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L Tsai

Publications and source records attributed to L Tsai.

At least 19 recordsLinked to original sources

Synthesis of 5-methylaminomethyl-2-selenouridine in tRNAs: 31P NMR studies show the labile selenium donor synthesized by the selD gene product contains selenium bonded to phosphorus.

An enzyme preparation from Salmonella typhimurium catalyzes the conversion of 5-methylaminomethyl-2-thiouridine in tRNAs to 5-methylaminomethyl-2-selenouridine when supplemented with selenide and ATP. Similar preparations from a Salmonella mutant strain carrying a defective selD gene fail to catalyze this selenium substitution reaction. However, supplementation of the deficient enzyme preparation with the purified selD gene product (SELD protein) restored synthesis of seleno-tRNAs. In the absence of the complementary enzyme(s), the SELD protein catalyzes the synthesis of a labile selenium donor compound from selenide and ATP. 31P NMR studies show that among the products of this reaction are AMP and a compound containing selenium bonded to phosphorus. The reaction is completely dependent on the addition of both selenide and magnesium. The dependence of reaction velocity on ATP concentration shows sigmoidal kinetics, whereas dependence on selenide concentration obeys Michaelis-Menten kinetics indicating a Km value of 46 microM for selenide.

Adenosine Monophosphate

ADHD and PDD.

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Attention Deficit Disorder with Hyperactivity

Effect of cortisol on the secretion of testosterone and estradiol-17 beta by human granulosa-luteal cell cultures. A model system for analyzing hormonal alterations in female athletes.

The combination of hypercortisolism and usually low androgen and estrogen levels is frequently observed in female long distance athletes. In order to find a useful model system for studying the underlying mechanisms, the following studies were performed. The effect of cortisol on the secretion of testosterone (T) and estradiol-17 beta (E2) by human granulosa-luteal cells was studied in vitro in cultures of cells recovered from mature follicles of gonadotrophin stimulated women (participating in the IVF program). Following 24 hours of culture in tissue culture medium without hormonal additives, the granulosa-luteal cells were incubated for 6 hours in media with addition of 4-androstene-3,17-dione (A-4) as precursor and hMG and cortisol in different combinations. The secretion of T was significantly stimulated by cortisol but not by human menopausal gonadotrophin (hMG). Cortisol, but not hMG, also increased the secretion of E2, although this effect was not statistically significant. These in vitro findings make a direct effect of cortisol upon ovarian sex steroid secretion less likely as the mechanism behind the subnormal sex steroid levels in female long distance athletes. Instead, inadequate gonadotrophic stimulation, related to hypothalamic amenorrhea, and/or a selective decrease in the adrenal secretion of precursor steroids, may be an explanation.

Estradiol

Mood disorder in a group of self-cutting adolescents.

This study describes the occurrence of mood disorder in a group of adolescent inpatients involved in self-cutting behavior. Although self-cutting is generally said to be associated with borderline personality disorder, a substantial number of patients who cut themselves in our study were clinically depressed. In addition, the self-cutting adolescents had a high prevalence of child abuse, past psychiatric history, and family psychiatric history. The incidents tended to spread by contagion and occur at times of change affecting the milieu. Clinical implications of these findings are discussed.

Adolescent

Okadaic acid interferes with lipoprotein-supported corticosterone production in adrenal cells.

Rat adrenocortical cells in culture respond to stimulation by ACTH alone (15 fold over basal) and to ACTH + added lipoproteins (as an exogeneous source of cholesterol), with an additional 25-30 fold rise in steroidogenesis. With the addition of okadaic acid (OKA, 100 nM), a potent protein phosphatase inhibitor, the lipoprotein-induced rise in steroidogenesis is blocked. If 20 alpha-hydroxycholesterol is provided instead of lipoprotein-cholesterol, OKA has no effect suggesting that OKA affects only actively transported cholesterol. Since the OKA block is preceded by specific morphological changes in the cell (i.e., the loss of Golgi-associated microtubules followed by the disruption of the Golgi apparatus itself), it is hypothesized that some OKA-sensitive phosphoprotein associated with the microtubule/Golgi network of adrenocortical cells is critical for lipoprotein-derived cholesterol uptake and/or transport during steroidogenesis.

Adrenal Cortex

Cortisol and androgen concentrations in female and male elite endurance athletes in relation to physical activity.

