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Influence of plasma proteinase inhibitors and the secretory leucocyte proteinase inhibitor on pancreatic elastase-induced degradation of some plasma proteins.

Pancreatic elastase-induced degradation of some plasma proteins was studied in an in vitro model. The digestion was correlated with the degree of saturation of the alpha 1-proteinase inhibitor (alpha 1PI) and also with varying amounts of secretory leucocyte proteinase inhibitor (SLPI). SLPI was found to inhibit pancreatic elastase showing a Ki of about 10(-7) M for the complex. On the addition of human pancreatic elastase to plasma cleavage of C3, kininogen, fibrinogen and fibronectin was observed when the alpha 1PI approached saturation. In the present in vitro model it was possible to block the cleavage of the four plasma proteins, mentioned above completely with SLPI. Addition of the inhibitor also decreased the consumption of alpha 1PI.

Blood Proteins↗

The effect of dietary copper on rat plasma apolipoprotein B, E plasma levels, and apolipoprotein gene expression in liver and intestine.

The plasma levels of apo B and apo E, and the level of hepatic and intestinal mRNA coding for these apolipoproteins were investigated in weanling male rats pair-fed for 6 wk with a control or copper-deficient diet. Plasma cholesterol, triglycerides, and phospholipids were significantly increased, and plasma apo B and apo E levels were also markedly increased in copper-deficient rats as compared to control rats. Copper deficiency significantly increased triglyceride levels and decreased cholesterol levels in the liver. No major differences in the levels of hepatic and intestinal apo B and apo E mRNA occurred between control and copper-deficient rats. These data imply that hypertriglyceridemia dn hypercholesterolemia owing to the copper deficiency are not accompanied by modifications in the gene expression at the mRNA level in the liver and intestine of the apolipoproteins studied.

Animals↗

The interaction of plasma renin activity and plasma atrial natriuretic peptide in 21-hydroxylase deficiency patients.

Plasma renin activity (PRA) determination is the main index used to evaluate the mineralocorticoid control in 21-hydroxylase deficiency (21-OHD). PRA values within or at the upper limit of the age-appropriate range, or values <5 or 10 ng/ml/h have been regarded as adequate control. Atrial natriuretic peptide (ANP) has opposite actions to those of angiotensin II/aldosterone, and could help to understand the hydrosaline homeostasis in 21-OHD. We studied the interaction between PRA and ANP levels in 10 controls and 26 patients with 21-OHD under corticoid treatment. Patients were divided into two groups according to PRA levels, < or > or = 5 ng/ml/h, irrespective of the clinical form of 21-OHD. Blood samples for determination of PRA and ANP levels were taken after 30 min in the sitting position (basal), after 30 min in the recumbent position and after 15 min of 20 degrees head-down tilting. ANP levels (pg/ml) in the basal, supine and after head-down tilting position were 25.9 +/- 1.6, 42.7 +/- 7.4 and 54.3 +/- 5.5 in controls; 28.5 +/- 2.1, 38.3 +/- 2.1 and 48.8 +/- 4.1 in the group with PRA levels <5 ng/ml/h, and 20.9 +/- 1.9, 26.6 +/- 2.5 and 34.6 +/- 3.1 in the group with PRA levels > or = 5 ng/ml/h, respectively. Basal and after head-down tilting ANP plasma levels were similar between the controls and the group with PRA levels <5 ng/ml/h. However, the group of patients with PRA levels > or = 5 ng/ml/h showed lower basal and stimulated ANP levels compared to the control group (p<0.05). The decreased plasma ANP levels in the basal condition and after head-down tilting indicate a chronic contraction of the extracellular volume in 21-OHD patients with increased PRA levels. Therefore, mineralocorticoid deficiency is counteracted by decreased ANP secretion in order to preserve fluid and electrolyte homeostasis.

Adolescent↗

Determination of oxaliplatin in human plasma and plasma ultrafiltrate by graphite-furnace atomic-absorption spectrometry.

A method for sensitive determination of the anti-cancer agent oxaliplatin in human plasma and human plasma ultrafiltrate (pUF) is presented. The method is based on the quantification of platinum by graphite-furnace atomic-absorption spectrometry, with Zeeman correction and an atomisation temperature of 2,700 degrees C. Sample pretreatment involves dilution of the samples with a solution containing 0.15 mol L(-1) NaCl and 0.20 mol L(-1) HCl in water. Validation was performed in accordance with the most recent FDA guidelines for bioanalytical method validation. All results were within requirements. The validated ranges of quantification were 0.10-400 micromol L(-1) for human pUF and 0.50-400 micromol L(-1) for plasma. The assay is now successfully used to support pharmacokinetic studies of cancer patients treated with oxaliplatin.

