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Biomedical subjects

F L Schuster

Publications and source records attributed to F L Schuster.

At least 19 recordsLinked to original sources

Beyond viruses: clinical profiles and etiologies associated with encephalitis.

BACKGROUND: Encephalitis is a complex syndrome, and its etiology is often not identified. The California Encephalitis Project was initiated in 1998 to identify the causes and further describe the clinical and epidemiologic characteristics of encephalitis. METHODS: A standardized report form was used to collect demographic and clinical data. Serum, cerebrospinal fluid, and respiratory specimens were obtained prospectively and were tested for the presence of herpesviruses, arboviruses, enteroviruses, measles, respiratory viruses, Chlamydia species, and Mycoplasma pneumoniae. The association between an identified infection and encephalitis was defined using predetermined, organism-specific criteria for confirmed, probable, or possible causes. RESULTS: From 1998 through 2005, a total of 1570 patients were enrolled. Given the large number of patients, subgroups of patients with similar clinical characteristics and laboratory findings were identified. Ten clinical profiles were described. A confirmed or probable etiologic agent was identified for 16% of cases of encephalitis: 69% of these agents were viral; 20%, bacterial; 7%, prion; 3%, parasitic; and 1%, fungal. An additional 13% of cases had a possible etiology identified. Many of the agents classified as possible causes are suspected but have not yet been definitively demonstrated to cause encephalitis; these agents include M. pneumoniae (n=96), influenza virus (n=22), adenovirus (n=14), Chlamydia species (n=10), and human metapneumovirus (n=4). A noninfectious etiology was identified for 8% of cases, and no etiology was found for 63% of cases. CONCLUSIONS: Although the etiology of encephalitis remains unknown in most cases, the recognition of discrete clinical profiles among patients with encephalitis should help focus our efforts toward understanding the etiology, pathogenesis, course, and management of this complex syndrome.

Adolescent↗

Axenic growth and drug sensitivity studies of Balamuthia mandrillaris, an agent of amebic meningoencephalitis in humans and other animals.

A cell-free growth medium for the opportunistic pathogenic ameba Balamuthia mandrillaris is presented. This represents an advance over the use of monkey kidney cells for growth of the amebas and can be helpful in isolation of these amebas from brain tissue from cases in which amebic meningoencephalitis is a diagnostic possibility, as well as for biochemical and molecular biological studies. Three isolates of Balamuthia have been cultured in this medium. The cell-free growth system was also used to screen cultures for sensitivity to a variety of antimicrobial agents. Of the various drugs tested, pentamidine isethionate was most effective against amebas (ca. 90% inhibition after 6 days of exposure), but the drug was amebastatic and not amebacidal in the axenic system at the highest concentration tested (10 micrograms/ml).

Amebiasis↗

Effects of magainins on ameba and cyst stages of Acanthamoeba polyphaga.

Amebic keratitis produced by Acanthamoeba spp. is an increasingly important ocular infection in extended-use contact lens wearers. Problems associated with the infection are compounded by the lack of effective and well-tolerated chemotherapeutic agents. The magainins, a group of naturally occurring and synthetic membrane-active peptide compounds, have been shown to be active in vitro against a clinical isolate of Acanthamoeba polyphaga. Two magainins tested extensively had minimal inhibitory and minimal amebicidal values of 20 and 25 micrograms/ml for magainin MSI-103 and 25 and 40 micrograms/ml for magainin MSI-94, respectively. Both amebastatic and amebicidal activities are enhanced by combining the magainins with silver nitrate (200 micrograms/ml) and/or other marginally effective antimicrobial agents. These combinations have activity against both trophic and cystic stages in the Acanthamoeba life cycle and have promise as antimicrobial agents in the treatment of amebic keratitis.

Acanthamoeba↗

Leptomyxid ameba, a new agent of amebic meningoencephalitis in humans and animals.

Amebae belonging to the order Leptomyxida are regarded as innocuous soil organisms incapable of infecting mammals. We report here the isolation of a leptomyxid ameba from the brain of a pregnant baboon (Papio sphinx) that died of meningoencephalitis at the San Diego Zoo Wild Animal Park. By using rabbit anti-leptomyxid serum in the immunofluorescence assay, we have identified the leptomyxid ameba in the brain sections of a number of human encephalitic cases from around the world as well as a few cases of meningoencephalitis in animals in the United States, which suggests that the leptomyxid amebae are potential etiologic agents of fatal meningoencephalitis in humans and animals.

Amebiasis↗

Morphological response of cultured cells to Naegleria amoeba cytopathogenic material.

