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Biomedical subjects

M Kogut

Publications and source records attributed to M Kogut.

At least 19 recordsLinked to original sources

In vitro interleukin-1 and tumor necrosis factor-alpha production by macrophages from chickens infected with either Eimeria maxima or Eimeria tenella.

We measured the in vitro production of interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF- ) by macrophages from chickens during and immediately following an infection with one of two different species of Eimeria, E. maxima and E. tenella. Quantitatively, the amounts of IL-1 produced during each infection were nearly identical regardless of the oocyst dose of each parasite. TNF production followed a biphasic pattern of increased production with the first peak associated with the pathogenesis of disease and the second peak associated with the development of protective immunity. These experiments together with others we have reported imply cells taken from chickens infected with coccidia have a greater capacity to produce cytokines upon stimulation in vitro than cells from non-infected birds. The production of significantly greater amounts of TNF during the days 3-6 after inoculation correlates with the appearance of the most characteristic local and systemic pathophysiological changes in the host induced by the coccidia. The excessive release of TNF in response to a heavy coccidial infection may account for many of the pathological features observed with avian coccidiosis probably through the release of other mediators.

Animals

Dynamics of cytokine production during coccidial infections in chickens: colony-stimulating factors and interferon.

We assayed two classes of immunoregulatory cytokines, colony-stimulating factors (CSF) and interferon (IFN), during and immediately after a primary coccidial infection in chickens. Coccidial infection induces significant alterations in serum colony-stimulating activity (CSA) and these alterations immediately precede the characteristic biphasic leukocytosis. CSA rose sharply during the first 24 h post-inoculation (PI), but returned to control levels by 48 h PI. At this time, we detected an increase in peripheral blood leukocytes which peaked at 96 h PI. A second phase of CSA increase began 96 h PI and peaked at 120-144 h PI which again preceded the second phase of leukocytosis. We also examined the production of IFN during the first 20 days PI. Splenic T cells from Eimeria maxima-infected chickens produced significantly less IFN on day 5 PI compared to T cells from the coccidia-free controls. By days 10 and 15 PI, there was no significant difference in IFN production between the T cells of infected and non-infected chickens. However, by day 20 PI, IFN production by the T cells of the infected birds produced significantly more IFN than the control T cells. The results of our studies indicated the differential production of two different cytokines by chickens during and following a primary coccidial infection. Based on these experiments, CSF may be some of the first cytokines produced during an E. maxima-infection, while IFN may be one of the later cytokines produced.

Animals

Use of 23Na nuclear magnetic resonance spectroscopy to determine the true intracellular concentration of free sodium in a halophilic eubacterium.

We present new data obtained by 23Na nuclear magnetic resonance spectroscopy, which can distinguish free intracellular sodium from cell-bound sodium, showing that the intracellular concentration of Na+ the halophilic eubacterium Vibrio costicola is only 5 to 20% of that in the extracellular medium. Previous methods could not distinguish free intracellular Na+ from that bound to cell structures, and it was believed that in halophilic eubacteria the total monovalent cation concentration inside matched that of the NaCl outside. Information obtained by the newer technology raises fundamental questions about the ways in which these organisms and others which live in hypersaline environments function and cope with osmotic stress.

Bacteria

The role of osmotic effects in haloadaptation of Vibrio costicola.

Growth rates of Vibrio costicola showed a broad optimum between 0.8 and 1.5 M-NaCl, and there was no growth above 3.3 M-NaCl in a peptone-based medium. The minimum requirement of 0.5 M-NaCl for growth in NaCl alone was reduced to 0.3 M-NaCl when the total solute concentration was raised to 0.5 to 1.0 M equivalent with sucrose or glycerol. Compared with equivalent NaCl concentrations, higher concentrations of sucrose were more inhibitory to growth, whereas glycerol had less effect. Increasing the medium NaCl concentration suddenly by 2- or 3-fold with either a constant starting, or final, salt concentration showed that, after the shift-up, the lag in growth, the rate of growth, and the inhibition of phospholipid synthesis depended both on the final NaCl concentration and the magnitude of the shift in salinity. The time-courses of phospholipid synthesis following a 2- or 3-fold shift-up in NaCl or sucrose media were very similar and exhibited a relative increase in phosphatidylglycerol synthesis over that of phosphatidylethanolamine. This 'switch-over' was not seen following shift-up in glycerol media when there was also a stimulation, rather than inhibition, of phospholipid synthesis. It is concluded that during phenotypic haloadaptation of V. costicola, osmotic effects play a significant part in the sensing of and response to raised external salinity.