Antineoplastic Agents↗

The effects of anti-androgenic and estrogenic disrupting contaminants on breeding gland (nuptial pad) morphology, plasma testosterone levels, and plasma vitellogenin levels in male Xenopus laevis (African clawed frog).

Nuptial pads are areas, digits, and forearm characterized by epidermal keratin hooks and dermal breeding glands opening on the epidermis surface in male frogs. The presence of the epidermal hooks and secretory activity of the breeding glands are androgen-dependent and considered functioning during mating. Chemical pollutants released into the aquatic environment by humans are suspected of disrupting the normal hormonal pathways and functioning. Among the best known of these endocrine-disrupting contaminants (EDCs) are compounds that mimic the steroid 17beta-estradiol. However, recently specific anti-androgenic activity by certain EDCs, including DDE and several fungicides, have been shown in mammalian studies. The African clawed frog, Xenopus laevis, has been proposed as an ideal species to be used as a model system for screening of endocrine disruptors. In this study we employed a newly developed ELISA for the measurement of plasma vitellogenin (VTG), normally produced in the liver of females under the influence of estrogens but also in males when exposed to exogenous estrogens. Furthermore, we investigated the potential of breeding (nuptial) gland activity to be employed as a biomarker system for screening for anti-androgenic activity by certain EDCs. Our results indicate that the pharmaceutic anti-androgen flutamide did significantly (p < 0.05) effect the androgen-dependent breeding (nuptial) glands and plasma testosterone concentrations in male X. laevis. Our results further confirm that the dicarboximide fungicide vinclozolin mimics the anti-androgenic action of flutamide. Vinclozolin, however, did not significantly effect the plasma testosterone concentration. Results also confirm the ability of 17beta-estradiol to stimulate production and release of VTG in the liver of male X. laevis, although we could not confirm the in vivo induction of VTG by estrogenic mimics octylphenol and nonylphenol. Although more dose-response research is needed, this preliminary study confirms the potential of VTG production and breeding gland activity as biomarkers to screen chemicals and environmental samples for estrogenic and anti-androgenic activity using X. laevis males.

Androgen Antagonists↗

Plasma renin activity and plasma aldosterone during the normal menstrual cycle.

Six normal women were studied under metabolic balance conditions during the menstrual cycle. Plasma renin activity (PRA) and plasma aldosterone were estimated throughout the cycle under standardized, uniform conditions with sensitive radioimmunoassays. There is a linear increase in PRA and plasma aldosterone during the luteal phase of the cycle provided ovulation occurs. If ovulation fails, no rise is seen in either parameter. Thus, a functioning corpus luteum seems to be essential for the production of these hormonal changes.

Adult↗

Correlation of nocturnal increase in plasma oxytocin with a decrease in plasma estradiol/progesterone ratio in late pregnancy.

OBJECTIVES: Our purpose was to determine whether plasma oxytocin concentrations show a daily rhythm. STUDY DESIGN: Ten women between 37 and 39 weeks of gestation volunteered for the study. They were admitted 1 to 2 days before the experiment. Three blood samples were taken with 2-minute intervals each time, at 8 AM, 4 PM, and 12 midnight. Oxytocin, 17 beta-estradiol, progesterone, and cortisol were measured by radioimmunoassay with highly specific antibodies. Statistical analysis of variance by Friedman's test was followed by multiple range testing, and p < 0.05 was considered significant. RESULTS: Significant daily rhythm was found for plasma cortisol, progesterone, and oxytocin and for the estradiol/progesterone ratio. Oxytocin showed a nocturnal peak and a strong negative correlation with the estradiol/progesterone ratio. CONCLUSIONS: The daily rhythm in plasma oxytocin parallels the rhythm in uterine activity (shown by others), suggesting a causal relationship between the two. Both may in turn be related to the ratio of circulating estradiol and progesterone.

Circadian Rhythm↗

Plasma membranes from rat intestinal epithelial cells at different stages of maturation. I. Preparation and characterization of plasma membrane subfractions originating from crypt cells and from villous cells.

