Terminal nerve gonadotrophin-releasing hormone (GnRH) neurones express multiple GnRH receptors in a teleost, the dwarf gourami (Colisa lalia).

Gonadotophin-releasing hormone (GnRH) peptide released from the terminal nerve (TN)-GnRH neurones of the dwarf gourami primarily modifies the electrical properties of various neurones, including the TN-GnRH neurones themselves. However, our knowledge on the expression of GnRH receptors (GnRHRs) in the TN-GnRH neurones is still limited. Here, we used the single-cell reverse transcriptase-polymerase chain reaction after whole-cell patch-clamp recording to study the distribution of various GnRHR types expressed in the individual TN-GnRH neurones. We found that TN-GnRH neurones express two of the three types of GnRHRs cloned in the dwarf gourami: GnRHR1-2 and -R2, but not -R1-1. Furthermore, in agreement with our previous findings, all TN-GnRH neurones contained mRNAs of salmon GnRH but not chicken GnRH-II.

Pharmacokinetics of ramipril in the elderly.

Ramipril (HOE 498) is a pro-drug of which the main metabolite (HOE 498 diacid or ramiprilat) is a potent angiotensin converting enzyme inhibitor. Thirteen healthy white volunteers (5 females and 8 males, ages 65 to 76 years) participated in a study to investigate the pharmacokinetics of HOE 498 in the elderly. After administration of 10 mg of HOE 498, sequential urine and serum specimens were obtained for assay of HOE 498 and metabolites (HOE 498-glucuronide, diacid, diacid-glucuronide, diketopiperazine and diketopiperazine acid). Side effects, clinical chemistry and hematology were monitored. HOE 498 reached peak concentrations of 62.4 +/- 23.3 ng/ml in serum after 0.7 +/- 0.3 hours. Serum levels decreased with an apparent half-life of 0.9 +/- 0.4 hours. The diacid was rapidly formed in serum, reaching peak concentrations of 40.6 +/- 14.0 ng/ml after 2.0 +/- 0.6 hours and declining with a half-life of 2.2 +/- 0.5 hours. A prolonged terminal phase of serum concentration versus time curve was observed at concentrations less than 1 ng/ml. The mean recovery of HOE 498 and metabolites in urine, up to 26 hours after administration, was 35 +/- 14% of the dose. The apparent half-lives, calculated from urine parameters, for HOE 498 and the diacid were 2.6 +/- 0.9 and 4.0 +/- 1.1 hours, respectively. The mean peak concentration and half-life of HOE 498 in serum are slightly higher in the elderly than in younger volunteers. Complete urinary collection was not possible, but urinary recovery did not seem different from younger volunteers.

Influence of renal function on the pharmacokinetics of ramipril (HOE 498).

The pharmacokinetics of ramipril (HOE 498) were studied after oral administration of a single 10 mg dose to 24 hypertensive patients with different degrees of renal function. The creatinine clearance ranged between 4.1 and 126 ml/min/1.73 m2 and was below 35 ml/min/1.73 m2 in 16 patients. Angiotensin converting enzyme activity and the concentrations of ramipril and its active diacid metabolite ramiprilat were measured in plasma up to 10 days after drug intake. Urine levels of ramipril, ramiprilat, their glucuronides and 2 major metabolites (a diketopiperazine and a diketopiperazine acid) were measured up to 4 days after medication. The plasma concentration-time curve of ramiprilat was polyphasic with an initial steep decline after the peak level and a subsequent very long terminal phase at low concentrations. Impaired renal function resulted in higher peak levels of ramiprilat, longer times to peak and a markedly slower decline of plasma ramiprilat levels. Hence, the duration of angiotensin converting enzyme inhibition was considerably prolonged in renal failure and depended on the severity of renal impairment. The urinary excretion of ramipril and its metabolites decreased with decreasing renal function and was linearly related to the creatinine clearance, suggesting an alternative pathway of elimination. The pattern of excretion rates of ramipril and its various metabolites was not affected by renal failure. In contrast to the marked changes in the renal elimination, no relevant differences were observed in the absorption of ramipril from the gastrointestinal tract. Systolic and diastolic blood pressure decreased in all groups. The single 10 mg dose of ramipril was well tolerated.

