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  • Immunocytochemistry  (41)
  • healthy volunteers  (37)
  • Column liquid chromatography  (26)
  • Springer  (104)
  • Elsevier
  • 1980-1984  (104)
  • 1970-1974
  • 1955-1959
  • 1984  (104)
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  • 1980-1984  (104)
  • 1970-1974
  • 1955-1959
Year
  • 1
    ISSN: 1432-041X
    Keywords: Major haemolymph proteins ; Development ; Cuticle ; Immunocytochemistry ; Ceratitis capitata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The developmental profile of the major haemolymph proteins (ceratitins) inCeratitis capitata was studied. Ceratitin concentration in the haemolymph decreases dramatically during the last days of pupal life, while the amounts of ceratitins in whole organism extracts remain unchanged. By electrophoretic, immunological and immunofluorescence techniques it was revealed that ceratitins are reabsorbed by the fat body and a fraction of them is deposited in the cuticle. The possible role of ceratitins is discussed.
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  • 2
    ISSN: 1432-2048
    Keywords: Immunocytochemistry ; Lectin (localization) ; Phaseolus (lectin) ; Phytohemagglutinin ; Seed (lectin)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have examined the properties and subcellular localization of phytohemagglutinin (PHA), the major lectin of the common bean (Phaseolus vulgaris.), in the axis cells of nearly mature and imbibed mature seeds. On a protein basis the axis contained about 15% as much PHA as the cotyledons. Localization of PHA was done with an indirect immunolabeling method (rabbit antibodies against PHA, followed by colloidal gold particles coated with goat antibodies against rabbit immunoglobulins) on ultra-thin cryosections which were embedded in plastic on the grids after the immunolabeling procedure. The embedding greatly improved the visualization of the subcellular structures. The small (4 nm) collodial gold particles, localized with the electron microscope, were found exclusively over small vacuoles or protein bodies in all the cell types examined (cortical parenchyma cells, vascular-bundle cells, epidermal cells). The matrix of these vacuoles-protein bodies appears considerably less dense than that of the protein bodies in the cotyledons, but the results confirm that in all parts of the embryo PHA is localized in similar structures.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    European journal of clinical pharmacology 26 (1984), S. 627-630 
    ISSN: 1432-1041
    Keywords: bupropion ; ethanol ; pharmacokinetic interaction ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary A study was performed to determine whether there is a pharmacokinetic interaction between alcohol and the novel antidepressant bupropion. In the first part 8 healthy male volunteers received single doses of 100 mg bupropion hydrochloride orally on 2 occasions accompanied by either ethanol in orange or plain orange drink according to a balanced cross over design. Plasma bupropion concentrations were determined by radioimmunoassay and kinetics analysed with the aid of NONLIN. Blood alcohol levels were assessed by breathalyser. The disposition of bupropion was adequately described by a 2 compartment model and kinetic parameters were not significantly altered by the presence of alcohol. In the second part of the study the same subjects received 40 ml ethanol in orange drink 3.5 h after ingestion of 100 mg bupropion or dummy tablet in a double blind cross over fashion. Bupropion did not affect alcohol kinetics. In contrast to many other psychotropic drugs there is no evidence for a kinetic interaction between bupropion and alcohol.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    European journal of clinical pharmacology 26 (1984), S. 635-637 
    ISSN: 1432-1041
    Keywords: theophylline ; inhalation ; saliva-serum distribution ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Six healthy volunteers received an iv infusion of 317 mg lysine theophylline (equivalent to 197 mg anhydrous theophylline) in order to calculate theophylline clearance by standard methods. They subsequently received a 20 minute inhalation of nebulised lysine theophylline. Serum and salivary theophylline concentrations were measured and all saliva was collected for the first hour. From these concentrations estimates were made of the distribution of theophylline into the blood and saliva with 40% to 94% identified in the blood. Very high salivary concentrations were reached during the inhalation phase with saliva: serum concentration ratios of between 60 and 1600.
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  • 5
    Electronic Resource
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    Springer
    European journal of clinical pharmacology 27 (1984), S. 115-117 
    ISSN: 1432-1041
    Keywords: benzodiazepine antagonist ; Ro 15-1788 ; healthy volunteers ; pharmacokinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The pharmacokinetics of the selective benzodiazepine antagonist Ro 15-1788 has been studied in 6 healthy male volunteers following a single intravenous dose of 2.5 mg. The drug was only slightly bound to plasma proteins (40±8%, mean±SD). A negligible amount (〈0.2% of the dose) of unchanged drug was recovered in urine. Hepatic elimination was rapid, as shown by a short t1/2 of 0.9±0.2 h, and high total plasma and blood clearances of 691±216 ml/min and 716±199 ml/min, respectively. The fast decline of plasma levels from about 60 to 2 ng/ml accounts for the short-lasting reversal of benzodiazepine-induced sedation by Ro 15-1788.
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  • 6
    ISSN: 1432-1041
    Keywords: nitroglycerin ; plasma concentration ; transdermal administration ; bioavailability ; pharmacodynamics ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary In 6 healthy volunteers, intravenous infusions of nitroglycerin 4.8 and 10.6 µg/min yielded mean steady-state plasma concentrations of 0.5±0.02 and 0.82±0.04 ng/ml as determined by a gas chromatographic/mass spectrometric method. The plasma concentrations reached in the same subjects 17 h after application of Nitroderm TTS 5 and 10 with in vivo release rates of 3.7 and 5.7 µg/min were 0.28±0.01 and 0.37±0.01 ng/ml, respectively. Thus, 75% of the quantity of nitroglycerin released by the systems passed into the circulation. The inter-individual and intra-individual variations in plasma concentrations were similar for both modes of administration. The nitroglycerin-induced morphological changes in the fingerpulse wave were clearly dose-dependent, but it seems that this pharmacodynamic parameter is determined less by the plasma concentration than by the nitroglycerin content of the vascular wall.
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  • 7
    Electronic Resource
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    Springer
    European journal of clinical pharmacology 27 (1984), S. 371-373 
    ISSN: 1432-1041
    Keywords: disodium chromoglycate ; nasal airway resistance ; platelet activating factor ; healthy volunteers ; rhinomanometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Thirteen healthy subjects participated in a double blind, randomized, cross-over investigation of whether or not intranasal disodium chromoglycate could block the change in nasal airway resistance induced by platelet activating factor (PAF). Placebo or active drug was given for 3 days before intranasal challenge with PAF. Nasal airway resistance before and at intervals after callenge was determined with a rhinomanometer. Pretreatment with disodium chromoglycate blocked the decrease in nasal airway resistance induced by PAF. This indicates an alternative mode of action of disodium chromoglycate in the treatment of allergic airway diseases.
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  • 8
    ISSN: 1432-1041
    Keywords: spironolactone ; canrenone ; metabolites ; pharmacokinetics ; single/multiple oral doses ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The pharmacokinetics of canrenone and ‘total metabolites’ after base hydrolysis was studied in eight young volunteers following single and multiple dose oral administration of spironolactone. The plasma levels of canrenone and ‘total metabolites’ were fitted to a two-compartment open model with a first-order absorption process. From our eight normal subjects studied, the harmonic mean of the distributive half-life (t1/2α) of canrenone was found to be 1.66 h, and the harmonic mean of the terminal elimination half-life (t1/2β) to be 22.6 h. Harmonic means of the distributive and elimination half-lives of ‘total metabolites’ after base hydrolysis were 2.48 h and 28.8 h respectively. The accumulation ratio of canrenone was 2.53, whereas that of ‘total metabolites’ was 1.89. Despite the fact that spironolactone has been shown to induce hepatic metabolism of other drugs, no evidence of autoinduction was noted in the present study, as plasma levels of canrenone and ‘total metabolites’ were found to obey a linear two-compartment model with reproducible absorption and disposition after single and multiple doses.
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  • 9
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    European journal of clinical pharmacology 27 (1984), S. 447-452 
    ISSN: 1432-1041
    Keywords: alaproclate ; antipyrine clearance ; serotonin reuptake inhibitor ; healthy volunteers ; antipyrine metabolism ; metabolite clearance ; alaproclate kinetics ; inhibition of drug metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Alaproclate, a selective serotonin reuptake inhibitor, presently undergoing clinical trial for the treatment of major depressive disorders, has been shown to inhibit hexobarbital metabolism in mice. In the present study the influence of oral alaproclate on the total plasma clearance of antipyrine and on the formation of its metabolites was investigated in 10 healthy volunteers. The peak level of alaproclate was reached after about 1.5 h, and after a distribution phase, its plasma elimination half-life was between 3.0 and 3.5 h. Antipyrine tests were performed before treatment, during the first four doses and after the seventh dose of alaproclate 200 mg/day. During treatment, total plasma antipyrine clearance and the clearance for production of all antipyrine metabolites were reduced by 30%, indicating non-selective inhibition of oxidative drug-metabolizing enzyme activity in man by alaproclate. After the last dose of alaproclate, antipyrine plasma clearance and the clearance to its metabolites returned to control values. In order to allow more detailed evaluation of the results, the time course of the clearances for production of metabolites was investigated. This revealed that the extent of inhibition of metabolite formation by alaproclate was dependent on the plasma alaproclate level, indicating a rapidly reversible inhibition.
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  • 10
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    Springer
    European journal of clinical pharmacology 27 (1984), S. 471-475 
    ISSN: 1432-1041
    Keywords: acyclovir ; A515U ; 6-deoxyacyclovir ; pharmacokinetics ; prodrug ; antiviral chemotherapy ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary A515U (6-deoxyacyclovir) is an analogue of acyclovir devoid of antiviral activity in vitro but which is well absorbed and undergoes conversion to acyclovir after oral administration to rats. The tolerance and pharmacokinetics of various doses of A515U have been studied in 8 healthy volunteers. Single oral doses of 25, 50, 100, 200 and 400 mg A515U and 400 mg acyclovir for comparison were administered to the volunteers at weekly intervals. Concentrations of the parent drug and acyclovir were determined in plasma and urine. The prodrug was well tolerated and did not cause adverse reactions or changes in haematological or biochemical variables. It was well absorbed and conversion to acyclovir was rapid and extensive at all doses. Plasma concentrations of acyclovir achieved with 50 mg A515U orally were comparable to and less variable than those produced by 400 mg acyclovir. A515U was rapidly cleared with a short plasma elimination half life of approximately 0.5 h. The attainment of high plasma concentrations of acyclovir by oral administration of a prodrug may represent an important advance in antiviral chemotherapy.
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  • 11
    ISSN: 1432-1041
    Keywords: benzodiazepine antagonist ; visual function ; intrinsic effect ; Ro 15-1788 ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Ro 15-1788 is a specific benzodiazepine antagonist, which has also been shown to have some agonist properties. Since benzodiazepine receptors are involved in the physiological mechanisms of vision, a possible intrinsic effect of Ro 15-1788 was sought in 6 healthy volunteers by study of psychophysical flicker thresholds, including critical fusion frequency and low frequency modulation threshold, and pattern reversal visual evoked response, using double blind cross-over methodology. In each session 2 tablets of Ro 15-1788 30 mg were administered. Using a two factorial univariate analysis of variancé, no change was detected in any of the parameters studied.
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  • 12
    Electronic Resource
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    European journal of clinical pharmacology 27 (1984), S. 535-538 
    ISSN: 1432-1041
    Keywords: pindolol ; healthy volunteers ; heart rate ; intrinsic sympathomimetic activity ; diurnal heart rate ; nocturnal heart rate ; rebound effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The effect on heart rate of pindolol 5, 15 and 30 mg/day, a beta-adrenoreceptor blocker possessing intrinsic sympathomimetic activity, administered to 8 healthy volunteers for 14 days was studied. Heart rate was continuously recorded over 24 h during placebo treatment before each sequence, every 2 days during treatment, and then on the 15th, 17th and 18th days. Pindolol in the three doses used had no significant effect on mean heart rate over 24 h. It tended to lower mean diurnal heart rate non-significantly between noon and 6 p.m. Pindolol raised nocturnal heart rate between midnight and 6 a.m. to a comparable extent at all the doses used. Sympathetic tone is at its lowest during that period, which makes it possible to detect the intrinsic sympathomimetic activity of pindolol. After cessation of treatment, a rebound effect was observed, cardioacceleration being most marked after 30 mg/day.
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  • 13
    ISSN: 1432-1041
    Keywords: zetidoline ; prolactin ; aldosterone ; dopamine ; healthy volunteers ; pharmacodynamic effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The neuroendocrinological effects of acute oral administration of 20 mg zetidoline, a new antipsychotic drug with antidopaminergic properties, were evaluated in 8 healthy volunteers, by a double-blind, crossover comparison with placebo. Zetidoline significantly increased serum prolactin (p〈0.01 at 1–3 h; p〈0.05 at 4–6 h). No significant change was observed in blood levels of aldosterone, renin, cortisol, growth hormone and electrolytes, or in blood pressure and heart rate. The data suggest that the drug increases prolactin through blockade of dopaminergic receptors. The lack of change in the aldosterone levels may be evidence against the hypothesis of dopaminergic control of aldosterone secretion.