Naegleria amoebae contain cytopathogenic material (NACM). The morphological response of cultured cells to this material follows a number of characteristics in common with those resulting from infectious agents. The cytopathologic changes varied depending on the strain of the cultured cells. Among those from 17 different vertebrate sources, both primary and continuous cell lines, some were destroyed completely by dilutions of NACM up to 10(-8) while others appeared unaffected by NACM at any concentration. The response had no apparent relationship to species, organ source, or passage level of the cells. The reaction was typified by a long latent period (4-10 days) during which the number of cells in the culture increased up to 10-fold, followed abruptly by a short period (less than 24 h) during which all of the cells were destroyed. The latent period was prolonged when the culture conditions were adverse, or when the amount of NACM in the inoculum was minimal. A high multiplicity of NACM in the inoculum lysed the entire culture, while dilutions near the end-point caused generalized or only focal changes of rounded cytopathic cells. The cytopathic effect could be maintained in cultured cells by serial passage, such that the total activity greatly exceeded what could be attributed to the original inoculum. These findings are consistent with the concept that NACM has properties of an infectious agent and that its quantity is enhanced and spread through the culture by cell-to-cell contact and by cell division.

Amoeba↗

Phenothiazine compounds inhibit in vitro growth of pathogenic free-living amoebae.

The phenothiazine compounds trifluoperazine dihydrochloride and chlorpromazine hydrochloride have in vitro activity against the pathogenic free-living amoebae Naegleria fowleri, Acanthamoeba culbertsoni, and Acanthamoeba polyphaga. Drug concentrations of 10 microM were amoebastatic; concentrations of 50 microM were either amoebastatic or amoebicidal. Concentrations of 100 microM were generally amoebicidal. The mechanism of drug action is unclear. It may reflect sensitivity of amoeba calcium regulatory protein to the phenothiazine compounds or may be due to the lipophilic action of the drugs on the amoeba plasma membrane. Accumulation of these drugs in the central nervous system makes them potentially useful chemotherapeutic agents in humans for treatment of amoebic meningoencephalitis caused by N. fowleri and Acanthamoeba spp.

Amebicides↗

Ultrastructure of the amoeboflagellate Tetramitus rostratus.

The life-cycle of the amoeboflagellate Tetramitus rostratus includes amoeboid, cyst, and flagellate stages. The ultrastructure of these three stages is illustrated, with particular emphasis on flagellate morphology. Amoeba morphology is typical of that of limax amoebas. Cysts, forming from trophic amoebas, are enclosed by a wall made up of two layers: ectocyst (ca. 70 nm), and endocyst (200 nm). The wall apparently forms from precursor material present in vesicles in the pre-cyst stage cytoplasm. Flagellate morphology is characterized by a well-defined top-shaped profile, maintained by microtubules under the plasma membrane. The flagellar apparatus or mastigont consists of four flagella, their basal bodies, sheaves of microtubules associated with two of the basal bodies, and several rhizoplasts (periodicity 20 nm). A deep, microtubule-supported, ventral invagination appears to function as a gullet. A small number of mitotic stages observed in amoeboid and flagellate individuals suggests similarity in the division process in both stages: intranuclear mitotic apparatus, nucleolus persisting through mitosis, no centrioles or basal bodies functioning as centrioles, difficulty in resolving chromosomes. The text compares ultrastructures of several amoeboflagellate organisms and evaluates the phylogenetic significance of those features common to different species. On the basis of this study, Tetramitus most closely resembles Naegleria spp.

Animals↗

Calcium regulation of flagellation in Naegleria gruberi.

The amoeba-to-flagellate transformation of Naegleria gruberi was studied, exploring the role of calcium in control of this pattern of morphogenesis. A direct and an indirect role for calcium are postulated based on experimental results. Direct inhibition in the presence of calcium is caused by ionophore A23187, substances with ionophore-like activity (amphotericin) and the hormone calcitonin, which facilitate calcium uptake into the cytosol. The indirect role is difficult to assess, but is believed to be related to calcium regulatory protein and its control of cyclic nucleotide levels in the cell, based on inhibition by trifluoperazine. Calcium flux was studied by addition of 45Ca2+ to cell cultures, and tracing its movement during the transformation period. Ultrastructural localization of calcium was attempted in amoeboid and flagellate stages, as well as in reverting flagellates. Deposits that might represent calcium were observed under the plasma membrane and, in calcium-induced reversion, in electron-dense spheres seen in the cytoplasm and at sub-membrane locations, suggesting expulsion of excess calcium.

