A double-labelling fluorescent assay for concomitant measurements of [Ca2+]i and O2. production in human macrophages.

To measure intracellular free Ca2+ concentration ([Ca2+]i) and superoxide (O2) production in human alveolar macrophages, we used the fluorescent Ca2+ indicator fura-2 and the O2-sensitive dye dihydrorhodamine-123, which becomes fluorescent in its oxidized form, rhodamine-123. We describe a new double-dye technique whereby the kinetics of both [Ca2+]i levels and O2. production can be monitored simultaneously. This technique was developed in the dimethylsulfoxide-differentiated monocytic-like U-937 cell line (not equal to U-937), validated by comparison with single dye measurements and applied to human alveolar macrophages. The chemotactic peptide N-formyl-L-Methionyl-L-Leucyl-L-Phenylalanine induced in both cell types a similar transient elevation in [Ca2+]i, followed within seconds by a sustained increase in O2 production, which was however 4-fold weaker in not equal to U-937 cells. These results indicate that O2 production is an early event following the stimulation of human alveolar macrophages. This new double-dye technique may be relevant to other O2 ion-producing cells and could help to define more precisely the kinetics of the events leading to this biological response.

Effects of acute doses of oxiracetam in the scopolamine model of human amnesia.

The scopolamine model of amnesia has been used to test the pharmacodynamic efficacy of oxiracetam in 12 healthy volunteers. The subjects were divided into four experimental groups, according to a double-blind cross over incomplete randomized block design. After a baseline neuropsychological examination, each subject received in two separate sessions one of the following treatments, as acute oral doses: oxiracetam 800, 1600, 2400 mg or placebo. One hour after treatment scopolamine hydrobromide (0.5 mg) was given subcutaneously. The cognitive performance was tested before and 1, 2, 3 and 25 h after scopolamine administration. Scopolamine caused a deterioration of performance of verbal episodic memory, semantic memory and attention tests. In comparison to placebo, oxiracetam improved the overall test performance, with a statistically significant difference at the dose of 1600 mg on delayed recall of word lists, and showed dose-related antagonism of scopolamine-induced effects also on semantic memory and attention. The efficacy of an acute dose of oxiracetam in reducing scopolamine-induced cognitive impairment supports the potential usefulness of this pharmacological model of amnesia for studying the effects of cognition enhancers in humans.

Clinical studies with oxiracetam in patients with dementia of Alzheimer type and multi-infarct dementia of mild to moderate degree.

The cognitive and behavioral effects and the safety of oxiracetam therapy during a placebo-controlled trial and the relevant follow-up up to 1 year in patients with senile dementia of Alzheimer type (SDAT) and multi-infarct dementia (MID) of mild to moderate degree were studied. Sixty male and female outpatients participated in the double-blind, placebo-controlled, parallel-group, randomized trial, comparing the effects of oxiracetam 800 mg b.i.d. and placebo during 90 days of treatment. At the end of therapy, statistical analysis evidenced significant improvements in the group receiving oxiracetam in respect to the placebo group on Mini Mental State Examination, Auditory Continuous Performance Test, Rey's 15 Words Test, Block Tapping Test, Mattis Word Fluency, Luria Alternating Series and Instrumental Activities of Daily Living. Afterwards, 29 of the 30 patients who received oxiracetam, participated in the open follow-up study, receiving 800 mg b.i.d. oxiracetam for a total standard period of 1 year. Statistical improvements in comparison to baseline were again found on the same tests of the first 90 days (except for Rey's 15 Words Test) and on the Memory item of the Inventory of Psychic and Somatic Complaints Elderly. During the late phase of the follow-up, statistically significant worsenings in comparison to baseline were observed on Digit Span Backward, Gibson's Spiral and some non-memory items of IPSC-E. Neither severe adverse events were observed during the whole study, nor changes in routine laboratory examinations. In conclusion, in the present population of patients with mild to moderate degree dementia, the safety of 1,600 mg/day of oxiracetam also up to 1 year of treatment was confirmed.(ABSTRACT TRUNCATED AT 250 WORDS)

Pharmacokinetics of rokitamycin after single administration to healthy volunteers.

The pharmacokinetics of rokitamycin tablets were studied in 12 healthy volunteers in a randomized cross-over design. The doses tested were 200 mg, 300 mg, 400 mg and 600 mg, as single oral administration. Rokitamycin was absorbed quickly with Tmax for all doses around 30 min after drug intake. Total AUC and Cmax values were linearly related to the administered dose. The buffer formulation determined a low interindividual variation. The overall findings show a good similarity with the data obtained in Japanese subjects. Tolerability was very good.

