Search PubMed⌕ Search

Biomedical subjects

A Flynn

Publications and source records attributed to A Flynn.

At least 73 records · Page 4Linked to original sources
PubMed1997

Continuous, pulsed or single acute irradiation of a transplanted rodent tumour model.

BACKGROUND: Recent advances in remote afterloading pulsed mode brachytherapy have provided a much needed tool for the radiation oncologist. It has the versatility of optimised physical dose distribution along with improved staff radiation protection and patient nursing. PURPOSE: This preliminary study was designed to explore the radiobiological equivalence between conventional continuous low dose rate tumour irradiation (CLDR) and the new technique of pulsed dose irradiation (PDR). MATERIALS AND METHODS: Subcutaneous isogenic sarcomas transplanted in female John's Strain Wistar rats were irradiated locally with acute, pulsed or continuous interstitial low dose-rate exposures at 9-11 mm mean diameter. RESULTS: As expected, single acute doses (5-40 Gy) were more effective (P < 0.01) in achieving tumour growth delay (1.4 days/Gy) than CLDR exposure (4-51 Gy) over 24-48 h (0.93 days/Gy). However, PDR treatment (8 hourly fractions/day) at high dose-rate (8-48Gy) over 8-72 h was significantly (P = 0.01) more effective (1.66 days/Gy) than CLDR but not acute exposures. CONCLUSIONS: These data suggest that, clinically a significantly improved therapeutic ratio may also be achievable with pulsed high dose rate brachytherapy, and that further radiobiological studies with in-vivo tumour models are needed.

Animals↗
PubMed1996

Insulin-stimulated phosphorylation of initiation factor 4E is mediated by the MAP kinase pathway.

The cap-binding initiation factor 4E (eIF4E) is regulated by phosphorylation and by the inhibitory binding protein 4E-BP1. Here we show that insulin-induced phosphorylation of eIF4E is not significantly affected by rapamycin, but is sensitive to wortmannin, which inhibits phosphatidylinositol 3'-kinase and blocks the activation of MAP kinase. Since PD098059, an inhibitor of MAP kinase activation, also blocks insulin-induced phosphorylation of eIF4E, the MAP kinase pathway seems to mediate this effect. Phosphorylated eIF4E can still bind to 4E-BP1. These data illustrate that (i) distinct signalling pathways mediate the phosphorylation of eIF4E and 4E-BP1 and (ii) phosphorylation of eIF4E, unlike that of 4E-BP1, does not lead directly to the release of 4E-BP1.

Androstadienes↗
PubMed1996

Both rapamycin-sensitive and -insensitive pathways are involved in the phosphorylation of the initiation factor-4E-binding protein (4E-BP1) in response to insulin in rat epididymal fat-cells.

There is mounting evidence that in fat and other insulin-sensitive cells activation of protein synthesis may involve the dissociation of a protein (4E-BP1) from eukaryotic initiation factor (eIF)-4E thus allowing formation of the eIF-4F complex. This study compares the effects of insulin and epidermal growth factor (EGF) on the phosphorylation of 4E-BP1 in fat-cells (followed by gel-shift assays and incorporation of 32P) and on its association with eIF-4E. Several lines of evidence suggest that mitogenactivated protein kinase (MAP kinase) is not involved in these effects of insulin. Insulin causes much more extensive phosphorylation and dissociation of 4E-BP1 from eIF-4E than EGF, although EGF activates MAP kinase to a much greater extent than insulin. Moreover, MAP kinase does not phosphorylate 4E-BP1 when it is complexed with eIF-4E. In contrast, insulin activates the 40S ribosomal protein S6 kinase (p70S6K) 18-fold compared with a 2-fold activation by EGF, and the time course of this activation is similar to the phosphorylation and dissociation of 4E-BP1. Rapamycin, a specific inhibitor of the activation of this latter kinase, inhibits dissociation of 4E-BP1 from eIF-4E in cells incubated with insulin but reveals a phosphorylated from of 4E-BP1 which remains bound to eIF-4E. It is concluded that in rat epididymal fat-cells, the effects of insulin on 4E-BP1 involves multiple phosphorylation events. One phosphorylation event is rapamycin-insensitive, occurs only on bound 4E-BP1 and does not initiate dissociation. The second event does result in dissociation and is blocked by rapamycin, suggesting that the p70S6K signalling pathway is involved: p70S6K itself is probably not involved directly as this kinase does not phosphorylate 4E-BP1 in vitro.

Adipocytes↗
PubMed1996

Insulin and phorbol ester stimulate initiation factor eIF-4E phosphorylation by distinct pathways in Chinese hamster ovary cells overexpressing the insulin receptor.

