (DDS) such as transdermal delivery system. A new HPLC method providing a detection limit of 0.75 ng, sensitive enough to quantify low concentrations of serum piroxicam down to 5 ng.ml-1 was reported in this paper. A Waters Model 481 instrument was used throughout the experiment. Isoxicam was proved to be the most suitable internal standard at maximum absorption wave length of 360 nm. A mixture of methanol and ammonium acetate 0.1 mol.L-1 (1:0.9 vol.vol-1) was selected as mobile phase with a flow rate of 1 ml.min-1. 0.025% tetramethyl ethylene diamine was added to ammonium acetate solution and adjusted to pH 4.5 with citric acid before mixing. Calibration curve was linear (r = 0.9999) in the concentration range of 10-5,000 ng.ml-1. The within-day and day-to-day precisions (CV) of this method were 2.88% and 2.89% respectively, with average recoveries of 96.0-102.4% (10-5,000 ng.ml-1). No interference was found in the body fluids of subjects who took piroxicam concomitantly with other commonly used non-steroidal anti-inflammatory medicines such as indomethacin, ibuprofen, naproxen, phenylbutazone and acetylsalicylic acid.

Animals↗
PubMed1990

Thermoregulation and body fluids: role of blood volume and central venous pressure.

Dehydration due to hyperthermia induces both hyperosmolality and hypovolemia. Hyperosmolality reduces evaporative cooling, and alters the thermal responsiveness of the hypothalamic center to changes in both the central and peripheral milieu. Hypovolemia also reduces the thermoregulatory response, but its effect is more variable. The potential sensor of hypovolemia is the CVP, which is influenced by redistribution of blood volume, changes in blood volume, and alterations in cardiac function. The control of CVP is related to the regulation of vascular compliance, stressed blood volume, and unstressed blood volume. Vascular compliance is also involved in the regulation of fluid shifts between the ISF and the intravascular fluid space, and buffers changes in the CVP. Regulation of fluid replacement after thermal dehydration can be considered both from the point of view of osmoregulation and volume regulation. In the rehydration process, control of plasma osmolality precedes blood volume regulation, which also suggests that changes in blood volume sensed as changes in the CVP are regulated within a narrow range by various mechanisms. These findings suggest a hierarchic structure for the homeostatic mechanisms related to thermoregulation, with higher priority being given to the maintenance of cardiac output and the cellular volume of the brain at the expense of peripheral circulation and cell volume.

Animals↗
PubMed1986

George E. Brown memorial lecture. Role of atrial peptides in body fluid homeostasis.

Extracts of mammalian atria, but not ventricles, induce marked diuresis, natriuresis, and reduction in blood pressure when infused systemically in rats and dogs. These extracts also inhibit aldosterone biosynthesis and renal renin release. Natriuretic peptides, 21 amino acids and longer, have been isolated from atria of rodents and man, and share a nearly homologous amino acid sequence at the carboxyterminus. Natriuretic activity resides in a 17-amino acid ring formed by a disulfide bridge, and the C-terminal Phe-Arg appears necessary for full biological potency. The deoxyribonucleic acid-encoding atrial natriuretic peptides have been cloned and the gene structure elucidated. Reduction of the diuretic and natriuretic responses to an acute volume load by right atrial appendectomy first suggested a role for atrial peptides in the physiological response to plasma volume expansion. Subsequently, release of peptides with natriuretic and spasmolytic properties from isolated heart preparations in response to right atrial distension was demonstrated by bioassay and radioimmunoassay. The presence of these peptides in normal rat and human plasma in concentrations of 20-100 pM, and the findings of increased levels in response to acute and chronic plasma volume expansion, rapid atrial tachyarrhythmias, systemic hypertension, congestive heart failure, and renal insufficiency imply that they play an important role in body fluid homeostasis. The mechanisms by which atrial peptides increase renal salt and water excretion are as yet unclear. Renal vascular effects have been consistently demonstrated, and limited evidence for direct actions on tubule ion transport has also been reported recently. In vitro, these peptides cause precontracted vascular and nonvascular smooth muscle to relax, mediated by a direct action on smooth muscle cells. Specific receptors for these peptides have been characterized in crude membranes prepared from whole kidney homogenates and adrenal glomerulosa cells, in intact glomeruli and cultured glomerular mesangial cells, and in intact bovine aortic smooth muscle and endothelial cells. Natriuretic peptides stimulate cyclic guanosine monophosphate accumulation in target tissues, and augment particulate guanylate cyclase activity in membrane fractions, suggesting that cyclic guanosine monophosphate is the second messenger mediating their cellular action.

Aldosterone↗
PubMed1980

Body fluid volumes and the response of renin and aldosterone to short- and long-term thiazide therapy of essential hypertension.

