Search PubMed⌕ Search

Biomedical subjects

M J Thomas

Publications and source records attributed to M J Thomas.

At least 109 records · Page 6Linked to original sources
PubMed1994

Chemical and biological characterization of oxo-eicosatetraenoic acids.

Eicosatetraenoates (ETEs) with 5-oxo residues are known to induce human neutrophil (PMN) Ca2+ transients and chemotaxis. We find that 5-oxoETE, 5-oxo-8-trans-ETE, 5-oxo-15-hydroxy-ETE, 5-hydroxy-ETE, 5-hydroxy-15-oxoETE, 5,15-dioxoETE, and 5,15-dihydroxy-ETE have respective relative potencies of 10, 5, 3, 1, 0.2, 0.1, and 0.02 in: a) causing PMN to mobilize Ca2+, aggregate, and release small amounts of granule enzymes and b) promoting large degranulation and oxidative burst responses in PMN co-challenged with platelet-activating factor, tumor necrosis factor-alpha, or ATP. Contrastingly, 12(R)-hydroxy-ETE, 12(S)-hydroxy-ETE, and 12-oxoETE induced PMN Ca2+ transients and aggregation [respective potencies (5-hydroxy-ETE = 1) of 0.1, 0.01, and 0.003] but did not effect degranulation, and 15-hydroxy-ETE, 15-oxoETE, and 15-oxo-11-trans-ETE were inactive in all assays. Finally, 5-oxo/hydroxy-ETEs desensitized PMN to themselves but not to 12-oxo/hydroxy-ETEs or leukotriene (LT)B4; 12-oxo/hydroxy-ETEs and LTB4 desensitized PMN to themselves and each other but not to 5-oxo/hydroxy-ETEs; 15-oxo/hydroxy-ETEs did not desensitize PMN; and a LTB4 receptor antagonist blocked responses to LTB4 and 12-oxo/hydroxy-ETEs but not to 5-oxo/hydroxy-ETEs. Thus, 5-oxo/hydroxy-ETEs act by a common, LTB4 receptor-independent mechanism that recognizes 5- but not 12- or 15-oxo/hydroxy-ETEs and prefers oxo over hydroxy residues at C5 whereas 12-oxo/hydroxy-ETEs act via a LTB4 receptor mechanism that recognizes 12- but not 5- or 15-oxo/hydroxy-ETEs and prefers hydroxy over oxo residues at C12.

Arachidonic Acids↗
PubMed1994

Fatty acid composition of low-density lipoprotein influences its susceptibility to autoxidation.

Low-density lipoprotein (LDL) oxidation was studied using copper or the water-soluble initiator azobis(2-amidinopropane) dihydrochloride (ABAP) to catalyze the reaction. These studies were carried out with purified, native LDLs that had a well-defined composition and which contained different concentrations of polyunsaturated fatty acids (PUFA) and alpha-tocopherol. The LDL was obtained from nonhuman primates fed diets enriched in cholesterol and one of four types of fatty acids: saturated (Sat), monounsaturated (Mono), omega-6 (omega-6FA), or omega-3 (omega-3FA) fatty acids. The PUFA concentration of the LDLs depended upon the diet and had the following order: omega-6FA > Sat approximately Mono approximately omega-3FA. Linoleic acid was the predominant PUFA in all of the LDLs. The rates of oxidation were linearly dependent upon the concentration of PUFA. When ABAP was used to initiate oxidation the lag time was linearly related to the amount of alpha-tocopherol. However, with copper catalysis no linear correlation was evident. If the different enrichments were analyzed independently, it was found that copper-catalyzed oxidation of LDLs enriched with omega-6 and omega-3 fatty acids showed a linear correlation between the lag time and the amount of alpha-tocopherol but that LDLs enriched with Sat or Mono fatty acids did not show a correlation. These results demonstrate that the rate of oxidation is dependent upon PUFA concentration and that the ability of alpha-tocopherol to inhibit oxidation depends upon the lipid environment and the mode of initiation.