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  • 14
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    European journal of clinical pharmacology 26 (1984), S. 463-470 
    ISSN: 1432-1041
    Keywords: dihydroergotamine ; 8′-hydroxy-dihydroergotamine ; plasma metabolites ; bioavailability ; receptor affinity ; healthy volunteers ; liver microsomal incubates
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The plasma concentrations and urinary excretion of dihydroergotamine and its metabolites have been measured after a single oral administration of 3 mg tritium-labelled drug to 6 male volunteers. The plasma level of non-volatile radioactivity declined biphasically with α- and β-phase half-lives of 2.1 h and 32.3 h, respectively. The peak plasma concentration was reached within 3.2h. Urinary excretion of total non-volatile radioactivity was low, amounting to 1.0% of the dose. The parent drug and four metabolites could be quantitated in urine and plasma samples. Metabolite 4 (8′-hydroxy-dihydroergotamine) was isolated from incubates of rat and monkey liver microsomal preparations. In human liver microsomal incubates, metabolite 4 was shown to be the primary metabolite of dihydroergotamine. In receptor binding studies performed with mammalian brain preparations, metabolite 4 had IC50-values at 6 monoaminergic binding sites similar to those of dihydroergotamine. Thus, it appears that the active principle consists at least of dihydroergotamine and its 8′-hydroxy derivative. As the concentration of metabolite 4 exceeded 5–7 times that of dihydroergotamine in urine and plasma, the bioavailability of dihydroergotamine should be reevaluated, taking into account the plasma concentrations of the parent drug and of its acitve metabolite, 8′-hydroxydihydroergotamine.
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  • 15
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    European journal of clinical pharmacology 27 (1984), S. 191-196 
    ISSN: 1432-1041
    Keywords: pizotifen 5-hydroxyindoleacetic acid ; homovanillic acid ; urinary excretion ; healthy volunteers ; migraine prevention
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary A single dose of 0.5 mg pizotifen or a placebo was administered to 10 healthy male volunteers in a double blind cross-over trial. 5-hydroxyindoleacetic acid (5-HIAA) and homovanillic acid (HVA) in hourly urine samples were determined by liquid chromatography with amperometric detection. The 5-HIAA levels were strongly correlated with the HVA levels in control samples (r=0.95, p〈0.001). Pizotifen produced a significant increase in the urinary 5-HIAA/HVA ratio over the 3 hours following absorption of the drug (+0.21, +0.18, +0.19, p〈0.05). The increase demonstrates an interaction between pizotifen and 5-HT metabolism, which may be involved in its antimigraine effect.
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  • 16
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    European journal of clinical pharmacology 27 (1984), S. 483-489 
    ISSN: 1432-1041
    Keywords: radiosensitiser ; pharmacokinetics ; healthy volunteers ; tumour patients ; Ro 03-8799
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary A new hypoxic cell radiosensitiser, Ro 03-8799 has been administered intravenously to human volunteers and its kinetic parameters derived from plasma and urine data. Good penetration of drug into tumour tissue is found, consistent with its large volume of distribution. The plasma clearance of this compound is rapid due to high metabolic and renal clearances. These parameters combine to produce an elimination half-life of 5.6 h, approximately half that of misonidazole, a well studied radiosensitiser. It is hoped that this decrease in total body exposure will also reduce the cumulative toxicity seen when misonidazole is administered repeatedly.
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  • 17
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    European journal of clinical pharmacology 27 (1984), S. 501-503 
    ISSN: 1432-1041
    Keywords: valpromide ; valproic acid ; pharmacokinetics ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The pharmacokinetics of valpromide, a primary amide of valproic acid, was investigated in 6 healthy, adult male volunteers, each of whom was given 900 mg as a marketed, enteric-coated tablet and a solution. Valpromide was biotransformed to valproic acid after the administration of the tablet and the solution with a bioavailability of 0.79±0.24 and 0.77±0.12, respectively, relative to a marketed tablet of valproic acid. The absorption of valpromide was not rate-limited by dissolution. As a solid, non-hygroscopic, neutral prodrug of valproic acid, valpromide may be a good alternative to valproic acid and sodium valproate.
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  • 18
    ISSN: 1432-1041
    Keywords: trimethoprim ; concentration ; urinary excretion ; healthy volunteers ; steady state ; pharmacokinetics ; serum creatinine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The steady state pharmacokinetics of trimethoprim was determined after 300 mg orally once daily to 6 healty volunteers for 9 days. The microbiological assay of plasma level was unreliable at trimethoprim concentrations 〉4 µg/ml, so results from an HPLC-assay are given. Steady state was present after 3 days. The plasma concentration peaked 1 to 4 h (mean 2.0 h) after the dose at a mean of 6.0 µg/ml (range 3.1–9.5 µg/ml); the minimum value was 1.5 µg/ml (range 0.6–2.9 µg/ml). The mean AUCss was 77 µg/ml · h and the mean plasma clearancess was 67 and 74 ml/min on Days 8 and 9. Renal clearance was about 60% of the plasma clearance. The average plasma half life was 10.6 h (range 8.7–15.3 h). Thus, there was considerable interindividual variation in all pharmacokinetic parameters. 72 h after the last dose trimethoprim was detectable in plasma in only 1 of the 6 subjects. The minimum urinary concentration of trimethoprim during treatment was always well above (range 22 to 220 µg/ml) the MIC values for most urinary tract pathogens. Therefore, a daily dose of 300 mg trimethoprim results in a therapeutic concentration in urine at steady state that lasts throughout the dosing interval and in most subjects probably lasts also for a further 24 h. Trimethoprim administration raised mean serum creatinine from 67 to 97 µmol/l, probably due to competitive inhibition of the tubular secretion of creatinine.
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  • 19
    ISSN: 1432-1041
    Keywords: acetylsalicylic acid ; platelet function ; salicylate ; controlled release formulation ; single dosing ; continuous dosing ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The extent to which a controlled release acetylsalicylic acid (ASA) formulation inhibited platelet function has been evaluated in single and chronic dosing studies. In the single dose study, the platelet inhibitory effect of the controlled release formulation was compared with that of an equivalent dose of soluble ASA and an equimolar dose of sodium salicylate (SA). In the chronic dosing study, ASA dose-response curves for platelet function, including cyclooxygenase activity, were determined for various doses (20–1300 mg) of the controlled release (enteric coated pellets) ASA formulation taken by volunteers daily for one week. Platelet function was assessed by the degree of inhibition of aggregation for several aggregating agents, and the degree of inhibition of activity of platelet cyclooxygenase quantified by the estimation of malondialdehyde (MDA) production. Plasma ASA and SA concentrations were also determined in each study. The controlled release product inhibited platelet function to the same extent as an equimolar dose of soluble ASA, but did so with much lower and sometimes undetectable peak systemic plasma ASA concentrations. SA, the direct metabolite of aspirin, did not have any effect on platelet function. The ASA dose-platelet function response curves obtained from chronic dosing with the controlled release formulation appeared to be similar to those reported previously for the soluble product. The inhibition of platelet function appeared to be unrelated to plasma ASA concentrations.
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  • 20
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    European journal of clinical pharmacology 27 (1984), S. 23-27 
    ISSN: 1432-1041
    Keywords: hypertension ; alpha2-adrenoceptors ; blood pressure ; clonidine ; guanfacine ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The time courses of the changes in plasma growth hormone and noradrenaline concentrations in response to 15 min infusions of clonidine 0.2 mgs and guanfacine 2 mgs, were studied in six normal volunteers, in a double-blind, randomised, crossover study. Plasma noradrenaline fell within 15 min of the commencement of drug administration, by 36±14% after clonidine (p〈0.05) and by 32±11% (p〈0.05) after guanfacine. Plasma growth hormone was not significantly elevated until the 30th minute to 12.0±4.7 lU/ml (p〈0.05) after clonidine and 14.7±11.5 lU/ml (p〈0.05) after guanfacine, having been undetectable prior to both drugs. The reduction in plasma noradrenaline by these α2-adrenergic agonists, prior to activation of central adrenoceptors as detected by changes in plasma growth hormone, is evidence for a peripheral component in their sympatholytic effect.
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  • 21
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    European journal of clinical pharmacology 27 (1984), S. 75-80 
    ISSN: 1432-1041
    Keywords: bupropion ; alcohol interaction ; healthy volunteers ; performance ; autonomic functions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The effects of bupropion and ethanol were examined alone and in combination in a placebo controlled, double-blind, crossover study in 12 healthy volunteers. Results were subjected to analysis of variance and differences ofp〈0.05 taken as significant. In the main study using the Wilkinson auditory vigilance test, no active treatment or combination of treatments produced significant change compared with placebo. However, when compared with bupropion 100 mg, vigilance was significantly impaired by 32 ml alcohol alone though not when combined with bupropion. No significant changes in reaction time or short term memory occurred. Visual analogue scales indicated that the subjects were mentally slower after alcohol 32 ml than after placebo. Combination of bupropion 100 mg with alcohol 32 ml abolished this difference. A similar pattern occurred with group ratings indicating mental sedation. Subjects were clearly able to differentiate between the 16 ml and 32 ml doses of alcohol when assessing their degree of inebriation. Combination of bupropion with alcohol made no difference to the ratings of inebriation. The top dose of alcohol tended to increase energy in the low frequency EEG bands. Combination of the top alcohol dose with bupropion, however, produced a significant reversal with lowered energy in the 4–7.5 Hz band. Combination of bupropion with alcohol failed to change the blood alcohol concentration achieved.
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  • 22
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    European journal of clinical pharmacology 27 (1984), S. 119-121 
    ISSN: 1432-1041
    Keywords: S-adenosyl-L-methionine ; pharmacokinetics ; protein binding ; dose-dependent kinetics ; healthy volunteers ; urinary excretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary S-Adenosyl-L-methionine (AdoMet) kinetics was studied in 6 male subjects given 100 and 500 mg i. v. Drug concentrations in plasma and urine were assayed using a radioenzymatic method. Pharmacokinetic parameters were estimated according to an open two-compartment model. The apparent volumes of distribution after the 100 and 500 mg doses were 407±27 and 443±36 ml/kg (mean±SEM), terminal half-lives 81±8 and 101±7 min and body clearances 3.7±0.5 and 3.1±0.2 ml/min per kg. Urinary excretion was 34±3 and 40±2% of the administered dose. The results demonstrate that drug disposition occurs more via metabolism than via renal excretion, and it is not dependent on the administered dose. Binding of AdoMet to serum proteins is negligible.
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  • 23
    ISSN: 1432-1041
    Keywords: nitroglycerin ; plasma concentration ; transdermal administration ; bioavailability ; pharmacodynamics ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary In 6 healthy volunteers, intravenous infusions of nitroglycerin 4.8 and 10.6 µg/min yielded mean steady-state plasma concentrations of 0.5±0.02 and 0.82±0.04 ng/ml as determined by a gas chromatographic/mass spectrometric method. The plasma concentrations reached in the same subjects 17 h after application of Nitroderm TTS 5 and 10 with in vivo release rates of 3.7 and 5.7 µg/min were 0.28±0.01 and 0.37±0.01 ng/ml, respectively. Thus, 75% of the quantity of nitroglycerin released by the systems passed into the circulation. The inter-individual and intra-individual variations in plasma concentrations were similar for both modes of administration. The nitroglycerin-induced morphological changes in the fingerpulse wave were clearly dose-dependent, but it seems that this pharmacodynamic parameter is determined less by the plasma concentration than by the nitroglycerin content of the vascular wall.
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  • 24
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    European journal of clinical pharmacology 27 (1984), S. 453-458 
    ISSN: 1432-1041
    Keywords: dipyrone ; metabolism ; metabolite pharmacokinetics ; acetylation polymorphism ; healthy volunteers ; dapsone phenotyping
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The pharmacokinetics of the dipyrone metabolites 4-methylaminoantipyrine (MAA), 4-aminoantipyrine (AA), 4-formylaminoantipyrine (FAA) and 4-acetylaminoantipyrine (AAA) were evaluated following the administration of a single oral 1.0 g dose of dipyrone to 23 healthy volunteers. Twelve were slow and 11 were rapid acetylators as previously determined by dapsone phenotyping. For MAA and FAA the mean peak plasma concentrations were 10.5±2.8 µg/ml and 2.1±0.8 µg/ml and the half-lives were 3.3±1.0 and 10.1±1.8 h, respectively. No significant difference was found between rapid and slow acetylators in MAA and FAA kinetics. For AA, the mean peak plasma concentrations were 2.7±0.6 and 1.6±0.7 µg/ml (p〈0.01), the peak times 6.7±2.1 and 3.1±1.1 h (p〈0.01) and the half-lives were 5.5±1.0 and 3.8±1.2 h in slow and rapid acetylators, respectively. For AAA, the mean peak plasma concentrations were 1.6±0.4 and 4.4±1.1 µg/ml (p〈0.01) and the peak time 16.1±5.1 and 10.0±2.6 h (p〈0.01) in slow and rapid acetylators, respectively. There was no difference in the elimination half-life between the two groups (10.6±2.2 h). Thus, it has been demonstrated that the AAA/AA ratio is an indicator of the acetylation phenotype, as it is closely correlated with that determined by dapsone (r=0.895, p〈0.0005).
