Modelling of oxygen transport in systemic circulation in a hyperbaric environment.

A mathematical model is developed for analysing the transport of oxygen in the systemic capillaries and surrounding tissue in a hyperbaric environment. The governing equation in the capillary describes the transport due to molecular diffusion (radial as well as axial) and the convective effect of the blood. The non-linear oxygen dissociation curve is represented by a linear function simulating the conditions of a hyperbaric environment. The corresponding equation in the tissue region is based on the transport of oxygen due to radial as well as axial diffusion and the zero order metabolic consumption rate. The equations in both the regions are connected through the interface conditions. An analytical solution of the resulting system of elliptic partial differential equations with the physiologically relevant boundary conditions is obtained by the method of eigenfunction expansion. It is found that the amount of oxygen decreases from the core of the capillary to the periphery of the tissue. It is shown that significant radial diffusion of oxygen takes place in the initial part of the tissue close to the arterial end. The accumulation of oxygen in the tissue has been examined in terms of various non-dimensional parameters. The physiological relevance of these parameters in determining the degree of accumulation of O2 in the tissue in a hyperbaric environment is discussed in the light of previous experimental studies.

Numerical simulation of systemic O2 and CO2 exchange in a hyperbaric environment.

The process of gas exchange in systemic capillaries and its surrounding tissue is simulated numerically in a hyperbaric environment, taking into account the molecular diffusion, convection, saturation of haemoglobin with O2 and CO2, and the metabolic activity in the tissue. Krogh tissue-cylinder is used as a geometrical representation of the capillary-tissue system. The resulting system of non-linear governing equations together with the physiologically relevant boundary conditions is solved numerically. It is found that the concentration of oxygen decreases from the axis of the capillary to the tissue periphery whereas the concentration of carbon dioxide increases. It is shown that very little CO2 is transported radially. The location of the vulnerable region from the point of view of CO2 accumulation is found to be the rim (r = R2, z = L) situated at the periphery of the tissue near the venous end of the capillary. It is also found that accumulation of O2 decreases whereas that of CO2 increases in a hyperbaric environment. Finally, it is surmised that one of the reasons in causing discomfort among divers could be excessive accumulation of CO2 in the tissue.

The process of gas exchange in systemic circulation in a hyperbaric environment: an analytical approach.

A mathematical model is proposed to deal with the simultaneous transport of oxygen (O2) and carbon dioxide (CO2) in systemic capillaries and the surrounding tissue in a hyperbaric environment. The transport in the capillary region depends on molecular diffusion (radial as well as axial), the convective effect of the blood, and the saturation of haemoglobin with O2 and CO2. The corresponding equation in the tissue region describes the transport of the species due to radial and axial diffusion in the tissue and consumption of O2 in the metabolic process. The production of CO2 inside the tissue is incorporated through the respiratory quotient. The saturation of blood with O2 and CO2 have been approximated by linear functions to simulate the conditions of the hyperbaric environment. The resulting system of governing equations with the physiologically relevant boundary conditions is solved analytically. The concentration of O2 is shown to decrease from the core of the capillary to the tissue periphery, whereas the concentration of CO2 increases. It is shown that very little of the CO2 is transported radially. The location of the vulnerable point from the point of view of CO2 accumulation is found to be the corner (x = R2, z = L) situated at the periphery of the tissue near the venous end of the capillary. The accumulation of O2 and CO2 in the tissue is discussed in terms of various dimensionless parameters. It is found that the accumulation of CO2 increases whereas that of O2 decreases in the hyperbaric environment. Finally, it is surmised that one of the major causes of discomfort among divers could be excessive accumulation of CO2 in the tissue.

Effect of ketoconazole in combination with other inhibitors of sterol synthesis on fungal growth.

The effect of combination of ketoconazole with other sterol synthesis inhibitors on fungal growth was tested against a variety of fungi selected for resistance to ketoconazole. All of the sterol inhibitors, at concentrations lower than their MICs, caused an increase greater than fourfold in the ketoconazole susceptibility of some fungi. Some of the sterol synthesis inhibitors showed this effect with ketoconazole at levels that may be achieved clinically.

Effect of ketoconazole on the fungicidal action of amphotericin B in Candida albicans.

