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Biomedical subjects

E Bucht

Publications and source records attributed to E Bucht.

53 records · Page 3Linked to original sources

Postmenopausal osteoporosis: response of immunoextracted calcitonin to a calcium clamp.

Fourteen postmenopausal females with osteoporosis (PMO) and 14 healthy postmenopausal females (PMF) were studied by means of the calcium clamp-technique. Plasma was collected before start and during the 1 hour calcium clamp. Calcitonin (CT) was immunoextracted from plasma with antiserum No. 1 directed against the mid- and carboxyterminal portion of CT. Thereafter radioimmunoassay was performed on the extracts with antiserum No. 2 directed against the carboxyterminal of CT. The detection limit of the assay was 0.8 pg/tube. There was no difference between patients and controls in the basal CT values and CT was detectable in all samples after immunoextraction. Plasma CT increased significantly during the calcium clamp in osteoporotic patients as well as in controls and the CT response did not differ between the groups. The results do not support the hypothesis of a diminished CT secretion as a contributing factor to the development of osteoporosis.

Aged↗

Serum calcitonin levels in anesthetized rats.

In order to determine whether a short period of general anesthesia influences the levels of serum calcitonin (CTs) and whole blood ionized Ca2+ and pH, 10 rats were sequentially anesthetized at 5-day intervals by halothane, ether, or CO2/O2, respectively, and retroorbital blood samples were collected during anesthesia. We noticed significant differences of serum calcitonin but the role of the anesthetics remains unclear, since other factors also could have accounted for the observed variations. Blood pH was strongly decreased by CO2/O2. Whole blood ionized calcium exhibited marked changes, but no correlation was found between whole blood ionized calcium and serum calcitonin.

Anesthesia, Inhalation↗

The influence of sex steroid hormones on plasma calcitonin response to the calcium clamp in normal subjects.

The influence of sex steroid hormones on plasma calcitonin levels in healthy subjects was studied in 15 males and 10 premenopausal (PREMF) and 12 postmenopausal females (PMF). A standardized ionized calcium stimulus was achieved by means of the calcium clamp technique, and a sensitive RIA was used to determine immunoreactive CT (iCT) in plasma. Plasma iCT levels increased in response to the calcium clamp with an initial peak at 15 or 30 min. The iCT levels then declined but remained at an elevated level for the rest of the 180-min infusion period in all three groups. In males a positive correlation was found between the serum testosterone levels at 0 min and the estimates for initial iCT response, i.e., the change between 0 and 15 min (r = 0.80, n = 11, p less than 0.01), between 0 and 30 min (r = 0.56, n = 15, p less than 0.05) and between 0 and the maximum value (r = 0.68, n = 15, p less than 0.01). In the two female groups no such correlation was found. Serum 17 beta-estradiol or dehydroepiandrosterone sulfate concentrations were not correlated either to basal iCT levels or to the iCT response to the calcium clamp. PMF had lower levels of 17 beta-estradiol than PREMF (p less than 0.001), while the testosterone levels were similar. The iCT levels and response to the calcium clamp showed no significant difference in the two groups. The results indicate that sex difference in the plasma CT levels may be related to different testosterone levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Evidence for precursors of calcitonin/PDN 21 in human milk.

Large amounts of immunoreactive PDN-21 (iPDN -21) were found in human milk in concentrations similar to those of immunoreactive calcitonin (iCT): 187 +/- 73 pM (mean +/- S.D.) vs 210 +/- 83 pM. (n = 17). Calcitonin (CT) was immunoextracted from milk by means of CT antibodies coupled to Sepharose 4B, and the extracts were gel-chromatographed on Sephadex G-75 after treatment with 6 M guanidine HCl. iCT and iPDN-21 in the fractions were determined with radioimmunoassay. The main part of iCT eluted as high molecular weight forms and these fractions also contained iPDN-21. Enzymatic cleavage of immunoextracted milk CT by trypsin demonstrated that iPDN-21 could be split apart from the high molecular weight forms and be recovered at the position of synthetic human PDN-21 on gel chromatography. iCT was eluted in the region of monomeric CT and as larger forms. Since PDN-21 constitutes the carboxyterminal of preprocalcitonin, our results indicate that human milk contains precursors of calcitonin.

Calcitonin↗

Melatonin, cortisol, prolactin, and calcitonin secretion in primary hyperparathyroidism before and after surgery.