Pharmacokinetics of cadralazine in hypertensive patients.

Pharmacokinetics of cadralazine, urinary recovery of its putative active metabolite 3-hydrazino-6-aminopiridazine derivative (ISF 2405) and clinical effects were assessed in a placebo-controlled trial in 8 hypertensive in-patients, after acute oral administration of cadralazine. After a 2-week placebo-washout period, the protocol envisaged two consecutive days of monitoring of blood pressure and heart rate. On the first day a placebo tablet was given (9 am), while on the second day patients received a 30 mg cadralazine tablet in single-blind conditions. Blood and urine samples were obtained during the active drug day until 12 h after administration. Concentration of cadralazine in plasma and urine was detected by a specific HPLC method, while the metabolite ISF 2405 was detected in urine by a GC-MS method. Cadralazine caused gradual and long-lasting pressure decrease, statistically significant in comparison to placebo between 3 and 12 hours from drug intake, accompanied by a significant increase in heart rate. Cadralazine by oral route was promptly absorbed with a mean peak time of 1.3 h. Thereafter it followed a monoexponential decay curve, with a plasma half life of 3.1 h. The relative different bioavailability of oral cadralazine among patients was not correlated with cardiovascular changes. Urine recovery of unchanged drug after 12 h was high, reaching 67.3%, while concentration of metabolite ISF 2405 was about 1/1000 of parent compound.

Oxiracetam pharmacokinetics following single and multiple dose administration in the elderly.

The kinetics of the nootropic drug oxiracetam was studied in 6 elderly female patients (age range 69-96 years) in good physical condition. After a single oral dose (800 mg), serum oxiracetam levels reached a peak (25 +/- 6 micrograms/ml) within 1-3 h and declined thereafter with a half-life of 3 to 6 h. Eighty-four percent of the administered dose was recovered in urine as unchanged drug within 24 h. During a maintenance regimen (800 b.i.d. at 8 a.m. and 2 p.m.) for 7 days oxiracetam did not show any accumulation in serum, trough serum levels before the morning and afternoon doses being 4.6 +/- 2.3 and 17.0 +/- 8.2 micrograms/ml respectively on day 4 and 5.7 +/- 3.3 and 17.9 +/- 7.7 micrograms/ml respectively on day 7. Comparison with previous pharmacokinetic data obtained in young subjects suggests that the clearance of the drug is lower in the elderly, probably as a result of the physiological decrease in renal function in old age.

Clinical and neuropsychological study with oxiracetam versus placebo in patients with mild to moderate dementia.

40 out-patients with a mild to moderate degree of dementia (11 less than or equal to MMSE less than 24) participated in a between-subjects (n = 20 + 20) double-blind placebo-controlled randomized trial comparing the effects of oxiracetam 800 mg bid and placebo during 90 days of treatment. At the end of therapy, statistical analysis (ANOVA) detected significant differences between groups: after oxiracetam treatment, improvements were observed on Mini Mental State Examination, Auditory Continuous Performance Test, Block Tapping Test, Word Fluency and Instrumental Activities of Daily Living. No side effects were observed. In conclusion, in the present population of patients with mild to moderate degree dementia, 1600 mg/day of oxiracetam was effective in enhancing both attentional activities and other, more complex, neuropsychological functions.

Controlled clinical trial of cadralazine as a second-step drug in the treatment of hypertension.

The antihypertensive efficacy of a new long-lasting vasodilator, cadralazine, and the diuretic chlorthalidone have been compared in hypertensive patients receiving concurrent treatment with atenolol. After a 4-week run-in period with atenolol alone 100 mg/day, two groups of 10 patients whose diastolic blood pressure exceeded 100 mm Hg were given for a period of 65 days either cadralazine 15 mg/day or chlorthalidone 25 mg/day, according to a randomized, double-blind, between-patients design. Compared to atenolol alone, both cadralazine and chlorthalidone induced a statistically and clinically significant decrease in blood pressure. The antihypertensive effect did not differ significantly between groups. Good compensation of the atenolol-induced decrease in heart rate was obtained with cadralazine, whereas during atenolol + chlorthalidone treatment at times the standing heart rate was significantly lower than during treatment with atenolol + cadralazine. Side-effects, many of which were already present during atenolol treatment, occurred with a similar frequency in both groups. It is concluded that atenolol + cadralazine and atenolol + chlorthalidone are equally well tolerated, acceptable and effective in the treatment of hypertension, but that further studies are warranted to explore the potential haemodynamic advantages of the cadralazine + atenolol combination.

