Gender differences in tumor necrosis factor alpha and leptin secretion from subcutaneous and visceral fat tissue.

Tumor necrosis factor alpha (TNFalpha) and leptin concentrations were determined in the abdominal subcutaneous and visceral (omental) adipose tissue of patients undergoing elective open-abdominal surgery and compared with their body mass index. The concentration of leptin did not differ significantly between women and men, being high in subcutaneous fat tissue and low in visceral fat tissue. TNFalpha concentration in subcutaneous fat tissue was approximately the same in both genders, but it was significantly lower in visceral fat tissue of women and unchanged in visceral fat tissue of men. A significant correlation between BMI and leptin was found in the two fat tissue compartments of both genders, but the correlation between BMI and TNFalpha was found only in subcutaneous fat tissue of women.

Bacterial immunomodulator Olimunostim inhibits degranulation of mastocytes in vitro.

A bacterial immunomodulator Olimunostim (OS) is one of the first Czech biologic response modifiers, which have been introduced into clinical practice. OS activates the mechanisms of non-specific immunity and is used for the most part in chronic infections. Since OS is indicated in patients suffering from infection complicated allergic diseases (asthma bronchiale, atopic eczema, etc.), (1) the concentration of histamine (Hi) and Hi-like compounds in OS, and (2) the possible role of OS in the process of histamine release from rat peritoneal mastocytes (PMC) were studied. The cells were preincubated with soluble fractions of OS. PMC degranulation was stimulated by the compound 48/80. The amount of Hi and Hi-like substances in the supernatant and in the pellet of PMC was determined by the fluorimetric method. Compounds of Hi nature were also evaluated in OS (lysate of S. aureus, K. pneumoniae, P. acnes) by original modification of high-performance liquid chromatography. The data provide the first evidence for a dose-dependent inhibitory effect of OS on the degranulation of rat PMC. Moreover, we found that 1 mg of OS itself contained 0.376 microgram of Hi and 0.142 microgram of 1-methylHi. Histamine was detected in P. acnes (0.672 microgram/mg), K. pneumoniae (0.136 microgram/mg), and S. aureus (0.175 microgram/mg), while methylHi was detected only in the case of K. pneumoniae (0.454 microgram/mg). The results of our in vitro study do not support the hypothesis of inducing type I allergic reaction by OS or facilitating OS in crossing the enteral barrier.

Mutants of dermatophytes resistant to ketoconazole.

Authors describe the frequency and properties of mutants resistant to ketoconazole, obtained from the dermatophytes Microsporum gypseum and Trichophyton mentagrophytes. Spontaneous mutants occurred with the frequency of 10(-9) to spore and nucleus. Ultraviolet radiation increased the frequency of mutants by two numeric orders. The resistance level was relatively small in both types of mutants (RL within the range of 2-5).

Effect of ketoconazole on potassium pool, c-AMP-release and ergosterol synthesis in resistant mutants of Microsporum gypseum.

The effect of ketoconazole (KK) on chosen biochemical indexes was studied in three mutants of dermatophyte Microsporum gypseum (MG-155 wild strain) resistant to KK. Fungistatic concentration of KK added to liquid cultivate medium increases significantly ergosterol pool and nonsignificantly demethylation activity, potassium content, cAMP release in three days old mycelium of mutants when compared to the cultivation without KK. On the contrary, there is a significant suppression of ergosterol level and demethylation activity followed by highly significant increasing of potassium content and cAMP release in the case of wild strain MG-155.

Histamine of mouse lungs after single exposure to nitrogen dioxide.

Pulmonary histamine and the weight of lungs were studied in mice, exposed to a single one-hour effect of high concentrations of edemagenic gas nitrogen dioxide in a metabolic chamber. Nitrogen dioxide concentrations were chosen according to the results of nitrogen dioxide analysis of the mining atmosphere immediately after the mining blasts. The results were estimated by the method of paired comparison with the findings registered in mice, exposed to the effect of air atmosphere under identical experimental conditions. The intervals following immediately the exposure and 5 hours after were chosen for the evaluation, with regard to the dynamics of the early and late stages of hypersensitivity of the first type. i) Immediately after the exposure to the concentrations of 43, 250, 387 and 540 mg.m-3, no significant differences were observed in the amount of pulmonary histamine. In concentrations higher than 43 mg.m-3, the weight of lungs increased (the proportion of pulmonary water and the dry tissue). ii) Five hours after the exposure (nitrogen dioxide concentrations 66, 130, 137 and 270 mg.m-3), pulmonary histamine decreased, at the concentration of 137 mg.m-3 in a significant way, on the other hand, it increased significantly at the concentration of 270 mg.m-3. Both concentrations higher than 130 mg.m-3 manifested an increased weight of lungs (increased proportion of dry tissue and pulmonary water). The obtained data do not allow to establish unambiguously the part of histamine on the pulmonary changes following the effect of nitrogen dioxide. The edemagenic effect of nitrogen dioxide estimated after one-hour influence can be considered as reversible up to five hours after the exposure in concentrations lower than 130 mg.m-3. The metodical part of the study gives detailed description of exposure technique and it brings a survey of methods of histamine determination in blood and tissues.

Sensitivity of Trichophyton mentagrophytes strains to three imidazole drugs.