Amidines↗
PubMed1994

Acinic cell carcinoma of salivary glands. Prognostic relevance of DNA flow cytometry and nucleolar organizer regions.

OBJECTIVE: To test the prognostic ability of flow cytometry and counts of silver-enhanced intranuclear nucleolar organizing regions (AgNORs) in acinic cell carcinoma. DESIGN: Using statistical methods to establish if analysis of DNA content, percentage DNA synthetic (S)-phase, and the AgNOR counts on 45 cases of acinic cell carcinoma with clinical follow-up ranging from 10 to 379 months are predictors of time to recurrence or time to death due to acinic cell carcinomas. MAIN OUTCOME MEASURES: Whether tumors with low AgNOR counts and diploid DNA are favorable acinic cell carcinomas and the converse, aneuploid DNA and elevated AgNOR count, predict unfavorable neoplasms. RESULTS: Tumors with a diploid DNA content had as unfavorable a clinical course as aneuploid acinic cell carcinomas. Similarly, S-phase and AgNOR count results showed considerable overlap when separated into carcinomas with or without local recurrence, metastasis, or death due to tumor. Statistical evaluation also failed to provide predictors of clinical course based on ploidy, percentage S-phase, or AgNOR counts. CONCLUSION: The results, although negative, are important in showing that data on DNA content, cell cycle, and nuclear limits useful in other neoplasms are of limited practical application in establishing predictors of time to recurrence or time to death in acinic cell carcinomas. Solving the enigmas, for clinicians and pathologists, associated with acinic cell carcinomas will require further information about the biology of this neoplasm.

Adult↗
PubMed1994

Relationship of clear cell oncocytoma to mitochondrial-rich (typical) oncocytomas of parotid salivary gland. An ultrastructural study.

Nosologically, the designation clear cell oncocytoma seems illogical and contradictory when the definition of an oncocytoma relies on mitochondrial-rich cells; its relationship to the granular (mitochondrial-rich) oncocytoma needs further clarification. Electron microscopy of six examples of oncocytoma from five patients allowed assessment of cellular features in three clear cell variants, two typical oncocytomas and one with a roughly equal proportion of clear and granular tumor cells. Ultrastructurally, in the clear cell types of oncocytoma, a considerable to extensive portion of the central cytoplasm was occupied by monoparticulate glycogen with margination of mitochondria and other organelles. The degree of extraction of the glycogen during fixation and processing accounts for variation in the extent of the clear cell component as phosphotungstic acid-hematoxylin staining reveals considerable mitochondria in many of the tumor cells in the clear cell variant of oncocytoma. Ultrastructural findings further support an interrelationship between clear and granular cells, as some typical oncocytes are evident in the clear cell variant of oncocytoma, and one oncocytoma in this series had a roughly equal number of glycogen-rich and mitochondrial-rich tumor cells both of which took part in the formation of microlumens. The designation clear cell oncocytoma is appropriate within the oncocytoma subgroup of salivary gland tumors.

Adenoma, Oxyphilic↗
PubMed1994

Lipoperoxidation in plasma and red blood cells of patients undergoing haemodialysis: vitamins A, E, and iron status.

In 14 patients undergoing haemodialysis, lipoperoxidation (LPO) processes were determined in plasma and red blood cells (RBC) before and after a dialysis session by determining (a) the direct substrate, polyunsaturated fatty acids (PUFA); (b) the end product of LPO, malondialdehyde (MDA); and (c) the hydrophobic antioxidant systems, vitamins A and E. In plasma before dialysis, linoleic and arachidonic acid, and the antioxidant vitamin E, were significantly lowered as compared to the healthy controls (p < 0.05). On the contrary, the free MDA level was enhanced (p < 0.05). These results were emphasized by a dialysis session. In RBC of these patients, no difference in linoleic acid, free MDA, or vitamin E level were observed before or after dialysis when compared to controls. However, only vitamin A was significantly higher in haemodialysis patients (before and after dialysis) and in renal failure patients (p < 0.05) than in the healthy control group. The present results suggest that increased RBC vitamin A may offer some degree of protection against oxidative stress in erythrocytes, but not in plasma where LPO is demonstrated.

