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Spontaneous formation of immunogenic conjugates by 3-hydroxy-para-aminobenzoic acid--a biotransformation product of procaine and para-aminobenzoic acid.

3-Hydroxy-para-aminobenzoic acid (3-HPABA), a biotransformation product of procaine and para-aminobenzoic acid, is capable of binding covalently to proteins by simple incubation at alkaline pH. When administered incorporated in Freund adjuvant to rabbits, 3-HPABA conjugates with normal rabbit serum proteins (3-HPABA-NRS) give rise to both hapten specific antibodies and anti-NRS antibodies. Cross immunological reactions were found between 3-HPABA and ortho-aminophenol (OAP) and 3-hydroxy-sulfanilamide (3-HS) immunological systems, but not between 3-HPABA and para-aminophenol (PAP) and 5-aminosalicyclic acid (5-ASA) immunological systems. The significance of these findings for the explanation of procaine allergy and the possible use of 3-HPABA conjugates for skin testing in procaine hypersensitivity is discussed.

4-Aminobenzoic Acid↗

Purification and characterization of carbonic anhydrase from the saliva of the rat.

Carbonic anhydrase purified from the saliva of the rat had kinetic properties identical with those of carbonic anhydrase II from rat red cells, but its molecular properties were distinctly different from the type II isozyme. Kinetic parameters were measured under steady state conditions by stopped-flow spectrophotometry and under equilibrium conditions by an 18O exchange method. The turnover number kcat for hydration of CO2 was 6.5 X 10(4) s-1 and the Michaelis constant was 4.2 mM at pH 7.5 and 25 degrees C, values which are equal to the steady state constants for red cell carbonic anhydrase II from the rat. Inhibition of the salivary isozyme by sulfanilamide (Ki = 3.7 microM) was nearly as efficient as inhibition of the erythrocyte isozyme II (Ki = 1.1 microM). The molecular weight for the salivary isozyme was 46,000 and the isoelectric point was 5.5. Salivary carbonic anhydrase had high mannose oligosaccharide components as measured by concanavalin A binding. The amino acid composition for the salivary isozyme was not similar to rat type II, but it was similar to that reported for membrane-bound carbonic anhydrase from bovine lung (Whitney, P.L., and Briggle, T.V. (1982) J. Biol. Chem. 257, 12056-12059). These observations suggest to us that salivary carbonic anhydrase is a secretory product.

Animals↗

Mechanisms of contact photosensitivity in mice. III. Predictive testing of chemicals with photoallergenic potential in mice.

The photoallergenic potential of various chemicals was assessed in mice. Mice were sensitized with applications of the test agent on the shaved abdomen; they underwent irradiation with black light on days 0 and 1. On day 5, ear lobes were painted with the chemicals and underwent irradiation with black light. The degree of the sensitivity was assessed by measuring the ear thickness 24 hours after challenge. Tetrachlorosalicylanilide, tribromsalan, bithionol, and chlorpromazine were photosensitizers. Hexachlorophene and sulfanilamide induced contact photosensitivity after pretreatment of mice with cyclophosphamide. Reactions of the recently discovered photosensitizers such as musk ambrette, 6-methylcoumarin, 7-methylcoumarin, and benzocaine were enhanced by cyclophosphamide pretreatment. Salicylanilide, salicylic acid, and 3,4-dichloroaniline failed to demonstrate photosensitization potential. This mouse model and cyclophosphamide pretreatment is a useful technique to determine photosensitization potentials for a variety of new chemicals.

Animals↗

[Autoimmune and diabetogenic effects of bucarban].

Experiments on animals were made to study the diabetogenous effect of repeated administrations of bucarban. The drug-induced immunization and insulin release were accompanied by the increased immune response of the humoral and cellular types specific for bucarban and insulin as well. The changes in the animals' immune status were accompanied by disorders of carbohydrate and lipid metabolism, characteristic for diabetes mellitus and by a lowering of the hypoglycemic effect of the antidiabetic drug in question. It was shown for the first time that the changes in carbohydrate, lipid and lipoprotein blood indicators seen in the animals given sulfanilamide antidiabetics occur in the presence of intense autoimmune response to the body own insulin and are undoubtedly related to this response.

Animals↗

A colorimetric assay of glutathione S-transferases using o-dinitrobenzene as a substrate.

