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PubMed1977

Serum myeloperoxidase and lactoferrin in neutropenia.

Radioimmunosorbent assays for determination of serum content of the neutrophil proteins myeloperoxidase and lactoferrin are described. Serial studies were performed in patients with neutropenia. In 2 cases of cyclic neutropenia the myeloperoxidase level showed slight variations within the normal range during the cycle while lactoferrin displayed a clear correlation with neutrophil counts. In 1 case with persistent agranulocytosis myeloperoxidase was normal but lactoferrin was extremely low. During the regeneration phase of drug-induced neutropenia neutrophil counts and serum lactoferrin increased in a parallel fashion. Since serum myeloperoxidase was normal during profounded neutropenia it is suggested to derive primarily from myeloperoxidase-rich granulopoietic precursor cells of the marrow. Serum lactoferrin on the other hand seems to derive from leakage of more granulopoietic cells of blood and marrow. Studies of neutrophil proteins of serum may aid in evaluation of neutropenic patients.

Adolescent↗
PubMed1980

Human renal cytoplasmic carbonic anhydrase. Tissue levels and kinetic properties under near physiological conditions.

The cytoplasmic form of human renal carbonic anhydrase, CA-C (carbonate dehydratase, EC 4.2.1.1), purified by affinity chromatography, was characterized kinetically at 37 degrees C in 25 mM N-methyl imidazole buffer, I = 0.15, pH 7.1. using a pH-indicator stopped-flow technique. Under these conditions the rate constants for the uncatalyzed hydration of CO2 and dehydration of H2CO3 were 0.12 . s-1 and 0.60 . s-1, respectively. The kinetic parameters for CA-C were found to be: Hydration reaction, Km = 11.8 mM, V/[E]0 = 10.6 x 10(5) . s-1, dehydration reaction Km = 70 mM, V/[E]0 = 5 x 10(5) . s-1. In the hydration reaction CA-C was non-competitively inhibited by acetazolamide, Ki = 16 nM, sulfanilamide, Ki = 8 micrometer, and chlorothiazide, Ki = 1 micrometer. The levels of immunoassayable CA-C in cortex, medulla and papilla of perfused donor kidneys were 1.3, 1.0 and 0.6 mg enzyme protein/g tissue protein respectively which corresponded well with the levels measured catalytically. The erythrocyte form, HCA-B, was also detected immunochemically (approximately 0.1 mg/g protein) but is thought to be a contaminant. Calculations indicated that the uncatalyzed hydration of CO2 in the tubular cells can support 17 or 0.7% of the rate of urine acidification, dependent on whether the cellular alkaline pH-disequilibrium during acid secretion is 0.1 or 0.01 pH units, respectively. CA-C accelerates the hydration rate 6800-fold which enables the cell to sustain high rates of proton generation, while maintaining near CO2-equilibrium and maximal buffering capacity. Even at an assumed pH-disequilibrium of only 0.01 pH-unit, CA-C is present in 50-fold excess of apparent physiological needs. When the enzyme is inhibited the rate of the uncatalyzed reaction increases, which partly overcomes the effect of inhibition.

Carbonic Anhydrases↗
PubMed1981

Immunoglobulin E anti-Staphylococcus aureus antibodies in atopic patients.

Sera from 56 patient and normal adults were examined to quantitate total immunoglobulin E (IgE) and IgE antibodies to Staphylococcus aureus and Staphylococcus epidermidis. Patients were divided into six groups based on clinical symptoms; a seventh group consisted of normal adults. Anti-S, aureus IgE binding was significantly higher in three groups of patients (those with eczema, those with or without series staphylococcal abscesses, and allergic patients with staphylococcal skin infections) than it was in the control group. Patients with high IgE due to allergies or parasitic infections without staphylococcal infections and patients with low or normal IgE and serious staphylococcal infection showed low levels of binding. The assay measured specific binding of IgE to bacterial antigens.