Glycerol

Haloadaptation: salt sensing and cell-envelope changes.

When moderately halophilic bacteria adapt phenotypically to altered salinity, they modify the composition of their cell envelopes. These alterations are discussed in the context of how external salt concentration could be sensed and the adaptive changes triggered. A general model for salt sensing and signal transduction is suggested.

Adaptation, Physiological

An investigation of mistranslation in vivo induced by streptomycin by an examination of the susceptibility of abnormal proteins to degradation.

Proteolysis rates in vivo were measured in Escherichia coli cultures during treatment with dihydrostreptomycin and under various other conditions. Dihydrostreptomycin treatment caused an increase in the proteolysis rate, compared to untreated controls. The proteolytic system in vivo responsible for the elevated proteolysis in the early stages of dihydrostreptomycin treatment, or that during canavanine and puromycin treatment, were not inhibited by addition of phenylmethanesulphonyl fluoride. This agent did inhibit proteolysis rates in cultures whose growth was inhibited by starvation, or had been completely stopped by dihydrostreptomycin. It seems, therefore, that the extremely high proteolysis rates in cultures at this stage of dihydrostreptomycin treatment were due to the action of two protease systems: the one concerned with the breakdown of abnormal proteins, and the other concerned with normal protein turnover and active during a non-specific decline of growth. The proteolytic rate at complete growth inhibition brought about by dihydrostreptomycin was intermediate between those induced by canavanine and puromycin at the same stage of treatment. This indicated a similar hierarchy in the extent and nature of abnormality in the proteins synthesized under these conditions. The relationship between the abnormality of proteins induced by dihydrostreptomycin and the importance of this in the antibiotic mechanism is discussed.

Bacterial Proteins

Survey of biochemical topics used by clinicians in teaching and practice.

In a pilot survey of biochemical topics, implied and overt, one of us (M.K.) attended 14 clinicians in 7 London medical schools during 4 half-day sessions, chosen at random, and recorded the activities and their relation to biochemistry. The sample was not intended to be representative but aimed at reasonable comprehensiveness. A list of twenty 'biochemical and related' topics was derived from these records, sent to the clinician for approval and comment, and used for rating the incidence of topics during the sampled sessions. Ratings by the two authors independently and the clinicians were combined. Collaboration between biochemist and clinicians was found crucial for the interpretation of clinical events in biochemical terms. Extension of this survey, using a questionnaire embodying the 20 biochemical categories, is to serve as a basis for defining objectives in biochemistry teaching for medical students.

Biochemistry

Effects of dihydrostreptomycin on ribosome function in vivo: lack of correlation between changes in ribosome patterns and growth.

In vivo treatment of susceptible Escherichia coli cultures with low concentrations of dihydrostreptomycin leads to a decline in polysomes and a corresponding increase in 70S particles which behave as run-off ribosomes, as well as free 30S and 50S subunits. We have examined the timing and extent of these effects on ribosomes and compared them to the effects of this antibiotic on growth and protein synthesis. We have shown that no changes in ribosome distribution are observed until growth inhibition by dihydrostreptomycin is almost complete. Thus, intracellular dihydrostreptomycin can inhibit growth and net protein synthesis without apparently affecting the ribosome cycle. Since it is known that the antibiotic combines with free 30S subunits, the question is how such combination can bring about the observed inhibition of protein synthesis and growth. We suggest that specific interaction of intracellular antibiotic with proteins of the 30S subunits allows repeated use of the ribosome cycle by such affected particles, but with selective misreading of certain amino acid codons as terminator codons, so that they produce incomplete polypeptide chains. The cumulative effect of such a mechanism would lead to eventual cessation of protein synthesis and growth.

Dihydrostreptomycin Sulfate