Conditions have been examined for the use of o-dinitrobenzene as a substrate for colorimetric assay of glutathione S-transferases. Activities can be determined by measuring nitrite released enzymatically from the substrate using a diazo-coupling method with N-(1-naphthyl)ethylenediamine dihydrochloride and sulfanilamide. The assay can be done in the presence of large amounts of reduced glutathione (GSH), cysteine, and protein, and is capable of quantitating the enzyme activity in about 30 micrograms (wet weight) of monkey liver, corresponding to 0.1 microgram of purified glutathione S-transferase from the same source. This method is suitable for assaying simultaneously a large number of samples with reasonable sensitivity and speed.

Animals↗

Evaluation of fluorometric determination-thin layer chromatographic identification of aminacrine hydrochloride in drug preparations.

Utilizing the fluorescent property of aminacrine hydrochloride and a filter fluorometer, a fluorometric method for aminacrine hydrochloride in drug combinations was developed, collaboratively studied, and adopted as official first action in the 11th edition of Official Methods of Analysis. Identity was confirmed by thin layer chromatography (TLC). Additional analytical work was undertaken with a grating fluorometer to support a change in the method status to official final action. The grating instrument recorded the aminacrine hydrochloride spectrum as opposed to the total fluorescence emission measured by the filter instrument. The spectrum of aminacrine hydrochloride showed that the molecule was exhibiting self-absorption of the emitted radiation even at concentrations of 10(-6)M and that the ratio of the 2 peaks in the emission spectrum varied with concentration. Additional analyses of an authentic cream preparation that also contained sulfanilamide gave an average recovery of 86.0% for aminacrine hydrochloride in 10 replicate portions. Because of these observations, the current Associate Referee's recommendation to delete the fluorometric procedure from the 13th edition of Official Methods of Analysis was adopted. A recommended deletion of the TLC identification test was also adopted.

Aminacrine↗

[Plasmid resistance of the enterobacteria isolated in intestinal disorders in children].

Resistance to antibiotics and sulfanilamides was determined in enterobacteria (E. coli, Pr. mirabilis, Sh. sonnei, Sh. flexneri) isolated from children with intestinal disorders. Elimination of the resistance determinants in polyresistant strains, conjugation of R plasmids (75 per cent in Pr. mirabilis, 70 per cent in E. coli, 78 per cent in Sh. sonnei, 69 per cent in Sh. flexneri), mobilization of nonconjugative plasmids on triple crossing and capacity of R plasmids for repression of the F-factor functions were studied. The data of the study are evident of the plasmid background in enterobacteria isolated from children with intestinal disorders and confirm the plasmid nature of the resistance in the predominating number of the strains. It is suggested that Proteus plays a significant role in the prevalence of plasmid resistance among enterobacteria.

Child↗

Nitrosation of orally administered drugs under simulated stomach conditions.

Of a total of 61 drugs considered to be potentially nitrosatable on the basis of their chemical structure, 7 have indeed been nitrosated in gastric juice at pH 2 and under simulated human stomach conditions, where the pH was varied from about 7 to 2 within 60 min. The 7 nitrosatable drugs were ethambutole, despiramine hydrochloride, clomipramine hydrochloride, phenoxymethyl penicillin, aminophenazone, piperazine diadipate and analgine. Inhibition of nitrosation under simulated stomach conditions was also investigated using ascorbic acid, sulfanilamide, p-aminosalicylic acid and sulfamic acid. The first three agents were effective inhibitors, whereas sulfamic acid stimulated nitrosation within a distinct pH range.

Animals↗

[The new features of meningococcal meningitis (author's transl)].

According to a previous pattern epidemics of meningococcal meningitis (MM) were localized in North tropical Africa, caused by A serotype, easily treated with sulfanilamides and prevented by a polyosidic vaccine, A type. Changes have occured : epidemics in Finland and Brazil, with an African A type germ, in place of the American, sulfa-resistant, C type; outburst of epidemics in South tropical Africa; presence of C type in Tchad, Niger and Nigeria. In the same time dramatic evolutions due to endotoxinic processes and drug-resistances are more frequently observed.

Adolescent↗

Current therapy of vulvovaginitis.