Adult↗
PubMed1984

IgE class immune complexes in Felty's syndrome: characterisation of antibody activities in isolated complexes.

By means of a double polyethylene glycol (PEG) precipitation and PRIST technique IgE was detected in 3% PEG precipitates and in the immune complex enriched fractions purified by solid-phase Clq adsorption from sera of 11 of 20 patients with Felty's syndrome. No correlation was found between the occurrence of complexed IgE and total protein content of the immune complex enriched material. IgE rheumatoid factor and anti-IgE antibody activity were detected in some of the immune complex fractions. Serum levels of complement C3, C4, and factor B were low in IgE immune complex positive cases. Only 4 of 20 patients with articular rheumatoid arthritis had IgE-containing immune complexes.

Aged↗
PubMed1980

A family study of spina bifida and anencephalus in Belfast, Northern Ireland (1964 to 1968).

The parents of 226 of the 360 patients with anencephalus or spinal bifida or both, born in Belfast 1964 to 1968, were visited to document the occurrence of these malformations among other relatives. The proportions of sibs with anencephalus and spina bifida were 10.41% for spina bifida index patients and 6.4% for anencephalus. For patients born after the index patients, the proportions were 12.19% and 6.35%, respectively. The overall incidence of either malformation among sibs was 8.87%. This estimate is higher than the 4 to 5% commonly reported and is probably related to the specific background of the Northern Ireland population, which is known to have the highest incidence of CNS malformations in the United Kingdom. The substantial size of this risk indicates the importance of amniocentesis for monitoring subsequent pregnancies of women who have had one child with a CNS malformation.

Anencephaly↗
PubMed1989

The source of immunoglobulin E in the fetus at term.

A high level of immunoglobulin E (IgE) in cord blood (CB) is well known to be predictive of the development of clinical allergy during early childhood. We have tried to ascertain the source of IgE in cord blood by carrying out a series of measurements of IgE concentrations in arterial and venous CB of newborns and comparing them to IgE concentrations in blood samples from both parents. We found that the concentrations of IgE in arterial and venous CB were almost completely identical in the 131 newborns examined whereas IgD concentrations were significantly higher in arterial than in venous CB. Moderate but significant positive correlations were found between IgE concentrations in maternal blood and CB; paternal blood and CB; and maternal and paternal blood. Our data suggest that CB-IgE is essentially a product of fetal lymphocytes. The positive correlations found between the IgE blood concentrations in the newborns, the mothers and the fathers, is most likely an effect of the amount and nature of the allergenic constituents in the family environment.

Amniotic Fluid↗
PubMed1995

The relationship between parental and children's serum IgE and asthma.

This paper examines the familial aggregation of physician-diagnosed asthma in relation to the age- and sex-standardized total serum IgE levels of children and their parents in a sample of the general population in Tucson, Arizona, that has been followed in a longitudinal study for over 20 yr. There were 591 nuclear families containing 1,177 children who provided information about the presence or absence of a physician diagnosis of asthma. The serum IgE data were less complete: both parents and one or more of their children in 251 of the nuclear families, containing 468 children, had serum IgE levels measured. There was a very strong tendency for asthmatic patients to have asthmatic children, but only a small part of this appeared to be related to the familial aggregation of total serum IgE. In the absence of an asthmatic parent, there was a slight but significantly higher prevalence of asthma in children of whom both parents had IgE levels in the highest tertile. Very high rates of children's asthma depended on there being an asthmatic parent who also had at least moderate levels of serum IgE. It was also shown that asthmatic children have considerably higher total IgE levels than would be expected on the basis of their parents' IgE levels alone. The data appear compatible with several familial-aggregation hypotheses and a strong environmental influence determining which children are likely to develop asthma. We speculate that the inflammation in the airways of asthmatic patients itself tends to increase the serum IgE level, possibly secondary to mediators that it generates.

