Richard E. Dinterman

The effect of T3 and reverse T3 administration on muscle protein catabolism during fasting as measured by 3-methylhistidine excretion☆☆☆

Since recent studies have indicated that measurement in urine of the amino acid, 3-methylhistidine, accurately reflects the extent of muscle catabolism, and because it has been suggested that thyroid hormones may influence muscle breakdown, especially during fasting, the effect of T3 and reverse T3 (rT3) administration on the excretion of 3-methylhistidine was examined in obese subjects during fasting. The mean (+/- SE) 3-methylhistidine excretion in patients fed an egg protein diet (devoid of meat protein) was 256 +/- 35 mumoles/day and decreased to 190 +/- 14 mumoles/day during fasting. T3 administration (100 microgram/day x 5 days) increased 3-methylhistidine excretion to 304 +/- 37 mumoles/day during its ingestion and to 485 +/- 46 mumoles/day in the T3 posttreatment interval. T3 doses of 10 microgram every 4 hr (q4h) for the first 6 days of fasting also appeared capable of increasing 3-mehis excretion whereas 5 microgram T3 q4h administered during the first 6 days of fasting did not increase 3-mehis excretion. Reverse T3 administration (80 microgram q6h) during fasting was associated with a mean 3-methylhistidine of 130 +/- 13 mumoles/day, a value no higher than in patients fasted alone. These observations suggest that: (1) skeletal muscle catabolism decreases during fasting: and (2) pathophysiologic doses of T3 (60 microgram/day or more), but not reverse T3, enhance muscle catabolism during fasting.

Role of the Liver in Regulation of Ketone Body Production during Sepsis

During caloric deprivation, the septic host may fail to develop ketonemia as an adaptation to starvation. Because the plasma ketone body concentration is a function of the ratio of hepatic production and peripheral usage, a pneumococcal sepsis model was used in rats to measure the complex metabolic events that could account for this failure, including the effects of infection on lipolysis and esterification in adipose tissue, fatty acid transport in plasma and the rates of hepatic ketogenesis and whole body oxidation of ketones. Some of the studies were repeated with tularemia as the model infection. From these studies, it was concluded that during pneumococcal sepsis, the failure of rats to become ketonemic during caloric deprivation was the result of reduced ketogenic capacity of the liver and a possibly decreased hepatic supply of fatty acids. The latter appeared to be a secondary consequence of a severe reduction in circulating plasma albumin, the major transport protein for fatty acids, with no effect on the degree of saturation of the albumin with free fatty acids. Also, the infection had no significant effect on the rate of lipolysis or release of fatty acids from adipose tissue. Ketone body usage (oxidation) was either unaffected or reduced during pneumococcal sepsis in rats. Thus, a reduced rate of ketone production in the infected host was primarily responsible for the failure to develop starvation ketonemia under these conditions. The liver of the infected rat host appears to shuttle the fatty acids away from beta-oxidation and ketogenesis and toward triglyceride production, with resulting hepatocellular fatty metamorphosis.

Further evidence that leukocytic endogenous mediator (LEM) is not endotoxin

Despite several similarities, the effects of leukocytic endogenous mediator (LEM), a small protein, were further differentiated from bacterial endotoxin, a complex lipopolysaccharide, on the basis of non-identical biological activities. When either substance was administered to normal rats, each produced significant depression in serum zinc and iron concentrations, as well as a flux of amino acids to the liver. However, only LEM produced these effects on host metabolism in rats made tolerant to endotoxin. The effects of LEM and endotoxin on the synthesis and/or release of acute phase serum glubulins were also compared. Endotoxin produced a significant increase in only the α2-macrofeto-protein of normal rats. By contrast, LEM produced significant increases in all the acute phase serum protein fractions measured in either normal or endotoxin-tolerant rats. The differences and relationships between LEM and endotoxin on host responses are discussed.

Glucose and alanine metabolism during bacterial infections in rats and Rhesus monkeys.