Dose linearity and other pharmacokinetics of cefodizime after single-dose intravenous administration.

Pharmacokinetics of cefodizime, a broad-spectrum cephalosporin antibiotic, were determined after intravenous (IV) administration of single doses of 1.0, 1.5, and 2.0 gm to 12 healthy male volunteers in an open study. In a separate pilot study, data were obtained after IV administration of 0.5 gm of cefodizime to six healthy male volunteers. Determinations of cefodizime in serum and urine using a microbiological assay agreed with determinations using high-pressure liquid chromatography (HPLC), which specifically measures the unchanged drug. Active metabolites of cefodizime were not detected. After IV administration of 1.0, 1.5, and 2.0 gm of cefodizime, initial mean serum concentrations were 215, 322, and 394 mg/L, respectively (HPLC determinations). A linear response to dosage was shown (coefficient of correlation r = .89), as was the case for area under the serum concentration-time curve to infinity, AUC infinity (r = .82), and 48-hour urinary recovery of cefodizime (r = .94). In all cases, the corresponding values obtained after the 0.5-gm dose showed that the linearity extended to this dose. The kinetics of single-dose administration of cefodizime corresponded to a two-compartment open model with an apparent steady-state volume of distribution of about 40 L. Volume of distribution, terminal elimination half-life, serum and renal clearance, and percent urinary recovery were not dose dependent. Cefodizime combined a long terminal elimination half-life (mean, 2.5 hours) with a high urinary recovery (80%). After IV administration of cefodizime, urinary concentrations of the unchanged drug remained above 5 micrograms/ml for at least 24 hours after drug administration and were dose dependent. Mean values for the 1.0-gm and 2.0-gm doses were, respectively, approximately 12 mg/L and 30 mg/L, several times the minimal inhibitory concentrations (MIC90) for most clinically relevant bacteria. These results suggest that once- or twice-daily IV dosing with cefodizime (depending on the dose regimen) would be suitable for clinical use. The drug was safe and well tolerated.

Multiple-dose pharmacokinetics of ofloxacin, a new broad-spectrum antimicrobial agent.

Twelve healthy male volunteers received 14 oral doses of ofloxacin (300 mg each), and the concentrations of the unchanged drug were measured at various times in serum and urine over a period of 15 days. Serum and urine ofloxacin concentrations were determined in specific assays using high-pressure liquid chromatography (HPLC); urine levels were also determined by means of a microbiological assay. A washout period of 72 hours followed the first and 14th doses, allowing comparison of serum pharmacokinetics at the beginning and end of the multiple-dose regimen. Doses 2 to 14 were administered at 12-hour intervals. Maximum serum concentration (Cmax), concentration at 12 hours after dosing (C12), and area under the serum concentration-time curve (0 to 72 hours) were all 1.3 to 1.7 times greater after the 14th dose than after the initial dose. A 1.6-fold increase in C12 was already evident after the third dose; C12 remained more or less constant thereafter. Thus it is concluded that ofloxacin rapidly attained steady-state serum levels under a multiple-dose regimen, at levels only slightly above those following a single dose. High serum Cmax levels (4.6 and 5.9 micrograms/ml after the first and 14th doses, respectively) and long serum half-lives (6.0 and 7.3 hours after the first and last doses, respectively) indicated long-lasting, clinically relevant serum ofloxacin concentrations after oral administration. Serum ofloxacin levels 24 hours after the last dose were in the range of 0.3 to 0.7 microgram/ml, above the minimal inhibitory concentration (MIC90) for most bacterial strains. Cumulative urinary recovery remained high throughout the study. After 14 doses of ofloxacin (total, 4.2 gm), 88% of the unchanged drug was recovered in the urine; urinary concentrations remained above 1 microgram/ml, far above the MIC90 for most bacterial strains, for at least 108 hours after the final dose. High renal clearance values relative to total clearance (98%) confirmed the importance of the renal elimination route. Determinations of ofloxacin in urine using a microbiological assay were in close agreement with the HPLC determinations for all samples obtained throughout the study. Thus no detectable appearance of active metabolites occurred under the multiple-dose regimen. Ofloxacin was well tolerated; mild, probably drug-induced side effects were reported by three subjects, but they did not require any countermeasures.