Amphotericin B-susceptible Candida albicans became resistant to the drug after growth in the presence of ketoconazole. Chromatographic analysis of cellular sterols showed that the organisms became depleted of ergosterol in parallel with the development of amphotericin B resistance. The implications of these findings are discussed in relation to combination chemotherapy with these two important antifungal agents.

Action of antifungal imidazoles on Staphylococcus aureus.

In Staphylococcus aureus, using the imidazoles miconazole and ketoconazole, detailed studies of minimal inhibitory concentrations, kinetics of growth, viability, and release of intracellular K+ confirm that the two imidazoles work differently in this bacterium. Miconazole is bactericidal at low concentrations and causes release of cellular K+. Ketoconazole has no bactericidal effect at any tested concentration and has little effect on K+ permeability of S. aureus; it slows growth at high concentration. This is reflected in a low minimal inhibitory concentration for miconazole and a high one for ketoconazole. The probable mechanisms of the bacteriostatic and bactericidal effects of the imidazoles are discussed in light of these results and the previously described antifungal mechanisms of the drugs. alpha-Tocopherol blocks the action of both imidazoles.

Heterogeneity of action of mechanisms among antimycotic imidazoles.

The three imidazole antimycotics clotrimazole, miconazole, and ketoconazole all inhibit the demethylation of lanosterol to ergosterol, resulting in inhibition of growth of Saccharomyces cerevisiae; this is a fungistatic action. At higher concentrations clotrimazole and miconazole are fungicidal, whereas ketoconazole is not. The fungicidal action reflects direct membrane damage by the imidazoles. Evidence for this is that ketoconazole is markedly less active than the other imidazoles in its ability to allow methylene blue entry into cells and to disrupt liposome model membranes. The possible clinical significance of these findings is discussed.

Mechanisms of action of the antimycotic imidazoles.

The mechanism of the antifungal action of the imidazole antimycotics, miconazole ano clotrimazole, on Saccharomyces cerevisiae was explored. When grown aerobically both drugs were fungistatic at low concentrations and fungicidal at high concentrations. When grown anaerobically the fungistatic effect was not seen, but killing still occurred at high concentrations. The fungistatic effect correlated with inhibition of ergosterol synthesis and elevated lanosterol/ergosterol ratios in the organisms. The fungicidal effect involved rapid membrane damage and was unrelated to the imidazole-induced block in ergosterol synthesis. These agents each have 2 distinct antifungal actions.

Effect of antiparkinsonian medication on plasma levels of chlorpromazine.

Twenty-one chronic schizophrenics were stabilized with chlorpromazine therapy at their therapeutic dosage for one month. Trihexyphenidyl hydrochloride or identical placebo was then added according to a double-blind, split crossover design. The duration of each half of the crossover was 15 days. Steady state blood samples were drawn three times weekly during the experimental period and the amount of chlorpromazine was determined. The results indicated there were no differences in the levels obtained between the trihexyphenidyl and the placebo phases. A two-hour postdrug blood sample was also drawn at the end of each phase and again, there were no differences between the two conditions. The importance of these results is discussed.

Detection of 3-hydroxy fatty acids at picogram levels in biologic specimens. A chemical method for the detection of Neisseria gonorrhoeae?

A method for the detection of 3-hydroxy dodecanoic acid at low picogram levels is described. The procedure involves preparation of a heptafluorobutryl derivative of the butyl ester of the fatty acid and its detection by gas-liquid chromatography using an electron capture detector. The method was adapted for use with biological specimens. Potential of the method for screening for gonococcal infection is discussed. Limitations of the method are that about 105 Neisseria gonorrhoeae cells are required for detection and that interfering substances are a major problem working at maximum sensitivity of the electron capture detector necessitating complex purification procedures. The method eliminates the need to maintain the viability of cells in specimens, thus facilitating collection and transport of specimens.

Effect of free fatty acids on liposome susceptibility to imidazole antifungals.

The presence of free fatty acids in liposome model membranes sensitizes these membranes to the action of the imidazole antifungals, clotrimazole, micronazole, and sulconazole. Unsaturation of the fatty acids is an important variable; the effect of linoleic and oleic acids is much greater than that of stearic acid. The imidazoles differ somewhat in action, with clotrimazole potency greatest both on membranes with and without fatty acids. Sulconazole has very little activity on membranes without fatty acids even at the highest concentrations tested. The data are discussed with reference to the susceptibility of various cells to the imidazoles and the specificity of imidazole action. A modification of the enzymatic method generally used for assay of marker glucose with liposome systems is also presented.