To investigate the effects of bacterial infection on glucose and alanine metabolism, a variety of studies were carried out in rat and monkey models. These included glucose turnover by a pulse-dose technique in infected rats; alanine and glucose production and utilization in control and septic monkeys; in vivo measurement of gluconeogenesis in rats, with and without an alanine load; the in vitro rate of glucose formation from various substrates by isolated liver perfusion and hepatic cells; and in vivo rates of oxidation of glucose labeled with 14C at the 1 or 6 carbon position. In rats, glucose turnover was markedly accelerated 24 hr after inoculation of either 10(4) or 10(7) Streptococcus pneumoniae. Glucose utilization and production were also accelerated during illness and early recovery from a pneumococcal infection in monkeys. The rates of gluconeogenesis as measured by either a pulse technique in rats or continuous infusion of labeled alanine in monkey were significantly elevated during pneumococcal septicemia. During the agonal stages (10(7) of the pneumococcal infection in rat, an alanine load resulted in a decreased rate of labeled glucose production and an increase in plasma glucose when compared to values of fasted control rats. However, early illness caused similar or increased rates of glucose production from alanine in vivo. Similar reduced rates of glucose formation were observed when the isolated livers or hepatocytes from rats during the agonal stages of infection were perfused with excess quantities of gluconeogenic substrates. Thus, in the rat, gluconeogenic capacity (ability to form glucose from excess substrates) appears to decrease only during the agonal stages of pneumococcal infection. During acute pneumococcal sepsis in the rhesus monkey, alanine production and utilization were significantly elevated and it was estimated that over 90% of the newly produced alanine was utilized for glucose synthesis. When arterial--venous differences were measured across the hindquarters, significantly more alanine was released, presumably from skeletal muscle of the septic monkey, compared to the recovery period or in the control groups. Thus, the increase in glucose synthesis in both rat and monkey appears to be correlated with substrate availability and kinetic rate, rather than gluconeogenic capacity of the liver. The major increase in glucose utilization during both S. pneumoniae and Francisella tularensis live vaccine strain (LVS) infections in rat was a progressive elevation in the rate of oxidation via the pentose phosphate shunt in the rat. Further, the rate of oxidation appeared to be correlated with the magnitude of the bacteremia, which is an indication of the severity of the infection...

Cutaneous absorption and decontamination of [3H]T‐2 toxin in the rat model

Cutaneous absorption and decontamination of [3H]T-2 mycotoxin using various treatment modalities incorporating water, detergent, sprays, and scrubbing of application sites were examined in the rat model at 5, 30, 60, and 1440 min (24 h) postexposure. Rats were killed immediately after treatment and radiolabeled T-2 remaining in full-thickness skin samples were determined. Absorption and decontamination were followed over time, and decontaminating treatment modalities were evaluated for efficacy. Less than 1% of the applied dose was absorbed in 5 min, and 50% was absorbed in 24 h. At 5 min, 99.5 +/- 0.05% of nonabsorbed (residual) [3H]T-2 was removed, and 58 +/- 5.2% of residual toxin was removed at 24 h with a 2.5% detergent/water spray. When treatment modalities were evaluated at 60 min, a 2.5% detergent/water scrub followed by a detergent/water spray produced optimal decontamination by removing 81 +/- 2.2% of residual toxin. All treatment modalities using detergent and/or water removed significant amounts of toxin (p less than or equal to .0001); a dry scrub was not efficacious. Treatment should be initiated as soon as possible after exposure for best results. However, the stratum corneum acts as a reservoir for the toxin, and decontamination should be carried out even if delayed several hours or days after exposure. Dermal absorption pharmacokinetics found in these studies are similar to those described for other low-molecular-weight compounds, and the decontamination results from T-2 toxin should be applicable to other, similar toxic substances.