Cadralazine, a new vasodilator, in addition to a beta-blocker for long-term treatment of hypertension.

51 hypertensive outpatients, whose diastolic blood pressure exceeded 100 mmHg after a 2-week period on atenolol alone (100 mg once daily) participated in this long-term study. They received, in addition to atenolol, the vasodilator cadralazine (ISF 2469; 10 to 30 mg once daily) for a standard period of 24 weeks, according to an open design. Cadralazine caused a progressive and important decrease in both systolic and diastolic blood pressure, from 173/111 mmHg (end of atenolol alone) to 154/99 mmHg (12th week, p less than 0.01/p less than 0.01; mean dose, 24.5 mg/day). At this time a diuretic was added as a third-step drug in 15/51 initial patients (29%), and final blood pressure in all patients was 150/96 mmHg (p less than 0.01/p less than 0.01), with positive results in 88% of the cases. During cadralazine treatment, heart rate was always significantly lower than before atenolol alone; the most common side effects, many of which were already present during treatment with atenolol alone, included headache, asthenia, dizziness, palpitation and flushing, and tended to disappear spontaneously as therapy progressed. Routine laboratory tests did not show important changes; sodium excretion was not reduced. In conclusion, the therapeutic efficacy of cadralazine, its low or absent salt and water retention effects, its good tolerability, and the high compliance obtained with once daily administration allowed the use of this vasodilator as a second-step drug for long-term treatment of hypertension.

Comparison of the antihypertensive activity of cadralazine (ISF 2469) and dihydralazine during chronic treatment.

The antihypertensive activity of a new vasodilating agent (ISF 2469) cadralazine (CD) was compared in a randomized, crossover, intrapatient study with that of hydralazine (HD) in 20 hypertensive patients whose diastolic blood pressure (BP) was greater than 95 mmHg during treatment with atenolol (AT), 100 mg 1 X daily. The initial dose of CD was 15 mg 1 X daily; after 15 days in case of poor response, the dose was increased to 20 mg 1 X daily. HD was given at a dose of 25 mg 3 X daily and was increased to 50 mg 2 X daily in case of poor response. BP values (standing) during AT were 174/108; they fell to 144/88 during CD and to 138/88 during HD. No significant difference was detected between the two drugs for both systolic and diastolic BP (supine and standing). Heart rate increased with both drugs, with a greater increase during CD. The difference was clinically nonsignificant. A total of 24 patients were enrolled, but 4 had to cease treatment because of side effects during HD. The overall prevalence of side effects was much higher during HD, especially during the first days of therapy. Also the severity of side effects was greater during HD. Our data show that CD has the same antihypertensive activity as HD with a lesser incidence of side effects and with a single dose/day administration in contrast with three administrations of HD. This can result in greater patient compliance.

Cadralazine (ISF 2469): dose-related antihypertensive activity after single oral administration to patients.

Cadralazine (ISF 2469) was administered to 24 hypertensive patients in single oral doses of 7.5, 10, 15, 20 and 30 mg, according to a single-blind, placebo-controlled, within-patient change-over design. The study was done in 2 stages: in the first a range including the upper and lower doses was studied (7.5, 15, 30 mg and placebo), and in the second the range of doses was restricted (10, 15, 20 mg and placebo). The drug produced a significant decrease in blood pressure in the supine and standing positions. The decrease became clinically important starting from the 15 mg dose. Its action was still significant 12 h after administration. A significant increase in heart rate was also observed. All the effects were correlated with the dose. Side effects occurred mainly after the 30 mg dose. Thus, cadralazine, in a single oral dose in man, showed good antihypertensive activity starting from the 15 mg dose, and its effect was dose-related, slow in onset and long-lasting.

Plasma FSH, LH and prolactin levels in postmenopausal women undergoing cyclofenil treatment.

Ten postmenopausal women were studied in an attempt to identify the mechanism of action of cyclofenil, a non-steroidal anti-estrogen which has proved to be effective in the climacteric syndrome. A double-blind inter-patient clinical investigation was undertaken, with patients assigned randomly to treatment for 10 days with cyclofenil, 400 mg/day, or else conjugated estrogens, 1.25 mg/day, always given orally at 8 a.m. Serum FSH, LH and PRL levels were determined daily 2 days before, during and for 2 days after treatment. On the first and tenth day of treatment, five blood samples were drawn between 8 a.m. and 4 p.m. to ascertain if there was any drug-induced variation in the hormonal secretory patterns. Furthermore, endometrial biopsies of all patients were taken before and immediately after the 10-day treatment. In 5 patients, endometrial biopsies were repeated after 3-6 months of therapy with cyclofenil. The results indicate that cyclofenil has two opposing actions on the hypothalamic-hypophyseal axis, one estrogen-like, in that it depresses serum FSH levels, and the other antiestrogen-like, in that it depresses serum PRL levels. They also show that at both the peripheral and the central level cyclofenil is a drug of first choice for postmenopausal women at risk for endometrial and mammary neoplastic pathology.