To determine the mechanism of the maturation of the brush border membrane in intestinal epithelial cells, purification of the plasma membrane from undifferentiated rat crypt cells and of the basal-lateral membrane from villous cells has been performed. The method is based on density perturbation of the mitochondria to selectively disrupt their association with the membrane. With both cell populations, two membrane subfractions displaying the same respective density on sucrose gradient have been obtained with an overall yield of 15--20% and a 10-fold enrichment of the plasma membrane markers 5'-nucleotidase and (Na+ + K+)-dependent, ouabain-sensitive ATPase chosen to follow their purification. The four fractions were constituted by sheets and apparently closed vesicles of various sizes. Each fraction was characterized by a distinct protein composition and different levels of enzyme activities. The cells, used for the preparation of the membranes, were isolated as a villus to crypt gradient. This separation and that of the membranes, led to the conclusion that the (Na+ + K+)-dependent ATPase is localized principally in the plasma membrane of all cells whatever their state of maturation, while 5'-nucleotidase is predominantly located in the basal-lateral membrane of the villous cells and may serve as a specific marker for the purification of this membrane. Finally it has been shown that aminopeptidase, dissacharidases and alkaline phosphatase do not appear simultaneously in the maturation process of the cells, alkaline phosphatase being absent from the crypt cells and aminopeptidase being the first to be synthesized. This enzyme seems to appear in the crypt cells membrane before being integrated into the mature brush border membrane.

Aging↗

Effects of clofibrate treatment on plasma triglyceride concentration, plasma post-heparin clearing factor lipase (lipoprotein lipase) activity and serum clearing factor lipase activating ability in maturity-onset diabetes.

The effects of clofibrate on plasma triglyceride concentration, plasma post-heparin clearing factor lipase activity and serum clearing factor lipase activating ability were studied in a group of maturity-onset diabetic patients. Significant falls in both triglyceride concentration and in activating ability occurred within 2 weeks of beginning clofibrate treatment and, when treatment was stopped after 4 weeks, these changes were reversed within a further 4 weeks. Plasma post-heparin clearing factor lipase activity, on the other hand, was significantly increased during clofibrate administration and fell again when the treatment was stopped. The possible interrelationships of these findings are discussed.

Blood Proteins↗

Plasma high density lipoprotein subfractions in subjects with different coronary risk indices as assessed by plasma lipoprotein concentrations.

The distribution of plasma high density lipoprotein (HDL) subfractions was determined in 2 groups of healthy male subjects with different coronary risk indices (CI) as assessed by the ratio of total plasma cholesterol/HDL-cholesterol. The subjects in the 2 groups were of similar age and fitness (as assessed by VO2max). The higher risk group (CI greater than 4.0) contained a lower relative concentration of a specific HDL subfraction, HDL2b, separated by gradient gel electrophoresis, and a lower level of an apo E-rich HDL fraction, isolated by affinity chromatography, than the lower risk group (CI less than 4.0). The concentration of total HDL-cholesterol was higher in the lower risk group due to a difference in HDL2-cholesterol when separation was achieved by polyanion precipitation, but not when separation was made by ultracentrifugation. These observations suggest that the level of these specific HDL subfractions might, when taken in conjunction with plasma cholesterol concentration, provide a better index of coronary risk than that of total HDL as conventionally employed.

Adult↗

Plasma lipoprotein lipids in relation to the MspI polymorphism of the apolipoprotein AII gene in Caucasian men. Lack of association with plasma triglyceride concentration.

Digestion of the human apolipoprotein (apo) A-II gene with the endonuclease MspI produces fragments of 3.0 or 3.7 kb, reflecting the presence or absence of a polymorphic site within an Alu sequence 3' to the gene. Patients with hypertriglyceridemia have been shown to have an increased prevalence of the 3.0 kb allele. To explore this observation further, plasma lipoprotein lipids were studied in a random sample of fasted middle-aged Caucasian men, of which 59 were 3.0 kb homozygotes, 24 were heterozygotes, and 2 were 3.7 kb homozygotes. After adjusting for the effects of age, height, weight, alcohol intake and cigarette consumption by covariance analysis, no statistically significant associations were present between genotype and the concentrations of triglyceride in whole plasma or the d less than 1.019 g/ml fraction of plasma (i.e., VLDL + IDL). Nor were the cholesterol concentrations in VLDL + IDL, low density lipoprotein (LDL, d = 1.019-1.063 g/ml), high density lipoprotein (HDL), HDL2 or HDL3 related to genotype. In an independent comparison of eight 3.0 kb homozygotes and eight 3.7 kb homozygotes (all Caucasians) drawn from a different community, genotype was unrelated to the triglyceride or cholesterol concentrations in VLDL (d less than 1.006 g/ml), IDL + LDL (d = 1.006-1.063 g/ml) or HDL, after adjustment for the effects of covariates. These results suggest that the MspI polymorphism of the apo A-II gene is not associated with genetic variation that significantly affects triglyceride transport in the majority of men.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein A-II↗

The effects of carbon dioxide inhalation on plasma MHPG, plasma hormones respiratory rate, and behavior in the rhesus monkey.