Trichomoniasis is reliably treated with a single 2-g dose of metronidazole; however, with this regimen simultaneous treatment of sexual partners is particularly important. Trichomoniasis in pregnant women, who should not receive metronidazole, might be treated initially with clotrimazole vaginal suppositories, which appear to cure about 50% of cases. Topical antifungal agents of the imidazole class are superior to polyenes in treating vulvovaginal candidiasis. Boric acid powder applied intravaginally in gelatin capsules for 14 days appears as effective as imidazoles. Nonspecific vaginitis is now recognized as involving infection with anaerobic bacteria of the vaginal flora as well as Gardnerella vaginalis. The condition is most successfully treated with a seven-day course of metronidazole, which probably acts by eradicating the anaerobes. In addition, metabolites of metronidazole may act directly on G. vaginalis. Sulfanilamide-aminacrine-allantoin preparations are much less effective than specific therapies and have no role in the treatment of vulvovaginitis.

Antifungal Agents↗

Gas-chromatographic determination of disopyramide in serum, with use of a nitrogen-selective detector.

In this procedure for disopyramide in serum, the drug is extracted into n-heptane/isobutanol (96/4 by vol), then back-extracted into 1 mol/L H2SO4. The acidic solution is made basic with sodium hydroxide, extracted with diethyl ether, and the extract evaporated. The residue is redissolved in ethanol and analyzed by gas-chromatography, with use of a nitrogen-selective detector. p-Chlorodisopyramide is used as internal standard. Concentration and instrument response for serum extracts are linearly related from 1 to 5 mg/L, the slope being 0.61, the y-intercept -0.10, the standard error of estimate 0.01, and the correlation coefficient 0.99. Within-run precision was 6 and 4% for 3 and 5 mg/L concentrations, respectively, with a between-run precision of 7% at the 3 mg/L concentration. Diazepam interferes, but procainamide, chlordiazepoxide, quinidine, lidocaine, propranolol, sulfanilamide, and many other basic drugs do not.

Chromatography, Gas↗

Determination of nitrate in vegetables by continuous flow: interlaboratory study.

A collaborative study for the determination of nitrate in leafy vegetables, such as endive, lettuce, spinach, and beetroot, by continuous flow (CF) was conducted by the Project Group on Collaborative Studies of the Inspectorate for Health Protection, Food Inspection Service, in The Netherlands. After extraction with water and filtration, samples were cleaned up by dialysis in the CF system. Extracted nitrates were reduced to nitrite in the system by metallic cadmium, and then the nitrite was reacted with sulfanilamide and N-1-naphthylethylenediamine to form a reddish-purple azo dye. This dye was measured colorimetrically at 530 nm. Fourteen vegetable samples (including 7 blind duplicates) containing nitrate at ca 900 to 5200 mg/kg were analyzed singly by the proposed procedure by 13 laboratories. The data were analyzed by the International Union for Pure and Applied Chemistry (IUPAC)-International Organization for Standardization (ISO)-AOAC protocol for statistics. One collaborator was identified as an outlier for all results. For all samples analyzed, the repeatability relative standard deviation values varied from 1.7 to 5.5%, whereas the reproducibility relative standard deviation values ranged from 3.3 to 5.9%.

Chromatography↗

[Antimicrobial action of the combination sulfamoxole/trimethoprim in vivo (author's transl)].

The experimental infections of the mouse by gram-positive and gram-negative germs are effectively treated with the combination of N1-(4,5-dimethyl-2-oxazolyl)-sulfanilamide (sulfamoxole) and 2,4-diamino-5-(3,4,5-trimethoxy-benzyl)-pyrimidine (trimethoprim) (CN 3123, Nevin, Supristol). The potentiating effect of both substances is demonstrated by the calculated index of the synergistic effect. The therapeutic efficacy of the combination sulfamoxole/trimethoprim is proved by two models of experimental urinary infections in the rat. In the malaria of chicken and mice a notable therapeutic success is observed.

Animals↗

[Clinical trial of the antibacterial combination sulfamoxole/trimethoprim (CN 3123). 1st communication. Efficacy activity spectrum--resistance in therapy (author's transl)].