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  • 25
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    European journal of clinical pharmacology 27 (1984), S. 495-497 
    ISSN: 1432-1041
    Keywords: ranitidine ; aldosterone secretion ; sodium depletion ; drug interference ; plasma renin activity ; biochemical parameters ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The effect of a 3-day oral course of ranitidine on plasma aldosterone level has been studied in 6 normotensive volunteers maintained in a state of sodium depletion. A significant fall in plasma aldosterone (p〈0.05–0.02), in both the overnight recumbency levels and in the levels obtained during a two hour period of ambulation was observed. The change took place in the absence of variation in plasma renin activity and potassium. Plasma cortisol and prolactin levels were lower after ranitidine at the beginning of the test but their values were not significantly different after ambulation during ranitidine therapy. Ranitidine appears to interfere with aldosterone secretion in vivo.
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  • 26
    ISSN: 1432-1041
    Keywords: cimotaxone ; MAO inhibitor ; plasma prolactin ; circadian rhythm ; healthy volunteers ; hypothalamic MAO ; prolactin secretion ; metabolism ; pharmacokinetics
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Prolactin (PRL) secretion is stimulated by serotonin (5-HT) and inhibited by dopamine (DA). 5-HT is generally recognized as a substrate for type A monoamine oxidase (MAO), whereas DA is considered as a substrate for either A or B, or both forms of MAO, depending on the species and tissues used. The effect of cimoxatone, a reversible, selective MAO-A inhibitor, on diurnal variation in plasma PRL level was investigated in healthy adults after a single 40 mg oral dose, as an indirect approach to investigating whether DA is preferentially a substrate for Type A or B MAO in man. The circadian rhythm in PRL, stress conditions and diet were taken into account in the present study, which was placebo-controlled. There was a slight but significant reduction in circulating PRL in the six subjects, which persisted for at least 9 h after cimoxatone. However, the duration of the decrease in plasma PRL was shorter than the inhibition of MAO-A. The results are not inconsistent with the presence of both forms of MAO in the human hypothalamus and with DA as a substrate for both forms in this region, if it is assumed that the hypothalamic concentrations of the drug during the period 0–9 hours was sufficiently high to inhibit DA deamination by both forms of MAO.
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  • 27
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    European journal of clinical pharmacology 26 (1984), S. 129-131 
    ISSN: 1432-1041
    Keywords: aprindine ; antiarrhythmic agent ; healthy volunteers ; plasma level ; oral administration ; pharmacokinetics ; urinary excretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The disposition of aprindine following a single oral dose can best be described by a two-compartment open model. The mean plasma half-life (t1/2β) increased from 8.0±2.1 h (SD) after a 25 mg dose to 9.4±2.9 h after 50 mg and to 15.8±2.6 h after 100 mg, with a decrease in total plasma clearance (Cl/F) and volume of distribution at steady state (Vdss/F) and during β-phase (Vdβ/F). The area under plasma concentration-time curve (AUC), maximum plasma concentration (Cmax) and the amount of unchanged aprindine excreted in the urine increased in a non-linear fashion with the increase in dose. The t1/2β after multiple oral doses showed a 3-fold increase over the single dose value. These results indicate that aprindine shows dose-dependent non-linear kinetics.
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  • 28
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    European journal of clinical pharmacology 26 (1984), S. 603-608 
    ISSN: 1432-1041
    Keywords: pinacidil ; hypertension ; pinacidil pyridine-N-oxide ; urinary excretion ; protein binding ; pharmacokinetics ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Preliminary investigation in 3 healthy volunteers suggested that intravenous pinacidil in a dose of 0.2 mg/kg had a potent but well-tolerated hypotensive action in the supine position. Facial flushing, uncomfortable chest sensation and distressing postural hypotension occurred at serum concentrations above 300 ng/ml. Pinacidil, 0.2 mg/kg, was given intravenously over 4 min to 15 healthy volunteers in the supine position. Maximum fall in mean arterial pressure (MAP) was 15.7±6.0 mmHg. Maximum rise in heart rate was 23.8±6.6 beats/min. Pinacidil serum distribution half-life ( $${\text{T}}_{{\raise0.7ex\hbox{${\text{1}}$} \!\mathord{\left/ {\vphantom {{\text{1}} {{\text{2}}\alpha }}}\right.\kern-\nulldelimiterspace}\!\lower0.7ex\hbox{${{\text{2}}\alpha }$}}}$$ ) was 13.4±8.5 min and elimination half-life ( $${\text{T}}_{{\raise0.7ex\hbox{${\text{1}}$} \!\mathord{\left/ {\vphantom {{\text{1}} {{\text{2}}\beta }}}\right.\kern-\nulldelimiterspace}\!\lower0.7ex\hbox{${{\text{2}}\beta }$}}}$$ ) was 2.13±0.49 h. The apparent volume of distribution (Vdβ) was 90.3±13.21 and total body clearance was 31.1±9.61/h. Pinacidil was approximately 40% bound to plasma protein over the concentration range 40–400 ng/ml. Urinary excretion of unchanged pinacidil accounted for 5.7 ± 1.3% of the administered dose over 24 hours and urinary excretion of the major metabolite, pinacidil pyridine-N-oxide, was 31.6±9.2% of the administered dose. It was concluded that intravenous pinacidil is a potent vasodilator hypotensive compound, with a duration of action between 1.5 and 2 h.
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  • 29
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    European journal of clinical pharmacology 26 (1984), S. 761-767 
    ISSN: 1432-1041
    Keywords: drug absorption ; intoxication ; activated charcoal ; disopyramide ; indomethacin ; trimethoprim ; healthy volunteers ; adsorption capacity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The efficacy of various charcoal-to-drug ratios for the absorption of drugs was studied in 6 healthy volunteers and in vitro at two pHs. Disopyramide 200 mg, indomethacin 50 mg and trimethoprim 200 mg were ingested on an empty stomach with 100 ml water. After 5 min the subjects ingested a charcoal suspension in 300 ml — 2.5 g, 10 g, 25 g or 50 g of Norit A, or 10 g of PX-21, or water 300 ml only. Increasing the dose of activated charcoal from 2.5 g to 50 g reduced the gastrointestinal absorption of disopyramide and indomethacin from 30–40% to 3–5%, and that of trimethoprim from 10% to 1% of the respective controls. Disopyramide and trimethoprim were best adsorbed by charcoal in vitro at neutral and indomethacin at acid pH, but saturation of the adsorption capacity was apparent at charcoal-to-drug ratios less than 7.5. Combining the in vitro and in vivo results it can be concluded that the dose of activated charcoal to be given in acute intoxication should be as large as possible, because the drug history is often unknown.
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  • 30
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    European journal of clinical pharmacology 27 (1984), S. 75-80 
    ISSN: 1432-1041
    Keywords: bupropion ; alcohol interaction ; healthy volunteers ; performance ; autonomic functions
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The effects of bupropion and ethanol were examined alone and in combination in a placebo controlled, double-blind, crossover study in 12 healthy volunteers. Results were subjected to analysis of variance and differences ofp〈0.05 taken as significant. In the main study using the Wilkinson auditory vigilance test, no active treatment or combination of treatments produced significant change compared with placebo. However, when compared with bupropion 100 mg, vigilance was significantly impaired by 32 ml alcohol alone though not when combined with bupropion. No significant changes in reaction time or short term memory occurred. Visual analogue scales indicated that the subjects were mentally slower after alcohol 32 ml than after placebo. Combination of bupropion 100 mg with alcohol 32 ml abolished this difference. A similar pattern occurred with group ratings indicating mental sedation. Subjects were clearly able to differentiate between the 16 ml and 32 ml doses of alcohol when assessing their degree of inebriation. Combination of bupropion with alcohol made no difference to the ratings of inebriation. The top dose of alcohol tended to increase energy in the low frequency EEG bands. Combination of the top alcohol dose with bupropion, however, produced a significant reversal with lowered energy in the 4–7.5 Hz band. Combination of bupropion with alcohol failed to change the blood alcohol concentration achieved.
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  • 31
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    European journal of clinical pharmacology 27 (1984), S. 23-27 
    ISSN: 1432-1041
    Keywords: hypertension ; alpha2-adrenoceptors ; blood pressure ; clonidine ; guanfacine ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The time courses of the changes in plasma growth hormone and noradrenaline concentrations in response to 15 min infusions of clonidine 0.2 mgs and guanfacine 2 mgs, were studied in six normal volunteers, in a double-blind, randomised, crossover study. Plasma noradrenaline fell within 15 min of the commencement of drug administration, by 36±14% after clonidine (p〈0.05) and by 32±11% (p〈0.05) after guanfacine. Plasma growth hormone was not significantly elevated until the 30th minute to 12.0±4.7 lU/ml (p〈0.05) after clonidine and 14.7±11.5 lU/ml (p〈0.05) after guanfacine, having been undetectable prior to both drugs. The reduction in plasma noradrenaline by these α2-adrenergic agonists, prior to activation of central adrenoceptors as detected by changes in plasma growth hormone, is evidence for a peripheral component in their sympatholytic effect.
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  • 32
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    European journal of clinical pharmacology 27 (1984), S. 119-121 
    ISSN: 1432-1041
    Keywords: S-adenosyl-L-methionine ; pharmacokinetics ; protein binding ; dose-dependent kinetics ; healthy volunteers ; urinary excretion
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary S-Adenosyl-L-methionine (AdoMet) kinetics was studied in 6 male subjects given 100 and 500 mg i. v. Drug concentrations in plasma and urine were assayed using a radioenzymatic method. Pharmacokinetic parameters were estimated according to an open two-compartment model. The apparent volumes of distribution after the 100 and 500 mg doses were 407±27 and 443±36 ml/kg (mean±SEM), terminal half-lives 81±8 and 101±7 min and body clearances 3.7±0.5 and 3.1±0.2 ml/min per kg. Urinary excretion was 34±3 and 40±2% of the administered dose. The results demonstrate that drug disposition occurs more via metabolism than via renal excretion, and it is not dependent on the administered dose. Binding of AdoMet to serum proteins is negligible.
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  • 33
    ISSN: 1432-1041
    Keywords: acetylsalicylic acid ; platelet function ; salicylate ; controlled release formulation ; single dosing ; continuous dosing ; healthy volunteers
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The extent to which a controlled release acetylsalicylic acid (ASA) formulation inhibited platelet function has been evaluated in single and chronic dosing studies. In the single dose study, the platelet inhibitory effect of the controlled release formulation was compared with that of an equivalent dose of soluble ASA and an equimolar dose of sodium salicylate (SA). In the chronic dosing study, ASA dose-response curves for platelet function, including cyclooxygenase activity, were determined for various doses (20–1300 mg) of the controlled release (enteric coated pellets) ASA formulation taken by volunteers daily for one week. Platelet function was assessed by the degree of inhibition of aggregation for several aggregating agents, and the degree of inhibition of activity of platelet cyclooxygenase quantified by the estimation of malondialdehyde (MDA) production. Plasma ASA and SA concentrations were also determined in each study. The controlled release product inhibited platelet function to the same extent as an equimolar dose of soluble ASA, but did so with much lower and sometimes undetectable peak systemic plasma ASA concentrations. SA, the direct metabolite of aspirin, did not have any effect on platelet function. The ASA dose-platelet function response curves obtained from chronic dosing with the controlled release formulation appeared to be similar to those reported previously for the soluble product. The inhibition of platelet function appeared to be unrelated to plasma ASA concentrations.
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  • 34
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    European journal of clinical pharmacology 27 (1984), S. 115-117 
    ISSN: 1432-1041
    Keywords: benzodiazepine antagonist ; Ro 15-1788 ; healthy volunteers ; pharmacokinetics
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The pharmacokinetics of the selective benzodiazepine antagonist Ro 15-1788 has been studied in 6 healthy male volunteers following a single intravenous dose of 2.5 mg. The drug was only slightly bound to plasma proteins (40±8%, mean±SD). A negligible amount (〈0.2% of the dose) of unchanged drug was recovered in urine. Hepatic elimination was rapid, as shown by a short t1/2 of 0.9±0.2 h, and high total plasma and blood clearances of 691±216 ml/min and 716±199 ml/min, respectively. The fast decline of plasma levels from about 60 to 2 ng/ml accounts for the short-lasting reversal of benzodiazepine-induced sedation by Ro 15-1788.
