Biomedical subjects
M Forte
Publications and source records attributed to M Forte.
Visceral leishmaniasis recrudescence in a patient with AIDS.
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Identification of macrotetrolide antibiotics in a screen to detect calcium channel blocking agents.
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[Treatment of dermatomycoses with a new antimycotic: itraconazole. Comparative studies of 2 posologic regimens].
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[Cutaneous and mucosal candidiasis. Incidence and identification of isolated species in selected case material].
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Contact dermatitis due to cyclopyroxolamine.
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Mutations resulting in resistance to polyene antibiotics decrease voltage-sensitive calcium channel activity in Paramecium.
In this report, the isolation of Paramecium tetraurelia mutants resistant to the polyene antibiotics amphotericin B and filipin are described. These antibiotics are known to specifically interact with membrane sterols to produce a cytotoxic effect. Four mutants resistant to amphotericin B and two mutants resistant to filipin have been isolated. In each case, an individual mutant shows resistance to both amphotericin B and filipin. Genetic analysis indicates that all 6 mutations map to the same complementation group and are not allelic to any of the 3 pawn mutations or to either of the two "barium shy" mutations. The behavioral analysis suggests that these mutations cause an alteration of normal ion channel function. Direct electrophysiological analysis of one mutant indicates mutations to polyene antibiotic resistance cause a specific decrease in voltage-sensitive Ca2+ channel activity.
Ketoconazole treatment in superficial mycoses.
Ketoconazole is a new antifungal drug, used for oral treatment, with a broad spectrum of activity. It is a member of the imidazole series and is active in superficial and deep mycotic diseases, in candidosis and pityriasis versicolor. This new imidazole derivative has been successfully administered in several cases of dermatophytosis, yeasts and pityriasis versicolor in the authors' clinic, as well as in many other research centres. Treatment has been given to patients with tinea corporis, tinea cruris, tinea pedis, with or without location in interdigital spaces, while the cases of tinea capitis are still under investigation. The following variants of candida infections have been treated: onychomycosis, intertrigo and mucocutaneous candidosis due to prolonged immunosuppressive therapy. No remarkable haematochemical disorders have been observed after treatment.
Muzolimine vs. furosemide in nephrotic syndrome: further support of different carrier(s) operating for enteral absorption.
The authors have compared during 4 weeks of study the effects of furosemide (F) by oral route or i.v. and muzolimine (M)/os in 10 patients with nephrotic syndrome (NS) and normal renal function. A satisfactory diuretic response was observed with i.v. F (100 mg) and M/os (30-60 mg) with respect to the basal condition (P less than 0.001), while F/os (100 mg) was ineffective in these patients. This behaviour may be explained by the different pharmacological properties of M vs. F, by gut alterations due to the oedema of mucosa or loss of some "carrier(s)" operating for intestinal absorption of these two high ceiling diuretics.
Voltage-dependent calcium channels from Paramecium cilia incorporated into planar lipid bilayers.
Two different divalent cation-selective channels from Paramecium cilia were incorporated into planar lipid bilayers. Both channels were much more permeable to divalent than univalent cations, and one of them discriminated significantly among the divalent cations. The selectivity and voltage dependence of the latter channel are comparable to those of voltage-dependent calcium channels found in a variety of cells. A combined biochemical, biophysical, and genetic study of calcium channels is now possible.
Characterization and purification of a soluble protein controlling Ca-channel activity in paramecium.
The analysis of the voltage-sensitive Ca++ channel of the unicellular eucaryote, Paramecium has been extended to a biochemical level based on recent observations that the transfer of cytoplasm from wild-type cells into mutants lacking Ca++-channel function ("pawn" in P. tetraurelia and "CNR" in P. caudatum) causes mutant cells to regain Ca++-channel function. We have microinjected various cytoplasmic fractions into mutant cells and measured the restored Ca++-channel function using a convenient behavioral assay. Following the "curing" activity, we characterized and purified the component from wild-type cytoplasm that can restore the function missing in cells carrying mutations in the cnrC gene. The curing factor is not an RNA, but a heat-labile, -SH-containing protein that appears to affect existing mutant channels on the ciliary membrane. We have purified this factor over 500-fold from the soluble cytoplasm using conventional techniques. The protein is of low apparent molecular weight (less than 30,000 daltons), acidic, soluble, and does not have the properties of calmodulin.
Differences in visual sensitivity among mindfulness meditators and non-meditators.
Tachistoscopic presentation of light flashes was used to test for differences in visual sensitivity among 3 groups of practitioners of Buddhist mindfulness meditation and non-meditator-controls. Meditation practitioners were able to detect light flashes of shorter duration than the non-meditators. There were no differences among the meditator groups. There were no differences among the groups in ability to discriminate between closely spaced successive light flashes. The lower detection threshold for single light flashes for the meditators may reflect an enduring increase in sensitivity, perhaps the long-term effects of the practice of mindfulness meditation on certain perceptual habit patterns. The lack of significant differences in the discrimination of successive light flashes probably reflects the resistance of other perceptual habit patterns to modification. The results support the statements found in Buddhist texts on meditation concerning the changes in perception encountered during the practice of mindfulness.
Visual sensitivity and mindfulness meditation.