Adult↗
PubMed1994

Rapid nuclear actions of growth hormone.

The mechanisms by which growth hormone (GH) initiates its effect on growth are largely unknown. In this report we examine the acute actions of GH with a focus on the intracellular signaling pathways leading from the cell-surface GH receptor into the nucleus, and culminating in the activation of specific target genes. We show that in vivo GH treatment leads to the rapid appearance of tyrosine-phosphorylated nuclear proteins and the equally rapid induction of c-fos and insulin-like growth factor I gene transcription. A model is proposed for a GH-activated intracellular signal transduction pathway.

Animals↗
PubMed1994

Rapid activation of rat insulin-like growth factor-I gene transcription by growth hormone reveals no alterations in deoxyribonucleic acid-protein interactions within the major promoter.

Insulin-like growth factor-I (IGF-I) is an important mediator of prenatal and postnatal growth, but little is known about the control of IGF-I gene expression. Previously, we demonstrated that GH rapidly stimulates hepatic IGF-I transcription in vivo in hypophysectomized (hypox) rats. In this study, we show that GH induces IGF-I gene transcription through the major promoter, promoter 1, and identify and characterize DNA-protein interactions throughout the promoter. In vitro deoxyribonuclease-I footprinting was used to analyze 1711 nucleotides of promoter 1 and the entire 328-nucleotide 5'-untranslated region of exon 1, using hepatic nuclear protein extracts from male juvenile hypox rats given a single ip injection of GH or saline 60 min before death. Fourteen DNA-protein binding sites were identified, with 6 located in the highly conserved 5'-untranslated region of exon 1. These latter sites were further characterized for specificity and regulation by GH, using gel mobility shift assays. Two of these DNA-protein interactions were also detected by in vivo dimethylsulfate footprinting. All DNA-protein binding was seen using hepatic nuclear protein extracts from hypox rats and did not change within 15, 30, 60, or 120 min after treatment with GH. Our results thus define a series of constitutive DNA-protein interactions within the major rat IGF-I gene promoter that may be involved in mediating GH-activated nuclear signals to initiate IGF-I transcription.

Amino Acid Sequence↗
PubMed1994

[Demonstration of the anti-lipid peroxidation effect of 3',5,7-trihydroxy-4'-methoxy flavone rutinoside: in vitro study].

Diosmin (DI) 3'5,7-trihydroxy-4'-methoxy flavone rutinoside is a member of the flavonoid family, some of whom have antioxidant or free radical scavenger properties. Paw oedema induced by doxorubicin in rats is reduced by DI as observed by SAUVAIRE et al (1989). This suggests a free radical scavenger activity for DI. In this work we demonstrate that such activity is able to protect isolated human LDL from oxidation in vitro. The study of the electrophoretic mobility of oxidized LDL and of total-MDA values as lipoperoxidation-marker indicates an oxidation inhibiting effect higher than 70% for 0.16 mM DI in LDL mixtures containing 50 mM Cu2+ or 4.3 mU/ml xanthine-oxidase and incubated during 20 and 6 hours respectively. Owing to the high level of the oxidizing conditions and the vitamin E (48 mM), vitamin A (1.4 mM) and beta-carotene (2.4 mM) content of the LDL mixtures, it is concluded that DI is clearly able to complement the antioxidant effect of isoprenoids which are naturally present in LDL mixtures.

Adult↗
PubMed1994

[In vivo study of the antilipoperoxidant effect of 3',5,7-trihydroxy-4'-methoxy flavone 7 rutinoside].