We have developed a one-dimensional isoelectric focusing technique to measure changes in the steady-state phosphorylation of the cap-binding initiation factor, eIF-4E. We have used a Chinese hamster ovary cell line transfected with the human insulin receptor (CHO.T cells) to study the regulation of eIF-4E phosphorylation by insulin and other stimuli. Exposure of CHO.T cells to insulin, phorbol ester or serum resulted in a rapid increase (up to twofold) in eIF-4E phosphorylation. As a control, we have also performed experiments with the parental cell line, CHO.K1 cells, in which both serum and phorbol ester, but not nanomolar concentrations of insulin, produce similar changes in eIF-4E phosphorylation. We have used two complementary approaches to study the role of protein kinase C (PKC) in these responses: a highly specific inhibitor of PKC and down-regulation of PKC by prior treatment of the cells with phorbol ester. In CHO.T cells, both approaches indicate that PKC is required for the response to phorbol ester but that insulin and serum each increase eIF-4E phosphorylation by a mechanism(s) independent of this protein kinase. Similarly, PKC is necessary for the effects of phorbol ester, but not of serum, on eIF-4E phosphorylation in CHO.K1 cells. These data indicate that multiple signal transduction mechanisms are involved in the modulation of eIF-4E phosphorylation and the implications of these findings are discussed.

Amino Acid Sequence↗
PubMed1996

Glucose stimulates the activity of the guanine nucleotide-exchange factor eIF-2B in isolated rat islets of Langerhans.

Over short time periods glucose controls insulin biosynthesis predominantly through effects on preexisting mRNA. However, the mechanisms underlying the translational control of insulin synthesis are unknown. The present study was carried out to determine the effect of glucose on the activity and/or phosphorylation status of eukaryotic initiation and elongation factors in islets. Glucose was found to increase the activity of the guanine nucleotide-exchange factor eIF-2B over a rapid time course (within 15 min) and over the same range of glucose concentrations as those that stimulate insulin synthesis (3-20 mM). A nonmetabolizable analogue of glucose (mannoheptulose), which does not stimulate insulin synthesis, failed to activate eIF-2B. The best characterized mechanism for modulating eIF-2B activity involves changes in the phosphorylation of the alpha-subunit of its substrate eIF-2. However, in islets, no change in eIF-2 alpha phosphorylation was seen under conditions where eIF-2B activity was increased, implying that glucose regulates eIF-2B via an alternative pathway. Glucose also did not affect the phosphorylation states of three other regulatory translation factors. These are the cap-binding factor eIF-4E, 4E-binding protein-1, and elongation factor eEF-2, which do not therefore seem likely to be involved modulating the translation of the preproinsulin mRNA under these conditions.

Animals↗
PubMed1996

Replacement of hairpin and loop implants by optimised straight line sources.

Remote after loading is desirable for all forms of brachytherapy but is difficult in the oral cavity because the tight curvature at the top of implanted loops impedes passage of the source. For this reason we investigated differential loading of straight catheters to simulate a conventional loop or hairpin. Using a pulsed brachytherapy remote after loader the top four dwell positions of straight catheters were given two to four times the dwell time of other source positions. This raised the reference isodose to cover the surface mucosa without significantly changing either the total volume treated or the volume receiving > 150% of reference dose when compared with conventional loop implants of equivalent source length and separation. This optimised straight line implant should therefore be amenable to remote afterloading and have similar dose/volume characteristics to loops.

Brachytherapy↗
PubMed1996

Effect of dietary calcium intake and meal calcium content on calcium absorption in the rat.

Fifty-four male and forty-five female 7-week-old rats, Wistar strain, average weights 190 g and 140 g respectively, were randomized by weight into three groups of eighteen rats each (males) and three groups of fifteen rats each (females) and fed on a semi-purified diet containing (per kg) 2 (low), 5 (normal) or 20 g (high) Ca as CaCO3 for 2 weeks. Each group was then further randomized into three groups of six rats each (males) and five rats each (females) and given a meal (10 g of the same diet) containing either 2, 5 or 20 g Ca as 47CaCO3. 47Ca was determined in quantitative daily collections of faeces over 7 d and fractional absorption of 47Ca estimated by extrapolating the linear portion (days 3-7) of the plot of log 47Ca retention v. time back to the time of isotope administration. Absorption of meal Ca was higher in males than in females and was affected similarly in males and females by previous dietary Ca intake and meal Ca content. Fractional absorption of meal Ca decreased with increasing previous dietary Ca intake and with increasing meal Ca content, and the combined effect of these two variables caused fractional Ca absorption to vary from 11-89%. Absolute absorption of meal Ca decreased with increasing previous dietary Ca intake and increased with increasing meal Ca content. The influence on Ca absorption of variations in meal Ca content (load effect) was greater than that of variations in previous dietary Ca intake (adaptive effect). These results show that previous dietary Ca intake and meal Ca content are both major determinants of Ca absorption from meals in intact rats fed in the normal way and that the rat responds to these factors in a manner similar to that reported for humans. This study provides further evidence of similarities between rats and humans in dietary Ca absorption.