The effects of inhalation of air and 3 concentrations of carbon dioxide (CO2) on plasma levels of the norepinephrine metabolite, MHPG, plasma hormones, and behavioral activation were assessed in eight chair-adapted Rhesus monkeys (Macaca mulatta). In comparison to air, inhalation of 5%, 7.5% and 10% CO2 for 180 minutes produced significant dose-dependent increases in respiratory rate, plasma MHPG, cortisol, growth hormone and prolactin. CO2 at the 7.5% concentration produced peak changes in behavior at 15, growth hormone at 30, and cortisol and MHPG at 180 minutes without producing changes in prolactin. The lack of previously reported CO2 induced changes in MHPG, growth hormone and prolactin in humans exposed to 7.5% CO2 for only 15 minutes, may therefore relate to the relatively short duration of CO2 exposure.

Administration, Inhalation↗

Role of the kidney in the plasma clearance of angiotensinogen in the rat: plasma clearance and tissue distribution of 125I-angiotensinogen.

We studied the tissue distribution and plasma clearance of angiotensinogen (AGN) in rats following an i.v. injection of 125I-labeled AGN. The plasma clearance rate of [125I]AGN fits a two-compartment model with half-lives of 10.2 +/- 1.5 min and 4.1 +/- 0.5 h in non-treated rats, and the half-life of slower phase significantly increased to 10.2 +/- 1.1 h following bilateral nephrectomy. Radioactivity was predominantly distributed in the kidneys (4.9%), and to a lesser extent in the liver (1.8%), testis (1.2%), spleen (0.61%), heart (0.35%), lung (0.18%), thymus (0.03%) and brain (0.03%). The subcellular distribution of radioactivity in the kidney was 64% in the soluble fraction and 33% in the crude mitochondrial-lysosomal fraction. Sodium dodecylsulfate-polyacrylamide gel electrophoresis revealed that the radioactivity in the soluble fraction consisted of proteins corresponding to intact [125I]AGN, whereas the mitochondrial-lysosomal fraction contained additional radioactive proteins with molecular weights between 18,000 and 29,000. When isolated kidney cells were incubated with [125I]AGN at 0 degree C, the radioactive binding was saturable and specific with a Kd value of 4.8 x 10(-11)M, whereas incubation at 37 degrees C resulted in the appearance of degraded products of [125I]AGN in the medium. These results suggested that circulating AGN is cleared mainly by the kidneys via receptor-mediated endocytosis, which may play an important role in regulating plasma level of AGN.

Angiotensinogen↗

Brain tryptophan, plasma free tryptophan and distribution of plasma neutral amino acids.

Although rat brain tryptophan is strikingly elevated following portacaval shunt, plasma total tryptophan is unchanged and plasma free tryptophan is not elevated to the same degree as brain tryptophan. Investigation of the concentrations of the neutral amino acids (phenylalanine, tyrosine, methionine, threonine, leucine, isoleucine, and valine) revealed that their distribution and the sum of their concentrations were altered following portacaval shunt, and that this pattern was similar to that seen in humans with cirrhosis of the liver. It is suggested that both the elevation in plasma free tryptophan and the decrease in the competing neutral amino acids, act together to increase the transport of tryptophan into brain when portal blood is diverted around the liver. The implications of these findings in therapy of hepatic coma is discussed.

Amino Acids↗

Plasma catabolism of human apolipoprotein E isoproteins: lack of conversion of the doubly sialylated form to the asialo form in plasma.