In 3-month-old Wistar rats carrageenan and CCl4 injected intraperitoneally induce an acute phase reaction which is characterized by a marked increase in alpha 1, alpha 2, beta serum globulins. This reaction corresponds to a large increase in these globulins in the first case and a smaller one in the second. A lipoperoxidant effect is demonstrated by the serum lipoprotein mobility as the lipoperoxidation index (in MDA units) or the decrease in serum vitamin A and E concentrations. This effect is also greater in the first case than in the second one. In the same way the lipoperoxidant effect is shown in liver microsomes but with a lower amplitude in the first case than in the second one. The treatment of rats by intraperitoneal injection of diosmine (150 mg/kg per week) during the 8 weeks which precede the injection of carrageenan or CCl4 results in: i) a marked decrease in the acute-phase reaction and a lower one in the lipoperoxidant effect, in serum; ii) a decrease in the CCl4 induced lipoperoxidant effect in liver microsomes. It may be concluded that diosmine, not injected at the same time as carrageenan or CCl4, but during the previous 8 weeks is sufficiently well distributed in the whole body to produce a marked inhibition of the acute phase reaction and a perceptible effect on lipoperoxidation. It may be considered an effective complement to the natural antioxidant defences of the organism (vitamins A and E).

Acute-Phase Reaction↗
PubMed1994

Human growth hormone.

The study of human growth hormone is a little more than 100 years old. Growth hormone, first identified for its dramatic effect on longitudinal growth, is now known to exert generalized effects on protein, lipid, and carbohydrate metabolism. Additional roles for growth hormone in human physiology are likely to be discovered in the areas of sleep research and reproduction. Furthermore, there is some indication that growth hormone also may be involved in the regulation of immune function, mental well-being, and the aging process. Recombinant DNA technology has provided an abundant and safe, albeit expensive, supply of human growth hormone for human use, but the pharmacological properties of growth hormone are poor. Most growth hormone-deficient individuals exhibit a secretory defect rather than a primary defect in growth hormone production, however, and advances in our understanding of the neuroendocrine regulation of growth hormone secretion have established the basis for the use of drugs to stimulate release of endogenously synthesized growth hormone. This promises to be an important area for future drug development.

Adolescent↗
PubMed1993

Synthesis and characterization of a new photocrosslinking CTP analog and its use in photoaffinity labeling E. coli and T7 RNA polymerases.

A new photocrosslinking CTP analog that functioned as a substrate during transcription was synthesized and used to photoaffinity label E. coli and bacteriophage T7 RNA polymerases. This analog, 5-((4-azidophenacyl)thio) cytidine-5'-triphosphate (5-APAS-CTP) contains an aryl azide group approximately 10 A from the nucleotide base and specifically replaced CTP during synthesis of RNA by both polymerases. Analog was placed at the 3' end or internally within RNA. Both polymerases inefficiently incorporated two 5-APAS-CMP molecules sequentially, as was found for the related 5-APAS-UMP. Analog was placed at the 3' end of RNA in transcription complexes paused at the site of Q-modification of E. coli RNA polymerase, downstream of the lambda PR' promoter (+16), a pause that requires specific DNA sequences but no apparent RNA hairpin. Crosslinking was examined in the presence and absence of the NusA protein, which enhances the transcriptional pause at this site and is required for Q modification of the polymerase. Crosslinking of the 3' end of the RNA to NusA was not observed, consistent with our earlier results involving a NusA-enhanced pause site downstream from an RNA hairpin.

Affinity Labels↗
PubMed1993

5-Oxo-eicosatetraenoate, a potent human neutrophil stimulus.

