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Biomedical subjects

J F Roberts

Publications and source records attributed to J F Roberts.

At least 37 records · Page 2Linked to original sources
PubMed1986

Macrophage uptake of a lipoprotein-sequestered toxicant: a potential route of immunotoxicity.

An experimental system was chosen to investigate the bioactivity of a lipoprotein-sequestered toxicant at the cellular level based on recent studies demonstrating receptor-mediated uptake of lipoproteins by macrophages. Rat peritoneal exudate cell suspensions (PEC) were exposed to DDT and lipoprotein-sequestered DDT, followed by measurement of DDT uptake, metabolism, and cellular toxicity. In vitro uptake assays demonstrated that PEC suspensions treated for 10, 20, and 30 min with 2.5 microM lipoprotein-sequestered DDT had approximately a twofold increase over the amount of DDT associated with PEC treated with 2.5 microM free DDT. PEC were assayed for DDT metabolites to serve as a measure of the cellular internalization of the toxicant after treatment in vitro for 18 hr with either 1.5 microM DDT or lipoprotein-sequestered DDT. Evidence of DDT metabolism was only observed with PEC which had been treated with lipoprotein-sequestered DDT. These cells contained significantly higher amounts of DDT metabolites as compared to cells treated with unsequestered DDT (over an eightfold difference). Assays measuring macrophage phagocytic activity indicated that macrophages treated for 4.5 hr in vitro with 2.5 microM lipoprotein-sequestered DDT showed significant inhibition in their ability to phagocytize yeast particles. These results suggest that serum lipoproteins may facilitate the cellular uptake of lipoprotein-sequestered toxicants leading to altered cellular function (phagocytosis).

Animals↗
PubMed1986

Target ricin by coupling to an anti-macrophage monoclonal antibody.

By altering the receptor binding specificity of the highly potent natural toxin ricin, a macrophage specific immunotoxin was developed. Ricin ordinarily does not demonstrate cell type specificity and is capable of binding and entering cells through galactose containing receptors resulting in rapid cell death. A murine anti-rat peritoneal macrophage IgGl monoclonal antibody, B-6, was developed to serve as a target specific carrier for ricin. By covalently binding monoclonal antibody B-6 and reversibly binding lactose to ricin, a new biologically active hybrid toxin possessing macrophage specificity was developed. When P3X63-Ag8.653 myeloma cells, which served as an nonspecific target cell type, and macrophages were treated with the ricin conjugate over a broad range of concentrations and various time periods, the conjugate demonstrated substantially greater toxicity toward macrophages than myeloma cells even though both cell types responded similarly to treatments with unconjugated ricin. It was also observed that ricin was considerably more toxic to macrophages when conjugated to monoclonal antibody B-6 than unconjugated ricin. Through ricin-antibody conjugation a high degree of specificity and toxicity can be attained potentially suitable for anti-tumor reagents and immuno-modulators.

Animals↗
PubMed1985

A rapid spectrophotometric method for assessing macrophage phagocytic activity.

The method described provides a rapid and inexpensive in vitro assay of phagocytosis by mononuclear phagocytes. This assay utilizes yeast cells, stained with congo red, as the target particle and quantitation is performed spectrophotometrically. An attractive feature of this assay is that phagocytic activity is assessed using large sample sizes, in this case approximately 1 X 10(6) macrophages per sample, resulting in a more accurate evaluation of phagocytosis than assays dependent on microscopic quantitation.

Animals↗
PubMed1985

In vitro effect of aflatoxin B1 on rat liver macrophages (Kuffer cells).

The effect of aflatoxin B1 on the uptake and incorporation of [3H]leucine and [3H]uridine and on phagocytosis of latex particles was studied using cultures of rat liver macrophages (Kuffer cells). Aflatoxin B1 inhibited the incorporation of both isotopes, but inhibition of uridine incorporation was greater than that of leucine, suggesting that RNA synthesis was a major site of inhibition. Aflatoxin B1 also inhibited phagocytosis of latex particles in a time- and dose-dependent manner.

Aflatoxin B1↗
PubMed1983

Pathogenesis of change in the upper respiratory tracts of turkeys experimentally infected with an Alcaligenes faecalis isolate.

