Rapid effect of triiodothyronine on the mitochondrial pathway in rat liver in vivo.

Intravenous infections of minute doses of triiodothyronine were administered to thyroidectomized rats 30 minutes before they were killed. Hepatic mitochondria were isolated rapidly and formation of adenosine triphosphate and consumption of oxygen were assessed by a 2-minute incubation. Hormone injection enhanced formation of adenosine triphosphate 114 to 217 percent over control values, with a proportionate increase in consumption of oxygen. The ratio of phosphate to oxygen was about 2.0, signifying tightly coupled oxidative phosphorylation. Stimulation was not abolished by injection of cycloheximide, puromycin, actinomycin D, or chloramphenicol 1 hour before the rats were killed. This signifies direct mitochondrial stimulation by triiodothyronine in the absence of protein synthesis.

Conversion of thyroxine to triiodothyronine by cultured human cells.

Human liver and kidney cells convert 6 to 10 percent of added thyroxine to triiodothyronine in vitro at 37 degrees C. This extent of conversion is ten times greater than that in control studies with killed cells. Conversion is evident within 10 minutes and appears to be maximal within 1 hour. Greater net triiodothyronine formation results if greater amounts of exogenous thyroxine are added to the system, with no plateau evident even at very high thyroxine concentrations. The addition of high concentrations of nonradioactive triiodothyronine resulted in no evident inhibition of the conversion.

Conversion of thyroxine to triiodothyronine in normal human subjects.

The conversion of thyroxine to triiodothyronine, previously demonstrated in athyreotic human subjects, has been investigated in normal subjects who were given intravenous injections of purified thyroxine labeled with carbon-14 in ring A and in the alanine side chain. Evidence for the conversion of T4 to T3 was provided by the finding of carbon-14 in the T3 fraction isolated from serums. It is estimated that an appreciable fraction of T4 may be transformed to T3 in normal man.

Mitochondrial thyroid hormone receptor: localization and physiological significance.

Binding studies of thyroid hormone to submitochondrial fractions from rat liver suggest that the component responsible for high-affinity, low-capacity (saturable) binding of hormones arises from the inner mitochondrial membrane. The partially purified component, approximately 150,000 daltons, appears to be half protein and half lipid, largely phospholipids, tentatively identified as lecithin, phosphatidyl ethanolamine, and cardiolipin. A similar hormone-binding macromolecule was found in mitochondria from rabbit kidney, from human liver and kidney, and from rat kidney, myocardium, skeletal muscle, intestinal mucosa, whole small intestine, adipose tissue, and lung. It was absent from mitochondria of adult rat brain, spleen, and testis, organs calorigenically unresponsive to thyroid hormones injected in vivo, but was present in mitochondria from brains of rats 12 days old and younger. The organ distribution of the hormone-binding protein and its presence in neonatal brain mitochondria supports the biological relevance of the mitochondrial component as a thyroid hormone receptor.

Thyroid hormone action: the mitochondrial pathway.

The subcellular compartments have been investigated to compare proteins capable of binding triiodothyronine and thyroxine; specific binders have been found in cytosol, nuclei, and mitochondria from rat liver and kidney. The binding protein from the inner mitochondrial membrane had the highest association constant (greater than 10(11) liters per mole), suggesting possible direct hormone action on the mitochondria. Binding of hormone analogs was found to be related to known physiological potency, and stereospecific discrimination between L- and D-thyroxine was observed. The saturable receptor was found in the mitochondrial membranes of rat liver, kidney, myocardium, and skeletal muscle but not in mitochondria from the unresponsive tissues: brain, spleen, and testis. Oxidative phosphorylation by mitochondrial vesicles from hypothyroid rats increased after the addition of physiological concentrations of triiodothyronine, which corroborated direct hormone action on mitochondria.

Determination of triiodothyronine concentration in human serum.

A simplified method has been described for the measurement of triiodothyronine (T3) in human serum. The sensitivity was sufficient for determinations on hypothyroid as well as normal and thyrotoxic sera. The values obtained have been in reasonable agreement with a double isotope derivative assay. The normal T3 concentration in human serum approximates 0.2 mug/100 ml; the mean +/-SD of 31 normal sera was 220 +/-27 ng/100 ml. Elevations were observed in sera from 40 patients with thyrotoxicosis (752 +/-282 ng/100 ml), and diminutions were found in sera from 10 hypothyroid patients (98+/-48 ng/100 ml). In rare instances thyrotoxicosis may be due to elevated serum T3 with normal thyroxine (T4) concentration. The incidence of this condition remains to be determined. In approximately half the cases with low serum T4 after (131)I therapy, the eumetabolic state may be maintained by normal or elevated T3 concentration. From these data and kinetic studies indicating a rapid turnover it may be inferred that T3 rather than T4 may be the more important hormone in health and in disease.