The aim of the present study was to determine the diurnal secretion of melatonin, cortisol, prolactin, and calcitonin during chronic parathyroid hormone-dependent hypercalcemia. Eight women, aged 40-76 years, with primary hyperparathyroidism (PHPT) were studied before and after surgical removal of a parathyroid adenoma. The hormone concentrations in blood were determined at 08, 12, 16, 22, 02, 04, and 06 h. Concomitantly, the excretion of melatonin and cortisol in urine between 07-19 h and 19-07 h, and the clearance of calcium and creatinine were measured. Nyctohemeral serum prolactin and calcitonin were unaffected by moderate parathyroid hormone-dependent hypercalcemia. In contrast, serum cortisol and melatonin were significantly higher during active disease than after surgical cure. Mean 24-h variation of serum cortisol was 349 +/- 34 nmol/liter vs. 223 +/- 17 nmol/liter and mean serum melatonin was 0.13 +/- 0.04 nmol/liter vs. 0.06 +/- 0.02 nmol/liter. Endogenous creatinine clearance was similar before and after surgery, while the clearance of melatonin and cortisol significantly increased after surgery, indicating an increased tubular reabsorption of both hormones during active disease. Fasting morning glucose concentrations were also significantly decreased after successful surgery, 6.1 +/- 0.6 vs. 5.2 +/- 0.5 mmol/liter. It is suggested that the relative hypercortisolism may be the cause of the glucose intolerance in primary hyperparathyroidism. Three to 4 months after surgical cure the serum melatonin levels were significantly lower than those seen in age-matched controls, indicating a melatonin insufficiency in patients successfully treated for PHPT. The meaning of this finding is not yet understood but might be of importance in the development of primary hyperparathyroidism.

Adenoma↗

Large forms of immunoreactive calcitonin in human seminal fluid exhibit PDN-21 immunoreactivity.

Semen samples were obtained from 4 healthy males on two occasions. One of the samples was liquefied at room temperature for 20 min while the other was diluted immediately with buffer to suppress proteolysis. A pool of immunoextracted calcitonin from liquefied and diluted samples, respectively, was subjected to gel chromatography in a separate experiment. In lyophilized samples the majority of immunoreactive calcitonin (CT) had an approximate molecular weight (Mr) of 10 kilodaltons (Kd) and immunoreactive PDN-21 was also present in the fractions. In diluted ejaculates CT was composed of larger species as well as the 10 Kd form. PDN-21 was present in fractions containing the 10 Kd form but could not be detected in the peaks of larger molecular weight, probably because of lower sensitivity of the PDN-21 assay compared to the CT assay. Our finding of CT and PDN-21 in the same fractions after immunoextraction with CT antibodies suggests the presence of proforms of CT in human seminal fluid.

Calcitonin↗

Immunoextracted calcitonin from human seminal plasma constitutes different forms.

Levels of immunoreactive calcitonin (iCT) in human seminal plasma were studied after extraction with immobilized antibodies. Immunoextraction of semen was necessary to abolish the high concentrations of proteolytic enzymes that can interfere in radioimmunoassay (RIA). Since proteolysis occurs already during semen liquefaction, iCT levels and chromatographic properties were compared in extracts from liquefied ejaculates, and samples were the proteolysis was suppressed by immediate dilution of the ejaculates with buffer. The levels in the two groups were 81 +/- 43 pg-equivalents ml-1 seminal plasma (mean +/- SD, n = 6) in liquefied samples and 120 +/- 42 pg-equivalents ml-1 (n = 7) in diluted samples, which is about ten times the iCT levels in extracted blood plasma from healthy males. Extracted liquefied seminal plasma from two vasectomized men contained 93 and 107 pg-equivalents of iCT ml-1 seminal plasma, respectively. Gel chromatography of undiluted semen samples disclosed that iCT was composed of forms larger than monomeric CT, the major part with an approximate molecular weight (MW) of 10,000. Two of the six subjects had a smaller amount of iCT eluting with a MW of about 40,000. In diluted samples, iCT seemed to be more heterogenous with approximate molecular weights ranging from 40,000 to fragments smaller than monomeric CT. In some ejaculates, monomeric CT seemed to be present. Undiluted samples from vasectomized men contained iCT with the same chromatographic pattern as in normal men.

Calcitonin↗

Immunoextracted calcitonin in milk and plasma from totally thyroidectomized women. Evidence of monomeric calcitonin in plasma during pregnancy and lactation.