The course of changes within the upper respiratory tracts of turkey poults experimentally infected with Alcaligenes faecalis was studied. The initial change observed (5 days post-inoculation) was colonization of the upper respiratory tract by the bacterium. Changes in the nasal turbinates and trachea were first apparent as a focal loss of cilia but subsequently developed into a general loss of cilia (11 days post-inoculation). Eventually, the entire ciliated epithelial layer in the cranial region of the trachea was lost (13 days post-inoculation). With the loss of cilia and ciliated cells, a highly viscous mucus was able to accumulate in the anterior one-half to two-thirds of the trachea. In addition, changes in the gross structure of the trachea (flaccid trachea) were observed in all poults inoculated with A. faecalis. There was an apparent gradation in the severity of these changes from severe in the cranial region of the trachea to mild in the region just anterior to the bronchial bifurcation. The observations resulting from A. faecalis infection indicated two major tracheal changes responsible for the chronic and sometimes severe nature of this disease. These changes included a loss of ciliary activity and a flaccid trachea which together resulted in the accumulation and stasis of mucus and tracheal collapse.

Alcaligenes↗
PubMed1981

Cytotoxic activity of pathogenic Alcaligenes faecalis in turkey tracheal organ cultures.

Turkey tracheal organ cultures were used to study the virulence of Alcaligenes faecalis isolants that have been shown to be pathogenic for turkey poults. Viable infected and noninfected tracheal rings were examined by phase-contrast microscopy, and fixed stained sections were examined by light microscopy. Alcaligenes faecalis at concentrations of 10(8) and 10(9) colony-forming units/ml caused ciliostasis, hydropic degeneration (characterized by blebbing of the plasma membrane, cellular swelling, and cytoplasmic vacuolation), and eventual sloughing of the ciliated epithelium. Only ciliated epithelial cells appeared affected. For comparison, other bacterial isolants not pathogenic for turkeys were tried in this system. These bacterial isolants included 3 isolants of A faecalis from human beings and isolants of Klebsiella pneumoniae, Escherichia coli, and A faecalis from turkeys. Inoculation of each of these bacterial cultures onto tracheal organ cultures failed to produce the lesions described.

Alcaligenes↗
PubMed1979

Metabolic changes in Crithidia fasciculata accompanying physiological adaptation to growth in the presence of carbonyl cyanide m-chlorophenylhydrazone.

1. Crithidia fasciculata adapted to growth in the presence of 10(-5) M carbonyl cyanide m-chlorophenylhydrazone (CCCP), an uncoupler of oxidative phosphorylation, maintained adenosine phosphate pools and an adenylate energy charge comparable to those of control cells. 2. CCCP-adapted cells in the presence of the uncoupler respire endogenous substrate at a greater rate than control cells and this effect of CCCP appears readily reversible. 3. CCCP-treated, adapted cells, supporting high endogenous respiration rates, were not responsive to added substrates which significantly stimulated the oxygen utilization of normal C. fasciculata. 4. CCCP-adapted cells, provided with [U-14C]-labeled proline, utilize this substrate at 67% the rate of control cells, but divide the isotopic label between CO2 and protein in a ratio identical to that of normal cells. 5. The transport of alanine and proline by adapted C. fasciculata was severely impaired, while the transport of tyrosine and leucine was unaffected.

Adaptation, Physiological↗
PubMed1978

The ribonucleic acids of Crithidia fasciculata.

Crithidia fasciculata ribosomes were found to be 80S and to dissociate into 58 and 41S subunits; on 5 to 50% sucrose gradients, rRNA was separated into 25, 18, and 5S components. The molecular sizes of the heavier rRNA species, estimated by polyacrylamide gel electrophoresis were 1.24 and 0.84 M (X 10(6) daltons). The 25S RNA has a tendency to interact with the 18S RNA to give a complex that is difficult to separate by sucrose gradient centrifugation. The 25S RNA is also unstable and dissociates into 0.73 and 0.57 M components. The 18S RNA has molecular size (0.84 M) higher than the 0.7 M reported for most eukaryotes, but similar to that of Euglena and Amoeba. Ribosomal RNA hybridized 0.29% of the nuclear DNA. Mitochondrial RNA, extracted by a rapid procedure was resolved into 16 and 5S components in sucrose gradients.

Animals↗