Apolipoprotein E (apoE) circulates as a mixture of sialylated and asialylated forms. In this study the catabolic fate and the plasma turnover rate of the different apoE forms have been investigated in vivo in humans. Asialo apoE (E) and doubly sialylated apoE (Ess) were isolated by preparative isoelectrofocusing from the VLDL of subjects homozygous for the E3 allele. 131E3 and 125E3ss were injected simultaneously into three hypertriglyceridemic subjects, and plasma samples were collected up to the sixth day. VLDL were isolated by ultracentrifugation, and the apoE forms were separated by isoelectrofocusing. Gel bands corresponding to E3 and E3ss were cut out and counted for the associated radioactivity. Residence times in plasma for 131E3 and 125E3ss were 0.95 +/- 0.16 and 0.74 +/- 0.16 days, respectively. As determined from the gel count distribution up to 24 hours, no conversion of the injected sialylated form to the correspondent asialylated form was detected.

Aged↗

The role of fatty acid saturation on plasma lipids, lipoproteins, and apolipoproteins: II. The plasma total and low-density lipoprotein cholesterol response of individual fatty acids.

Regression analyses were performed on individual data from our two previous clinical investigations to establish the cholesterolemic effects of individual fatty acids. Our principal objective was to determine the effects of stearic acid on plasma total cholesterol (TC) and low-density lipoprotein (LDL) cholesterol levels. Our second objective was to determine the variation in the cholesterolemic response of individuals to changes (delta) in the major dietary fatty acids. The best-fitting linear regression equations relating delta TC and delta LDL (mg/dL) were as follows: delta TC = 2.3 delta C14:0 + 3.0 delta C16:0 - 0.8 delta C18:0 - 1.0 delta polyunsaturated fatty acids (PUFA) and delta LDL = 2.6 delta C14:0 + 2.9 delta C16:0 - 0.5 delta C18:0 - 0.7 delta PUFA, where delta fatty acid = change in intake expressed as percent of calories. Based on these equations, in which stearic acid has a significant, negative regression coefficient, and the other regression models analyzed, it appears that stearic acid has an independent cholesterol-lowering effect. Using the equation we developed, 75% of the actual cholesterolemic responses were within +/- 10 mg/dL of the predicted response. In summary, we have developed a predictive equation (similar to those developed by both Keys and Hegsted) to estimate changes in plasma TC and LDL cholesterol levels of young men in response to changes in dietary fatty acids. However, our predictive equation separates stearic acid from the other long-chain saturated fatty acids (SFA) and indicates that it has an independent cholesterol-lowering effect. Thus, stearic acid is a unique long-chain SFA because of its effect on plasma cholesterol level.

Apolipoproteins↗

Schedule-induced polydipsia experience decreases plasma corticosterone levels but increases plasma prolactin levels.

To determine the neuroendocrine pattern of response to excessive drinking induced by exposure of rats to an intermittent distribution of food (schedule-induced polydipsia, SIP), the present experiment investigated changes in plasma corticosterone, prolactin and catecholamines in chronically catheterized rats that had developed or not this form of adjunctive behaviour. It was found that rats that engage in excessive drinking displayed decreased plasma levels of corticosterone and increased levels of prolactin during the course of a SIP session. There was, however, no differences between groups in plasma catecholamine levels. The difference observed between SIP-pos and SIP-neg rats were entirely condition-specific, since they disappeared in the absence of access to water.

Animals↗

Quantitation of the three normally-occurring plasma fibrinogens in health and during so-called "acute phase" by SDS electrophoresis of fibrin obtained from EDTA-plasma.

Normal human plasma contains fibrinogens of different molecular weight. To quantitate these fibrinogens, plasma was clotted at pH 6.4 with 7.5 NIH U/ml thrombin in the presence of 8 mM EDTA. The clots were dissolved in 6.6 M urea and submitted to SDS electrophoresis on gels containing 3% polyacrylamide/0.5% agarose, and the fractions quantitated by densitometric scanning. The reproducibility of this method was high with variation coefficient 1.5%. Three main fibrinogen fractions were found: High molecular weight fibrinogen (HMW, mw 340 000), low molecular weight fibrinogen (LMW, mw 300 000) and LMW' (mw 280 000). In addition 5 weak bands could be seen. In plasma from 123 healthy subjects of both sexes, aged 1-93 years, HMW constituted 69.7% +/- 5.1, LMW 26.5% +/- 4.8 and LMW' 3.8% +/- 1.8 of the total fibrinogen. Women and older subjects displayed a small, but statistically significant reduction in the relative amounts of HMW (2-4%). Following surgery and extensive acute myocardial infarction the HMW/LMW ratio changed. The substantial increase in total fibrinogen regularly recorded was mainly due to HMW that reached maximal values after 3-4 days. LMW remained unchanged the first 2 days and then displayed a slight increase with a delayed maximum (8-11 days).

Adolescent↗