Regulation of CDC25B phosphatases subcellular localization.

The CDC25B dual specificity phosphatase is involved in the control of the G2/M transition of the cell cycle. Subcellular localization might represent an important aspect of the regulation of its activity. We have examined in transiently transfected asynchronous HeLa cells the localization of HA-tagged CDC25B proteins and found that they are nuclear or cytoplasmic suggesting the existence of an active shuttling. Accordingly, localization analysis of deletion and truncation proteins indicates that CDC25B contains a putative nuclear localization signal located between residues 335 and 354. We also demonstrated that a short 58 residues deletion of the amino-terminus end of CDC25B is sufficient to retain it to the nucleus. Mutational analysis indicates that a nuclear export sequence is located between residues 28 and 40. In addition, treatment of the cells with the exportin inhibitor, Leptomycin B, has the same effect. The mutation of Ser-323, a residue that is essential for the interaction with 14-3-3 proteins, also abolishes cytoplasmic staining. The subcellular localization of CDC25B is therefore dependent on the combined effects of a nuclear localization signal, a nuclear export signal and on the interaction with 14-3-3 proteins.

Use of formoterol dry powder administered for three months via a single-dose inhaler in 1,380 asthmatic patients.

Formoterol is a long-acting beta 2-adrenoceptor agonist available in a single-dose breath-actuated device (Foradil) for asthma treatment. Since efficacy and ease of use are key factors for compliance to therapy, the aim of this study was to assess correct use, efficacy and safety of Foradil in a 3-month, open, uncontrolled, multicenter trial. This study was performed on 1,380 patients with moderate or severe persistent asthma treated with inhaled corticosteroid (age: 48.4 +/- 16.2 years; FEV1: 65.4 +/- 19.4% of normal). During the study, compliance was over 90%. More than 90% of the patients used the inhaler correctly and found it easy or very easy to use. The mean increase in peak expiratory flow rate (PEFR), 30 to 60 min after inhalation, was 52.3 and 36.7 l/min for the morning and the evening respectively (p = 0.0001). This increase was already significant 5 min after inhalation, confirming the fast onset of action of formoterol. Mean predose PEFR and daytime/nocturnal symptom scores improved during the length of the study. Rescue short-acting beta 2-agonist consumption was more than three times reduced. At study completion, formoterol dosage was 12 micrograms and 24 micrograms twice daily for 71.5% and 28.5% of the patients respectively. Physicians judged the overall efficacy as good or very good in 87.1% of the patients, and they estimated the tolerability as very good or good in 92.6%. Drug-related adverse events were similar to those of other beta 2-agonists. In conclusion, this study demonstrated the ease of use of this formoterol single-dose dry powder inhaler, and confirmed the good efficacy and safety profile of this long-acting bronchodilator in asthma.

Therapeutic equivalence of diclofenac sustained-released 75 mg tablets and diclofenac enteric-coated 50 mg tablets in the treatment of painful osteoarthritis.

Efficacy and tolerability of diclofenac sustained-released (SR) 75 mg tablets taken b.i.d. were compared with that of enteric-coated diclofenac sodium 50 mg tablets given t.i.d. in a seven-day, randomised, double-blind, double-dummy, parallel groups study in 294 outpatients suffering from painful femorotibial or hip osteoarthritis. Primary efficacy criteria were spontaneous joint pain assessed on serial visual analogue scales during the first 36 hours and after 48 hours of treatment. The two treatments had equivalent efficacy since all the 90% confidence intervals of differences between means for pain intensity between the two groups were included within the interval (-10 mm; +10 mm). Rates of overall efficacy judged good to excellent ranged from 74.3-78.5% in both groups. One or more drug-related adverse events, mainly gastrointestinal, was reported by 24.5% and 27.2% of patients in diclofenac SR 75 mg and diclofenac 50 mg groups, respectively. Percentage of good compliance (i.e. > 90%) was higher with diclofenac SR 75 mg (p < 0.001).

[Controlled study of treatment of residual depression by clomipramine versus placebo]. / Etude contrôlée du traitement des dépressions résiduelles par la clomipramine versus placebo.

