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[Rapid diagnosis of mycobacteria, especially Mycobacterium tuberculosis and Mycobacterium avium complex, using MB Check system].

Fourty-five sputum specimens were subjected to isolation for mycobacteria either MB Check system (MB method; F. Hoffmann-La Roche Ltd., Basel, Switzerland) or 3% Ogawa egg medium (Ogawa method). Test sputum was treated with 4 volumes of 4% NaOH for 1-2 min and 0.1 ml of the resulting mixture was inoculated onto 3% Ogawa egg medium. The remaining portion of the mixture was neutralized with IN HCl, diluted with 1/15 M phosphate buffer (PB; pH 6.8), and subsequently centrifuged at 3,000 rpm for 20 min. The sediment was suspended in 1.5 ml of PB and 0.5 ml each was inoculated into MB Check M bottle (20 ml) supplemented with M supplement (1 ml). In MB method, bacterial growth was measured on Middlebrook 7H11 agar medium and Middlebrook 7H11 agar medium containing NAP (p-nitro-alpha-acetylamino-beta-hydroxy-propiophenone). Among 45 sputum specimens, the number of positive specimens for mycobacterial growth in the above two cultivation methods and time required for growth were as follows: 3% Ogawa egg medium; 12 specimens (26.7%) gave positive growth, including 7 of M. tuberculosis complex strains on 14-35 days (average 22 days) and 5 of M. avium complex strains on 14-21 days (average 18 days); MB method; 15 of specimens (33.3%) gave positive growth, including 8 of M. tuberculosis complex strains on 7-21 days (average 15 days), 6 of M. avium complex strains on 7-14 days (average 11 days) and 1 of M. scrofulaceum strain on 28 days. There was no specimen which was positive for mycobacterial growth on 3% Ogawa egg medium but negative on MB medium.

Culture Media↗

[Rapid diagnosis of mycobacteria, especially Mycobacterium tuberculosis and Mycobacterium avium complex, using BACTEC 460 TB system].

Fourty-five sputum specimens collected at the National Sanatorium Hiroshima Hospital were subjected to cultivation using either BACTEC 460 TB System (BACTEC method; Becton Dickinson Co., Towson, Md., U.S.A.) or 3% Ogawa egg medium. Test sputum was treated with four volumes of 4% NaOH for approximately two minutes, after which 0.1 ml of the treated sputum was immediately inoculated onto 3% Ogawa egg medium. After neutralizing the remaining pretreated sputum with 1N HCl, and diluting with 1/15 M phosphate buffer PB; pH 6.8), it was then centrifuged at 3,000 rpm for 20 min and the sediment was suspended in 1.5 ml of PB. Volumes of 0.5 ml each were inoculated into BACTEC 12B medium (4 ml), containing PANTA for prevention of contamination and POES for promoting the growth of mycobacteria. In the BBCTEC method, bacterial growth was measured in terms of increases in the Growth Index (GI) values which were determined by the amount of 14CO2 released from the 14C-labelled palmitate during cultivation at 37 degrees C (positive growth; GI greater than or equal to 50). Moreover, rho-nitro-alpha-acetylamino-beta-hydroxy-propiophenone (NAP)-sensitivity testing was done by transferring a part of the BACTEC 12B culture showing positive growth to a NAP vial, and thereafter subjected to further cultivation.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗

Supersensitivity to catecholamines after impairment of extraneuronal uptake or catechol-O-methyl transferase.

In cat papillary muscle and nictitating membrane block of extraneuronal catechol-O-methyl transferase (COMT) by 3',4'-dihydroxy-alpha-methyl propiophenone (U-0521) or of extraneuronal uptake by hydrocortisone causes supersensitivity to catecholamines whenever the experimental conditions result in a high sensitivity of the organ to catecholamines. After block of monoamine oxidase the extraneuronal O-methylation of (-)-[3H]norepinephrine by the isolated nictiating membrane is due to two O-methylating systems (Km and Vmax: 7.5 muM and 0.73 nmoles - g-1 - min-1, and 131 muM and 8.5 nmoles - g-1 - min-1, respectively). Hydrocortisone (28 muM) blocked the activity of the high affinity system without affecting the low affinity system. Apparently, there exists an extraneuronal compartment of high affinity that has a hydrocortisone-sensitive uptake mechnism; this compartment influences the concentration of catecholamines below the Km of this compartment. Supersensitivity ensues when either uptake or enzyme is blocked. Since the sensitivity effects of U-0521 and hydrocortisone are not additive, the high affinity compartment must a) metabolize most of the catecholamine transported into the compartment, and b) have a limited storage capacity for catecholamines after block of COMT.

Animals↗

Antiarrhythmic effects of two new propafenone related drugs. A study on four animal models of arrhythmia.