Practitioners of the mindfulness form of Buddhist meditation were tested for visual sensitivity before and immediately after a 3-mo. retreat during which they practiced mindfulness meditation for 16 hr. each day. A control group composed of the staff at the retreat center was similarly tested. Visual sensitivity was defined in two ways: by a detection threshold based on the duration of simple light flashes and a discrimination threshold based on the interval between successive simple light flashes. All light flashes were presented tachistoscopically and were of fixed luminance. After the retreat, practitioners could detect shorter single-light flashes and required a shorter interval to differentiate between successive flashes correctly. The control group did not change on either measure. Phenomenological reports indicate that mindfulness practice enables practitioners to become aware of some of the usually preattentive processes involved in visual detection. The results support the statements found in Buddhist texts on meditation concerning the changes in perception encountered during the practice of mindfulness.
Characterization of cytoplasmic factors which complement Ca2+ channel mutations in Paramecium tetraurelia.
The analysis of Ca2+-channel function in the single-celled eukaryote Paramecium can be extended to a biochemical based on recent observations that transfer of cytoplasm from wild-type cells into mutants lacking Ca2+-channel function ("pawn" mutants) causes the mutant cells to regain Ca2-channel activity. Using a convenient behavioral assay for Ca2+-channel function, we have used microinjection of cytoplasmic fractions into mutant cells to enrich for and characterize those components from wild-type cytoplasm which can "cure" cells carrying mutations in the 3 different pawn genes affecting Ca2+-channel activity (pwA,pwB, and pwC). In each case, the curing factor appears to be a protein component of an intracellular membrane. They are distinguishable on the basis of thermal, pH and divalent ion sensitivities. In addition, the factor curing the pwC mutational defect has been purified more than 180-fold. Furthermore, the pwB curing activity appears to be amplified during sequential transfer between pwB cells.
Mutant analysis shows that the Ca2+-induced K+ current shuts off one type of excitation in Paramecium.
Two mutants of Paramecium tetraurelia, called "pantophobiacs," were found to lack most of the slow Ca2+-induced K+ outward current. Passive properties, the transient Ca2+ inward current, and the fast depolarization-induced K+ outward current remain normal. The mutant defect reduces the ability to shut off a normal, excited state of the membrane and results in repeated, long backward swimming instead of the wild-type jerks in response to a variety of ions, to heat, and to touch.
Microinjection of cytoplasm as a test of complementation in Paramecium.
Mutants in Paramecium tetraurelia, unable to generate action potentials, have been isolated as cells which show no backward swimming in response to ionic stimulation. These "pawn" mutants belong to at least three complementation groups designated pwA, pwB, and pwC. We have found that microinjection of cytoplasm from a wild-type donor into a pawn recipient of any of the three complementation groups restores the ability of the pawn to generate action potentials and hence swim backward. In addition, the cytoplasm from a pawn cannot restore a recipient of the same complementation group, but that from a pawn of a different group can. Electrophysiological analysis had demonstrated that the restoration of backward swimming is not due to a simple addition of ions but represents a profound change in the excitable membrane of the recipient pawn cells. Using known pawn mutants and those which had previously been unclassified, we have been able to establish a perfect concordance of genetic complementation and complementation by cytoplasmic transfer through microinjection. This method has been used to classify pawn mutants that are sterile or hard-to-mate and to examine the ability of cytoplasms from different species of ciliated protozoa to restore the ability to swim backward in the pawn mutants of P. tetraurelia. A cell homogenate has also been fractionated by centrifugation to further purify the active components. These results demonstrate that transfer of cytoplasm between cells by microinjection can be a valid and systematic method to classify mutants. This test is simpler to perform than the genetic complementation test and can be used under favorable conditions in mutants that are sterile and in cells of different species.
Mutational alteration of membrane phospholipid composition and voltage-sensitive ion channel function in paramecium.
A behavioral mutant of Paramecium tetraurelia (baA) has been isolated that has an abnormal response when placed in solutions containing Ba2+. This mutant is shown here to have a dramatic alteration of the sphingolipid and phosphonolipid composition of its ciliary membrane. This biochemical defect is present in independently isolated alleles at baA locus and segregates in crosses with the behavioral phenotype. Electrophysiologically, the mutation reduces significantly conductance of both voltage-sensitive Ca2+ channels and voltage-sensitive K+ channels. When the mutant is grown in sterol-supplemented medium, its behavior, electrophysiological properties, and lipid composition are hardly distinguishable from wild type grown under similar conditions. This mutant then, provides strong evidence that membrane lipids significantly influence the function of the membrane molecules responsible for the generation of action potentials.
Ionic channels of Paramecium: from genetics and electrophysiology to biochemistry.
This paper reviews the combined genetical, electrophysiological and biochemical analysis of excitation that has been carried out in Paramecium. Paramecium cells display graded Ca++ action potentials in response to a variety of stimuli. These action potentials regulate the orientation of the ciliary beat hence the cell's swimming behaviour. A large array of mutants displaying altered behaviour have been isolated and mapped to over 20 loci. Detailed electrophysiological analyses have been carried out on several classes of mutants revealing defects in specific ion channels in some cases. Mutants have proven very useful to analyze channel properties, to unravel interactions between channels and to discover the function of these channels in a variety of cellular processes. Some important channels are located in the ciliary membranes and cilia as well as ciliary membranes can now be purified in high purity and reasonable yield. These fractions have been used recently in a variety of biochemical approaches to gain insight into the molecular components of the excitation machinery. Specific alterations in some minor membrane proteins have been found in two mutants as well as a specific defect in sphingolipids in a third mutant. Those alterations had to be distinguished from large scale variations in membrane proteins and lipids that occur in this organism in response to modifications in growth conditions. Several other recent biochemical developments are reviewed and the advantages as well as the difficulties of the genetic approach to the molecular study of biological processes are discussed.