Preparation and properties of thyroxine-binding alpha globulin (TBG).

Thyroxine-binding alpha globulin (TBG) in human serum was isolated from Cohn fractions IV-5,6 and IV-4 by (1) chromatography on carboxymethyl (CM) cellulose, (2) gel filtration on Sephadex G-200, (3) chromatography on diethylaminoethyl-Sephadex, (4) a novel procedure of "double-gel" electrophoresis, and (5) preparative polyacrylamide gel electrophoresis. The protein was homogeneous by analytical disc gel electrophoresis, immunoelectrophoresis, and ultracentrifugal analyses (sedimentation velocity and sedimentation equilibrium), and after addition of thyroxine-(125)I showed a constant specific radioactivity on polyacrylamide electrophoresis. The sedimentation and diffusion coefficients were s(20, w), 3.0 x 10(-13) sec, and D(20, w), 8.05 x 10(-7) cm(2).sec(-1), and the molecular weight obtained by sedimentation equilibrium was 36,500. Gel filtration studies on Sephadex G-200 demonstrated that the protein had the same elution volume as that of native TBG in serum, apparently excluding the possibility of a subunit of the native protein. Chemical composition was ascertained by amino acid and carbohydrate analyses. The maximal thyroxine (T4)-binding capacity measured by reverse flow paper electrophoresis was 15,000 mug per g of protein, representing more than 2100 times that of the starting material, or about 5000 times that of whole serum. Based on the molecular weight obtained, the TBG preparation could bind 0.7 mole T4 per mole of protein, suggesting a single binding site. The association constant for T4 was estimated to be of the order of 10(10) by competitive binding studies employing TBG and T4-binding prealbumin (TBPA).

Adenovirus infection after pediatric bone marrow transplantation.

Retrospective analysis of 206 patients undergoing 215 consecutive bone marrow transplants (BMT) at St Jude Children's Research Hospital between November 1990 and December 1994 identified 6% (seven male, six female) with adenovirus infection. The affected patients had a median age of 7.9 years (range 3-24 years) at time of transplantation. Although transplants were performed for hematologic malignancies, solid tumors or nonmalignant conditions, only patients with hematologic malignancies had adenoviral infections. Adenovirus was first detected at a median of 54 days (range -4 to +333) after BMT. Adenovirus developed in eight of 69 (11.6%) patients receiving grafts from matched unrelated or mismatched related donors, in four of 52 (7.7%) receiving grafts from HLA-matched siblings, and in one of 93 (1.1%) receiving autografts. The most common manifestation of adenovirus infection was hemorrhagic cystitis, followed by gastroenteritis, pneumonitis and liver failure. The incidence of adenovirus infection in pediatric BMT patients at our institution is similar to that reported in adult patients. Using univariate analysis, use of total body irradiation and type of bone marrow graft were significant risk factors for adenovirus infection. Only use of total body irradiation remained as a factor on multiple logistic regression analysis.

Thyroid hormone action: effect of triiodothyronine on mitochondrial adenine nucleotide translocase in vivo and in vitro.

Adenine nucleotide translocase (AdNT) levels were measured as the exchange of extramitochondrial against intramitochondrial adenosine diphosphate (ADP) in liver, spleen, and testes mitochondria isolated from normal and hypothyroid rats using the "back-exchange" and atractyloside-stop method of Pfaff and Klingenberg. The results provide confirmation of previous reports that mitochondria from hypothyroid rats show a markedly diminished AdNT activity, which is restored to normal levels within 72 hours by intraperitoneal injection of 10 to 20 micrograms triiodothyronine (T3)/100 g body weight. The latter dose was found in dose-response studies to result in maximal stimulation of AdNT in liver mitochondria. Qualitatively similar results on AdNT activity were obtained in liver mitochondria within 30 to 60 minutes following intravenous injection into hypothyroid rats of a more physiological dose of T3 (40 ng/100 g body weight). AdNT in mitochondria isolated from spleen and testes (organs that do not exhibit a calorigenic response after administration of thyroid hormone to the whole animal) failed to respond to thyroidectomy and to administration of T3. More recently, we have observed that in vitro replacement of T3 also stimulates AdNT activity in hypothyroid liver mitochondria. The enzyme adenosine triphosphate (ATP) synthase was examined as another possible candidate for direct hormonal stimulation of mitochondria. Simultaneous determinations on the same rats after intraperitoneal injection of T3 (20 micrograms/100 g body weight) showed little or no effect on ATP synthase until after 37 to 85 hours, whereas enhanced activity of the translocator was regularly observed at 17 hours.(ABSTRACT TRUNCATED AT 250 WORDS)

Atrophie blanche. An unusual ulcerative state of the lower limb.

A case of atrophie blanche, an unusual ulcerative state of the lower limb, is presented herein.