David L. Bunner

Alteration of multiple cell membrane functions in L-6 myoblasts by T-2 toxin: an important mechanism of action.

Recent studies suggest that T-2 toxin interacts with cell membranes and alters membrane function. This study was done to assess the effect of T-2 toxin on a broad range of cell membrane functions in L-6 myoblasts. The following parameters were assessed after exposure to T-2 toxin for 10 min: (1) the uptake of calcium, rubidium, and glucose; (2) the uptake of leucine and tyrosine and incorporation into protein; (3) the uptake of thymidine and incorporation into DNA; and (4) residual cellular lactate dehydrogenase (LDH) as a measure of cell membrane integrity. The effects of T-2 toxin on these parameters were: (1) The minimal effective concentration (MEC) of T-2 toxin that caused a reduction in the uptake of calcium and glucose was 4 pg/ml. The uptake of rubidium was increased at 0.4 pg/ml and then reduced at 4 pg/ml and higher concentrations. (2) The MEC for reduction of the uptake of leucine and tyrosine and their incorporation into protein was 4 pg/ml. (3) Thymidine uptake and incorporation into DNA showed a biphasic response with an increase at 0.4 pg/ml and a reduction at 4 pg for uptake and 40 pg/ml for incorporation. (4) Intracellular LDH was reduced at 4 ng/ml. (5) Calcium efflux was reduced after 1-, 5-, and 15-min exposures to T-2 toxin in a concentration of 40 ng/ml. All of the changes noted, including protein synthesis inhibition, were present to a significant degree within 10 min of exposure to T-2 toxin. This time interval is too short to attribute all of these effects directly to protein synthesis inhibition since most short-lived proteins have half-lives measured in hours. In conclusion, T-2 toxin appears to have multiple effects on cell membrane function at very low concentrations (0.4 pg/ml to 4 ng/ml), which are independent of protein synthesis inhibition. These likely include effects either direct or indirect on amino acid, nucleotide, and glucose transporters, as well as calcium and potassium (rubidium) channel activities.

Circadian Growth Hormone and Prolactin Blood Concentration During a Self-Limited Viral Infection and Artificial Hyperthermia in Man

Growth hormone and prolactin blood concentrations were measured in five human volunteers over 28-hour periods including 24 hourly samples (0800 to 0800 hours) followed by an oral glucose tolerance test (0800 to 1100 hours) both preexposure and during the peak febrile phase of a self-limited viral infection, Sandfly fever. Several months after recovery, three of the subjects were studied for 24-hour periods while they sat in a tub of water at 41 degrees C for 2 hours from 1300 to 1500 hours. During all studies, mealtimes (0800 hours, 1130 hours, 1630 hours) and dark phase (2300 to 700 hours) were fixed. Growth-hormone concentrations were strikingly elevated throughout the 24-hour study done during the febrile period of Sandfly fever infection (P less than .01) except for the period of normal nocturnal release when they were not significantly different from the baseline study. No additional nocturnal surge was noted the already elevated growth-hormone values during the viral-induced fever. Growth-hormone values tended to decline slowly during the night but increased considerably during the glucose-tolerance test the following morning. These changes were similar to responses previously reported in patients with cases of malnutrition. A clear-cut increase in growth-hormone concentrations (P less than .001) was also seen during a brief 2-hour period of artificial hyperthermia, suggesting that elevated body temperature alone may explain part of the increase in growth-hormone values seen during the fever of infection. A nocturnal surge of growth hormone was still seen in the artificial hyperthermia study, albeit somewhat delayed.(ABSTRACT TRUNCATED AT 250 WORDS)

Glucose-dependent insulin inhibition of ketone body formation from long-chain fatty acids in the perfused livers of fasted rats

The data presented in this report show a direct effect of insulin on impairment of ketone body production in perfused livers from fasted rats. The data also show that physiologic levels of insulin alone or glucose alone are not sufficient to cause an impairment in ketogenesis. Only when insulin and glucose are both present at levels seen in infected rats is ketone body production impaired.

The hemostatic derangement produced by T-2 toxin in cynomolgus monkeys.

T-2 toxin, a mycotoxin produced by several strains of the genus Fusarium, has been implicated as a cause of serious illness in both animals and man. Hemorrhage is a feature of the syndromes which have been described. An LD20 dose of T-2 was administered im to adult cynomolgus monkeys. This resulted in prolongation of prothrombin and activated partial thromboplastin times and a decrease in multiple coagulation factors. These changes were detected within hours of toxin administration, were maximal at 24 hr, and returned to normal over the next 3 days. Fibrin-fibrinogen degradation products were not detected at any time point. Repeated phlebotomy produced a significantly greater increase in platelet count in control monkeys, which could be taken as evidence for an effect of toxin on platelet kinetics. In treated animals, the hematocrit level declined by about 10%, but a similar decrease occurred in control animals. The white blood cell count increased 4 to 5 times over pretreatment values. Despite the changes in multiple laboratory parameters, treated monkeys did not exhibit clinical evidence of hemorrhage. In three animals which died as a result of toxicosis, necropsy revealed mild petechial hemorrhage involving the colon and heart, as well as necrosis of lymphoid tissues.

No effect of modulators of reactive oxygen-induced pathology on microcystin-LR intoxication

Because reactive oxygen species are formed during the metabolism of several toxins that cause similar pathologic changes, we hypothesized that compounds that alter the concentration of reactive oxygen species would alter the toxic effects of the peptide-hepatotoxin produced principally by Microcystis aeruginosa. Pretreatment with alloxan, butylated hydroxyanisole or desferrioxamine did not alter the severity of microcystin-LR intoxication in fed mice. Furthermore, fasting mice for 24 hr before testing, which unmasks lipid peroxidation in paracetamol intoxication, did not alter the effect of butylated hydroxyanisole pretreatment.