Mycobacterial infections of animals: pathology and pathogenesis.
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Biomedical subjects
Publications and source records attributed to M E Hines.
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The isolation of drug residues, environmental contaminants or naturally occurring component molecules from biological materials is often a complex undertaking. We report here the development and application of a simple approach to the disruption of biological samples that also allows for the rapid fractionation and isolation of the sample's natural components or incurred residues. This process, called matrix solid-phase dispersion (MSPD), combines the use of mechanical forces generated from the grinding of samples with irregular shaped particles (silica or polymer based solid supports) with the lipid solubilizing capacity of a support-bound polymer (octadecylsilyl or others) to produce a sample column material from which dispersed sample matrix components can be selectively isolated. The factors governing this process and examples of its various applications are presented.
A 12-year-old female sarus crane (Grus antigone) developed a recurrent proliferative lesion in the subcutaneous tissue of the tarsometatarsus, which failed to respond to medical and surgical therapy. The crane was killed and microscopic examination of the tissues taken at necropsy revealed a myxomatous, poorly-differentiated sarcoma with metastasis to the liver and kidney. Immunohistochemical staining for muscle actin, smooth muscle myosin and vimentin were positive, indicating that the primary and metastatic tumours were leiomyosarcomas. Location of the metastatic lesions in only the portal venous system of the liver and veins of the kidney indicated that the route of metastasis was the portal circulation via the ischiatic vein, caudal mesenteric vein and both the renal-portal shunt and hepatic portal vein. This is the first report of metastatic subcutaneous leiomyosarcoma in an avian species.
Extra-adrenal paragangliomas associated with vertebral pain and clinical neurological abnormalities as a result of metastasis to the vertebral column were diagnosed in four dogs of different breeds by light microscopy. All were males (two intact and two neutered) aged 8 years. Metastatic neoplasms occurred as extradural masses with associated bone lysis at vertebrae C4 (2 cases), T12-L1 (1 case) and L4 (1 case). The neoplastic cells exhibited similar morphology with little variation between cases. All neoplasms showed cytoplasmic granules after staining with the Churukian-Schenk modification of the Pascual argyrophil stain for neurosecretory granules or for membrane bound electron-dense granules (dense-core granules). On immunohistochemical examination the neoplastic cells gave positive results for neuron-specific enolase and negative results for chromogranin and epithelial membrane antigen. Multiple organ metastasis and metastasis to bone have been reported previously, but these cases were unusual due to the involvement of the spine as an apparent predilection site for metastasis, and the sex (male) and age of the animals affected.
Glycolipid fractions from Mycobacterium avium serovar 2 (Mycobacterium paratuberculosis 18) inhibited the killing of Candida albicans by activated bovine peripheral-blood-derived macrophages. Fractions were derived by using the matrix solid-phase dispersion technique, which is a new method of simultaneous lysis and partial fractionation of components of bacterial cells. Further purification of active fractions was performed by concanavalin A affinity chromatography, centrifugal filtration, and differing solvent solubility. Three different fractions were isolated and partially characterized. Two of these fractions have characteristics typical of glycolipids, and the third fraction has characteristics compatible with a peptidoglycolipid. This peptidoglycolipid fraction has been purified and named MIF-A3.
Because of the rising incidence of clinical mycobacterial infections and the difficulty in identification and characterization of mycobacteria at the subspecies and serovar levels, a technique for differentiation that could be performed quickly and with relatively little equipment and expense was developed. Lysis and fractionation of mycobacteria by matrix solid-phase dispersion followed by thin-layer chromatography were used to produce chemotype profiles of the lipid and glycolipid components of each isolate. Organisms tested included Mycobacterium scrofulaceum, Mycobacterium phlei, Mycobacterium smegmatis, Mycobacterium flavescens, Mycobacterium kansasii, Mycobacterium bovis, 11 isolates of Mycobacterium gordonae, 10 serovars of Mycobacterium avium, and four strains of Mycobacterium paratuberculosis. A relative retention (Rx) value was established for each visible band on the chromatographs by comparison with a band common to all organisms tested. The chemotype profiles produced were different for every separate species and serovar sampled. These findings suggest that matrix solid-phase dispersion and subsequent thin-layer chromatography may have the sensitivity and flexibility to characterize and identify mycobacteria at the species and subspecies levels and potentially to identify clinical isolates of mycobacteria that have been difficult to identify by standard means. The technique has certain advantages over current biochemical, immunologic, and genetic methods because it is relatively simple to perform, is inexpensive, and requires a small amount of bacterial sample.