Determination of plasma catecholamines by means of radioenzymatic labelling and high pressure liquid chromatographic separation.

0.05 ml plasma samples are incubated with 3H-S-adenosylmethionine and catechol-O-methyl-transferase. The resulting methodoxy derivatives are extracted, the extracts separated by high pressure liquid chromatography and the metanephrine fractions collected. Evaluation is performed by liquid scintillation counting of radioactivity in the respective fractions. The following performance criteria are presented: precision, accuracy, detectability (40 pg/ml for adrenaline and noradrenaline, 130 pg/ml for dopamine), linearity and day-by-day variation. Comparison with a standard method shows an excellent correlation for adrenaline and noradrenaline.

The effect of the converting enzyme inhibitor HOE 498 on the renin angiotensin system of normal volunteers.

The converting enzyme inhibitor HOE 498 was evaluated in 12 normotensive male volunteers aged 21 to 26. The efficacy of single 5, 10 or 20 mg oral doses in blocking the pressor response to exogenous angiotensin I was tested in 3 of the subjects. All 3 doses of HOE 498 reduced the pressor response to exogenous angiotensin I to below 50% of control within 1,5 h following administration of the drug. Plasma renin and converting enzyme activity, blood angiotensin I, as well as plasma angiotensin II and aldosterone were measured serially before and up to 72 h following oral administration of a single dose of 2.5, 5, 10 or 20 mg of HOE 498 to groups of 5 volunteers each. As expected, blood angiotensin I levels and plasma renin activity rose while plasma converting enzyme activity, plasma angiotensin II and aldosterone concentration fell after administration of the drug. While the dose of 2.5 mg did not reduce plasma converting enzyme activity below 20% of control, the higher doses all resulted in plasma converting enzyme inhibition exceeding 90%. No side-effects were observed. It is concluded that in normal volunteers HOE 498 is an effective potent and long-acting converting enzyme inhibitor. Based on these preliminary findings it is expected that 5 mg HOE 948 will turn out to be adequate for therapeutic use.

Fluorometric determination of clobazam, a 1,5-benzodiazepine, in human plasma.

A fluorometric procedure for clobazam, a 1,5-benzodiazepine, based on a fluorophore formed upon irradiation of the drug using short wavelength UV light (254 nm) for 35 min is presented. Fluorescence is linear over a 100-6400-ng/ml range using excitation and emission wavelengths of 350 and 400 nm, respectively. Application of the method to the determination of clobazam in spiked human plasma samples revealed that the drug can be determined at nanogram per milliliter levels with an accuracy of 1-5%. The procedure is specific for clobazam in samples containing its major plasma metabolite, N-desmethylclobazam, and also in samples containing 1,4-benzodiazepines and other selected drugs. A plasma level-time profile after oral administration of a single 40-mg dose of clobazam to a healthy adult male is also illustrated.

Positive inotropic effect of the inhibition of cyclic GMP-stimulated 3',5'-cyclic nucleotide phosphodiesterase (PDE2) on guinea pig left atria in eu- and hyperthyroidism.