Lipid composition and sensitivity of Prototheca wickerhamii to membrane-active antimicrobial agents.

The lipid composition of Prototheca wickerhamii ATCC 16529 is presented and discussed in relation to the unique susceptibility of the organism to drugs of three membrane-active antimicrobial classes: the polyenes, the polymyxins, and the imidazoles. The presence of ergosterol in the neutral lipid fraction of the membrane is likely responsible for the exquisite susceptibility to amphotericin B. The presence of a large quantity of free fatty acids in the membrane appears responsible for imidazole susceptibility. The membrane determinants of polymyxin B susceptibility are less well defined.

Polymyxin B-penicillin antagonism in Proteus mirabilis.

Polymyxin B protects Proteus mirabilis from usually lethal penicillin concentrations.

Phospholipids and fatty acids of Neisseria gonorrhoeae.

The phospholipids and fatty acids of two strains of Neisseria gonorrhoeae of different penicillin susceptibilities were examined. The phospholipids, which comprise about 8% of the dry weight of the cells, consisted of phosphatidylethanolamine (70%) and phosphatidylglycerol (20%); small amounts of phosphatidylcholine and traces of cardiolipin were also present. Growing and stationary-phase cells were similar in content and composition of phospholipids except for phosphatidylcholine, which increased two- to fivefold in the stationary-phase cells. The fatty acids of the phospholipids were characterized by two major acids, palmitic and a C16:1, with myristic and a C18:1 acid present in smaller amounts. The fatty acids present in purified phospholipid fractions varied considerably in relative proportions from fraction to fraction. No significant difference in the composition of phospholipids from the two strains was evident. Large amounts of beta-hydroxy lauric acid were detected only after saponification of the organisms. Differences in the lipid composition between the gonococcus and other gram-negative bacteria are discussed.

Detection of agents that alter the bacterial cell surface.

Proteus mirabilis is not killed by polymyxin B, normal serum, or sodium deoxycholate. Exposure to polymyxin B renders the cells susceptible to killing by the latter two agents. The data suggest that this synergism is due to polymyxin B-induced surface changes. The results point out an inadequacy of existing methods of screening for antibiotics; they fail to detect agents which, while showing no in vitro effect on growth, may alter a resistant organism so that it becomes more susceptible to other antimicrobials or host defense mechanisms. A method is described which can be used to detect such cell surface-modifying agents.

Effect of polymyxin B on antibiotic-resistant Proteus mirabilis.

The surface properties of polymyxin B-resistant Proteus mirabilis are markedly altered by the antibiotic. The effects include the development of susceptibility to surface-active agents such as deoxycholate or tris(hydroxymethyl)aminomethane and a marked increased osmotic fragility. However, cell wall impermeability to various agents such as erythromycin, actinomycin D, bacitracin, lysozyme, and triphenyltetrazolium chloride apparently remains intact. These results support the concept that the actions of the polymyxins on gram-negative bacteria are multiple; the action(s) of the antibiotic responsible for lethality in vitro, presumably at the level of the cytoplasmic membrane, may represent only a portion of the potential therapeutic effects of the antibiotic in vivo.

Mechanism of polymyxin B resistance in Proteus mirabilis.

The lipids from three types of organisms-a Proteus mirabilis wild type highly resistant to polymyxin B, a polymyxin B-sensitive mutant derived from the wild type, and the wild type grown in the presence of sulfadiazine resulting in phenotypic conversion to polymyxin B sensitivity-were examined to determine the nature of polymyxin B resistance. The phospholipid compositions were nearly identical; each organism contained similar small amounts of N-methyl phosphatidylethanolamine in addition to comparable quantities of phosphatidylethanolamine, phosphatidylglycerol, and cardiolipin. the fatty acid compositions were similar in the exponential phase of growth; in the stationary phase, sulfadiazine markedly inhibited the synthesis of cyclopropane fatty acids. Liposomes prepared from the dried lipids of the three types of organisms were extensively and similarly disrupted by the polymyxin. These findings suggest that polymyxin B resistance in P. mirabilis is determined by the cell envelope which prevents access of the antibiotic to the susceptible lipid target sites.