Use of Lipid Calories during Pneumococcal Sepsis in the Rhesus Monkey

Abstract : A number of studies in patients and monkeys have suggested that intravenous infusions of lipids can be utilized as a major source of calories during sepsis and/or trauma. Others, however, have observed that lipid calories were not as effective as glucose calories in sparing body protein in severely septic and/or burned patients. In trying to resolve this controversy, a parenteral nutrition model was utilized during pneumococcal sepsis in the rhesus monkey. Earlier studies indicated that when monkeys were infused with 0.55 grams amino acid nitrogen and 13 kilocalories per kilogram per day, they lost approximately 13% of their body protein as a result of the clinical illness associated with pneumococcal sepsis. Addition of 85 kilocalories per kilogram per day of either dextrose or lipid emulsion (Intralipid) effectively prevented this infection-related loss of body protein. The present study was designed to determine whether the elevated plasma insulin concentrations associated with glucose infusions are necessary for protein sparing or would interfere with utilization of lipid calories. In this study, septic and control monkeys were infused with a hypocaloric amino acid-dextrose solution to which was added lipid emulsion. If septic monkeys were totally maintained on a hypocaloric infusion of amino acids and 32 kilocalories per kilogram per day from dextrose, they lose 6.8 + or - 1.0% of their body protein over the 6-day experimental period. Addition of lipid calories (55 kilocalories per kilogram per day) to this mixture resulted in a gain of 1.1 + or - 0.7% of body protein. In both experimental groups plasma insulin was approximately 100 to 200 microunits per milliliter, which resulted in a marked inhibition of ketogenesis and lipolysis. Despite elevated plasma insulin, the septic monkey appeared able to utilize the fatty acids from the infused lipid effectively as a calorie source to spare body protein. Since lipid emulsion contains approximately 13% glycerol...

Protein-sparing therapy during pneumococcal infection in rhesus monkeys.

A model was developed in the rhesus monkey to determine if the marked wasting of body proteins associated with sepsis could be prevented by an intravenous supply of various nutritional substrates. All monkeys were given a basic infusion of 0.5 gm of amino acid nitrogen/kg body weight via an indwelling catheter in the jugular vein. Three groups were given diets with no added calories, 85 calories/kg from dextrose or 85 calories from lipid. In each group, six monkeys were inoculated with 3 x 10(8) Streptococcus pneumoniae and four with heatkilled organisms. In the monkeys infused with the amino acids alone, pneumococcal sepsis resulted in a fourfold increase in loss of body proteins compared with calorie-restricted controls. Addition of 85 calories/kg/day of either dextrose or lipid reduced body wasting associated with infectious disease. The calories from lipid were utilized bythe septic host as a source of energy, with a slightly reduced efficiency when compared with the isocaloric infusion of dextrose. The nitrogen sparing of the fat emulsion could not be accounted for by its glycerol content. Therefore, the septic monkey seemed to utilize fatty acids as an energy substrate. It appears that the carbohydrate calories tend to favor the synthesis of peripheral proteins (associated mainly with skeletal muscle), while lipid calories favor synthesis of visceral proteins such as plasma albumin and acute-phase proteins.

Penetration of [3H]T-2 mycotoxin through abraded and intact skin and methods to decontaminate [3H]T-2 mycotoxin from abrasions.

Penetration of 50 muCi of [3H]T-2 mycotoxin through abraded and intact skin was studied in anesthetized rats sacrificed at 5, 15, 30, 45, 60 and 90 min post-exposure. The greatest penetration was through abraded skin (49 +/- 7%) at 90 min post-exposure, whereas penetration through intact skin (2 +/- 3%) was substantially less (P less than 0.0015). Methods to decontaminate [3H]T-2 mycotoxin from abraded skin over time were studied. Treatment of [3H]T-2 contaminated abrasions by applying Trau + Medic dressing, applying Charcoal Cloth-Anti-bacterial Field Dressing (Charcoal Dressing), or swabbing with povidone-iodine 30 min post-exposure removed 17-32% of the applied [3H]T-2. Immediate blotting with immediate removal of the dressings absorbed 103 +/- 4% (Trau + Medic) and 87 +/- 4% (Charcoal Dressing) of the applied [3H]T-2, while immediate blotting and leaving the dressing in place for 30 min removed 91 +/- 5% (Trau + Medic) and 76 +/- 3% (Charcoal Dressing). It appears that immediate blotting with either dressing followed by immediate removal before application of a clean dressing is an effective method for decontaminating [3H]T-2 from abrasions.