The significance of PDE2 on the atrial inotropy was studied in eu- and hyperthyroidism. The contractile force was measured and negative inotropic capacity of N6-cyclopentyladenosine (CPA) was determined on left atria isolated from 8-day thyroxine- or solvent-treated guinea pigs, in the presence or absence of EHNA (adenosine deaminase and PDE2 inhibitor) or NBTI (nucleoside transporter inhibitor). EHNA was administered to inhibit PDE2, while NBTI was used to model the accumulation of endogenous adenosine. The reduction of the contractile force caused by EHNA was smaller in the thyroxine-treated atria than in the solvent-treated samples. Contrary, NBTI induced a decrease in the contractile force without significant difference between the two groups. In addition, EHNA enhanced the efficiency of CPA in thyroxine-treated atria and did not affect it in solvent-treated samples, while the response to CPA was decreased by NBTI in all atria, especially in hyperthyroidism. On the basis of greater retention of the contractile force and sustained/enhanced responsiveness to CPA in the presence of EHNA we conclude that PDE2's inhibition has a significant positive inotropic effect in guinea pig atria and this effect is proven to be augmented in hyperthyroidism.

Pharmacokinetic investigations on diminazene and rolitetracycline in comparison to a combination of both.

Serum level tests were carried out on healthy cows with a combination drug of one part Berenil and two parts Reverin. When compared with the commercial preparations Berenil (Diminazene) and Reverin (Rolitetracycline), the kinetic behaviour of Reverin in the combination preparation was identical. In the case of Berenil, administered in the form of the combination drug, the second slow elimination phase with a 63 hour half-life, as found after the administration of Berenil only, could not be observed. Eight hours after administration of the combination only minimal Berenil serum levels were detectable, after 24 hours the levels were lower than the limit of detection. This has a practical bearing on the question of residues.

Fast camera studies at an electron cyclotron resonance table plasma generator.

A simple table-size ECR plasma generator operates in the ATOMKI without axial magnetic trap and without any particle extraction tool. Radial plasma confinement is ensured by a NdFeB hexapole. The table-top ECR is a simplified version of the 14 GHz ATOMKI-ECRIS. Plasma diagnostics experiments are planned to be performed at this device before installing the measurement setting at the "big" ECRIS. Recently, the plasma generator has been operated in pulsed RF mode in order to investigate the time evolution of the ECR plasma in two different ways. (1) The visible light radiation emitted by the plasma was investigated by the frames of a fast camera images with 1 ms temporal resolution. Since the visible light photographs are in strong correlation with the two-dimensional spatial distribution of the cold electron components of the plasma it can be important to understand better the transient processes just after the breakdown and just after the glow. (2) The time-resolved ion current on a specially shaped electrode was measured simultaneously in order to compare it with the visible light photographs. The response of the plasma was detected by changing some external setting parameters (gas pressure and microwave power) and was described in this paper.

[Analytical methods I]. / Analytische Methoden I

The most frequently used methods such as spectrophotometry, fluorometry, etc., for the analysis of drugs in biological fluids are compiled. The usefulness of quantitative chromatographical procedures is also mentioned. Reliability criteria of these assays are extensively discussed and some results are compared to those found by other methods like labelling and microbiology.

Interference by the new antibiotic cefpirome and other cephalosporins in clinical laboratory tests, with special regard to the "Jaffé" reaction.

The new cephalosporin, cefpirome, was investigated for its possible interference in clinical laboratory tests, especially the determination of creatinine. Tests for bilirubin, cholesterol, protein, urea, and uric acid were also studied. A selection of commercially available compounds such as cefoperazone, cefazolin, cefamandole, cefoxitin, latamoxef, ceftriaxon, cefotiam, cephaloridine, cephalotin, cefotaxime, cefazedone and cefuroxime, and cefodizime (a compound under development) were also included in these investigations. Interference was found only with the "Jaffé method" for the determination of creatinine. Pronounced interactions with alkaline picrate were observed with cefpirome, cefoxitin, cephalotin and cephaloridine, whereas the other compounds showed only weak reaction or no reaction at all. The cephalosporins did not interfere in any of the other tests mentioned above. Care must therefore be taken in the determination of creatinine in samples from patients under treatment with these drugs. The assay methods of choice at present are enzymatic or specific HPLC assays.

Physico-chemical and anlytical studies of penbutolol.