LG 6-101 (1-[3-(2-methoxy-3-(2-methylpropylamino)-propoxy)-4-methyl- 2-thienyl]-3-phenyl-1-propanone, hydrochloride) and LG 6-102 (2-(2-methoxy-3-propylamino-propoxy)-3-phenyl-propiophenone, hydrochloride) are two new antiarrhythmic drugs. They are structurally related to propafenone which is a widely used class 1 antiarrhythmic drug with relatively low bioavailability. Both substances were characterized in four animal models of arrhythmia and compared to propafenone. In isolated guinea pigs left auricles LG 6-101 and LG6-102 were about twice as effective as propafenone regarding the prolongation of the functional refractory period but did not decrease contractility more than propafenone. LG 6-101 was significantly more effective (p less than or equal to 0.002) than propafenone or LG 6-102 in delaying the onset of ventricular premature beats in ouabain induced arrhythmias in guinea pigs. In aconitine induced arrhythmias in rats, LG 6-101 and LG 6-102 did not differ in their antiarrhythmic effects from propafenone, whereas the protection against cardiac arrest was significantly (p less than or equal to 0.003) better for LG 6-101 than for propafenone or LG 6-102. In arrhythmias induced by occlusion of the left descending coronary artery in rats the drugs tested showed as good antiarrhythmic effects as propafenone. In this model the size of the ischemic area was also measured and LG 6-101 was the most effective drug in that respect. These results suggest that both LG 6-101 and LG 6-102 are potent antiarrhythmic substances which in some models were more effective than propafenone.

Aconitine↗

Liquid chromatographic determination of bendiocarb in technical materials and wettable powder formulations: collaborative study.

A liquid chromatographic method for determination of bendiocarb in technical materials and wettable powders was tested by 12 collaborators. Bendiocarb is dissolved in acetonitrile containing 0.1% propiophenone as internal standard. This solution is analyzed on a liquid chromatograph utilizing a reverse phase (C18) column. The compound is detected at 254 nm and peak area is used for quantitation. The 3 different materials studied contained 20, 80, and nominally 100% bendiocarb. Each was examined in duplicate to provide the necessary matched pairs. Collaborators approved of the ease and simplicity of the method and, in particular, the way the method can be applied to automatic injection assemblies. The statistical data show acceptable precision of the method: Reproducibility coefficients of variation were 20% material, 2.04%; 80% material, 1.02%; and nominal 100% material (technical product), 0.64%. The method has been adopted official first action.

Carbamates↗

Accumulation of 3H-adrenaline by rabbit aorta.

The accumulation of (-)-3H-adrenaline (3H-A) by rabbit isolated aorta was studied. In all experiments, monoamine oxidase and catechol-O-methyltransferase were inhibited by treatment with pargyline and 3',4'-dihydroxy-2-methyl-propiophenone (U-0521), respectively. The relationship between the accumulation of 3H derived from 3H-A (10(-8) M) and the duration of incubation (0-3 h) was linear. The 3H-accumulation after 3 h incubation was 22.5 ml X g-1. In reserpine-treated tissue, the 3H-accumulation levelled off after 30 min and was 8.5 ml X g-1 after 3 h. The concentration of 3H-A or (-)-3H-noradrenaline (3H-NA) and the 3H-accumulation (ml X g-1) were inversely related. At 10(-8) M, the 1-hour accumulation of 3H derived from 3H-A and 3H-NA was 7.8 and 15.2 ml X g-1, respectively. With increasing concentrations (3 X 10(-8)-10(-4) M), the accumulation values approached each other. At 10(-4) M, the accumulation was 2.3 and 2.8 ml X g-1 for 3H-A and 3H-NA, respectively. The accumulation of 3H derived from 3H-A (10(-8)-10(-4) M) by reserpine-treated tissue also showed an inverse relationship with concentration: 5.4 ml X g-1 (10(-8) M) and 2.6 ml X g-1 (10(-4) M). The accumulation of 3H derived from 3H-A (10(-8) M; 1 h) was dependent on the bath temperature (1-37 degrees C). The accumulation increased continuously from 1.1 ml X g-1 (1 degree C) to 11.1 ml X g-1 (37 degrees C). Storage of tissue (0-5 days in salt solution without equilibration with 95% O2/5% CO2; 4 degrees C) did not affect the accumulation of 3H derived from 3H-A (10(-8) M; 1 h). Thereafter (7-14 days), the accumulation decreased. The inhibitory potency (IC50;-log M) of desimipramine, cocaine, (-)-propranolol, (-)-isoprenaline, and (+/-)-normetanephrine on accumulation of 3H derived from 3H-A (10(-8) M; 1 h) was found to be 8.26; 6.50; 5.48, 4.88, and 4.02, respectively. The maximal degree of inhibition was almost the same for these drugs, while that of clonidine and corticosterone was 50 and 20%, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

[Experimental observations on the iodination of phenols in the preparation of intermediates of pharmaceutical interest].