The clinical trial of the combination N1-(4,5-dimethyl-2-oxazolyl)-sulfanilamide (sulfamoxole) and 2,4-diamino-5-(3,4,5-trimethoxy-benzyl)-pyrimidine (trimethoprim) (CN 3123, Nevin, Supristol) showed that, when given in the ratio of 5:1 and in a suitable dosage computed from pharmacokinetic information, it exhibited antibacterial activity and had a therapeutic effect in about 90% of the patients. In view of the numerous types of pathogen identified in the clinical trial, CN 3123 can be described as a broad-spectrum chemotherapeutic agent. CN 3123 was also effective against infections caused by sulfonamide-resistant or antibiotic-resistant pathogens.

Adolescent↗

[Clinical trial of the antibacterial combination sulfamoxole/trimethoprim (CN 3123)/Third communication: Clinical results of a field trial with CN 3123 in bronchopulmonary and ENT (ear/nose/throat) infections (author's transl)].

The following conclusions emerged from an analysis of the therapeutic results documented during the clinical trial of N1-(4,5-dimethyl-2-oxazolyl)-sulfanilamide (sulfamoxole) and 2,4-diamino)-5-(3,4,5-trimethoxy-benzyl)-pyrimidine (trimethoprim) (CN 3123, Nevin, Supristol) in bacterial infections of the respiratory tract and ENT organs: 1. In bacterial infections of the bronchopulmonary system, the antibacterial activity of CN 3123 was demonstrated in 94% of the patients, in that 75% of patients were cured and there was a definite improvement in 19%. 2. In bacterial infections of the ENT organs, the antibacterial activity of CN 3123 was demonstrated clinically in 95% of the patients, in that 89% of the patients were cured and there was a definite improvement in 6%. 3. The clinical assessment of the activity of CN 3123 correlates closely with the results of bacteriological checks.

Acute Disease↗

[Clinical trial of the antibacterial combination sulfamoxole/trimethoprim (CN 3123)/Fourth communication: Clinical results with CN 3123 in infections of the gastrointestinal tract, the skin and gynecological adnexes (author's transl)].

The antibacterial activity of N1-(4,5-dimethyl-2-oxazolyl)-sulfanilamide (sulfamoxole) and 2,4-diamino-5-(3,4,5-trimethoxy-benzyl)-pyrimidine (trimethoprim) (CN 3123, Nevin, Supristol) was studied in various bacterial infections in the fields of internal medicine, dermatology and gynecology. It was found that: 1. CN 3123 exhibits antibacterial activity in bacterial infections of the intestinal tract, particularly in salmonellosis and shigellosis. 2. CN 3123 is effective against bacterial infections of the female genital organs. 3. CN 3123 is very effective in patients with bacterial infections of the skin. 4. The antibacterial activity of CN 3123 in these indications is comparable to its action on bacterial infections of the urinary organs and respiratory tract and in ENT infections.

Adolescent↗

[Clinical trial of the antibacterial combination sulfamoxole/trimethoprin (CN 3123). Fifth communication: Comparison of two double-blind studies with CN 3123 undertaken for demonstration of its efficacy (author's transl)].

Two double-blind trials which, independently, revealed the same conclusion that there is no significant difference between the therapeutic effect of N1-(4,5-dimethyl-2-oxazolyl)-sulfanilamide (sulfamoxole) and 2,4-diamino-5-(3,4,5-trimethoxy-benzyl)-pyrimidine (trimethoprim) (CN 3123, Neven, Supristol) and a reference preparation, were compared with each other in respect of rating the therapeutic result. The authors discuss the greater occurrence of Proteus pathogens at one trial centre, and its bearing on the rating of the therapeutic effect.

Bacterial Infections↗

[Clinical trial of the antibacterial combination sulfamoxole/trimethoprim (CN 3123)/Sixth communication: Results of compatibility of CN 3123 (author's transl)].

1. During the clinical trial of N1-(4,5-dimethyl-2-oxazolyl)-sulfanilamide (sulfamoxole) and 2,4-diamino-5-(3,4,5-trimethoxy-benzyl)-pyrimidine (trimethoprim) (CN 3123, Nevin, Supristol) side-effects were reported in only 56 (=4.1%) of the 1371 patients who took part. 2. The numerous laboratory parameters studied in the whole group and in the group which received long-term treatment did not show any significant changes due to CN 3123. 3. The frequency and the type of side-effects which occurred under the combined preparation CN 3123 were the same as those known to be caused by sulfonamide treatment. 4. The good tolerance, which is a familial feature of sulfonamide treatment, is not impaired when sulfamoxole is combined with trimethoprim.

Adolescent↗