The choice of a new treatment strategy for a patient suffering from residual depression in sometimes difficult; in reality, any change in the prescription involves a risk of losing any benefit, even partial, that has recently been obtained; consequently, can one be sure that a new antidepressant treatment will be beneficial? To attempt to find the answer to this question, the Laboratories Ciba-Geigy have conducted a controlled study versus placebo to evaluate the efficacy of clomipramine in residual depression which has been progressing for more than a year. 207 patients were pre-included in the 7-day wash-out phase and treated with placebo. During this period, a clinical examination and laboratory work-up made it possible to exclude patients with a curable cause of partial resistance to antidepressants (e.g., hypothyroidism) or who were "placebo-responders". After this run-in, 181 patients were included if they complied with the criteria characterizing a major depressive episode in partial remission (according to the DSM III-R), present for at least one year and treated throughout this period with at least two antidepressants at effective dosages. In addition, these patients had to have a score between 15 and 25 on the Montgomery and Asberg Depression Rating Scale (MADRS). The patients included were randomized into two groups, receiving either an clomipramine 75 tablet or a placebo tablet for two weeks; the dosage could be increased to two tablets as at D14 if necessary and maintained for the next six weeks. The only authorized concomitant treatments were tranquillizers (lorazepam or bromazepam) and/or non-barbiturate hypnotics (zopiclone or flunitrazepam).(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of verapamil and propranolol on bronchoconstriction induced by leukotriene D4 in the isolated perfused guinea pig lung]. / Effets du vérapamil et du propranolol sur la bronchoconstriction induite par le leucotriène D4 au niveau du poumon isolé et perfusé de cobaye.

The effects of verapamil and propranolol on the LTD4-induced bronchoconstriction were studied in the isolated, perfused and ventilated guinea pig lungs. Verapamil (4.10(-5) et 1, 2.10(-4)M.1(-1], a calcium antagonist, produces a significative inhibition of LTD4-induced bronchoconstriction. On the other hand, propranolol (10(-7)M and 10(-4)M) does not increase the constrictor effects of LTD4 in guinea-pig isolated lung. These first results obtained in guinea-pig lung, confirmed the protective effect of verapamil, effect previously demonstrated in vivo or in vitro by using lung parenchymal strips. We have also confirmed the difference between the in vivo and in vitro effects of propranolol.

Influence of solute polarity in column-switching chromatography for the assay of drugs in plasma and urine.

The polarity of a drug is one of the most important parameters for the elaboration of switching systems. If the polarity of the drug is low or medium, "reversed-phase" chromatography is well adapted. The plasma or urine sample is diluted with water, centrifuged and injected first into a column of medium polarity (C2, CN or diol bonded phases). The compounds of interest are stopped on the top of the column and rinsed with water, then eluted and chromatographed on a C8 or C18 analytical column. A third column of still lower polarity can be added to improve the specificity of the system. In each successive step, the polarities of the mobile phases and columns should be decreased to reconcentrate the sample and reduce the band broadening that occurred in the previous step. Compounds of high polarity show almost no retention on reversed-phase columns, and normal-phase chromatography should be used. Aqueous solutions cannot be injected into polar bonded-phase columns as they lead to excessive band broadening. This problem can be solved by diluting plasma or urine with a large volume of a water-miscible organic solvent and injecting the clear supernatant. The compounds to be assayed are first reconcentrated on a polar column (NH2 or N(CH3)2 bonded phase) and then eluted. The selected "heart cut" of the eluate is chromatographed on another, more polar column. The influence of the polarity of drugs on the choice of switching systems is exemplified by assay methods for drugs of low, medium and high polarity.

Determination of cefsulodin, cefotiam, cephalexin, cefotaxime, desacetyl-cefotaxime, cefuroxime and cefroxadin in plasma and urine by high-performance liquid chromatography.

Closely related methods for the determination of several cephalosporins in plasma and urine are described. Deproteinized plasma or diluted urine is directly injected on a RP-8 or RP-18 bonded-material column. Chromatography is performed either in the reversed-phase or the ion-pair mode. The limits of sensitivity range from 0.4 to 2 mumol of cephalosporins per liter of plasma, and from 20 to 100 mumol per liter of urine. The sensitivity may be improved two to five times by using precolumn loading, direct sample clean-up and automatic injection. The stability of the cephalosporins in plasma, urine and water and the reproducibility and accuracy of the methods are reported.