5-Oxo-eicosatetraenoate (5-oxoETE), a newly defined arachidonate metabolite, resembled 5-hydroxyeicosatetraenoate (5-HETE) in stimulating neutrophils to mobilize Ca2+ an in promoting PMN degranulation responses to other agents. It was, however, 10-fold stronger than 5-HETE and, like leukotriene (LT) B4, had intrinsic PMN degranulating effects. Nonetheless, 5-oxoETE and 5-HETE desensitized PMN to themselves or each other but not to LTB4; LTB4 desensitized to itself but not to 5-oxoETE or 5-HETE; and an antagonist blocked LTB4 but not 5-oxoETE or 5-HETE. 5-OxoETE and 5-HETE thus induce diverse PMN responses using a shared, down-regulatable, and receptor-like mechanism that does not involve LTB4 receptors; 5-oxoETE is the preferred natural agonist for this mechanism.

Arachidonic Acids↗
PubMed1993

Effects of the excitatory amino acid receptor antagonists kynurenate and indole-2-carboxylic acid on behavioral and neurochemical outcome following experimental brain injury.

The overactivation of the NMDA receptor is thought to be a major contributor to the pathophysiologic sequelae of traumatic brain injury (TBI), which commonly includes memory dysfunction. Uniquely, potentiation of the NMDA receptor is dependent on the binding of glycine to a distinct site on the receptor. Despite the potential role of the NMDA receptor in the development of post-TBI cognitive deficits, no studies to date have evaluated the efficacy of NMDA receptor antagonists on memory dysfunction after brain injury. Moreover, glycine site antagonists have not been employed as potential therapeutic agents in models of TBI. In the present study, we evaluated the effects of two glycine site antagonists, kynurenate (KYNA) and indole-2-carboxylic acid (I2CA), on memory and motor dysfunction, cerebral edema formation, and changes in regional total tissue brain [Na], [K], [Ca], [Mg], and [Zn], following lateral fluid-percussion brain injury in the rat. We found that both KYNA (300 mg/kg) and I2CA (50 mg/kg and 20 mg/kg) administered 15 min postinjury dramatically attenuated trauma-induced cognitive dysfunction (p < 0.01). In addition, both compounds improved neurologic motor deficits measured at 2 weeks following injury (p < 0.05). KYNA (300 mg/kg) reduced edema in the cortex, hippocampus, and thalamus, while I2CA (20 mg/kg) reduced edema formation only in the thalamus. Differential effects of KYNA and I2CA on cation concentrations were also noted. KYNA attenuated the postinjury increase in regional tissue [Ca]; however, it had little effect on other cation concentrations. I2CA reversed the postinjury regional increases in [Na] and decreases in [K], [Mg], and [Zn], but had little effect on [Ca] changes. These results indicate that KYNA and I2CA may have differential, but beneficial effects on both behavioral and neurochemical sequelae of TBI.

Animals↗
PubMed1992

Superoxide generation by the human polymorphonuclear leukocyte in response to latex beads.

In this study, superoxide formation was not immediately detected when polymorphonuclear leukocytes were treated with linoleyl alcohol-coated, carboxy-modified latex beads. However, all other measures of neutrophil activation were present. Superoxide was not detected until 30 min after the initial exposure to beads. However, an O2(-)-producing, NADPH-dependent oxidase is active 15 min after exposure. When polymorphonuclear leukocytes are pretreated with cytochalasin B, superoxide production is detected immediately after exposure to the beads. Superoxide secretion after treatment with linoleyl alcohol-coated latex beads is compared with the response to other latex beads. The results imply that neutrophils form a phagolysosome around linoleyl alcohol-coated latex beads that is tightly sealed and does not allow superoxide to escape into the medium where it could be detected by the reduction of ferricytochrome c.

Cells, Cultured↗
PubMed1992

Urate causes the human polymorphonuclear leukocyte to secrete superoxide.

The human polymorphonuclear leukocyte generates O2-. and H2O2 when it is treated with uric acid. A transition metal catalyzed reaction between O2-. and H2O2 can give the hydroxyl radical and myeloperoxidase forms hypochlorous acid from H2O2 and chloride. Therefore, the uric acid-induced secretion of oxidants may be responsible for a large part of the inflammation associated with gout.

Free Radicals↗