A study has been made on the iodination of phenols by elemental iodine in Na2CO3 solution. The results attained by applying the suggested procedure to a number of practical cases have been very encouraging. This method gives rise to the iodine derivatives of aldehydes (2-hydroxy- and 4-hydroxy-3,5-diiodobenzaldehyd), ketones (2-hydroxy-5-iodo-, 2-hydroxy-3,5-diiodo- and 4-hydroxy-3,5-diiodo-acetophenone, 4-hydroxy-3,5-diiodo-propiophenone), acids (3,5-diiodo-4-hydroxybenzoic acid, 3-formyl-4-hydroxy-5-iodobenzoic acid), aminoacids (3-iodo- and 3,5-diiodo-tyrosine), esters (methyl ester of 2-hydroxy- and 4-hydroxy-3,5-diiodobenzoic acid, methyl ester of 3-iodo-5-chloro-, 3-iodo-5-bromo- and 3,4,5-triiodosalicylic acid, dimethyl- and diethyl-ester of 4-hydroxy-5-iodo-isophthalic acid), amides (2-hydroxy-3,5-diiodobenzamide) and benzoxazole derivatives (2-(2'-hydroxy-3',5'-diiodophenyl)benzoxazole).

Chemical Phenomena↗

Diethylstilbestrol: placental transfer, metabolism, covalent binding and fetal distribution in the Wistar rat.

The disposition of [14C]diethylstilbestrol (DES) was investigated in the near-term pregnant Wistar rat. Both constant maternal i.v. infusions and direct fetal i.p. injections of [14C]DES were utilized as exposure routes. Maternal femoral vein infusions (total dose: 0.3-0.4 mg of DES/kg of maternal b.wt.) resulted in a stable maternal plasma level of DES during a 3-hr period. At 3 hr, fetal plasma levels were 2 to 3 times higher than maternal plasma DES. The chorioallantoic placenta and visceral yolk sac concentrated DES to levels 5 to 7 times that in the maternal plasma. DES was highly concentrated in the extraembryonic membranes and did maintain high fetal plasma levels of DES. Fetal reproductive tract had total 14C-activity levels 20 to 25 times higher than those in maternal plasma. The extractable 14C-activity in the reproductive tract was 20% DES, 15% glucuronides and 65% oxidative metabolites (beta-dienestrol, 4-OH-propiophenone, omega-OH-DES + unknowns). Fetal liver contained DES (approximately 2%), glucuronides (75%) and oxidative metabolites (approximately 20%). 14C analysis of the whole fetus indicated that the DES was 75% and glucuronides was 25%. Maternal and fetal liver had similar metabolites. Direct fetal injections of [14C]DES resulted in similar metabolite profiles in fetal tissues. The degree of covalent binding of [14C]DES to cellular constituents was tissue-specific, especially the fetal reproductive tract. Thus, DES and its oxidative metabolites were concentrated in the fetus and selected fetal tissues. It is proposed that both the estrogenic activity of DES and the potential reactivity of its metabolites may be responsible for the toxicity expressed in the fetus. Inasmuch as direct fetal injections of [14C]DES resulted in high tissue localization of both oxidative and conjugative metabolites within the fetus, it is suggested that fetal tissues may metabolize DES.

Animals↗

[Anti-calcium effects of 4-hydroxypropiophenone derivatives--potential beta-adrenoreceptor blockers].

The present paper carries out the pharmacological evaluation of 4-(2-hydroxy-3-isopropylaminopropoxy)-3-(alkoxymethyl) propiophenones with an ethoxy, propoxy and butoxy-group, whose structures are typical of the blockers of beta-adrenergic receptors. In the above-mentioned compounds the anticalcium effect on the frequency and the amplitude and the negative chronotropic and the negative inotropic effects were evaluated by means of the method of spontaneously pulsing guinea-pig atria within a concentration range of 4-16 microgram.cm-3. The obtained results confirmed a significant anticalcium effect on the heart rate in the compounds with a propoxy and a butoxy group, and in the standard verapamil. In all three compounds as well as in the standard verapamil, no anticalcium effect on the amplitude was found. The results of this membrane efficacy are in agreement with the preceding evaluation of the antidysrhythmic and anti-isoprenaline activity with the most marked effect in the compound with a propoxy group. On the basis of these results it is possible to conclude that in these derivatives of 4-hydroxypropiophenone also the anticalcium effect can participate, besides the beta-adrenolytic and the membranostabilizing effects, in the antidysrhythmic activity.

Adrenergic beta-Antagonists↗