Non-randomized comparative study of irradiation alone or in combination with surgery in stage Ib, IIa and "proximal" IIb carcinoma of the cervix.

252 evaluable patients were treated in the Centre Claudius Regaud from January 1974 to December 1983 for stage Ib, IIa or proximal IIb carcinoma of the uterine cervix. This retrospective analysis compares results obtained either by radio-surgical combination therapy (113 patients = RS group) or by exclusive irradiation (139 patients = RT group). The comparison of the two groups in terms of patient age, obesity, associated vascular pathology and previous abdomino-pelvic surgery favored the RS group significantly. The distribution according to clinical stage also significantly favored the RS group. The proportion of patients with stage IIb disease was 12% in the RS group as opposed to 25% for the RT group. Despite unfavorable patient and tumor characteristics, therapeutic results in the RT group were similar to those of the RS group. Pelvic recurrences developed in 18/110 (16%) and 18/139 (13%) of the patients in the RS and RT groups, respectively. Distant metastases occurred in 5/92 (5%) patients in the RS group and 13/121 (11%) patients in the RT group, but the difference was not significant (p less than 0.1). Five year corrected actuarial disease-free survival was 82% in both groups. There were no major early complications in the RT group while four were found in the RS group, of which three were fatal. 2% of patients had major late complications in the RS group versus 6% in the RT group and none were lethal. 25% of the RT group patients had a moderate or mild complication versus 10% in the RS group but 2/3 of these complications recovered without sequellae.

Partial biochemical characterization of cell surface hydrophobicity and hydrophilicity of Candida albicans.

Hydrophobic yeast cells of Candida albicans are more virulent than hydrophilic yeast cells in mice. Results of experiments performed in vitro suggest that surface hydrophobicity contributes to virulence in multiple ways. Before definitive studies in vivo concerning the contribution of fungal surface hydrophobicity to pathogenesis can be performed, biochemical, physiological, and immunochemical characterization of the macromolecules responsible for surface hydrophobicity must be accomplished. This report describes our initial progress toward this goal. When hydrophobic and hydrophilic yeast cells of C. albicans were exposed to various enzymes, only proteases caused any change in surface hydrophobicity. Hydrophobic cell surfaces were sensitive to trypsin, chymotrypsin, pronase E, and pepsin. This indicates that surface hydrophobicity is due to protein. Papain, however, had no significant effect. The hydrophobicity of hydrophilic cells was altered only by papain. The proteins responsible for surface hydrophobicity could be removed by exposure to lyticase, a beta 1-3 glucanase, for 30 to 60 min. When 60-min lyticase digests of hydrophobic and hydrophilic cell walls were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a 12.5% resolving gel, each protein population contained a single unique protein that was not evident in the other protein population. However, when the cell wall surface proteins of hydrophobic and hydrophilic cells were first labeled with 125I and then removed by lyticase and analyzed by SDS-PAGE, at least four low-molecular-mass (less than 65 kilodaltons) proteins associated with hydrophobic cells were either absent or much less abundant in the hydrophilic cell digests. This result was seen for both C. albicans strains that we tested. When late-exponential-phase hydrophilic cells were treated with tunicamycin, high levels of surface hydrophobicity were obtained by stationary phase. These results indicate that the surface hydrophobicity of C. albicans reflects changes in external surface protein exposure and that protein mannosylation may influence exposure of hydrophobic surface proteins.

Inhibition of monoamine oxidase by 7-chloro-4-nitrobenzofurazan.

7-Chloro-4-nitrobenzofurazan (NBD-Cl) is a potent inhibitor of both types of monoamine oxidase (MAO). NBD-Cl competitively inhibited the oxidative deamination of kynuramine catalyzed by human placenta MAO-A, the oxidative deamination of benzylamine catalyzed by bovine liver MAO-B, the oxidative deamination of serotonin catalyzed by rat brain MAO-A, and the oxidative deamination of phenylethylamine catalyzed by rat brain MAO-B. In addition, a time-dependent inactivation of MAOs by NBD-Cl has been demonstrated upon incubation of the enzyme preparations with NBD-Cl at pH 9, but not at pH 7.5. The time-dependent inhibition of MAO by NBD-Cl could be prevented by the addition of 4-nitrophenyl azide, an active site-directed label of MAO, during incubation of the enzyme with NBD-Cl. On the basis of these findings, it is suggested that at pH 9, NBD-Cl modifies one (or more) essential lysine residue(s) in the active sites of the two types of MAO.

Formulation of vaccine adjuvant muramyldipeptides. 3. Processing optimization, characterization, and bioactivity of an emulsion vehicle.

An efficacious vaccine adjuvant which elicits both cell-mediated immunity (CMI) and humoral immune response was developed using [thr1]-Muramyldipeptide (MDP) in an oil-in-water emulsion vehicle containing poloxamer 401, polysorbate 80, and squalane. Processing optimization was performed to increase the physical stability of this adjuvant emulsion which, when prepared by conventional mixing methods, demonstrated good bioactivity but poor physical stability. Various manufacturing methods were compared with a microfluidization process, which produced the most stable and elegant emulsion vehicle. The microfluidized emulsion also elicited equivalent biological response in the animal model tested.

In vitro and in vivo studies evaluating a liposome system for drug solubilization.

A liposome system was developed which demonstrates suitability as an intravenous drug carrier for a lipophilic drug compound (RS-93522, a dihydropyridine CA2+ channel blocker). An aqueous phospholipid suspension was employed as a nontoxic solubilizing vehicle for this drug. The liposome formulation, composed of a 3% mixture of dioleoylphosphatidylcholine and dioleoylphosphatidylglycerol, produced a physically and chemically stable preparation which solubilized the lipophilic drug compound at a concentration 500 times above its intrinsic aqueous solubility. Characterizing the liposome-drug system by gel filtration chromatography showed that the drug comigrated with the lipid constituents of the liposome. Further in vitro studies established that the liposome-RS-93522 formulation allowed for rapid and complete transfer of the drug from the liposome to bind with albumin when added to human serum. In vivo studies with rats were performed in which the pharmacokinetics of the liposomal-RS-93522 system were compared to those of a cosolvent-solubilized RS-93522 solution. This study showed that the pharmacokinetic profiles of the two solutions were identical. All the evidence indicates that a liposome formulation of this type does not alter the distribution of the drug in serum and is, therefore, not likely to affect the intrinsic pharmacological or toxicological parameters of the drug relative to the conventional solvent/excipient-containing formulation. This liposome system demonstrates utility as a biocompatible, nontoxic drug delivery vehicle.

Characterization of the shear sensitivity property of petrolatum.

The effects of temperature and mechanical shear on the microcrystalline structure of petrolatum are examined; syneresis determinations and rheological methods are utilized as indicators of the integrity of this structure. Two simple and practical methods for determining the shear sensitivity of petrolatum are presented. The rheological technique of spring relaxation is evaluated and shown to be more valuable than the continuous rheogram data in predicting the microcrystalline structure of petrolatum.

Malaria resistance: artificial induction with a partially purified plasmodial fraction.

Plasmodium berghei were released from mouse erythrocytes by passage through a French pressure cell. The released organisms were washed and disintegrated; the soluble portion was chromatographed on a Sephadex (G-200) column. The void-volume eluate contained an erythrocyte-free plasmodial fraction which behaved as a vaccine, preventing parasitemia, anemia, and death in mice subsequently challenged with living Plasmodium berghei.