In the present paper physico-chemical and analytical studies on penbutolol sulfate (Hoe 893d) are reported. In addition to an interpretation of ultraviolet and fluorometric spectra, data are given regarding the dissociation constant, solubility, distribution, and protein binding. The substance and its major metabolte, as well as their glucuronides, are detectable by means of a selective fluormetric method. Moreover, mention is made of the results of a human pharmacokinetic study.

[A new fluorometric method for the determination of drugs in biological material (author's transl)]. / Eine neue fluorometrische Methode zur Bestimmung von Pharmaka in biologischem Material

A description is given of a new method for fluorometric determination of drugs in biological material. The assay is based on photochemical transformation of non-fluorescent compounds in fluorophors by short-wave UV light. The pharmacokinetics of the 2-[4-(4'-chlorophenoxy)-phenoxy]-propionic acid (HCG 004) serves as an example to demonstrate the usefulness of this method; reference is also made to analogously successful studies of other drugs.

High performance liquid chromatography in research of pharmacokinetics and metabolism.

High performance liquid chromatography in connection with monochromatic UV-detection has proved to be a powerful tool for separation and quantitative determination of drugs and their metabolites in body fluids. Serum samples from volunteers medicated with the psychotropic drugs fosazepam and nomifensine are analysed by this method. Separation times are less than 5 min; the detection limits are within the range of 30-50 ng/ml serum. The reliability of the method is discussed: The results are compared to those from the same serum samples obtained by measurement of total radioactivity and by quantitative mass spectrometry in order to confirm the accuracy of the method. Examples of the pharmacokinetic profile of the two drugs and their metabolites in serum are presented, based on analysis by the methods described.

Quantitative determination of clobazam in serum and urine by gas chromatography, thin layer chromatography and fluorometry.

The procedures available for determination of clobazam (Frisium, Hoechst) are gas chromatography, fluorometry, and thin-layer chromatography. The study presents detailed descriptions of analytical procedures appropriate for routine determinations in serum and urine, and results from human trials. Moreover, the physicochemical properties of clobazam, viz., solubility, distribution, and protein binding are given.

Avoidable mortality. Is it an indicator of quality of medical care in eastern European countries?

Age-standardized time trends (1979-1988) for avoidable mortality in two Eastern European countries (Hungary and Czechoslovakia) and selected developed countries (England and Wales, France, Italy, Japan, Portugal and USA) have been analysed. Mortality from both all avoidable causes and all other causes declined in the selected developed countries during the period of observation, the decline in rates for avoidable causes was faster than that for all other causes. In Hungary and Czechoslovakia the death rates from both groups of causes increased in the first part of the period studied and a decline in mortality from both types of causes could be observed from 1985. As a consequence, the difference in avoidable mortality between the Eastern European countries and the developed countries increased by the end of the observation. Studies on mortality from individual amenable causes showed that the death rates are usually much higher in Hungary and Czechoslovakia than in the developed countries and the differences did not diminish during the period of study. In Hungary and Czechoslovakia the bad pattern of mortality from conditions amenable to medical interventions is believed to reflect, at least in part, the crisis in the health services which these countries have experienced for the past decades.

Direct assay of conjugates of penbutolol and its metabolites in urine.

The metabolites of 1-tert.-butylamino-3-(2-cyclopentylphenoxy)propan-2-ol (penbutolol, Betapressin), penbutolol 2-glucuronide, 4'-OH-penbutolol 2-glucuronide, 4'-OH-penbutolol 4-sulfate and 1"-dehydropenbutolol 2-glucuronide were determined in urine by high-performance liquid chromatography (HPLC). The compounds were determined after direct injection, that is to say without prior cleavage of the conjugates to the corresponding aglycones. In the case of the glucuronides, the urine was injected into the HPLC system without further sample preparation. The sulfate was determined after ion-pair extraction. Fluorimetric detection was employed. Depending on the compound, the detection limits lay between 0.07 and 0.3 micrograms/ml. This method was used to determine the cumulative urinary excretion of a subject.