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  • 35
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    European journal of clinical pharmacology 27 (1984), S. 335-339 
    ISSN: 1432-1041
    Keywords: transdihydrolisuride ; dopamine agonist ; pharmacokinetics ; pharmacodynamics ; prolactin levels ; side-effects ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary Plasma levels and urinary excretion of the dopamine agonist, transdihydrolisuride (TDHL), were measured by radioimmunoassay in healthy male volunteers given TDHL 50 µg i.v. and oral doses of 200, 400 and 800 µg. Plasma prolactin was also measured by radioimmunoassay. Following i.v. injection, the concentration of TDHL declined with a half-life of 37±19 min. The total clearance was 38±27 ml/min/kg and the apparent volume of distribution was 1.3±0.4 l/kg. The bioavailability of oral TDHL was proportional to the dose; after 200, 400 and 800 µg the bioavailability was 20±25%, 31±24% and 48±26%. TDHL was almost totally metabolized and less than 0.5% of the dose was excreted unchanged in urine in 24 h. Plasma prolactin levels were depressed by 66±15%, 75±11% and 80±7% after TDHL 200 µg, 400 µg and 800 µg. The effect lasted for more than 12 h after the lowest dose and for more than 24 h after 400 and 800 µg. Side effects, mainly nausea and headache, only occurred at the two highest dose levels.
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  • 36
    ISSN: 1432-1041
    Keywords: molsidomine ; vasodilators ; pharmacokinetics ; pharmacodynamics ; dose-response relationship ; haemodynamics ; healthy volunteers
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary In 12 healthy male volunteers, molsidomine 1, 2 and 4 mg i.v. increased resting heart rate and decreased systolic blood pressure, the latter still being affected after 8 hours. After single oral doses of 1 and 2 mg, systolic pressure tended to be reduced for 90 minutes and exercise heart rate tended to be increased. After oral treatment with 2 mg molsidomine three times daily for 1 week, the pharmacokinetic parameters and the effects on heart rate and blood pressure after the final dose were not different from those after the first dose. The terminal half-life was independent of dose and route of administration. Clearance and distribution volume were not dose-dependent. The bioavailability of a 2 mg oral dose of molsidomine was 44%. Inter-individual variation in heart rate, blood pressure and pharmacokinetics was observed.
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  • 37
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    European journal of clinical pharmacology 27 (1984), S. 303-306 
    ISSN: 1432-1041
    Keywords: frusemide ; renal function ; ageing ; healthy volunteers
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The effect of IV frusemide was studied in six healthy young (mean age 26.5 years, range 21–33) and six healthy old (mean age 72.8 years, range 66–80) volunteers. A 24-h urine collection before frusemide showed no difference in volume and sodium excretion, although the old excreted less potassium. Creatinine clearance was significantly reduced in the older subjects. After frusemide, 20 mg IV, the pattern of sodium and water excretion over a 5-h period was different in the two groups. The peak effect was greater in the young and occurred within the first 30 min, but was delayed to between 30 and 60 min in the old. Thus in the young the time for 50% of the total sodium and water to be excreted was half that in the old. This delay in sodium and water excretion was related to baseline creatinine clearance. However, the total water, sodium and potassium excreted in the 5 h after frusemide did not differ in the two groups. These results suggest that the renal effects of frusemide are different in healthy elderly subjects as compared to the young. The delayed and reduced peak response is consistent with fewer nephrons in the elderly kidney.
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  • 38
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    European journal of clinical pharmacology 27 (1984), S. 319-324 
    ISSN: 1432-1041
    Keywords: theophylline ; enprofylline ; terbutaline ; haemodynamic response ; adenosine antagonism ; healthy volunteers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The haemodynamic response to two xanthines, enprofylline and theophylline, was studied in 6 healthy male volunteers at rest, during exercise and in combination with the beta2-agonist, terbutaline. At rest the haemodynamic effects of both xanthines were small and were qualitatively different from each other. While theophylline exerted a “pressor” response, enprofylline seemed to have arterial dilating ability. During exercise both xanthines as compared to placebo were associated with a higher heart rate and in general with increased systolic blood pressure. In combination with terbutaline enprofylline and theophylline both increased systolic blood pressure more than placebo, i. e. they augmented the positive inotropic effect of terbutaline. The systolic blood pressure was higher after theophylline than enprofylline despite their equipotent bronchodilator activity. This may reflect different inotropic effects of the xanthines as well as a difference in their influence on the response to adenosine.
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  • 39
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    European journal of clinical pharmacology 27 (1984), S. 325-328 
    ISSN: 1432-1041
    Keywords: theophylline ; sustained release ; pharmacokinetics ; chronic administration ; healthy volunteers ; plasma levels ; GCMS assay ; stable isotope technique
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    Topics: Chemistry and Pharmacology , Medicine
    Notes: Summary The pharmacokinetics of a new sustained-release preparation of theophylline (Dilatrane à Action Prolongée capsules filled with homogenous microgranules) has been after its studied administration to 7 healthy volunteers at 8 p.m. in order to achieve therapeutic levels at night and in the morning. In separate trials the test dose of 500 or 600 mg was administered for 7 days, once daily at 8 p.m. Plasma theophylline levels were measured by capillary gas chromatography with a mass specific detector after pentylation, using internal standards labelled with stable isotopes (15N-1,3 and 13C-2 theophylline). The new sustained-release preparation showed a monophasic regular absorption phase with very low interindividual variability. After administration, the plasma level stayed within 80% of the peak levels for 8.5±1.5 h. There was a good correlation between the dose and the steady state plasma level (r=0.9587; p〈0.05). This preparation can be chronically administered once daily day at 8 p.m. in order to achieve a therapeutic level during the night and the morning, and to provide sufficient protection during the nycterohemeral period, with a once dose a day schedule.
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  • 40
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    Cell & tissue research 237 (1984), S. 195-202 
    ISSN: 1432-0878
    Keywords: Pituitary ; Prolactin cells ; Estrogen ; Heterogeneity ; Immunocytochemistry
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    Topics: Biology , Medicine
    Notes: Summary This study was conducted to determine the functional and/or developmental relationships among three heterogeneous types of prolactin cells (I, II and III) in rats. Rats were injected subcutaneously daily with estradiol or testosterone propionate on days 10–20 after birth. Estradiol increased the proportion of cell types II and III, increased serum PRL levels 12-fold in males and 15-fold in females, and increased pituitary levels of prolactin 12-fold in males and 5-fold in females. Testosterone mainly increased the proportion of the Type-II cells, decreased serum levels of prolactin in males only, and did not change pituitary levels of prolactin. In a second experiment, treatment of rats with nafoxidine for five days after E2 treatment (days 10–20 after birth) increased the proportion of Type-I cells and decreased the proportion of Type-III cells and decreased serum and pituitary levels of prolactin by 50% in females and by 15 and 45% in males. In a third experiment utilizing adult male rats, estradiol and testosterone were found to modulate the relative ratios of the different types of PRL cells as they did in immature animals. The data taken as a whole suggest the possibility of an estrogen-stimulated conversion of one cell type to another, which may be a reflection of prolactin secretory activity.
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  • 41
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    Cell & tissue research 237 (1984), S. 245-252 
    ISSN: 1432-0878
    Keywords: Pineal organ ; Interstitial cells ; Astrocytes ; Immunocytochemistry ; Rat ; Mouse
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    Topics: Biology , Medicine
    Notes: Summary Antigenic markers characteristic of astrocytes and their differentiative states (i.e., glial fibrillary acidic protein (GFAP), vimentin, and M1 and C1 antigens) were investigated in the pineal gland of mouse and rat using double immunolabeling techniques. In both species the socalled interstitial cells as characterized by TEM were shown to be astrocytes, since they expressed vimentin, but neither fibronectin (a marker for fibroblasts and endothelial cells) nor the neuron-specific L1 antigen or tetanus toxin receptors. Subpopulations of vimentin-positive pineal astrocytes were also GFAP- and C1- antigen-positive. M1- antigenpositive cells were not detected. It is concluded that a considerable proportion of interstitial cells in the pineal gland of rat and mouse are immature astrocytes which, in contrast to other parts of the central nervous system, persist into adulthood.
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  • 42
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Reissner's fiber ; Ependyma ; Secretory process ; Comparative analysis ; Immunocytochemistry ; Vertebrates
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    Topics: Biology , Medicine
    Notes: Summary The subcommissural organs (SCO) of 76 specimens belonging to 25 vertebrate species (amphibians, reptiles, birds, mammals) were studied by use of the immunoperoxidase procedure. The primary antiserum was obtained by immunizing rabbits with bovine Reissner's fiber (RF) extracted in a medium containing EDTA, DTT and urea. Antiserum against an aqueous extract of RF was also produced. The presence of immunoreactive material in cell processes and endings was regarded as an indication of a possible route of passage. Special attention was paid to the relative development of the ventricular, leptomeningeal and vascular pathways established by immunoreactive structures. The SCO of submammalian species is characterized by (i) a conspicuous leptomeningeal connection established by ependymal cells, (ii) scarce or missing hypendymal cells, and (iii) a population of ependymal cells establishing close spatial contacts with blood vessels. The SCO of most mammalian species displays the following features: (i) ependymal cells lacking immunoreactive long basal processes, (ii) hypendymal secretory cells occurring either in a scattered arrangement or forming clusters, (iii) an occasional leptomeningeal connection provided by hypendymal cells, and (iv) in certain species numerous contacts of secretory cells with blood vessels. In the hedgehog immunoreactive material was missing in the ependymal formation of the SCO, but present in hypendymal cells and in the choroid plexuses. The SCO of several species of New-and Old-World monkeys displayed immunoreactive material, whereas that of anthropoid apes (chimpanzee, orangutan) and man was completely negative with the antisera used.
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  • 43
    ISSN: 1432-0878
    Keywords: Subcommissural organ ; Ependyma ; Comparative aspects ; Immunocytochemistry ; Secretory process ; Blood vessels
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    Topics: Biology , Medicine
    Notes: Summary In 76 specimens (amphibians, reptilians, mammals) belonging to 25 different vertebrate species, the region of the subcommissural organ (SCO) was investigated with the use of a primary antiserum raised against an extract of bovine Reissner's fiber+the immunoperoxidase procedure according to Sternberger et al. (1970). In the SCO of a toad (Bufo arenarum) and several species of reptiles (lacertilians, ophidians, crocodilians), the ependymal cells were the only type of secretory cell displaying vascular contacts, whereas in mammals ependymal and hypendymal cells established intimate spatial contacts with blood vessels. In Bufo arenarum, but especially in the reptilian species examined, the ependymo-vascular relationship was exerted by a population of ependymal cells having a rather constant location within the SCO and projecting to capillaries that showed a remarkably constant pattern of anatomical distribution. In the SCO of mammals the modality and degree of the structural relationships between secretory cells and blood vessels varied greatly from species to species. In the SCO of the armadillo and dog the secretory tissue was organized as a thick, highly vascularized layer with most of the cells oriented toward the capillaries. A rather opposite situation was found in the SCO of New-and Old-World monkeys, where vascular contacts were restricted to a few ependymal cells.
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  • 44
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    Cell & tissue research 237 (1984), S. 521-524 
    ISSN: 1432-0878
    Keywords: α-Melanocyte-stimulating hormone ; α-MSH-like peptide ; Immunocytochemistry ; Hypothalamus ; Lizard (Lacerta muralis)
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    Topics: Biology , Medicine
    Notes: Summary The distribution of α-melanocyte-stimulating hormone (α-MSH) was studied in the brain of the lizard Lacerta muralis by means of immunocytochemical staining methods. α-MSH-like containing cells were found in the ventro-lateral preoptic area and the paraventricular and supraoptic nuclei. Some scattered cells staining for α-MSH were also detected in the mesencephalo-diencephalic boundary region, while numerous α-MSH-like nerve fibres were localized in the medial eminence. No reaction was observed after the use of antiserum preabsorbed with synthetic antigen. These findings suggest that an α-MSH-like peptidergic system could possibly be involved in the hypothalamo-hypophysial regulation and/or play a role as neurotransmitter in this animal.
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  • 45
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    Cell & tissue research 238 (1984), S. 203-205 
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Hypothalamus, human ; Immunocytochemistry ; Pituitary stalk
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    Topics: Biology , Medicine
    Notes: Summary In order to study the distribution of neuropeptide Y-like immunoreactivity in the human hypothalamus, an immunocytochemical localization of this peptide was performed. Using antibodies developed against synthetic porcine neuropeptide Y (NPY), we have been able to localize immunoreactivity in neuronal cell bodies located exclusively in the infundibular nucleus. Immunostained fibers were found in several regions in the hypothalamus with a high concentration in the periventricular areas. Fibers were also found in the neurovascular zone of the median eminence, the pituitary stalk and the posterior pituitary. These results suggest that immunoreactive material related to porcine NPY is present in the human hypothalamus, with a distribution similar to that observed in the rat.