Amoeba↗

Ultrastructural observations of experimental Naegleria meningoencephalitis in mice: intranuclear inclusions in amebae and host cells.

Primary amebic meningoencephalitis was experimentallly produced in mice through intranasal instillation of pathogenic Naegleria fowleri. Experimental animals had a 64% mortality with average time of onset of symtoms of death occurring on the 7-8th day following inoculation. Ultrastructural studies of the olfactory lobes from brains of dead (or sacrificed) animals revealed major concentrations of amebae in the perivascular regions; amebae were also seen to be under attack by host polymorphonuclear leukocytes, and in the lumina of blood vessels. Amebae in brain tissue contained 30 nm intranuclear particles arranged in clusters. In the brains of some mice, dead presumably as a result of amebic meningoencephalitis, particles and crystalloids were observed in the nuclei of degenerating cells of the central nervous system. Some alternatives are examined to explain a possible relationship between ameba intranuclear particles and mouse brain cell intranuclear inclusions.

Amebiasis↗

5-Bromodeoxyuridine-induced formation of virus-like particles in Naegleria gruberi EGs.

Exposure of axenic cultures of the amoeboflagellate Naegleria gruberi EGs to the thymidine analogue 5-bromo-2'-deoxyuridine (BrdU) resulted in the induction of virus-like particles (VLP) and various structures associated with their development and presumed transmission. Previously, VLP induction could be accomplished only by growing amoebae in the presence of living bacteria as a food source. Addition of excess thymidine along with BrdU did not block induction of particles. This account demonstrates that the EGs-VLP system responded to BrdU as do a number of mammalian cell lines harbouring latent viruses, and provides the basis for future work on the infectivity of the VLPs for other amoebae as well as tissue culture cells.

Bromodeoxyuridine↗

The nature of a cytopathogenic material present in amebae of the genus Naegleria.

The Naegleria ameba cytopathogenic material (NACM) obtained from free-living and/or pathogenic amebae has been partially isolated and characterized. The evidence indicates that the material is a protein or has a protein component, and is in the estimated molecular weight range of 50,000 daltons. The partially isolated NACM, like its counterpart in the amebic lysates, is capable of sustaining itself in tissue cultures at a temperature which otherwise would inactivate it, and in serial passages through multiple dilutions. No distinctions between the NACMs from amebae of different Naegleria species or strains have been found in the biological response to the various treatments studied.

Amoeba↗

Ultrastructure of mitosis in the amoeboflagellate Naegleria gruberi.

Naegleria gruberi is an amoeboflagellate found in soil; mitosis is restricted to the amoeboid phase of its life-cycle. Ultrastructural examination of mitotic stages has confirmed some aspects of karyokinesis reported in earlier light-microscopic studies and expanded on other features of nuclear division described in electron-microscopic studies of Naegleria. The nuclear envelope remained intact throughout division, the nucleolus persisted, and centrioles were not found. Chromosomes were indistinguishable at the ultrastructural level, nor was any evidence detected of sites of microtubular attachment to possible chromosomes. An interzonal body, formed during separation in two of the nucleolus, was not an invariable feature of mitosis. The same was true of the polar caps, which appeared to be little more than the ends of the mitotic spindle. It is suggested that, in line with comparable situations in other protists, expansion of the nuclear envelope is chiefly responsible for separation of the nucleus into two daughter nuclei.

Cell Nucleolus↗

In vitro effects of amphotericin B on growth and ultrastructure of the amoeboflagellates Naegleria gruberi and Naegleria fowleri.

In vitro effects of the polyene antibiotic amphotericin B (AmB) on growth, viability, and ultrastructure of amoeboflagellates of the genus Naegleria were examined. The strains studied were the nonpathogenic Naegleria gruberi EG(B) and the Carter and TY strains of the pathogenic Naegleria fowleri. AmB was amoebicidal at all concentrations used (0.25, 0.50, and 1.0 mug/ml) when the drug was added to cultures in lag phase, as determined by viability testing, but was mainly inhibitory when added to log-phase cultures. The drug produced ultrastructural modifications at all concentrations (0.05 to 1.0 mug/ml). These changes included distortion of nuclear shape, increase in cytoplasmic membranes (both rough and smooth endoplasmic reticulum), decrease in number of food vacuoles, absence of pseudopod formation, mitochondrial abnormalities, increase in autophagic vacuoles, and blebbing of the plasma membrane. These alterations of amoebic ultrastructure became more pronounced with increased time in AmB and with increase in AmB concentration in the growth medium.

Amoeba↗