Indoprofen in the treatment of osteoarthrosis. A controlled trial in comparison with ibuprofen.

A controlled research has been carried out in 39 patients with osteoarthrosis in order to evaluate the efficacy of Indoprofen, a new analgesic-antiinflammatory compound. This product, a phenylpropionic acid derivative, was given by mouth at the dose of 600 mg daily for 7 days. For comparison a treatment with 900 mg daily of Ibuprofen for the same length of time was given to patients according to a double-blind cross-over experimental design. Significant decrease in overall pain intensity was observed together with significant improvement in motility. Indoprofen at the daily dose of 600 mg was comparable in efficacy with 900 mg of Ibuprofen in the relief of pain related to osteoarthrosis. Toleration of the drugs was very good.

Comparative study of the ventilatory responsiveness to CO2 in bronchitic and emphysematous patients with chronic respiratory failure.

The comparative ventilatory responsiveness to CO2 was studied in 13 chronic bronchitics and 14 emphysematous patients, all exhibiting respiratory insufficiency and with FEV1 less than 1,500 ml. The gas inhaled was enriched with oxygen (F1O2 =0.6) and contained 6% CO2, and measurements were taken when the patients had reached a stable state. The ratio delta VE/deltaPaCO2, which represents the ventilatory responsiveness to CO2, was higher in the emphysematous patients (1.18+/-0.51 liters-min-1. Torr-1) than in the bronchitics (0.76+/-0.34, p less than 0.025), but the deviation on either side of the mean was large in each group. The ventilatory responsiveness to CO2 was proportional to the initial PaCO2, FEV1, total airways resistance, total pulmonary work and especially to inspiratory mechanical work done on the lung ( r=-0.73, p less than 0.001). The difference in ventilatory responsiveness to CO2 between the bronchitic and emphysematous patients may be explained by the difference in energy expended in breathing. It was not possible to exclude an effect due to a difference in sensitivity within the respiratory centers, since inspiratory mechanical work was not measured during CO2 inhalation. It was thought likely that mechanical factors play a triggering role, in that they cause a fall in ventilatory responsiveness to CO2 and hypercapnia, the latter causing the central hyposensitivity which maintains the retention of CO2.

[Value of pulmonary hemodynamic exploration during muscular exercise in chronic bronchitis?]. / Que peut apporter l'exploration hémodynamique pulmonaire au cours de l'exercice musculaire dans la bronchite chronique

The value of pulmonary haemodynamic tests during physical exercise in chronic bronchitis was shown by the comparison of two groups of patients. In the first group (n=24) the PAP during exercise is lower than 30 torr. In the second it was over 30 torr. The PAP at rest was always lower than 20 torr. The load was 40 to 50 watts, i.e. an average O2 consumption of 500-600 ml.mm-1 m-2. The cardiac output doubled on average in exercise. Both groups varied markedly in their PAP at rest: 13.6 +/- 1.7 torr for the first group and 15.8 +/- 2.4 for the second (p less than 0.001). In fact differences in pressure during exercise (I=25.0 +/- 3.4 torr; II=39.6 +/- 7.4 torr, p less than 0.001) could be explained mainly by the differences of pulmonary vascular resistances (I=0.91 +/- 0.37; II=1.47 +/- 0.61, p less than 0.005): they tended to fall during effort in the first group and increased slightly in the second; and by the much higher increase in the pulmonary "capillary" pressure during exercise in the second group (I=12.5 +/- 4.4 torr; II=19.7 +/- 72 torr, p less than 0.001). The cardiac output during rest and exercise was equal in both groups. The haemo-dynamic "recovery delay" was much higher in the second group. The spirographic shortage was on the whole identical in both groups. PaO2 on average was higher in group I (p less than 0.05) where it improved during exercise (p less than 0.01). The PaO2 of the second group did improve during exercise. The haemodynamic differences were concomitant with the differences in gas exchanges during effort, of well known prognostic significance. As the "foretelling" of PAP in effort from the PAP at rest was quite poor, it appeared that haemodynamic test in effort has a real value in contributing efficiently to the differenciation of the degree in severeness. The threshold of 30 torr for PAP in exercise (and for the load mentioned above) seemed a good discriminating factor.