The level of immunoreactive calcitonin in the first produced breast milk was in totally thyroidectomized (TX) women 713 +/- 307 pg-eq/ml (mean +/- SD, N = 7) and in control women 772 +/- 329 pg-eq/ml (N = 33), i.e. about 45 times higher than in plasma (see below). On gel chromatography of immunoextracted milk from TX women, immunoreactive calcitonin appeared as high molecular weight forms in the same pattern as in milk from healthy women when the same antiserum was used for immunoextraction and radioimmunoassay (RIA). In another series of experiments, a new antiserum raised in rabbits was used for measurement of immunoreactive calcitonin after immunoextraction with 1. Plasma levels of immunoreactive calcitonin in the TX women during pregnancy were 16 +/- 6 pg-eq/ml (N = 6) and during lactation 14 +/- 7 pg-eq/ml (N = 5). Immunoreactive calcitonin was undetectable (less than 8 pg/ml) in plasma from those TX women in whom lactation had stopped (N = 5). Immunoextraction and gel chromatography of plasma collected during pregnancy and lactation from the TX women showed that the immunoreactive calcitonin present eluted in the region of monomeric calcitonin with both antiserum 1 and 2. In conclusion, high concentrations of high molecular weight forms of of immunoreactive calcitonin have been demonstrated in milk from TX patients, most probably devoid of any calcitonin-producing thyroid C-cells. This points to a local production site in the mammary gland.(ABSTRACT TRUNCATED AT 250 WORDS)

Calcitonin↗

Plasma calcitonin response to a calcium clamp in endogenous Cushing's syndrome.

Basal and calcium (Ca) stimulated plasma CT concentrations have been studied in 6 premenopausal females with Cushing's disease and 3 premenopausal females with Cushing's syndrome due to an adrenocortical adenoma. Thirteen healthy premenopausal females served as a reference group. A standardized Ca-stimulus was applied to all subjects by means of the 'calcium clamp' technique. Plasma immunoreactive CT (iCT) was determined by RIA using a carboxyl terminal antiserum with 8 pg-eq/ml as detection limit. The patients with Cushing's disease had normal basal plasma iCT levels and their iCT response during the 2 h calcium clamp was within the reference range in 5 out of 6 patients, the remaining patient showed a slightly exaggerated response. The patients with adrenal adenoma had elevated plasma iCT levels in the basal state but a normal response during the calcium clamp. It is therefore less likely that the bone resorption often seen in Cushing's syndrome is due to CT deficiency.

Adenoma↗

Plasma calcitonin response to a calcium clamp. Influence of sex and age.

The influence of sex and age on basal and calcium stimulated immunoreactive plasma calcitonin (iCT) were studied in 36 healthy subjects. An identical calcium stimulus, "calcium clamp", was applied to 10 young males, 13 premenopausal females of similar ages and 13 postmenopausal females. A significantly greater iCT response was found in males compared with both female groups. No age correlation in basal or calcium stimulated iCT levels was observed despite 35 years difference in mean age between the pre- and postmenopausal groups. A positive correlation between the induced change in whole blood ionized calcium (B-Ca++) and the iCT response, recorded after 15 min, was seen in young subjects of both sexes. Basal iCT and B-Ca++ were similar in all three groups. No correlation between basal iCT and basal B-Ca++, or between basal iCT and age was found. The total amounts of infused calcium per kg body weight were equal. The results indicate that male subjects have a greater secretory capacity of the thyroid C-cell than females. No reduction of the secretory capacity was observed with increasing age in females, neither was any difference found between pre- and postmenopausal females of the iCT-response.

Adult↗

Decreased plasma calcitonin response to a calcium clamp in primary hyperparathyroidism.

In order preoperatively to evaluate the calcium induced plasma calcitonin (CT) response in patients with mild primary hyperparathyroidism (PHPT), 7 postmenopausal (PM) females with PHPT and 14 PM healthy subjects underwent a 2 h calcium infusion. The infusions were performed by means of the 'calcium clamp' technique previously described by us in order to obtain an identical calcium stimulus in all subjects. Immunoreactive calcitonin (iCT) was determined by means of a sensitive RIA utilizing rabbit antibodies directed against the carboxy-terminal amino acid sequence of the CT molecule. All patients with PHPT had normal basal plasma iCT levels: 10 +/- 5 pg-eq/ml (mean +/- SD) as a group and not different from the control group: 13 +/- 5 pg-eq/ml. In the control group significantly increased plasma iCT was seen at 30, 60, 90 and 120 min in contrast to that of the PHPT group where a blunted response was observed. The results indicate a diminished CT reserve in these patients which may imply an impaired defence against the PTH mediated bone resorption seen in patients with PHPT.

Adenoma↗

Gel chromatography of immunoextracted plasma calcitonin in response to the calcium clamp in healthy males.