To evaluate the effect of variations in physical activity on selected hormone concentrations in male versus female athletes, fasting serum concentrations of cortisol (C), total-testosterone, free-testosterone, non sex hormone binding globulin (SHBG) bound testosterone, dehydroepiandrosterone, 4-androstene-3,17-dione and SHBG were studied. The tests were performed in nine male and seven female elite endurance athletes during the off-season (test 1), early in the competition season (test 2) and at the end of the competition season (test 3). The C concentration increased significantly during the competition season in women but not in men. Further, the mean C concentrations at test 3 as well as the mean level during the whole observation period (tests 1, 2, 3) were significantly higher in women than in men. No significant changes were found in androgen concentrations or androgen:cortisol ratios within the two groups. The differences between the sexes in C response may indicate different adaptive mechanisms to similar physical stress.

Adult

Depression in autistic disorder.

Depressive illness in a patient with Down's syndrome and autism responded to fluoxetine. The importance of diagnosing superimposed depression in people with developmental disorders is emphasised.

Adolescent

Uptake and utilization of lipoprotein cholesteryl esters by rat granulosa cells.

Earlier studies have shown that rat granulosa cells grown in serum-free medium are exquisitely responsive to exogenously provided lipoprotein cholesterol. In this study we compare the amount of cholesterol (cholesteryl ester) actually delivered from various homologous and heterologous cholesterol-rich lipoproteins and examine the intracellular pathways used in the delivery system. Granulosa cells were incubated for 5 or 24 h with 125I-labeled human (h) HDL3, rat (r) HDL or hLDL equipped with non-releasable apoprotein and cholesteryl ether tags which accumulate within cells, even after degradation. We show that all the tested lipoproteins were similarly efficient in cholesteryl ester delivery; i.e., based on cholesterol: protein ratios of the starting ligands, each delivered approximately the same cholesteryl ester mass and evoked a similar progestin response. However, each lipoprotein was processed quite differently by the granulosa cells: hHDL3-cholesteryl ester was taken up almost exclusively by an non-endocytic pathway, hLDL-cholesteryl ester almost exclusively by an endocytic pathway and rHDL-cholesteryl ester by both pathways. In general, there was no correlation between the total amount of lipoprotein bound or apoprotein internalized and/or degraded by the cells with the amount of cholesteryl ester received or the level of the progestin response. Hormone stimulation upregulated the preferred pathway for each lipoprotein.

Animals

Restoration of anabolic deficit and muscle glycogen consumption in competitive orienteering.

Consumption and restoration of muscle glycogen and changes in anabolic and catabolic steroid hormones were analyzed in five male elite orienteers during and after an orienteering competition. The magnitude of glycogen consumption and pronounced increase in serum-cortisol during the orienteering race reflect the great muscular output demands during forest running. The free testosterone/cortisol ratio was normalized to the initial level within four hours post-exercise. Synchronously, only 25% of the muscle glycogen loss was restored. Within 24 hours post-exercise all runners showed normalized levels of testosterone, cortisol and free testosterone/cortisol ratio. The glycogen content was also restored except in one of the runners. We conclude that daily orienteering competitions per se do not seem to create risks for developing a state of hormonal imbalance or significant decrease in glycogen when the carbohydrate supply is appropriate.

Adult

In vitro incorporation of selenium into tRNAs of Salmonella typhimurium.

Broken-cell preparations of Salmonella typhimurium rapidly incorporated 75Se from 75SeO3(2-) into tRNA by an ATP-dependent process. Selenium incorporation in the presence of 50 microM 75SeO3(2-) (0.8-1 pmol per A260 unit) was enhanced by the selenocysteine precursor, O-acetyl-L-serine (to 3.7 pmol per A260 unit). This increase in incorporation was a function of O-acetyl-L-serine concentration. Neither O-acetyl-L-homoserine nor O-phospho-L-serine stimulated the incorporation of selenium into tRNA. The incorporation of 75Se from 75SeO3(2-) was decreased by adding L-selenocysteine but not by adding the D isomer. When homologous bulk tRNA was added to the broken-cell preparations, an increased rate of 75Se labeling was observed. The supernatant fraction of the broken-cell preparation contained all of the enzymes required for this process. Reversed-phase HPLC analysis of labeled bulk tRNA digested to nucleosides showed the presence of a labeled compound that coeluted with authentic 5-methylaminomethyl-2-selenouridine.

Adenosine Triphosphate

Biosynthesis of selenium-modified tRNAs in Methanococcus vannielii.