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To derive a method for determining malignant potential of plasma cell tumors, a retrospective analysis of the DNA ploidy and relative p62c-myc oncoprotein content using bivariate flow cytometry was performed on 23 formalin-fixed, paraffin-embedded tissues from 23 dogs. The samples included one tissue each from 17 males and six females 2 to 16 years of age (mean = 7.5 years). Twelve breeds were represented, including three Cocker Spaniels, three Golden Retrievers, and five of mixed breed. Ten of the samples were histologically classified as malignant-plasma cell tumors, and ten specimens were classified as benign. Three samples of plasmacytic inflammation, from two Cocker Spaniels and one Shih Tsu, were included as controls. The ploidy and relative c-myc content data obtained were compared with the histologic grade. A significant difference in ploidy was found between benign and malignant tumors (P < or = 0.05). Five of nine malignant plasma cell tumors were aneuploid; the remainder were diploid (4/9) or tetraploid (1/9). Only one of the benign plasmacytomas was aneuploid (1/10), whereas six were diploid (6/10), and three were tetraploid (3/10). All of the controls were diploid (3/32). When relative amounts of p62c-myc from malignant and benign tumors were compared by flow cytometry, a greater significant difference was established (P < or = 0.01) than bu using aneuploidy alone. Relative values of p62c-myc content ranged from 219 to 553 units in 8/10 malignant plasma cell tumors and from 86 to 392 units in 3/10 benign plasmacytomas.(ABSTRACT TRUNCATED AT 250 WORDS)
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Consistent with previous reports by neuropsychiatrists, the results of the present investigation confirmed the existence of a neurobehavioral disorder characterized by the subjective experiencing of multiple cognitive, affective, and psychosensory phenomena similar to those associated with more classic partial seizure disorders. According to the literature, such patients typically respond favorably to anticonvulsants although they lack the customary motor manifestations of complex partial seizure (CPS) disorders and typically do not have stereotyped spells. This neuropsychiatric syndrome has recently been termed epilepsy spectrum disorder (ESD). In the present study, 30 patients with ESD were matched with equal numbers of treatment-refractory CPS patients and normal controls. All subjects were administered a standardized interview consisting of 35 cognitive, affective, and psychosensory partial seizure-like symptoms. The results indicated that ESD patients endorsed significantly more partial seizure-like symptoms than did CPS patients and controls. Relatively low levels of symptom endorsement by an unmatched psychiatric comparison sample indicated that the high levels of symptom endorsement by ESD patients could not be attributed to the presence of psychiatric dysfunction per se. Analysis of responding to 'foil' items unrelated to partial seizures indicated that high levels of symptom endorsement by ESD patients did not merely reflect a deviant response. Although ESD patients seldom present themselves at tertiary care epilepsy centers, the study of such patients is likely to be of relevance to mainstream epileptology.
A novel method for the lysis and subsequent fractionation of bacterial constituents from Mycobacterium paratuberculosis strain 19698 (M. paratuberculosis) and Escherichia coli strain DH5 alpha utilizing the technique of matrix solid-phase dispersion (MSPD) is described. Bacteria were blended with octadecylsilyl (C18) derivatized silica to obtain cellular lysis. The blended material was used to prepare a column which was sequentially eluted with solvents of increasing polarity. Fractionation of cellular components was confirmed by analysis of the solvent extracts. The possible applicability of the MSPD technique as a general method for the lysis and fractionation of bacterial components is proposed.
