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Biomedical subjects

P E Duffy

Publications and source records attributed to P E Duffy.

10 recordsLinked to original sources

Immunoaffinity chromatography using electroelution.

Immunoaffinity chromatography and electroelution of protein from solid-phase matrices are two powerful tools often used to purify proteins. In this study, we combined these two techniques and found that antigen was effectively recovered from immunoaffinity resins by electroelution. Yields ranged from 90.5% to 62.8%, with a mean of 74.0 +/- 7.4% (mean +/- standard deviation). A major portion of the eluate, 79.4 +/- 13.1%, was concentrated in a volume of 100 microliters and 94.0 +/- 2.0% was recovered in 200 microliters, even when 1 ml of resin was used. Electroelution had no major effect on the electrophoretic mobility of the antigen. Two distinct antigens, a relatively hydrophilic 230-kDa protein and a hydrophobic 28-kDa protein were successfully electroeluted. In addition two types of immunoaffinity resins, monoclonal antibody covalently linked to CNBr-activated Sepharose or immobilized protein A, were found to be compatible with this method.

Animals

Murine typhus identified as a major cause of febrile illness in a camp for displaced Khmers in Thailand.

Scrub and murine typhus have been identified as causes of illness among the 238,000 displaced Khmer people residing in temporary settlements on the Thai side of the Thai-Cambodian border. Still, the true extent of the problem and the relative frequency of infection with scrub typhus as compared to murine typhus are unknown. We evaluated consecutive patients with unexplained pyrexia (documented fever, no exclusionary diagnosis, and constitutional symptoms) in 1 temporary settlement over 1 month. Laboratory studies included culture of blood and assay of paired sera for rickettsial IgM and IgG antibody, for dengue IgM and IgG antibody, and for leptospiral IgM and IgG antibody. Among 37 patients (27 adults and 10 children), 28 (75%) had a rickettsiosis (26 cases of murine typhus and 2 cases of scrub typhus). No case of enteric fever, dengue, or leptospirosis was diagnosed. The illnesses of 9 patients were not identified. Signs and symptoms did not distinguish confirmed rickettsial infections from undiagnosed illnesses. The 1 month attack rate of rickettsial infection was 29/100,000 for children and 185/100,000 for adults. Murine typhus was a major cause of febrile illness in this settlement.

Adult

Disseminated Mycobacterium chelonae subspecies chelonae infection with cutaneous and osseous manifestations.

A 75-year-old man who had been receiving corticosteroids for treatment of chronic obstructive pulmonary disease presented with nodulopustular skin lesions, bone pain, and constitutional symptoms. Evaluation revealed a disseminated infection with Mycobacterium chelonae subspecies chelonae, with cutaneous and osseous involvement documented by histopathologic studies and cultures. The bone involvement is a novel observation for this subspecies. The patient was successfully treated with a three-drug regimen of tobramycin sulfate, erythromycin stearate, and ciprofloxacin hydrochloride. We present a discussion of the case in the context of the literature.

Aged

Glial fibrillary acidic protein in ependymomas and other brain tumors. Distribution, diagnostic criteria, and relation to formation of processes.

Glial fibrillary acidic protein (GFAP) was studied in ependymomas by the three-step immunoperoxidase method and compared to results in astrocytomas and normal tissues. An order of reactivity for GFAP is presented. Diagnostic criteria, based on GFAP content, are proposed. In ependymomas GFAP-positive cells give rise to only some of the tumor cells, usually those forming tubules and perivascular arrangements. It is hypothesized that the same GFAP-positive cells may form tubules at their luminal poles and may produce perivascular arrangements at their other poles. The aberrant regulation associated with neoplastic transformation in glia is often, but not always, reflected in increased GFAP content. In both astrocytes and ependymal cells GFAP may have a similar function since in both cells the increase in GFAP appears to be related to the development of fibrillary processes.

Astrocytes

Identification of glial fibrillary acidic protein by the immunoperoxidase method in human brain tumors.

The immunocytochemical localization of glial fibrillary acidic portein within glioma cell bodies and their processes by the immunoperoxidase method is demonstrated to be of diagnostic value. This method has advantages over "special" stains because it is not so dependent upon color alone, and because it identifies a specific protein in the cells. The immunoperoxidase method using antiserum to glial fibrillary acidic protein is shown to be useful for the differentiation of mixed glial and mesenchymal tumors, and for the diagnosis of tumors in which a glial or mesenchymal cell origin is in doubt.

Brain Neoplasms

Murine influenza virus encephalomyelitis. III. Effect of defective interfering virus particles.

Defective interfering influenza virus particles lessened the yield of infective virus recovered from brain after intracerebral challange of 3-week old mice, but failed to affect the appearance of disease or lethality. In 7-week old mice, the presence of defective interfering influenza virus particles reduced both lethality and virus yield. Viral inocula containing significant numbers of defective particles appeared to diminish later inflammatory changes, nerve cell degeneration and the appearance of viral antigen in deeper cerebral regions. The protective effect exerted by defective virus particles does not appear related to interferon production in brain.

Age Factors

Dopamine-containing neurons of the substantia nigra and their terminals in the neostriatum.

Tne ultrastructural and fluorescence histochemical characteristics of the mature rabbit substantia nigra and neostriatum have been reviewed as a frame of reference for the developmental study. Biochemical investigations were reported on neostriatal dopamine concentrations and the relative uptake and accumulation of 3H-dopamine by this tissue from fetal to adult stages, to provide quantitative data for correlation with the fluorexcence information. The development of the neurons of the substantia nigra and their axons which project to the neostriatum has been presented from their appearance at day 14 of gestation to their maturation in early postnatal life. The initial bipolar neuroblasts, which develop in the midline of the caudal mesencephalon, are fluorescent as soon as they emerge from the ependymal zone. Their fluorescent axons, which form the nigroneostriatal pathway, reach the telencephalon at day 16 of gestation and ramify extensively in the putamen by day 20, but do not enter the caudate nucleus until several days later. Some of the early fluorescent axonal profiles in the putamen are extremely large. Electron microscopic study of theis stage suggests that the large fluorescent profiles may correspond to axonal growth cones or early synapses. A distinct substantia nigra, pars compacta and reticulata, can be recognized by fluorescence microscopy by day 20 of gestation. Electron microscopy reveals that the young neurons are multipolar with numerous developing dendrites, some of which exhibit early synaptic junctions. The subsequent maturaition of these cells and the neuropil is described. The fluorescent axons of the substantia nigra grow into the putamen and caudate nucleus in a nonuniform manner forming fluorescent islands throughout the neostriatum in late fetal life. Occasionally, minute beaded fluorescent axons are found. These profiles might correspond to some of the axons with varicosities "en passage" revealed by electron microscopy. In an attempt to identify further the dopamine-containing axon, the ultratructure of adult neostriatum incubated in 5-hydroxydopamine was reported. Axonal varicosities "en passage" containing a dense "tag" in the vesicles were found. Most of the tagged boutons did not exhibit synaptic contacts. The possible significance of these finding s as related to dopamine secretion are discussed.

Animals

Infiltrating lipoma: a cause of monomelic hypertrophy.

Muscle biopsy and computerized tomography indicated that unilateral enlargement of the gastrocnemius muscle in a 12-year-old girl was the result of an infiltrating lipoma. This rare condition may cause diagnostic confusion before EMG and muscle biopsy studies are performed, but it can be readily distinguished from other causes of unilateral muscle enlargement using these methods.

Child