Plasma volume (PV), extracellular fluid volume (ECV) serum electrolytes, renin and aldosterone were measured before and after 1 week and 4 months of hydrochlorothiazide (HCT) treatment, 50 mg twice daily, in nine male patients with uncomplicated essential hypertension. All studies were carried out under strictly standardized conditions in a metabolic ward. After 1 week of HCT treatment, significant reductions were found in PV and ECV, but after 4 months only ECV was significantly reduced. During HCT therapy, renin and aldosterone were permanently elevated whereas serum sodium and potassium were lowered. After 1 week, renin was inversely correlated with PV and ECV and directly correlated with heart rate. After 4 months, renin was inversely correlated with serum sodium. These results indicate a permanent decrease in ECV during long-term HCT therapy. It is further suggested that the mechanisms responsible for the renin response during short- and long-term HCT treatment are different, changes in body fluid volumes and increased neural activity being responsible for the initial rise in renin, and serum sodium being the predominant factor during chronic treatment.

Adult↗
PubMed1980

Use of monoclonal antibodies to detect antigens of Toxoplasma gondii in serum and other body fluids.

Monoclonal antibodies to Toxoplasma gondii were used in an enzyme-linked immunosorbent assay to detect antigens of the parasite in toxoplasma lysate, in peritoneal fluid of mice, and in sera from humans acutely infected with T. gondii. Four of the six monoclonal antibodies were able to detect antigens of toxoplasma in these specimens. Control sera from individuals not infected with T. gondii and from individuals chronically infected with the parasite were negative in the enzyme-linked immunosorbent assay. Sera from individuals not infected with T. gondii but with positive titers from rheumatoid factor were also negative; 2 or 10 sera from individuals not infected with T. gondii but with positive titers for antinuclear antibodies reacted with the monoclonal antibodies. When the results of the enzyme-linked immunosorbent assay with monoclonal antibodies and with the F(ab)2 fraction of an immunoglobulin G from a rabbit infected with T. gondii were compared, it was noted that the F(ab)2 was more active in detecting parasite antigens than were the monoclonal antibodies. Thus, although monoclonal antibodies can be used to detect antigens of T. gondii in sera and other body fluids, polyvalent antibody (such as the F(ab)2 fraction) appears to be more satisfactory for this purpose.

Animals↗
PubMed2006

Exaggerated response of arginine vasopressin-enhanced green fluorescent protein fusion gene to salt loading without disturbance of body fluid homeostasis in rats.

We examined the effects of chronic salt loading on the hypothalamic expressions of the enhanced green fluorescent protein (eGFP), arginine vasopressin (AVP) and oxytocin (OXT) genes in AVP-eGFP transgenic rats that expressed eGFP in the hypothalamic AVP-containing neurones. In these rats, salt loading for 5 days caused a marked increase of the eGFP fluorescence in the magnocellular divisions of the paraventricular nucleus (PVN), the supraoptic nucleus (SON) and the internal layer of the median eminence. Expression of the eGFP gene was increased seven- to eight-fold in the PVN and SON of salt-loaded rats in comparison with euhydrated rats. By contrast, none of these changes were observed in the suprachiasmatic nucleus. The expression of the AVP and OXT genes was increased 1.5- to two-fold in the PVN and SON of salt-loaded nontransgenic (control) and transgenic rats. There were no differences in the expression levels of the AVP and OXT genes in the PVN and SON between nontransgenic (control) and transgenic animals under normal conditions and after salt loading. In the posterior pituitary gland, the intensity of the eGFP fluorescence did not change after salt loading for 5 days, but increased after 10 days of salt loading. Upon salt loading, significant increases in the plasma AVP concentrations, plasma osmolality and plasma Na+ were observed. Furthermore, there were no significant differences in changes of water intake, food intake, urine volume, urine osmolality, urine Na+ concentrations, and the body weights in both models under normal or salt-loaded conditions. Our results show that the response of the AVP-eGFP fusion gene to chronic salt loading is exaggerated, and humoral responses such as AVP and OXT and the body fluid homeostasis are maintained in AVP-eGFP transgenic rats. The AVP-eGFP transgenic rat gives us a new opportunity to study the dynamics of the AVP system in vivo.

Animals↗
PubMed2006

Analysis of body fluid mixtures by mtDNA sequencing: An inter-laboratory study of the GEP-ISFG working group.