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  • 46
    ISSN: 1432-0878
    Keywords: Peptidergic neurotransmission ; Lymnaea stagnalis ; Immunocytochemistry ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Three neuronal systems of the pond snail Lymnaea stagnalis were immunocytochemically investigated at the ultrastructural level with the unlabeled peroxidase-antiperoxidase technique. Preliminary electrophysiological and cell-filling investigations have shown that a cluster of neurons which reacts positively with an antiserum against the molluscan cardio-active peptide FMRFamide, sends axons to the penis retractor muscle. In this muscle anti-FMRF-amide (aFM) positive axons form neuro-muscular synapses with (smooth) muscle fibers. The morphological observations suggest the aFM immunoreactive system to be involved in peptidergic neurotransmission. In the right parietal ganglion a large neuron (LYAC) is penetrated by aFM positive axons which form synapse-like structures (SLS) with the LYAC. The assumption that the SLS represent the morphological basis for peptidergic transmission is sustained by the observation that iontophoretical application of synthetic FMRFamide depolarizes the LYAC. The axons of a group of pedal anti-vasopressin (aVP) positive cells run in close vicinity to the cerebral ovulation (neuro-)-hormone producing cell system (CDC system) Synapses or SLS between the two systems were not observed. The fact that (bath) application of arg-vasopressin induces bursting in the CDC, may indicate that the vasopressin-like substance of the aVP cells is released non-synaptically.
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  • 47
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    Cell & tissue research 238 (1984), S. 497-502 
    ISSN: 1432-0878
    Keywords: D2 glycoprotein ; Adrenal gland ; Immunocytochemistry ; Ultrastructure ; Cell adhesion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of the glycoprotein D2 in rat adrenal gland was investigated using immunohistochemical methods, and D2 localization in cultures of adult bovine chromaffin cells was studied by immunofluorescence. D2 was found to be situated on nerve fibers passing through the adrenal cortex and in the medulla zone, and also on the surface of all chromaffin cells. In addition, it was strongly expressed on the surface of glial (Schwann) cells. Cortical cells were unreactive to the antiserum. In cultures, all adrenalin and noradrenalin [dopamine-β-hydroxylase (DBH)-positive] cells were surface labelled for D2. A less frequent second cell type was recognized in vitro which was DBH negative but D2 positive. Such cells were presumed to be Schwann cells. These data are discussed in terms of the developmental origin of the cells and with regard to the putative functional rôle of D2 in cell adhesion phenomena.
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  • 48
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    Cell & tissue research 235 (1984), S. 211-214 
    ISSN: 1432-0878
    Keywords: Adenohypophysis ; Pars distalis ; Immunocytochemistry ; Amphibia ; Gymnophiona
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The indirect immunofluorescence method was used to identify and locate LTH-, STH-, LH-, TSH-, ACTH- and MSH-immunoreactive cells in the pituitary of Typhlonectes compressicaudus (Gymnophiona). The present study defines the histological and histochemical staining properties of each cell type identified.
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  • 49
    ISSN: 1432-0878
    Keywords: Caldesmon ; Actin ; Immunocytochemistry ; Small intestine ; Smooth muscle ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin-interacting protein) (Sobue et al. 1982) and of actin was studied in the rat's small intestine by means of light-microscopic immunocytochemistry. Positive immunostaining for caldesmon was seen in smooth muscle cells of the intestinal wall, and of blood vessels, and in the apical portion of the absorptive epithelial cells. The immunoreactivity in goblet cells was difficult to recognize. The positive reaction to immunostaining for actin showed almost the same pattern as that for caldesmon. These results suggest that this calmodulin-binding protein may play an important role in the control of actin-myosin interaction in smooth muscle cells and in non-muscle cells.
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  • 50
    ISSN: 1432-0878
    Keywords: FMRFamide ; Bovine pancreatic polypeptide ; Immunocytochemistry ; Peptidergic neurons ; Leptinotarsa decemlineata
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Particular neurons in the nervous system of the Colorado potato beetle, Leptinotarsa decemlineata, are recognized by antisera against bovine pancreatic polypeptide and FMRFamide. Both antisera react with the same neurons. Solid phase absorptions showed that antiserum against bovine pancreatic polypeptide cross-reacts with FMRFamide, whereas antiserum against FMRFamide cross-reacts with bovine pancreatic polypeptide. Some of the immunoreactive neurons have axons branching extensively within the neuropile, which suggests that the peptide is used as transmitter. In the corpus cardiacum, a neurohaemal organ in insects, numerous immunoreactive axon terminals are present. Here, the peptide material is presumably released as a hormone.
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  • 51
    ISSN: 1432-0878
    Keywords: Corticotropin-releasing factor ; Immunocytochemistry ; Hypothalamus ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The corticotropin-releasing factor (CRF)-containing neurons were investigated in the brain of the domestic fowl by means of the peroxidase-antiperoxidase technique at the light-microscopic level. The detection of CRF-immunoreactivity was facilitated by silver intensification. CRF-containing perikarya were found in the paraventricular, preoptic and mammillary nuclei of the hypothalamus and in some extrahypothalamic areas (nuclei dorsomedialis and dorsolateralis thalami, nucleus accumbens septi, lobus parolfactorius, periaqueductal gray of the mesencephalon, nucleus oculomotorius ventralis). Immunoreactive nerve fibers and terminals were demonstrated in the external zone of the median eminence and the organum vasculosum of the lamina terminalis. These results indicate that an immunologically demonstrable CRF-neurosecretory system also exists in the avian central nervous system.
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  • 52
    ISSN: 1432-0878
    Keywords: Thyroid ; Immunocytochemistry ; Caldesmon ; Actin ; Endocytosis ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The distribution of caldesmon (a calmodulin-binding, F-actin interacting protein; Sobue et al. 1982) and actin was studied in the rat thyroid gland by means of light-microscopic immunocytochemistry, and the fine-structural distribution of actin filaments was examined by use of heavy meromyosin (HMM). Caldesmon and actin were demonstrated in the apical cytoplasm of almost all the follicle epithelial cells in normal as well as TSH-treated animals. Immunoreactivities for both caldesmon and actin showed almost the same pattern in localization. The smooth muscle cells of the blood vessels were also positive for caldesmon and actin. By electron microscopy, numerous actin filaments decorated by HMM and running perpendicularly or randomly to the apical surface were recognized in the apical cytoplasm of the follicle epithelial cell. These results suggest that caldesmon and actin, in conjugation with calmodulin, play a role in the regulation of cellular activity such as exocytosis and endocytosis in the apical portion of the follicle epithelial cell.
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  • 53
    ISSN: 1432-0878
    Keywords: Urotensin ; Caudal neurosecretory system ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The localization of urotensin I has been investigated in the caudal neurosecretory system of the white sucker (Catostomus commersoni). The peptide is present in all the cells of the system both large and small, in the large axons passing to the urophysis, and in fine beaded fibres not only within the urophysis but also in a fine plexus lateral to the large cells in the spinal cord proper. The possibility that the caudal neurosecretory system is not a functionally uniform system but rather a collection of dissimilar cells of different synaptic inputs with a common entity, urotensin I, is discussed. Moreover, the feasibility of a urotensin I feedback loop is described.
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  • 54
    ISSN: 1432-0878
    Keywords: Pituitary rat ; LH cells ; FSH cells ; ACTH cells ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By means of immunocytochemistry, the development of ACTH-, LH- and FSH cells was examined in the anterior pituitary of 5-day-old neonatal, 15-day-old immature and adult rats. ACTH-positive cells are angular and the periphery of these cells is strongly reactive with anti-ACTH serum. In contrast, LH- and FSH-immunopositive cells are ovoid elements, ranging in cell size and intensity of staining. Angular cells, in which only the cell periphery reacted with anti-LHβ serum, were observed in neonatal and immature rats; however, these cells were not stained with either anti-FSHβ serum or anti-ACTH serum. Observation of serial semithin sections revealed that ACTH-immunopositive cells do not react with either anti-LHβ or anti-FSHβ serum. Finally, it was observed that ACTH cells and LH cells are both functionally differentiated already in 5-day-old neonatal rats.
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  • 55
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    Cell & tissue research 235 (1984), S. 77-83 
    ISSN: 1432-0878
    Keywords: Pituitary (rat) ; LH cells ; FSH cells ; Rapid freeze-substitution ; Immunocytochemistry ; Ferritin antibody
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Rapid freeze-substitution fixation was employed in immunocytochemical studies on the localization of LH and FSH in the typical gonadotrophs of the anterior pituitary in the untreated male rat; a modification of a recently described ferritin antibody method (Inoue et al. 1982) was used in these studies. It was shown that rapid freeze-substitution fixation provides good preservation not only of the ultrastructure but also of the antigenicity. Both LH and FSH were clearly demonstrated in the same gonadotrophic cells, but the subcellular localization of these gonadotrophins differed: (i) LH was mainly located in small secretory granules, 250–300 nm in diameter; (ii) FSH was mainly present in large secretory granules, up to 500 nm in diameter. In the pituitary gland of the adult male rat, all gonadotrophs that react to antibodies against gonadotrophins are characterized by small and large secretory granules. Other types of cells of the anterior pituitary containing either small secretory granules or resembling corticotrophs with secretory granules assembled at cell periphery did not react to either anti-LH beta or anti-FSH beta serum. For light microscopy, the peroxidase antibody method was used. All of the gonadotrophin-positive cells contain both LH and FSH. None of the pituitary cells reacted to antibody against only one gonadotrophin. However, some cells are “LH-rich” while other cells are “FSH-rich”.
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  • 56
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    Cell & tissue research 235 (1984), S. 159-169 
    ISSN: 1432-0878
    Keywords: Anterior pituitary ; Gonadotropic cells ; Immunocytochemistry ; Testosterone binding ; Cryo-ultramicrotomy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Several attempts have been made to localize steroids by means of immunocytological techniques. However, these methods were found inadequate for detecting steroids bound to their receptors. To localize endogenous testosterone (T) in its target cells at the ultrastructural level, an immunocytological technique was performed on ultrathin sections obtained by cryo-ultramicrotomy. T was detected in the pituitary glands obtained from intact male or female rats and castrated rats, but not in castrated + adrenalectomized rats. Animals were also injected either with testosterone, with other steroids (estradiol, progesterone, corticosterone) or with an androgen antagonist (cyproterone acetate). In addition, some ultrathin sections were preincubated either with phosphate buffers of various pH, corticosterone, cyproterone acetate solution, or with T solution. The content of T in the pituitary before and after fixation was measured by radioimmunoassay; it decreased after fixation. T immunoreactivity was localized in the gonadotropic cells only, both in the male and female rats. At the subcellular level, the immunoreactivity was detected in the cytoplasmic matrix and in the nucleus. Immunoreactive T disappeared 1) in rats after castration+adrenalectomy; by means of radioimmunoassay no T was measured in these pituitary glands; 2) in rats injected with 25 (μg/rat of cyproterone acetate; 3) after preincubation of pituitary sections on a drop of cyproterone acetate (1 × 10-6 M). The immunocytological reaction was not modified when the rats were injected with estradiol, progesterone or corticosterone (1 mg/rat), or after preincubation of the sections with corticosterone (1 × 10-3 M), or a buffer solution at pH 7.6. Lower or higher pH values led to a strong decrease in the immunoreactivity. After injection of T (15 μg/rat) the immunocytological reaction was more abundant in the nucleus and less in the cytoplasm. The immunoreactivity was again observed when the sections were preincubated with cyproterone acetate solution and then with T solution. These data suggest that T can be detected by means of immunocytochemistry. It is probably bound to a specific binding site.
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  • 57
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Supraoptic nucleus (SON) ; Oxytocin neurones ; Neuronal appositions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pre-embedding immunocytochemistry was performed on vibratome sections of the hypothalamus of lactating rats using antiserum directed against oxytocin. Electron microscopy revealed that numerous immunopositive somata and dendrites in the supraoptic nucleus were in direct apposition, without glial interposition; a number of them were also bridged by “double” synapses. The observations support the contention that the ultrastructural reorganisation of the nucleus apparent at lactation affects the magnocellular neurones secreting oxytocin.