A radioimmunoassay (RIA) was developed for immunoreactive calcitonin (iCT) in human plasma. Antibodies against synthetic human calcitonin (hCT) coupled to bovine serum albumin (BSA) were raised in rabbits and were directed against the carboxy terminal part of CT. The detection limit of the assay was 8 pg/ml. In 7 males the iCT response to a calcium-clamp was studied. Blood was collected at 0, 30 and 60 min after the start of the calcium infusion. iCT was measured directly in plasma and in extracts obtained after purification of plasma iCT by means of immobilized CT antibodies. There was a good correlation between iCT in plasma samples and extracts, r = 0.993, n = 14 (P less than 0.001). Dilution curves of extracts and plasma were parallel with the hCT standard curves. Gel chromatography of the extracts on Sephadex G-50 and G-75 disclosed heterogeneity of iCT in normal plasma during basal conditions as well as during calcium stimulation. Thirty min after the start of the calcium clamp all molecular forms, most likely constituting monomeric and dimeric CT and larger forms, were increased, while after 60 min iCT seemed to constitute predominantly forms larger than monomeric CT. Basal levels of unextracted iCT in healthy males (n = 44, 37 +/- 10 years) were 15 +/- 9 pg-equivalents/ml (mean +/- SD) which was higher than in females (n = 40, 32 +/- 9 years) 11 +/- 4 pg-equivalents/ml (P less than 0.05).

Adult↗

Calcitonin-like immunoreactivity in human milk, longitudinal alterations and divalent cations.

The concentration of immunoreactive calcitonin (iCT) in human milk was 10-40 times the serum level. Thus in milk samples collected from 29 mothers within one week and from 46 mothers three months post partum the levels of iCT were respectively 2.46 +/- 0.99 ng/ml (mean +/- SD) and 0.58 +/- 0.08 ng/ml. Eight of the 29 mothers delivered milk at two occasions, 1-2 and 4-6 days after delivery. There was a significant decline of iCT with about 20% during this period. One mother delivered milk for two months. During the first week after delivery the iCT decreased from 1.4 to 0.5 ng/ml and after that the iCT content remained rather constant. The level of ionized calcium (Ca++) was correlated to iCT the first week post partum (r = 0.72 p less than 0.01) but not in milk samples obtained after three months. The concentration of magnesium was weakly correlated to iCT during the first week post partum (r = 0.37 p less than 0.05). There was no correlation between iCT and the total calcium level nor to the concentration of zinc. The high concentration of iCT in milk compared to serum suggests a local production of iCT within the mammary gland or a specific transport and concentration mechanism.

Calcitonin↗

Heterogeneity of immunoreactive calcitonin in human milk.

Immunoreactive calcitonin (ICT) has been detected in human milk by radioimmunoassay (RIA), using antibodies raised against synthetic human calcitonin (hCT). The level of ICT was 10-40 times higher than the reference level of human serum. Gel chromatography of human milk disclosed a molecular weight heterogeneity of ICT, with at least two forms larger than the monomer (Mr greater than 70000 and 30000 approximately). The 30000 peak constituted the major fraction of ICT. No detectable amount of ICT was coeluted with [125I]CT. Reduction with disulphide cleaving agents, denaturation or acidification did not alter the gel chromatographic properties of ICT, while tryptic digestion caused partial degradation of ICT and formation of new ICT molecules with an approximate molecular weight of 6000.

Calcitonin↗

Quantitation of phenobarbital and its main metabolites in human urine.

A method for the quantitative determination of phenobarbital and free and conjugated p-hydroxyphenobarbital in urine samples is described. The method includes initial extraction, purification on a small chromatographic column and finally determination by gas chromatography. The barbituric acids are methylated by trimethylanilinium hydroxide which serves as a "flash heater" methylating agent. The conjugate of p-hydroxyphenobarbital, which appears to be a glucuronide, is hydrolysed with hydrochloric acid.

Child↗

Immunoextracted calcitonin in human gastric secretion.

Immunoreactive calcitonin (iCT) has been demonstrated in human gastric juice after immunoextraction with immobilized antibodies and subsequent radioimmunoassay. The basal levels were 4.5 +/- 3.1 (mean +/- SD) pg-Eq/ml gastric juice; range 1.2-9.1 pg-Eq/ml; n = 7, and after stimulatory gastric secretion test with pentagastrin 0.3 +/- 0.2 pg-Eq/ml; range 0.1-0.7 pg-Eq/ml; n = 7 (p less than 0.01). The main fraction of iCT from gastric juice eluted in the same region as synthetic human calcitonin (hCT) on Sephadex G-75 gel chromatography. Reverse phase chromatography in a fast protein liquid chromatography (FPLC) system revealed a slightly less hydrophobic character of the iCT from gastric juice compared to synthetic monomeric hCT. The results were further confirmed by using an additional antiserum. In plasma, the calcitonin (CT) levels were after immunoextraction at the basal state 6.6 +/- 1.7 pg-Eq/ml (mean +/- SD); range 5.1-10.1 pg-Eq/ml; n = 7 and after pentagastrin stimulation 9.4 +/- 5.4 pg-Eq/ml; range 6.3-18.5 pg-Eq/ml; n = 7.

Adult↗