The mitochondrial DNA (mtDNA) working group of the GEP-ISFG (Spanish and Portuguese Group of the International Society for Forensic Genetics) carried out an inter-laboratory exercise consisting of the analysis of mtDNA sequencing patterns in mixed stains (saliva/semen and blood/semen). Mixtures were prepared with saliva or blood from a female donor and three different semen dilutions (pure, 1:10 and 1:20) in order to simulate forensic casework. All labs extracted the DNA by preferential lysis and amplified and sequenced the first mtDNA hypervariable region (HVS-I). Autosomal and Y-STR markers were also analysed in order to compare nuclear and mitochondrial results from the same DNA extracts. A mixed stain prepared using semen from a vasectomized individual was also analysed. The results were reasonably consistent among labs for the first fractions but not for the second ones, for which some laboratories reported contamination problems. In the first fractions, both the female and male haplotypes were generally detected in those samples prepared with undiluted semen. In contrast, most of the mixtures prepared with diluted semen only yielded the female haplotype, suggesting that the mtDNA copy number per cell is smaller in semen than in saliva or blood. Although the detection level of the male component decreased in accordance with the degree of semen dilution, it was found that the loss of signal was not consistently uniform throughout each electropherogram. Moreover, differences between mixtures prepared from different donors and different body fluids were also observed. We conclude that the particular characteristics of each mixed stain can deeply influence the interpretation of the mtDNA evidence in forensic mixtures (leading in some cases to false exclusions). In this sense, the implementation of preliminary tests with the aim of identifying the fluids involved in the mixture is an essential tool. In addition, in order to prevent incorrect conclusions in the interpretation of electropherograms we strongly recommend: (i) the use of additional sequencing primers to confirm the sequencing results and (ii) interpreting the results to the light of the phylogenetic perspective.

Blood↗
PubMed1994

Sodium appetite, thirst, and body fluid regulation in humans during rehydration without sodium replacement.

After a 7-h H2O and Na+ depletion period (DP), produced by intermittent light exercise (8 bouts) at 35 degrees C, we examined thirst and taste palatability responses to 10 different NaCl solutions during 23 h of rehydration (RH) at 25 degrees C. During DP, net H2O and Na+ loss were 27.2 +/- 2.9 ml/kg and 3.29 +/- 0.45 meq/kg, respectively. Plasma osmolality (POsm) and plasma Na+ concentration ([Na+]p) increased significantly during DP by 3.4 +/- 1.2 mosmol/kgH2O and 3.0 +/- 1.0 meq/kgH2O, respectively. Plasma volume (PV) decreased by 6.5 +/- 1.9%. Thirst rating, renal fractional reabsorption of H2O, and plasma arginine vasopressin concentration (PAVP) increased as POsm increased. This increased thirst was accompanied by increased palatability ratings to H2O. During RH, subjects drank deionized H2O ad libitum and ate a Na(+)-free diet for 23 h. POsm and [Na+]p returned to control levels within 1 h RH and remained at or below the control thereafter. PV remained reduced by approximately 5% throughout RH. The increased thirst and PAVP returned to their respective control levels within 1 h of RH as POsm decreased, but thirst rating increased against between 17 and 23 h of RH without increase in POsm or PAVP. Palatability ratings to a 1 M NaCl solution at and after 3 h RH and palatability ratings to 0.3 M at 17 and 23 h RH were significantly higher than control. Plasma aldosterone concentration (PAldo) increased after DP, decreased with drinking, and increased again between 6 and 23 h of RH, accompanied by a marked decrease in fractional Na+ excretion to < 0.07%. Thus both Na+ preference and thirst in humans are influenced by body fluid and electrolyte status. The increased Na+ palatability (Na+ appetite) was preceded by osmotically induced thirst, and accompanied by nonosmotically driven thirst [extracellular fluid (ECF) thirst] and increased PAldo. The "Na+ appetite" and "ECF thirst" along with increased renal Na+ retention could contribute to ECF volume regulation after thermally induced H2O and Na+ depletion.

Adult↗
PubMed1988

[Studies on concentrations in body fluids of 16-membered ring macrolide antibiotics upon oral administration to healthy volunteers. Effects of their metabolites on bioassay values].

Plasma and urinary antibiotic concentrations measured by 3 different bioassay methods were compared to each other in vitro, after oral administrations of rokitamycin (RKM) tablet, midecamycin acetate (MOM) tablet or josamycin (JM) tablet at a dose of a potency of 600 mg to each of nine healthy volunteers. Method I (RKM bioassay method) was an agar-well method using the agar medium described in the Minimum Requirements for Antibiotic Products of Japan (MRAPJ medium, pH 6.5), method II (MOM bioassay method) was an agar-well method using mycin-assay agar (pH 8.0) and method III (JM bioassay method) was an agar-well method using nutrient agar (pH 7.8). Micrococcus luteus ATCC 9341 was used as the test organism in all the assay method. With any of the bioassay methods tested, plasma concentrations of RKM were determined to be higher than those of MOM or JM. When the three different bioassay methods were compared, method II and III gave consistently higher values than method I for plasma concentrations of any of the drugs tested. These higher values obtained by method II and III appeared to be due to their inability to correctly assay potencies of metabolites of these antibiotics. Using in vitro assay method, potencies of metabolites were found to be estimated higher than they should when method II and III were used. Method I, on the other hand, determined relative potencies of these antibiotics and their metabolites correctly, reflecting differences in MIC values of original antibiotics and their metabolites. Method I, therefore, should be used as the method of preference to determine concentrations of these drugs in body fluids.

Administration, Oral↗