Animal Feed↗
PubMed1996

Microsurgical correction of bilateral facial contour deformities.

Bilateral facial contour deformities are difficult reconstructive problems to correct. This paper summarizes our experience with five adult patients who were operated on for the correction of bilateral contour deformities. The etiologies of the contour deformities were one case of systemic lupus erythematosus, three cases of hemangioma/lymphangiomas, and one case of systemic lipodystrophy. Patient ages at the time of surgical correction ranged from 19 to 65 years. In two patients, staged microvascular free-tissue transfers were performed. In three patients, a single free flap was used for reconstruction. Seven microvascular free flaps were used to reconstruct facial contour: three superficial inferior epigastric flaps and four inframammary extended circumflex scapular flaps. In three patients, recipient sites were dissected by means of a limited preauricular incision, and the superficial temporal artery and vein were utilized as recipient vessels. The two remaining patients underwent free-tissue transfer to branches of the external carotid system. There were no flap losses in this series. Follow-up ranged from 1 to 4 years. A stable restoration of facial contour was achieved in all five patients. Microsurgical correction of soft-tissue contour problems is possible in bilateral deformities, and superior results are easier to achieve than in unilateral facial contour deformities. Either single or staged reconstructions can restore facial contour depending on the distribution of soft-tissue requirements.

Adult↗
PubMed1996

The role of eIF4 in cell proliferation.

Translational control has a key role in regulating cell growth. The mRNA binding translation factors (eIF4s) selectively modulate the translation of different mRNAs based on their differing properties, especially the extent of inhibitory secondary structure in their 5' untranslated regions. The mRNA 5' cap is recognized by eIF4E, which can then recruit other translation factors including the helicase cIF4A. Overexpression of eIF4E leads to cell transformation or disordered growth. It also enhances the translation of certain mRNAs, including that for cyclin D1. Decreased expression of eIF4E leads to reduced rates of cell growth. Transformed cells express elevated levels of eIF4E. eIF4E is subject to phosphorylation, which is increased by agents that stimulate translation. However, the role of eIF4E phosphorylation and the signalling pathways through which it is controlled remain to be elucidated. Oncogenic ras leads to increased phosphorylation of eIF4E, and the ability of ras to transform cells is diminished when eIF4E expression is decreased, implying that eIF4E is a key mediator of ras mediated transformation. The recent discovery of inhibitors of eIF4E (eIF4E binding proteins, 4EBP1/2) adds another regulatory component to the system. Although a role for 4EBP1 in cell transformation (or rather in impeding it) has yet to be established, this is an exciting and likely possibility. It should also be noted that 4EBP1 seems to be under the control of the p70S6K pathway, which has a role in cell cycle progression.

Animals↗
PubMed1995

Serine 209, not serine 53, is the major site of phosphorylation in initiation factor eIF-4E in serum-treated Chinese hamster ovary cells.

Ser-53 has previously been considered the major phosphorylation site in eukaryotic initiation factor (eIF)-4E, and this appeared to be supported by studies using a S53A mutant. Recently, however, several lines of evidence have indicated that Ser-53 might not be the true phosphorylation site. This prompted us to re-examine the phosphorylation site in eIF-4E using factor purified from 32P-labeled, serum-treated Chinese hamster ovary cells. Isoelectric focusing and phosphoamino acid analysis indicated the existence of a single phosphorylated serine. Edman degradation of the major radiolabeled tryptic product from 32P-labeled eIF-4E showed that the phosphorylated site was positioned three residues from the N terminus of this peptide. There are three serines in the sequence of eIF-4E that are three residues away from a tryptic cleavage site (i.e. lysine or arginine). 32P-Labeled eIF-4E was digested with trypsin, Lys-C, or trypsin followed by Glu-C and subjected to two-dimensional mapping; the data obtained eliminated two of these potential sites, leaving Ser-209. Comigration of the synthetic peptide SGS(P)209TTK with the radiolabeled tryptic product on (i) reverse-phase chromatography and (ii) two-dimensional mapping at different pH values confirmed that Ser-209 is the major phosphorylation site in eIF-4E in serum-stimulated Chinese hamster ovary cells.

Amino Acid Sequence↗
PubMed1995

Pseudoseizures.

Explore the source record for details and available documents.

Conversion Disorder↗
PubMed1995

Susceptibility of beta-lactoglobulin and sodium caseinate to proteolysis by pepsin and trypsin.