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  • 58
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    Cell & tissue research 235 (1984), S. 427-432 
    ISSN: 1432-0878
    Keywords: DPP IV ; Salivary glands ; Ultrastructural localization ; Immunocytochemistry ; PAP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructural localization of dipeptidyl peptidase IV (DPP IV) (EC 3.4.14.5) in rat submandibular and parotid glands was studied immunocytochemically by the peroxidase-antiperoxidase (PAP) method, using a monospecific antiserum against rat kidney DPP IV. There were no differences in the immunocytochemical localization of DPP IV between submandibular and parotid glands. In these glands, DPP IV was primarily found to be associated with the luminal and intercellular canalicular plasma membranes of acinar cells and with the luminal plasma membranes of intercalated and striated duct cells. Occasionally, immunoreaction of DPP IV was detected in cytoplasmic vesicles (vacuoles), lysosomes, and multivesicular bodies in some acinar cells as well as in ductal epithelial cells. Furthermore, the reaction product was also found within the lumina of peri-acinar and peri-ductal capillaries and in the cytoplasm of some fibroblasts in the interstitial connective tissue. These data suggest that DPP IV in the submandibular and parotid glands may play some role in the secretion or reabsorption processes of secretory proteins and peptides in these glands.
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  • 59
    ISSN: 1432-0878
    Keywords: S-100 protein ; Thymus ; Interdigitating cells ; Immunocytochemistry ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The present study deals with the localization and ultrastructure of S-100-immunoreactive cells in the human thymus. These immunoreactive cells are distributed mainly in the medulla with some scattered elements in the cortex. Electron-microscopic observation revealed that the cells are characterized by an irregularly shaped nucleus, tubulovesicular structures in the cytoplasm and characteristic interdigitations of the plasma membrane. The cells often embrace lymphocytes with their branched processes. On the basis of these morphological features, the immunostained elements were identified as interdigitating cells (IDCs). The immunocytochemistry for S-100 visualizes the precise distribution and extension of the IDCs under the light microscope and indicates that the IDCs form no structural networks such as those established by the thymic epithelial cells. Since the IDCs in human lymph nodes have also been reported to contain S-100-like immunoreactivity, S-100 protein can be regarded as a useful marker for identifying the IDCs in the human thymus and other lymphoid organs.
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  • 60
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; FMRF-amide ; Neurotransmitter ; Palaemon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of a specific antiserum against the molluscan cardio-excitatory tetrapeptide FMRF-amide in combination with the PAP-method it was possible to obtain positive immunocytochemical reactions in several neurosecretory regions of the eyestalk of the prawn Palaemon serratus. FMRF-amide-like material was found in perikarya and nerve fibers of the medulla terminalis and in neurons in the lamina ganglionaris. The immunoreactivity observed in the glandular tissue located at the basal insertion of the eyestalk muscles must be ascribed to a non-specific reaction. The identification of immunopositive nerve fibers, ending on a nerve bundle in the medulla terminalis, and the fact that immunoreactive material was absent in the neurohemal sinus gland seem to indicate a neurotransmitter/neuromodulator function.
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  • 61
    ISSN: 1432-0878
    Keywords: Fetal porcine pituitary ; ACTH, MSH, β-LPH, α- and β-endorphin, GH, PRL ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The aim of the present study on the fetal porcine pituitary was (1) to detect by means of the immunoperoxidase technique the earliest stages of cells producing polypeptide hormones: β-MSH, ACTH, β-LPH, α- and β-endorphin, growth hormone (GH) and prolactin (PRL), (2) to study the development of the synthesis and the storage of these hormones during fetal life, and (3) to detect whether several hormones can be located in one and the same cell. The corticotropic cells were revealed as the earliest functional elements of the fetal anterior pituitary. Our results indicate clearly that ACTH, β-MSH, β-LPH, α- and β-endorphin appear at 34 days in the same regular, round or ovoid cells; no differences in the time of their appearance could be observed. The ACTH-cells, irregular or angular in shape and endowed with cytoplasmic processes such as described in the adult pituitary, were not seen until day 50. The first GH-cells were detected between 40 to 45 days of fetal life. From day 45 to 90, the GH-cells greatly increased in number and in staining intensity of their progressively extending cytoplasmic area, but they displayed the same regular and round shape. The PRL-cells were the last cell type to appear in the fetal pituitary. The first PRL-cells, small in size and round or ovoid in shape with a high nucleus/cytoplasm ratio, were detected at day 70. At day 80, the PRL-cells increased in size and staining intensity. They displayed an irregular elongated or stellated shape and cytoplasmic processes resembling those characteristic of the adult pituitary. These data suggest that in the fetal porcine pituitary: (1) ACTH, β-LPH and related peptides are synthesized and stored in the same cells, and (2) PRL and GH appear in individual cellular elements.
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  • 62
    ISSN: 1432-0878
    Keywords: Anterior pituitary, porcine ; Gonadotropic hormones (FSH, LH) ; Immunocytochemistry ; Cellular compartments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Pre- and post-embedding immunocytochemical methods based on the use of specific antibodies against β-subunits of porcine LH and FSH were applied to determine the changes occurring in the anterior pituitary of the pig after gonadectomy. The results showed that (1) the total number of immunoreactive gonadotropes increased from 21–25% in control animals to 24–37% in castrated animals; (2) all gonadotropes contained both LH and FSH; (3) several types of immunoreactive LH/FSH cells were revealed; and (4) the two immunocytochemical methods used with dispersed cells localized the hormones in the same subcellular sites. However, the staining intensity in the different locations varied depending on the method applied. With the post-embedding method, a dense reaction product was found in the secretory granules but the cisternae of RER and the Golgi saccules were always slightly reactive. After the pre-embedding method, the staining intensity in the RER-cisternae and in the Golgi saccules was greatly increased. Thus, the two methodological approaches used in this study have permitted to visualize immunocytochemically the gonadotropic hormones not only at the sites of their storage but also along the intracellular pathway of the secretory material, i.e., at the site of its synthesis and during its passage via the Golgi zone.
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  • 63
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    Cell & tissue research 236 (1984), S. 171-180 
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Ultrastructure ; Supraoptic nucleus ; Neuropil ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The neuropil located ventral to the SON was investigated by the use of immunoperoxidase staining for neurophysins, oxytocin and vasopressin, and electron miroscopy. The study was performed in six groups of rats: 1) control; 2) infusion of isotonic saline into the CSF; 3) infusion of hypertonic saline into the CSF; 4) drinking hypertonic saline for 4 days; 5) same as group 4 but injection of colchicine into the CSF on second day of dehydration; 6) salt loading for 3 months. In the control rats the ventral neuropil contained a few immunoreactive processes, the general morphology of which was completely different from that of the neurosecretory axons emerging from the SON at its dorsal aspect. In rats of groups 3 to 6 the ventral processes (VP) became loaded with neurosecretory granules, whereas the perikarya and axons were depleted. Based on their general morphology and reactivity pattern it is suggested that the VP are dendrites. Most of these “dendrites” were embedded in a glial cushion formed by the processes of a particular type of marginal glia. Some of these “dendrites” enveloped an arteriole penetrating the optic tract. All VP were rich in synaptic contacts. The possibility that the VP of neurosecretory cells may be functionally related to the subarachnoid CSF and the arteriolar blood flow is discussed.
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  • 64
    ISSN: 1432-0878
    Keywords: ACTH cells ; Immunocytochemistry ; Morphometry ; Spontaneous hypertension
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary ACTH-immunoreactive cells in the anterior pituitary of 4-week-old spontaneously hypertensive rats (SHR) were studied with immunocytochemical and morphometric techniques. The results were compared with data from age-matched normotensive Wistar-Kyoto rats (WKY). No significant differences were found in volume density and average size of ACTH-immunoreactive cells between these two strains. However, SHR showed a significantly larger anterior lobe (2 P 〈 0.01) than WKY, indicating that the total number of ACTH-immunoreactive cells in the anterior pituitary is greater in SHR than in WKY. These data are in agreement with radioimmunological determinations showing a significantly elevated content (2 P 〈 0.01) but only a moderately higher concentration (0.05 〈 2 P 〈 0.10) of ACTH in the anterior pituitary of SHR as compared to WKY. The present results suggest an enhanced availability of ACTH in the anterior pituitary of 4-week-old SHR, a fact which could explain the markedly enhanced stress-induced release of ACTH previously found in these animals. This study further supports the hypothesis that, among other factors, an instability of the hypothalamo-pituitary-adrenal axis may contribute to the development of genetically programmed hypertension.
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  • 65
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    Cell & tissue research 236 (1984), S. 517-525 
    ISSN: 1432-0878
    Keywords: Bone marrow ; Actin ; Cell motility ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Blood cells proliferate extravascularly in the bone marrow and enter the circulation by migrating through endothelial cells of venous blood sinuses. This migration, or diapedesis, was suspected to involve actin. To test for the presence and distribution of actin, sections of rat bone marrow were examined by indirect immunocytochemistry. Affinity purified rabbit antichicken gizzard actin antibody, and goat-antirabbit IgG-FITC, or goat antirabbit IgG colloidal gold probes were used. The migrating cell contacts the endothelial cell and forms a podosome (a cortical bleb). Immunocytochemistry shows this region to contain actin. As diapedesis proceeds the podosome deforms, then breaches the endothelial cell. At this time the anterior portion of the leukocyte shows heavy labeling for actin. When the migratory cell traverses approximately half of its length through the endothelial cell, actin appears prominent in the caudal region of the cell. The immunocytochemical data suggest that actin is nonrandomly distributed in leukocytes undergoing diapedesis and may be a component of the force-generating mechanism responsible for this transcellular migratory event.
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  • 66
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Goat ; Folliculo-stellate cell ; GH cell ; S-100 protein ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Goat pituitary glands were immunohistochemically studied with antisera for bovine S-100 protein, rat LHβ, FSH, TSHβ, prolactin, ovine GH, and porcine ACTH1–39 by use of the superimposition technique on adjacent sections. Folliculo-stellate (F-S) cells were divided into two categories on the basis of ultrastructural properties: One consisted of a mass of agranular cells in which the pseudolumina were equipped with microvilli and cilia. Elongate gap junctions were often observed among these cells. The other was a group of granulated cells with or without pseudolumina. In this group the gap junctions were shown to be disintegrated. The dense granules 150–250 nm in diameter began to accumulate in the cells. However, neither type of these F-S cells was immunostained for S-100 protein. On the other hand, numerous polygonal, elongate, irregular or stellate cells containing S-100 protein were distributed throughout the gland. Most of them were immunohistochemically identical with the GH cells laden with the secretory granules 250–450 nm in diameter, but some of them were identical to TSH and prolactin cells which immunostained faintly for S-100 protein. This appears to be the first demonstration of GH cells intensely immunostained for S-100 protein.
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  • 67
    ISSN: 1432-0878
    Keywords: Hagfish ; Brain ; Pituitary ; FMRF-amide ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Paraffin sections of brain and pituitary of the hagfish Eptatretus burgeri were immunostained with an antiserum to FMRF-amide. Immunoreactivity was visible in a large number of neurons in the posterior part of the ventromedial hypothalamus and in long neuronal processes extending cranially from the hypothalamus to the olfactory system and caudally to the medulla oblongata. FMRF-amide-like immunoreactivity was also found in cells of the adenohypophysis. These observations suggest that the hagfish possesses a brain FMRF-amide-like transmitter system and pituitary cells containing FMRF-amide-like material. Antisera to ACTH, α-MSH and pancreatic polypeptide gave no immunoreaction in hagfish brain or pituitary.
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  • 68
    ISSN: 1432-0878
    Keywords: Crustacean hyperglycemic hormone ; Astacus leptodactylus ; Immunocytochemistry ; Quantitative electron microscopy ; Secretory cycle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical staining demonstrates striking differences in staining intensity among individual crustacean hyperglycemic hormone (CHH)-producing cells in the eyestalk of the crayfish Astacus leptodactylus. Based on these differences we arbitrarily subdivided the CHH-cells into three categories representing increasing immunoreactivity respectively: + cells, + + cells, and + + + cells. Electron microscopic investigations reveal that these differences in immunostaining are correlated with differences in the numerical density of the neurosecretory granules in the cytoplasm and that these may reflect differences in activity among the CHH-cells. Morphometric analyses at the light- and electron-microscopic levels indicate that the three distinguished categories of immunopositive cells represent different stages in the CHH-synthesizing process of the cells. The results of the present study demonstrate the application of the PAP-technique at the light-microscopic level as a method to obtain information pertaining to the dynamics of secretory activity of the CHH-cells.
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  • 69
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    Cell & tissue research 237 (1984), S. 321-327 
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Immunocytochemistry ; Respiratory tract ; Ear, middle
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Nerve fibres displaying neuropeptide-Y (NPY) immunoreactivity are abundantly distributed in the respiratory tract of cats, guinea-pigs, rats and mice. Fine beaded NPY fibres were seen in whole-mount spreads of the middle-ear mucosa. In the nasal mucosa and in the wall of the Eustachian tube NPY fibres were numerous around arteries and arterioles but sparse in the vicinity of veins; single fibres were found close to the acini of seromucous glands. In the tracheobronchial wall NPY fibres occurred in the proximity of blood vessels, in the subepithelial layer and in the smooth muscle. Surgical and chemical (6-hydroxydopamine treatment) sympathectomy resulted in disappearance of adrenergic and NPY-containing nerve fibres in the nasal mucosa. Sequential staining with antibodies against dopamine-β-hydroxylase (DBH) and NPY revealed that DBH and NPY occur in the same perivascular nerve fibres in the nasal mucosa. The distribution of NPY fibres in the respiratory tract suggests multiple functions of NPY, such as regulation of local blood flow, glandular secretion and smooth muscle activity.