Adolescent↗
PubMed2006

Glucocorticoid is involved in food-entrainable rhythm of mu-opioid receptor expression in mouse brainstem and analgesic effect of morphine.

The repeated manipulation of feeding schedule has a marked influence on the chronopharmaological aspects of many drugs. In this study, we investigated the role of endogenous glucocorticoid in the mechanism by which restricting the feeding time modulates the analgesic effect of morphine. Male ICR mice were housed under a light-dark cycle (light on from 07:00 to 19:00) with food and water ad libitum or under repeated time-restricted feeding (feeding time from 09:00 to 17:00) for 2 weeks before the experiment. Under the ad libitum feeding, mRNA levels of mu-opioid receptor and its binding capacity in mouse brainstem increased around the early dark phase, following the 24-h variation in circulating glucocorticoid levels. As a consequence, potent analgesic effects of morphine were observed in mice injected with the drug during the dark phase. Daily restricted feeding modulated the time-dependency of mu-opioid receptor function, accompanied by the alteration of the rhythm in circulating glucocorticoid levels. Under the time-restricted feeding, potent analgesic effects of morphine were found in mice injected with the drug during the light phase. Because the manipulation of feeding schedule was unable to produce the food-entrainable rhythm in the expression of mu-opioid receptor in the brainstem of adrenalectomized mice, endogenous rhythm of glucocorticoid secretion seems to be involved in the mechanism by which the time-restricted feeding modulates the analgesic effects of morphine.

Analgesics, Opioid↗
PubMed2006

Use of insulin immunoassays in clinical studies involving rapid-acting insulin analogues: Bi-insulin IRMA preliminary assessment.

BACKGROUND: In clinical studies involving rapid-acting analogues (RAAs), insulin immunoreactivity is frequently measured, including endogenous, regular insulin (RI) and RAA immunoreactivities. Such a procedure implies equivalent cross-reactivities of all insulins present in serum. Commercially available human insulin immunoassays have been widely used, but their limitations (including hemolysis and anti-insulin antibodies) were not fully investigated. The aims of our study were to compare cross-reactivities of RI and RAAs in buffer and in serum and to investigate insulin immunoassay pitfalls. METHODS: Cross-reactivities were assessed using Bi-insulin IRMA (Schering Cis-Bio International) in phosphate-buffered saline (PBS)-1% bovine serum albumin (BSA) and in pools of sera spiked with RI and RAAs (lispro and aspart). To investigate the influence of hemolysis, a pool of sera spiked with RAA was mixed with a concentrated hemolysate (final hemoglobin concentration 10 g/L) and incubated for 3 h at room temperature. To determine interference by anti-insulin antibodies, insulin was removed using charcoal from 18 sera with anti-insulin antibodies and from 17 sera without detectable anti-insulin antibodies. These insulin-free samples were then spiked with RI and RAAs and the immunoreactivity was determined. RESULTS: Compared with buffer, cross-reactivity in serum for RI, lispro and aspart was lower (35%, 29% and 26% lower, respectively). Hemolysis degraded almost all RI and RAAs contained in the serum (>or=95%). Anti-insulin antibody interference was significant for RI and RAAs (p<or=0.004) and correlated with anti-insulin antibody level in the serum (p<or=0.001). CONCLUSIONS: In serum, RI and RAA cross-reactivities are slightly lower than in buffer. For RAA assessment, hemolysed samples should be discarded and anti-insulin antibodies should be removed from samples before immunoreactivity measurements.

Antibodies↗
PubMedDate not supplied

Urinary tract infection in full-term newborn infants: risk factor analysis.