A total of 84 strains of the dermatophyte Trichophyton mentagrophytes were studied for their sensitivity to ketoconazole, miconazole, and clotrimazole. Minimal inhibitory concentration was used for determination of sensitivity. Variability in the sensitivity of strains to the tested imidazole drugs ranged within one to two orders (10(-2)-10(0) micrograms/ml). By means of UV radiation ketoconazole-resistant mutants were prepared: their frequency was 1.3 x 10(-7) mutants per spore and nucleus. A repeated exposition to the mutagen lead to increased resistance. Correlation was also studied between inhibitory effect of ketoconazole on mycelial growth and that on ergosterol biosynthesis but no significant results were obtained.

Study of lipids in dermatophytes. II. The effect of ketoconazole in the exponential stage of growth.

There was studied the effect of ketoconazole on the growth, the amount of ergosterol and the relative distribution of fatty acids in the sensitive strain MG-155 of M. gypseum and its two ketoconazole-resistant mutants UV-1 and UV-2 (induction by UV radiation) in the exponential stage growth. After three-day cultivation in the medium with ketoconazole (0.64 microgram.ml-1) there appeared 40% inhibition of growth in MG-155, 10% inhibition in UV-2 and only 4% inhibition UV-1. The amount of ergosterol decreased in MG-155, in both mutants the amount of ergosterol increased by the effect of ketoconazole (by more than 50%). The main saturate fatty acids are palmitic acid (10% and stearic acid (4%). The major fraction of unsaturate fatty acids is formed by linoleic acid (50%) and by oleic acid (4%). Ketoconazole does not affect the quality of fatty acids spectre, it changes only the size of the relative distribution of individual fatty acids. In this point of view, the mutant UV-2, and namely the mutant UV-1 (the higher degree of resistance) differ from the initial sensitive strain MG-155. Ketoconazole provokes in mutants a higher reduction of stearic acid fraction, of saturate fatty acids C greater than 18 and of triunsaturate fatty acids and it causes only minimum (4% for UV-1) decrease of oleic acid. The results of cultivations with postponed application of ketoconazole to the medium (on the 2nd the 1st day before the mycelium harvest) show in a decisive way that effect of ketoconazole is fully developed only on condition that ketoconazole is added to the medium simultaneously with the inoculum. There are discussed several conclusions with regard to the findings in the identical strains in stationary stage of growth.

Study of lipids in dermatophytes. I. The effect of ketoconazole on fatty acids of Microsporum gypseum and its mutants.

There was studied the spectre of fatty acids and its changes induced by ketoconazole in the strain M. gypseum and its two mutants (UV-1, UV-2). The main saturate fatty acid in MG-155 and in both mutants is palmitic acid, the main unsaturate fatty acid is linoleic acid. In comparison with MG-155, UV-1 and UV-2 manifest a lower amount of stearic and oleic acids and a higher amount of stearic acid) in MG-155 and UV-1, an increase of saturate fatty acids is observed in UV-2. The unsaturate fatty acids affected by ketoconazole increase in MG-155 and UV-1, they decrease in UV-2 (the changes are most marked in linoleic acid). Oleic acid, as the main representant of monounsaturate fatty acids, decreases in MG-155 and in both mutants. There is discussed the relationship of these changes in fatty acids distribution to the eventual damage of membrane function, to the growth characteristics and to the metabolism of ergosterol.

Adaptation of Microsporum gypseum to ketoconazole.

The mycelium of the strain MG-155 of the dermatophyte Microsporum gypseum was repeatedly cultivated in three subinhibitory concentrations of ketoconazole: 1, 2, and 6 micrograms/ml. The adaptability of the fungus to this antifungal drug was estimated on the basis of the sensitivity of spores and the sensitivity of the mycelium. In the first phase, the mycelium grew on the medium with ketoconazole in consequence to the physiological adaptation (without the change of genome). In the following phase of the adaptation training, there occurred a manifestation of resistant mutants. After twenty-eight transfers, only a slight increase of resistance was found (resistance level = 2), however, this resistance was of a constant character. Basing on the data of ergosterol amount, there is discussed the biochemical mechanism of the described changes in the sensitivity of the fungus to ketoconazole.

Ketoconazole-resistant mutants of Microsporum gypseum. I. Isolation of mutants, biochemical methods.

The ketoconazole-resistant mutants were isolated from four strains of the dermatophyte Microsporum gypseum. The frequency of spontaneous mutants ranged within 10(-8) to 10(-9) (per spore and nucleus). The method was established for obtaining the spontaneous mutants manifested as the faster growing sectors. By means of mutagenic effect of UV radiation, the frequency of mutants increased by one order. The method used for determination of ergosterol the biosynthesis of which could be associated with the mechanism of resistance to ketoconazole is described and evaluated.

Ketoconazole-resistant mutants of Microsporum gypseum. II. Characteristics of mutants.

The presented study deals with the properties of two wild strains of dermatophyte Microsporum gypseum and of eleven mutants resistant to ketoconazole. The growth rate of spontaneous mutants is greater than that of the wild strain, the group of UV-induced mutants manifests in general lower growth rate. The resistance level varies in the interval 1.1-1.6 (spontaneous mutants), resp. 2.7-5.5 (UV-induced mutants). The phenomenon of cross-resistance to other imidazole drugs (miconazole, clotrimazole) has been observed in mutants. There is further given the characteristic of ergosterol synthesis influence by the three used antifungal drugs. The data obtained are discussed from the point of view of possible biochemical processes resulting in the formation of resistance to ketoconazole.