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  • 70
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Biogenic amines ; Vasotocin ; Lymnaea stagnalis
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    Topics: Biology , Medicine
    Notes: Summary Consecutive sections of certain neurons in the central ganglia of the pond snail Lymnaea stagnalis appear to be immunoreactive to anti-dopamine and anti-serotonin. The Cerebral Giant Neurons stain in addition with antivasotocin. The observations indicate the presence of two biogenic amines within the same neuron and in addition their co-existence with a biologically active peptide.
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  • 71
    ISSN: 1432-0878
    Keywords: Retinal photoreceptors ; Opsin ; Optic nerve ; Immunocytochemistry ; Cyclostome, Myxine glutinosa
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    Topics: Biology , Medicine
    Notes: Summary Opsin-immunoreactive sites in the eye and optic nerve of the hagfish, Myxine glutinosa, were studied by use of light-microscopic pre- and postembedding peroxidase-antiperoxidase or avidin-biotin-peroxidase techniques, and the immuno-electron-microscopic protein A-gold method. At the light-microscopic level, a strong opsin immuno-reaction was obtained on the outer segments of the photoreceptor cells with sheep and rat antibodies against bovine (rhod)opsin. These outer segments were located in the marginal photoreceptor space and in follicles of the retina, as well as in the tubular lumen of the optic nerve. Ultrastructurally, two classes of outer segments can be distinguished; most of them exhibited a strong antiopsin reaction, while certain elements lacked immunoreactivity with the antisera employed. The protein A-gold particles marked opsin-immunoreactive sites on the photoreceptor membranes. The presence of opsin-immunoreactive material in the retina and optic nerve of the hagfish strengthens the view that this primitive eye lacking a cornea, lens and vitreous body is engaged in light perception. The morphological similarity between the eye and pineal tissue is discussed in connection with the absence of a pineal organ in this species.
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  • 72
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    Cell & tissue research 238 (1984), S. 113-119 
    ISSN: 1432-0878
    Keywords: Crustacea ; Gastrointestinal hormones ; Neuropeptides ; Immunocytochemistry ; Cuticle ; Cancer magister
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A gastric peptide from the Dungeness crab (Cancer magister), extracted and characterized previously (Larson and Vigna 1983b), was localized in the foregut (stomach) of this species by immunocytochemistry using antisera specific for the bioactive carboxy-terminal amino acid sequence common to gastrins and cholecystokinins (CCKs). Immunoreactivity was found in all gastric epithelial cells and in the procuticle. Electron microscopy revealed an absence of peptidergic secretory granules in the gastric epithelial cells. The pattern of immunostaining suggests that the gastric epithelial cells secrete this peptide apically where it is incorporated into the cuticle lining the lumen. Specific immunostaining could not be demonstrated in various neural ganglia or in the hypodermis. The distribution of this peptide is different from that of gastrin/CCK in vertebrates and other invertebrates. This suggests that the crab gastric peptide is sufficiently similar to gastrin/CCK to react with C-terminal specific antisera, but may be anatomically, functionally, and possibly phylogenetically otherwise unrelated.
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  • 73
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    Cell & tissue research 238 (1984), S. 289-296 
    ISSN: 1432-0878
    Keywords: Somatostatin (SRIF) ; Brain ; Pituitary gland ; Immunocytochemistry ; Teleosts
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    Topics: Biology , Medicine
    Notes: Summary Immunocytochemical investigations show that somatostatin (SRIF)-like immunoreactive material is present in the brain and the pituitary of nine different species of teleosts. In the brain, immunoreactive perikarya and fibers are observed in the preoptic periventricular nucleus, the entopeduncular nucleus, the anterior periventricular nucleus, and the nucleus lateralis tuberis. In the pituitary, SRIF-like-immunoreactive fibers occur in the proximal pars distalis (PPD), which contains the growth hormone (GH)-secreting cells. Nerve fibers are scattered among GH cells (cyprinids), or end on the basal lamina at the neuroglandular interface of the PPD (eel, salmonids). In the eel, the proximal neurohypophysis does not penetrate deeply into the PPD that is very poorly vascularized. In some species, e.g. Myoxocephalus, SRIF-like immunoreactive fibers are also observed in the caudal neurohypophysis, and even among MSH cells of the pars intermedia. In long-term starved carps and eels, the amount of SRIF-like material in the pituitary is clearly reduced. A possible role of SRIF in the concomitant stimulation of GH cells is discussed.
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  • 74
    ISSN: 1432-0878
    Keywords: Immunocytochemistry ; Slug brain ; Somatostatin-like material ; Maturation hormone
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    Topics: Biology , Medicine
    Notes: Summary Immunocytological tests reveal the presence of a somatostatin-like substance in perikarya and axons in the brain of the giant slug Limax maximus L. Controls carried out on adjacent sections with absorbed antiserum or different antibodies raised against several biologically active peptides of vertebrates (ACTH-17-39, α- and β endorphin, α- and β MSH, methionin-enkephalin, TRH) demonstrate the specificity of the “staining”. However, some cells are both somatostatin- and FMRF-amide-positive. In the cerebral ganglia, the right Z-area cells, responsible for the synthesis of the maturation hormone (MH) are strongly somatostatin-positive. These results suggest a similarity between the MH and the somatostatin-like material contained in the Z-area cells. The simultaneous presence of two peptides in one and the same cell, the nature (elementary granules or soluble product) of the material, and its site of release are discussed.
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  • 75
    ISSN: 1432-0878
    Keywords: FMRF-amide ; Immunocytochemistry ; Nervous system ; Microstomum lineare (Turbellaria)
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    Topics: Biology , Medicine
    Notes: Summary The central nervous system (CNS) and the peripheral nervous system (PNS) of the flatworm Microstomum lineare were studied by means of the peroxidase-antiperoxidase (PAP) immunocytochemical method, with the use of antisera to the molluscan cardioactive peptide FMRF-amide. FMRF-amide immunoreactive perikarya and nerve fibres are observed in the CNS and the PNS. In the CNS, immunoreactive perikarya and nerve fibres occur in the brain, in the epithelial lining and the mesenchymal surroundings of the ciliated pits, and positive fibres in the longitudinal nerve cords. In the PNS, immunoreactive fibre bundles with variocosities occur in the pharyngeal nerve ring, in symmetrical groups of perikarya on each side of the pharynx, and in the mouth area. Positive perikarya and meandering nerve fibres appear in the intestinal wall. A few immunoreactive cells and short nerve processes are observed at the male copulatory organ and on both sides of the vagina. Some immunoreactive peptidergic cells do not correspond to cells previously identified by histological techniques for neurosecretory cells. The distribution of immunoreactivity suggests that the FMRF-amide-like substance in CNS and PNS in this worm has roles similar to those of the brain-gut peptides in vertebrates. The status of FMRF-amide-like peptides as representatives of an evolutionarily old family of peptides is confirmed by the positive immunoreaction to anti-FMRF-amide in this primitive microturbellarian.
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  • 76
    ISSN: 1615-6102
    Keywords: Bauhinia purpurea ; Colloidial gold ; Golgi apparatus ; Immunocytochemistry ; Lectin ; Lowicryl K4M ; Protein body
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    Topics: Biology
    Notes: Summary The seed lectin of the tree legume,Bauhinia purpurea alba, was localized by electron microscopic immunocytochemistry. The pattern of lectin deposition and site of intracellular localization was examined in mid- to late-maturation seeds. The seed tissue was embedded in Lowicryl K4M, the use of which with seed tissues is discussed. Immunocytochemical labeling was accomplished with colloidal gold coupled to a second antibody. The immunocytochemical reaction was specific and sensitive. Protein bodies, Golgi apparatus and Golgi secretion vesicles were densely labeled. Golgi apparatus was oriented such that Golgi secretion vesicles were in close proximity to the protein bodies. The entire Golgi apparatus was labeled with no concentration gradient across the Golgi stack. These observations indicate that the final site of lectin deposition is the protein body, and that the Golgi apparatus plays an essential role in the deposition process.
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  • 77
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Aromatic acid isomers ; Low capacity ion-exchange resin ; Microbore packed columns
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    Topics: Chemistry and Pharmacology
    Notes: Summary A low-capacity, anion-exchange, porous-polymer resin was used as the packing material in a microbore column. Using a phosphate buffer containing various organic modifiers, retention behavior of substituted benzoic acid isomers with amino, chloro, methyl, nitro and carboxylic groups was investigated. The effect of ethanol, acetonitrile and sodium dodecylsulfate used as organic modifiers on the retention behavior at various eluent pH values was studies and optimum separations of the positional isomers reported.
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  • 78
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    Chromatographia 18 (1984), S. 668-672 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Mixed eluents ; Preferential sorption of eluents ; Pressure effects
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    Topics: Chemistry and Pharmacology
    Notes: Summary Pronounced base line perturbations were repeatably observed in LC both in alternating recycling and in backflushing experiments when binary eluents and refractometric detectors were used. The perturbations which we call eigenzones appeared immediately after operating the recycling or the backflushing valve, and their size and shape depended on the nature of both the column filling and the mixed mobile phase as well as on the pressure drop across the column. Experimental results presented confirm that the primary causes of the eigenzones were the pressure changes in the columns which influenced the sorption equilibrium of mixed eluent components within the column packing. Some practical consequences of this effect are briefly discussed.
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  • 79
    ISSN: 1612-1112
    Keywords: Polycyclic aromatic hydrocarbons ; Column liquid chromatography ; Polyphenyl bonded phases ; Retention and carbon content
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    Topics: Chemistry and Pharmacology
    Notes: Summary The relationship between the retention of PAHs having various molecular sizes and the carbon contents (%) of bonded stationary phases with various types of ligands were investigated. The examined stationary phases were phenyl, diphenyl, triphenyl and benzyl-bonded silicas. The data indicated that sometimes it is not true that the retention of any solute relates linearly to the carbon content of the stationary phase. As the result, it is suggested that the pore size distribution of the silica support, the bulkiness of the ligand bonded to the silica and the molecular size of solute should also be considered before any precise statement can be made.
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  • 80
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    Chromatographia 18 (1984), S. 175-182 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Fourier transform IR ; Polar compounds ; Coal products
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    Topics: Chemistry and Pharmacology
    Notes: Summary Fourier-transform infrared spectrometry (FTIR) was utilized as a detection system for high-performance liquid chromatography of polar compounds on a normal bonded-phase system. In addition to FTIR, UV (254 nm) and refractive index detection were employed in the separation of model compounds containing oxygen and nitrogen atoms in their molecules (phenols, alcohols, neutral and weakly basic nitrogen compounds). A polar amino-cyano (PAC) bonded-phase column was employed with a binary mobile phase (98:2 CDCl3:CH3CN). The separations and the information gained from each detector is discussed, including interactions between sample material and mobile phase. A polar fraction of a coal-derived product is also chromatographed under identical conditions. The results based on FTIR detection and chromatographic retention behavior suggest the exclusive presence of hindered and unhindered phenols.
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  • 81
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    Chromatographia 18 (1984), S. 211-215 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Alkylbenzenes ; PAHs ; Retention prediction ; Microcomputer
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    Topics: Chemistry and Pharmacology
    Notes: Summary The microcomputer-assisted retention prediction system in C18 reversed-phase HPLC is described. The system is based on the use of the hydrophobic parameter and the correlation factor of alkylbenzenes and polycyclic aromatic hydrocarbons (PAHs) which are highly correlated to their retention in reversed-phase HPLC. The system is evaluated by comparing the retention data between measured and predicted values. One of the typical examples applied for PAH analysis in the extracts of diesel particulate matter shows the high potential of the system investigated.
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  • 82
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    Chromatographia 18 (1984), S. 216-220 
    ISSN: 1612-1112
    Keywords: Cation exchangers ; Column liquid chromatography ; Exchange capacity, determination ; Organic peroxidation
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    Topics: Chemistry and Pharmacology
    Notes: Summary Two oxidation procedures for the synthesis of silicabased aliphatic sulfonic acid cation-exchangers for HPLC are described. Several reaction conditions for oxidizing a terminal-bonded thiol with organic peroxides are provided along with their yields, capacities, reproducibility, and chromatographic behavior. Also, three methods for determining exchange capacity are discussed, and their relationship to the capacity factors of charged solutes is compared. The maximum capacity determined by the titration of the bonded silica was 0.34 meq/gram. The general problems of synthesizing cation-exchangers, measuring their efficiencies and exchange capacities, and evaluating their overall performances are outlined.