OBJECTIVE: To analyze the correlation of risk factors to the occurrence of urinary tract infection in full-term newborn infants. PATIENTS AND METHODS: Retrospective study (1997) including full-term infants having a positive urine culture by bag specimen. Urine collection was based on: fever, weight loss > 10% of birth weight, nonspecific symptoms (feeding intolerance, failure to thrive, hypoactivity, debilitate suction, irritability), or renal and urinary tract malformations. In these cases, another urine culture by suprapubic bladder aspiration was collected to confirm the diagnosis. To compare and validate the risk factors in each group, the selected cases were divided into two groups: Group I - positive urine culture by bag specimen collection and negative urine culture by suprapubic aspiration, and Group II - positive urine culture by bag specimen collection and positive urine culture by suprapubic aspiration. RESULTS: Sixty one infants were studied, Group I, n = 42 (68.9%) and Group II, n = 19 (31.1%). The selected risk factors (associated infectious diseases, use of broad-spectrum antibiotics, renal and urinary tract malformations, mechanical ventilation, parenteral nutrition and intravascular catheter) were more frequent in Group II (p<0.05). Through relative risk analysis, risk factors were, in decreasing importance: parenteral nutrition, intravascular catheter, associated infectious diseases, use of broad-spectrum antibiotics, mechanical ventilation, and renal and urinary tract malformations. CONCLUSION: The results showed that parenteral nutrition, intravascular catheter, and associated infectious diseases contributed to increase the frequency of neonatal urinary tract infection, and in the presence of more than one risk factor, the occurrence of urinary tract infection rose up to 11 times.

Catheterization↗
PubMed1977

Two-site assay of bovine parathyroid hormone.

A two-site assay has been developed for bovine PTH. This technique involves reaction of the antigen with two antibody molecules with the purpose of increasing specificity. In paratice the hormone was extracted form plasma samples on to plastic tubes coated with antibody specific for PTH (1-34). Uptake was then measured using 125I-labelled antibodies specific for PTH (53-84). In this way a sensitive assay was obtained for PTH (1-84) which did not recognize molecular fragments. This technique was used in conjunction with immunoradiometric assays specific for either NH2-terminal (1-34) or CO2H-terminal (53-84) molecular fragments to study the clearance of infused PTH in the cow. Preliminary results support the hypothesis that the intact molecule is rapidly degraded in the peripheral circulation with the preferential disappearance of an NH2-terminal fragment. Studies on endogenous secretion during calcium and EDTA infusions indicated that there was little intact hormone present at the times when CO2H-terminal immunoreactivity was readily measured.

Animals↗
PubMed1994

Assay systems for the growth hormone-binding protein.

The first method used for detection of growth hormone-binding protein (GHBP) in biological fluids was based on the incubation of the sample with radiolabeled GH followed by separation of bound and free GH by gel exclusion chromatography. Recently, other methods have been developed which are faster and easier to use. These methods include variants of the original binding/column assay (e.g., separation of bound and free GH is obtained by immunoprecipitation, charcoal adsorption, ion exchange chromatography, or HPLC), and a ligand-mediated immunofunctional assay (LIFA), in which a monoclonal antibody is used to capture the GHBP on a microtiter plate; all binding sites are saturated with GH and an anti-GH antibody is used to detect the amount of GH (endogenous and exogenous) bound to the GHBP. To permit comparison of results obtained by different methods we have cross-validated the LIFA with two different binding assays: (i) the original long column assay (column assay), and (ii) an assay based on immunoprecipitation (RIPA) of the GH/GHBP complex with an anti-GHBP antibody.

Adolescent↗
PubMed1976

[Simultaneous radioimmunassay for urinary thyroxine (T4) and triioldothyronine (T3) (AUTHOR'S TRANSL)].