Selenium-containing nucleosides are natural components of several tRNA species in Methanococcus vannielii. In the present study, the incorporation of selenium from 75SeO3(2-) into these macromolecules was investigated in sonic extracts of M. vannielii. Nucleoside analysis of the 75Se-labeled tRNAs from these in vitro reaction mixtures demonstrated that the selenium was present in 75Se-labeled nucleosides identical to the two naturally occurring 2-selenouridines produced in vivo. Incorporation of selenium into these nucleosides was ATP-dependent and was maximal after 20 min. Addition of O-acetylserine enhanced the activity 2- to 3-fold, implicating a role for selenocysteine in the reaction. Added L-selenocysteine could function as a selenium donor, but the D isomer and DL-selenomethionine were inactive. RPC-5 chromatography of bulk tRNA isolated from M. vannielii grown on 75SeO3(2-) separated five major species of seleno-tRNAs. The amino acid-accepting activity of these tRNAs was investigated.

Amino Acids

Effects of interleukin-2 and interleukin-2-activated cells on in vitro myelopoiesis.

Lymphokine-activated killer (LAK) cells from human peripheral blood mononuclear cells cultured with recombinant interleukin-2 (IL-2) have been used clinically in adoptive immunotherapy for cancer patients. To study the influence of LAK cells and IL-2 on haematopoiesis, an in vitro assay system for colony formation of granulocyte-macrophage progenitor cells (GM-CFC) was used. LAK cells from cultures of either human peripheral blood (PB) or human bone marrow (BM) mononuclear cells were both inhibitory to allogeneic BM-derived GM-CFC. Inhibitory activity could be transferred with supernatants from co-cultures of LAK cells and BM targets, but also from the IL-2 activated PB- or BM-derived cells alone. The inhibitory activity from the initially non-cytotoxic/non-inhibitory BM population was rapidly induced by IL-2 activation, and preceded the generation of cytotoxic LAK cells in the culture. These experiments show that inhibition of haematopoietic progenitor cells by IL-2 is not dependent on generation of cytotoxic LAK cells, but rather the result of IL-2-induced cytokine production. We conclude that the synergistic action of interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha) may contribute to inhibition, but that also other cytokines are responsible for the observed inhibition of BM-derived GM-CFC.

Bone Marrow Cells

Derivatization of gamma-glutamyl semialdehyde residues in oxidized proteins by fluoresceinamine.

Oxidative modification of proteins is implicated in a number of physiologic and pathologic processes. Metal-catalyzed oxidative modification usually causes inactivation of enzymes and the appearance of carbonyl groups in amino acid side chains of the protein. We describe use of fluoresceinamine to label certain of those carbonyl groups. Fluoresceinamine reacted with those carbonyl groups to form a Schiff base which was reduced by cyanoborohydride to yield a stable chromophore on the oxidized residue. The high molar absorbtivity of the fluorescein moiety conferred high sensitivity upon the method. Labeled peptides were readily identified after tryptic digestion of oxidized glutamine synthetase. Further, acid hydrolysis of labeled glutamine synthetase allowed isolation of the derivatized, oxidized residue. The oxidized amino acid was identified as gamma-glutamyl semialdehyde. During metal-catalyzed oxidation, the inactivation of glutamine synthetase paralleled the appearance of gamma-glutamyl semialdehyde.

Aldehydes

Effect of IFN-gamma treatment and in vivo passage of murine tumor cell lines on their sensitivity to lymphokine-activated killer (LAK) cell lysis in vitro; association with H-2 expression on the target cells.

Interferon-gamma (IFN-gamma) treatment or in vivo passage of the murine YAC-1 lymphoma resulted in reduced sensitivity to in vitro lysis by syngeneic murine spleen cells cultured in rIL-2 (LAK-cells). IFN-gamma treatment also rendered the murine B16 melanoma less sensitive to lysis by syngeneic LAK cells, whereas in vivo passage did not alter LAK sensitivity. The reduction in sensitivity to lysis correlated with enhanced expression of cell surface H-2 on the target cells. The possible role of H-2 was studied with a beta 2-microglobulin-deficient, and thus H-2-deficient, variant of the YAC-1 lymphoma. This variant line remained H-2 negative even after IFN-gamma treatment or in vivo passage, and was highly sensitive to LAK-cell-mediated lysis, even after IFN-gamma treatment or in vivo passage. The present results are discussed in relation to IFN-gamma and in vivo induced modulation of MHC class-1 molecules on target cells and the possible consequences for interaction with activated as well as "natural" effector cells.

Animals