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  • 83
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    Chromatographia 18 (1984), S. 369-373 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion exchange ; dynamic ; Metal ions ; Equilibria effects
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    Topics: Chemistry and Pharmacology
    Notes: Summary Equilibria occurring during the dynamic ion exchange separation of metal ions were examined, and their effect on the chromatography of the metal ions was evaluated. Bonded reversed-phase silica, pure organic reversed phase, and silica columns were used to differentiate between the role of silica and the reverse phase. The results show that system peaks had the most important influence on the applicability of dynamic exchangers to quantitative metal ion determinations. The response of system peaks to sample composition (pH, ionic strength, matrix effects) was complicated and could not be predicted easily. Equilibria effects that can cause analytical errors were identified and recommendations for elimination of such errors are given.
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  • 84
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Amines ; Phenols ; Fourier transform I.R.
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    Topics: Chemistry and Pharmacology
    Notes: Summary Several chromatographic parameters have been examined as to their effect on detectability for microbore high-performance liquid chromatography coupled with Fourier-transform infrared spectrometry. Detection limits for several compound classes are presented as amount injected. Improvements in detectability for this technique are discussed.
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  • 85
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polystyrene packings ; Pre-column derivatization ; Thiamine compounds ; UV and fluorescence detection
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    Topics: Chemistry and Pharmacology
    Notes: Summary The chromatography of thiochrome and related phosphate esters was studied on two different polystyrene packing materials. The adsorbents were compared with respect to capacity factors, plate heights and resolution factors measured with different mixtures of phosphate buffer (pH 8.5) and methanol. In contrast with derivatized silica, these phases were found to be stable using an alkaline mobile phase and sample. Fast separations were possible with good peak symmetry at all organic modifier concentrations.
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  • 86
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    Chromatographia 18 (1984), S. 445-448 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Amino-bonded stationary phase ; Ion-exchange ; Fermentation solution ; Biomass
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    Topics: Chemistry and Pharmacology
    Notes: Summary In this work the analytical separation of biomass hydrolysis and fermentation products is described. Aminobonded phase and ion exchange materials (HPX 87H, HPX 42A, μ Spherogel Carbohydrate) were used as HPLC stationary phases for the determination of monomeric and oligomeric carbohydrates, carbohydrate degradation and fermentation products. The HPX 42A and the amino-bonded phase columns are especially suitable for the analysis of the gluco oligomers. The HPX 87H column separates the monomeric sugars and their degradation and fermentation products (e.g. furfurals and ethanol) very well. Using the μ Spherogel Carbohydrate column the lower gluco-oligomers as well as the monomeric sugars and their degradation and fermentation products, can be determined within 35 minutes.
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  • 87
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Chiral bonded phase ; Racemate resolution
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    Topics: Chemistry and Pharmacology
    Notes: Summary A chiral stationary phase with an immobilized, optically active diamine was prepared for the separation of enantiomers. The synthesis of the phase was carried out by bonding (−)trans-1,2-cyclohexanediamine to microparticulate silica gel through the coupling agent 3-glycidoxypropylsilane. The resolution of the racemic compounds catechin, 2,2′-dihydroxy-1,1′binaphthyl and trans-1,2-cyclohexandiol, is reported.
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  • 88
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    Chromatographia 18 (1984), S. 495-498 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; PAHs ; Molecular shape recognition
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    Topics: Chemistry and Pharmacology
    Notes: Summary The molecular-shape recognition of polycyclic aromatic hydrocarbons has been studied on various stationary phases in reversed-phase high-performance liquid chromatography. The examined stationary phases were phenyl-, diphenyl-, triphenyl- and benzyl-bonded silicas. The results of regression analysis inidcated that triphenylsilica is the best packing material to recognize the difference in the molecular-shape of structural isomers. This fact was confirmed by the separation of 4-ring isomers.
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  • 89
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention data ; Substituted benzene derivatives ; Correlations
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    Topics: Chemistry and Pharmacology
    Notes: Summary The capacity factors of substituted benzene derivatives on bonded phases of various chain lengths in the reversed-phase mode have been measured and correlation of retention data between four stationary phases has been investigated by linear regression analysis. As a result, a difference in the retention mechanism between phases of shorter chain length and octadecylsilica was observed. In addition, it is also apparent that the multicombination treatment between retention data and descriptors such as π, HA, HD and σ promises the possibility of predicting the retention of substituted benzene derivatives on various reversed-phase stationary phases.
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  • 90
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Polyamines ; Dansyl derivatives ; Peak compression
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    Topics: Chemistry and Pharmacology
    Notes: Summary Analytical conditions for the high-speed, reversed-phase, liquid chromatographic of a diamine (putrescine) and polyamines (spermine and spermidine) were determined. Various elution modes were employed using the same mobile phase constituents: 20mM sodium heptane sulfonate and 20mM acetic acid as solvent A and, pure acetonitrile as solvent B. Samples were derivatized with dansyl chloride before injection. Under isocratic conditions, the separation of the three polyamines was achieved in 7 min. The use of a linear elution gradient led to the same analytical time but with a better resolution of the putrescine peak from non-polyamine frontal peaks. For these measurements, the sample size was 5μl. This volume was increased to 20μl and the use of a steep gradient combined with the peak-compression technique allowed a fast analysis in 2–3 minutes, which may be compared with a 30 min run time necessary when a conventional column is used.
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  • 91
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    Protoplasma 122 (1984), S. 86-90 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Non-articulated laticifers ; Articulated laticifers ; Asclepiadaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary sensitive immunocytochemical method for the identification of laticifers has been developed. Frozen sections of various laticifer-bearing plant material, mounted on slides, were first flooded with the IgG fraction of rabbit anti-latex antiserum, prepared using whole latex ofAsclepias syriaca, then flooded with fluorescein-conjugated IgG fraction goat anti-rabbit IgG to visualize laticifers. Positive fluorescence was observed for laticifers in shoots and embryos ofA. syriaca andStapelia bella and embryos ofA. tuberosa. Laticifers did not fluoresce in shoots ofA. tuberosa andEuphorbia tirucalli, in embryos ofE. marginata, or in petioles ofMusa paradisiaca andCichorium intybus. Controls prepared with uninjected rabbit serum were negative (no fluorescence).
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  • 92
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    Protoplasma 121 (1984), S. 25-33 
    ISSN: 1615-6102
    Keywords: Immunocytochemistry ; Colloidal gold ; Wheat germ agglutinin ; Lectin ; Cryosections
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The chitin-binding lectin wheat germ agglutinin (WGA) is found at the periphery of wheat embryos, and a similar lectin is present at the root tips of older plants (Mishkind et al. 1982). Although a ferritin-conjugated secondary antibody is adequate for localizing WGA in embryos, native electron-opaque particles make the electron microscope identification of added label equivocal in other wheat tissues. As reported here, however, unambiguous ultrastructural localization of WGA-like lectin in adult wheat roots can be obtained with rabbit anti-WGA followed by colloidal gold-labeled goat anti-rabbit (GAR) IgG. Colloidal gold (CG) was prepared by the reduction of gold chloride with citrate, ascorbate or phosphorous. GAR IgG, prepared from serum by antigen affinity chromatograhy, was adsorbed to the gold particles to produce a stabilized suspension of GAR-CG. Localization was performed on 8–12 μM frozen sections of tissue fixed in 4% paraformaldehyde, 0.3% glutaraldehyde, and 0.75% acrolein in phosphate-buffered saline containing 1M sucrose. Localization with GAR-CG was first compared to that ascertained in embryos using other probes and was then extended to the roots of adult plants. An advantage of the GARCG method is that it permits the visualization of antigen at both the light and electron microscope levels in the same section. At the light level, the anti-WGA-GAR-CG complex appears as a red stain that is localized in specific tissues of embryos and in the caps and outer layers of adult roots. Sections in which lectin was detected at the light microscope level were embedded in plastic and sectioned for subcellular examination. Electron dense gold particles indicative of WGA are found at the periphery of protein bodies in wheat embryos and in vacuoles of the roots of adult plants. Sections incubated with control IgG lack reaction product.
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  • 93
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    Chromatographia 18 (1984), S. 574-578 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Microbore column ; Flow rate stability ; Performance ; Quantitative analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Some quantitative implications of the use of microbore columns in column liquid chromatography are investigated. Although for several pumps the flow rate stability at 50 μl min−1 is slightly worse than that of pumps operating at 1 ml min−1, the quantitative performance with respect to repeatability and reproducibility of response factors equals that of conventional liquid chromatography. Thermostatting is strongly recommended for microbore column operation.
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  • 94
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    Chromatographia 18 (1984), S. 62-64 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Reversed phase ; Microparticulate packing ; Amino acids
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    Topics: Chemistry and Pharmacology
    Notes: Abstract A procedure is described by which 20 commonly occurring amino acids may be separated by reversed-phase liquid chromatography at room temperature in a total time, including derivatisation with o-phthalaldehyde/2-mercaptoethanol reagent, of approximately 15 minutes. Further, an increase in column temperature enables increased resolution of certain amino acids to be obtained with small but acceptable losses in fluorescence intensity. The only major drawback to the method is that the derivatisation reagent does not react with proline or hydroxyproline.
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    Chromatographia 18 (1984), S. 15-17 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Ion-pair ; Dynamic equillibrium ; Aqueous and aqueous alcohol eluents
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    Topics: Chemistry and Pharmacology
    Notes: Summary This work describes the dynamic equilibrium of an ionpair, reverse-phase liquid chromatography system. A given adsorbed quantity of an anionic surfactant can lead to different capacity factors and selectivity for anionic solutes.
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  • 96
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Retention correlation ; Silica ; Normal and reversed phase
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    Topics: Chemistry and Pharmacology
    Notes: Summary The correlation between the retention data of polyaromatic hydrocarbons (PAH) obtained in normal-and reversed-phase liquid chromatography is investigated in order to determine the dominant factors controlling the retention. It is clear that the separation of PAHs on various chemically-bonded packing materials in normal- and/or reversed-phase modes is primarily controlled by the molecular structure and shape. The π-electron interaction between the solute and the stationary phase also contributes to the retention, although pure silica shows a somewhat different behavior.
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  • 97
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    Chromatographia 18 (1984), S. 600-602 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Sugars ; Copper (II) complex ; UV detection
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    Topics: Chemistry and Pharmacology
    Notes: Summary A sensitive method of sugar analysis by HPLC is described in which a copper (II) modified silica gel as stationary phase is used. Detection is based on UV absorption of a complex formed between the sugar, copper, and ammonia.
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  • 98
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    Chromatographia 18 (1984), S. 695-697 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Semi-microbore columns ; Reverse-phase ; 3 μm packing
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    Topics: Chemistry and Pharmacology
    Notes: Summary The preparation and use of semi-microbore (2mm i.d.), reverse-phase columns in HPLC has been investigated and they have been compared with 5 mm bore analytical columns. Injection valve design, detector cell volume, column loading and column linking are considered for both semi-microbore and analytical systems. Mobile phase flow-rate, detector time constant and column lifetime are also evaluated for small-bore systems.
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  • 99
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Quantitative determination of L-alanine ; DABS-Cl ; Tryglycine sulphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary A method for the quantitative determination of L-alanine in triglycine sulphate crystals, in the presence of excess amounts of glycine, is described. L-alanine is determined after derivatization with 4-N,N-dimethyl-aminoazobenzene-4′-sulfonyl chloride (DABS-Cl) and in the presence of DABS-valine used as the internal standard. The results obtained indicate that the method permits the determination of alanine in triglycine sulphate crystals at concentrations of about 1%.
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  • 100
    Electronic Resource
    Electronic Resource
    Springer
    Chromatographia 18 (1984), S. 73-80 
    ISSN: 1612-1112
    Keywords: Column liquid chromatography ; Microbore columns ; Performance ; High-speed separation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary Microbore columns of 1 mm i.d. have turned out to be very suitable for the achievement of efficient columns. The packing procedure for stainless steel 1 mm i.d. columns from 5 to 100 cm in length was studied. Stationary phases used were: pure silica gel, octyl, octadecyl and amino bonded silicas. The main parameters (slurry composition, packing system, choice of materials) are discussed. Short columns packed with 3 or 5μm particles allow high speed separations. A separation in 18 seconds is described. Very high plate numbers can be obtained with long columns. With 7–8μm particles, a 1 m column can produce 50,000 plates (h=3). Columns can be joined without loss of efficiency. 270 000 theoretical plates were obtained on a 6 m adsorption column with a test mixture. In reversed-phase chromatography, bile acid sodium salts can be separated on a 1 m column. In adsorption chromatography, details are given of the separation of a polystyrene oligomer sample, as well as a light and a heavy petroleum distillate samples on a 2 m column with refractive index detection in the last-case.
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