A radioimmunoassay for the determination of thyroxine (T4) and triiodothyronine (T3) in urine was developed. The extraction of a sample, the incubations with T3-und subsequently T4-antibody and the elutions of the respective bound fractions are performed all on the same Sephadex column. This principle can be applied to as many as 120 simultaneous determinations. The interassay coefficients of variation were 20.1% for T3 and 10.6% for T4, respectively. The recovery of standards added to urine was 107 +/- 8% (mean +/- SD) for T3 and 102 +/- 8% for T4. The 24 h collections of urine from healthy controls contained 1.70 +/- 0.40 mug T3 and 1.44 +/- 0.51 mug T4 (mean +/- SD, N=20). Three hourly fractions of the urinary excretion collected from 6 a.m. through 9 p.m. showed a significant (p less than 0.0005) elevation of the T4 excretion as compared to the night fraction, whereas an increased T3 excretion was only observed from 6-9 P.M. The T4 excretion was reduced in primary hypothyroidism (0.48 +/- 0.47 mug per 24 h, p less than 0.0005), whereas the T3 excretion was not that markedly reduced (1.30 +/-0.80 mug/d), pointing to the known residual T3 secretion. A reduced T4 excretion (0.85 +/- 0.34 mug/d, p less than 0.00005) contrasted with the normal T3 excretion in patients with endemic goiter, indicating the known preferential T3 secretion in iodine deficiency states. T3 thyrotoxicosis was accompanied by an elevated T3 excretion with normal urinary T4. Normal excretion of T4 and T3 were observed in patients with persistantly suppressed serum TSH levels after successful treatment for hyperthyroidism.

Circadian Rhythm↗
PubMed1979

Evidence of a high-activity C type of carbonic anhydrase in human ciliary processes.

Carbonic anhydrase activity was found in the ciliary process of fresh human donor eyes, originating from an enzyme antigenically similar to the erythrocyte high-activity isoenzyme HCA C. It was sensitive to inhibition by acetazolamide and resistant to inhibition by halides like HCA C. The enzyme is probably identical with HCA C. Its tissue concentration was one fifth to one tenth of that in the human kidney. The erythrocyte low-activity isoenzyme HCA B was also found in the processes as a contaminant.

Acetazolamide↗
PubMedDate not supplied

[The formation of the influenza virus aggregates, enriched with hemagglutinin].

The formation of electrostatic aggregates was studied by analysis of two types of virus-containing liquids: initial warm liquid collected at temperature 37 degrees and the same liquid stored over the night at temperature 4 degrees C. The formation of virus aggregates was revealed at 4 degrees C. The aggregates formed at temperature 4 degrees C had a relatively high HA/NP ratio in comparison with unassociated virus analyzed at 37 degrees. HA-enriched aggregates were found in the precipitate formed under short-term high-speed centrifugation as well as in "heavy arm" of the virus profile in the saccharose gradient. Aggregates formed at 4 degrees C dissociated at 37 degrees. The ability to form aggregates is reversible and correlates with the virus concentration. It is shown also that virus containing liquid contains heterogenic structures with molecular weight under 2000 kD having HA involved in the forming aggregates enriching HA. Possible nature of low-molecular HA-containing structures involved in the aggregates and nature of relations stabilizing aggregates are discussed.

Animals↗
PubMed2004

[Preclinical diagnosis of renal damage in essential hypertension].

AIM: To study renal functional reserve and 24-h beta 2-microglobulinuria (B2-MGU) in patients with different stages of essential hypertension (EH) and to evaluate acceptability of these parameters as early markers of preclinical renal impairment in EH. METHODS: We examined 150 patients (mean age 42.1 +/- 3.4 years) with EH without overt clinical manifestations of hypertensive nephropathy. Renal functional reserve (RFR) was determined with the use of acute oral protein loading and was defined as an increase in endogenic creatinine clearance in percent. B2-MGU was evaluated by radioimmunoassay (RIO-BETA-2-MICRO kit). Radioimmunoassay detection of microalbuminuria (MAU) was performed with the use of the standard kit Albumin-RIA. RESULTS: RFR appeared to be significantly lower in patients with EH compared to the control group, and its level significantly decreased with aggravation of EH. Both B2-MGU and MAU were significantly higher in patients with EH and significantly increased with aggravation of EH. CONCLUSION: We suggest that, apart from MAU, RFR abnormalities and excessive B2-MGU should be used as markers of preclinical renal impairment in EH.

Adult↗