Microbial Fe reduction in acetate- and succinate-containing enrichment cultures initiated with an estuarine sediment inoculum was studied. Fe reduction was unaffected when SO(4) reduction was inhibited by MoO(4), indicating that both processes could occur independently. Bacterially produced sulfide precipitated as FeS but was not completely responsible for Fe reduction. The separation of oxidized Fe particles from bacteria by dialysis tubing demonstrated that direct bacterial contact was necessary for Fe reduction. Fe reduction in cultures amended with NO(3) was delayed until NO(3) and NO(2) were removed. However, bacterial attachment to oxidized Fe particles in NO(3)-amended cultures occurred early during growth in a manner similar to NO(3)-free cultures. During late stages of growth, bacteria not attached to Fe particles became pale and swollen, while attached cells remained bright blue when examined by 4',6-diamidine-2-phenylindole epifluo-rescence microscopy. The presence of added oxidized Mn had no effect on Fe reduction. The results suggested that enzymatic Fe reduction was responsible for reducing Fe in these cultures even in the presence of sulfide and that cells incapable of Fe reduction became unhealthy when Fe(III) was the only available electron acceptor.
The distribution of methanogenic and sulfate-reducing bacteria was examined in sediments from three sites off the coast of eastern Connecticut and five sites in Long Island Sound. Both bacterial groups were detected at all sites. Three distributional patterns were observed: (i) four sites exhibited methanogenic and sulfate-reducing populations which were restricted to the upper 10 to 20 cm, with a predominance of sulfate reducers; (ii) three sites in western Long Island Sound exhibited a methanogenic population most abundant in sediments deeper than those occupied by sulfate reducers; (iii) at one site that was influenced by fresh groundwater, methanogens and sulfate reducers were numerous within the same depths; however, the number of sulfate reducers varied vertically and temporally with sulfate concentrations. It was concluded that the distributions of abundant methanogenic and sulfate-reducing bacteria were mutually exclusive. Methanogenic enrichments yielded all genera of methanogens except Methanosarcina, with the methanobacteria predominating.
The recovery of aerobic heterotrophic bacteria from Bermuda carbonate sediments on metal-supplemented media varied as much as 44-fold over a 15-cm depth. Distributional relationships with sulfate-reducing bacteria and sediment character indicated that metal tolerance was a function of metal bioavailability.
Twenty-five head-injured patients with localized theta bursts on standard or 24 h ambulatory EEG were administered a standardized interview for neuropsychiatric symptoms associated with complex partial seizures (e.g. olfactory hallucinations, memory gaps) and a battery of neuropsychological tests. Although the formal neuropsychological test performances of these patients were relatively normal they reported an abundance of seizure-like behavioural symptoms. While the frequency of these symptoms was high, they did not occur in a stereotyped complex or sequence. These findings suggest that localized theta bursts may be diagnostic of an underlying neuroelectrical disorder.
A biopsy taken from a mass on the dorsal surface of the bill of an adult female mandarin duck (Aix galericulata) was diagnosed as a malignant melanoma by light microscopy. Two months later, the tumor had enlarged considerably; the duck developed severe dyspnea and was euthanatized. At necropsy, there were metastases to lymphoid tissues in the lower regions of the neck. To our knowledge, this is the first report of a malignant melanoma in a mandarin duck.
A 2-week-old emu chick (Dromaius novaehollandiae) of approximately 200 g body weight was presented for necropsy with a history of weakness, diarrhea, pallor of the head, and acute death. Hemorrhagic enteritis with mild hepatomegaly was noted on gross examination. Microscopic examination revealed necrohemorrhagic enteritis with intralesional intranuclear basophilic viral inclusion bodies in intestinal epithelial cells; splenic lymphoid necrosis and fibrin exudation; hepatocellular vacuolar change; and multiple clusters of small gram-negative bacilli in the liver, spleen, yolk sac, and intestine. Transmission electron microscopy of negatively stained fecal specimens and thin sections of small intestine revealed clusters of viral particles consistent with adenovirus and rotavirus. Attempts at viral isolation from pooled tissue specimens were unsuccessful. Escherichia coli was isolated from specimens of liver and intestine and from an abdominal swab.