The susceptibility of beta-LG and sodium caseinate to proteolysis by pepsin and trypsin was investigated using SDS or urea-PAGE. The effects were studied of heat, urea, and 2-mercaptoethanol on proteolysis. Native beta-LG was resistant to hydrolysis by pepsin or trypsin because of its compact globular structure. Heat treatment of beta-LG solutions at 90 to 100 degrees C for 5 or 10 min caused changes in the structure or conformation of the protein that rendered it accessible to pepsin and enhanced the extent of proteolysis by trypsin. The susceptibility of beta-LG to proteolysis by pepsin was markedly increased in the presence of urea (3 to 6 M), and the effect was reversible after removal of urea by dialysis. Proteolysis by trypsin was also increased by the presence of 2% 2-mercaptoethanol. Sodium caseinate was very accessible to pepsin without pretreatment and was extensively hydrolyzed at pH 1 to 5 in the presence of 5 M urea (which prevented the protein from precipitation in the isoelectric region); optimal pH was about 2. The activity of pepsin on sodium caseinate at pH 2 was not significantly affected by urea concentration up to about 8 M. The results indicated that the changes in conformation and structure of beta-LG that were induced by heating, reduction, or urea rendered the protein susceptible to peptic hydrolysis.

Animals↗
PubMed1994

The RNA-binding properties of protein synthesis initiation factor eIF-2.

Protein synthesis initiation factor eIF-2 bound ATP in the presence or absence of Mg2+ ions. ATP impaired the binding of GTP or GDP to eIF-2. However, excess GTP did not significantly decrease the binding of ATP to eIF-2, suggesting eIF-2 has distinct ATP and GTP binding sites. Highly purified eIF-2 can bind mRNA, and this did not require the mRNA to be capped. mRNA binding was saturable, and maximal binding corresponded to about 0.4 mol mRNA bound per mol eIF-2. GTP, and, at lower concentrations, GDP, inhibited the binding of mRNA to eIF-2. In addition, ATP and other nucleoside triphosphates decreased mRNA binding. The implications of these findings for the structure and function of eIF-2 are discussed. Preparations of eIF-2 deficient in the beta-subunit showed reduced ability to bind mRNA, suggesting that while it is not essential for mRNA binding, this subunit is involved in the interaction. Consistent with this is the observation that ultraviolet crosslinking of mRNA to eIF-2 resulted primarily in labelling of the beta-subunit. Subsequent analysis revealed that mRNA was cross-linked to the C-terminal region of eIF-2b which contains a putative Zn-finger structure.

Adenosine Triphosphate↗
PubMed1994

Consumption of thermally-oxidized sunflower oil by chicks reduces alpha-tocopherol status and increases susceptibility of tissues to lipid oxidation.

The effect of heated sunflower oil consumption on alpha-tocopherol status, fatty acid composition and oxidative stability of chicken tissues was investigated. Chicks were fed on diets containing (g/kg): fresh sunflower oil (FSO) 40, heated sunflower oil (HSO) 40 or heated sunflower oil (40) supplemented with alpha-tocopheryl acetate (HSE) to a similar alpha-tocopherol concentration as the FSO diet. Concentrations of alpha-tocopherol in tissues of chicks fed on HSO and HSE were significantly lower than those of chicks fed on FSO. Significant correlations were observed between plasma alpha-tocopherol concentration and the alpha-tocopherol concentrations of other tissues (r > or = 0.67, P < 0.005) and between log plasma alpha-tocopherol and plasma thiobarbituric acid-reacting substances (TBARS) concentrations (r -0.851, P < 0.001). The concentrations of TBARS in tissues of chicks fed on the various diets were generally very similar before stimulation of peroxidation with Fe-ascorbate. Susceptibility of tissues to Fe-ascorbate-induced lipid peroxidation was increased by feeding HSO. Supplementation with alpha-tocopheryl acetate reduced susceptibility to lipid oxidation to varying degrees, depending on the tissue. The results suggest that chronic ingestion of oxidized lipids may compromise free-radical-scavenging activity in vivo by depleting alpha-tocopherol in the gastrointestinal tract, or possibly in plasma and other tissues.

Animals↗
PubMed1993

The role of the beta-subunit of initiation factor eIF-2 in initiation complex formation.

The functional properties of preparations of protein synthesis initiation factor eIF-2 which lack the beta-subunit (as confirmed immunologically) were compared with those of the heterotrimeric factor. The former can bind guanine nucleotides but not initiator tRNA, and also exhibits a substantially reduced rate of initiation factor eIF-2B-mediated GDP/GTP-exchange.

Animals↗
PubMed1993

Influence of heated vegetable oils and alpha-tocopheryl acetate supplementation on alpha-tocopherol, fatty acids and lipid peroxidation in chicken muscle.

1. Chicks were fed on diets containing fresh, heated or alpha-tocopheryl acetate-supplemented heated vegetable oils. The effects on alpha-tocopherol status, and on the fatty acid composition and oxidative stability of thigh and breast muscle were determined. 2. Plasma alpha-tocopherol was significantly correlated with alpha-tocopherol concentrations in thigh and breast muscle. 3. The fatty acid profiles of muscle lipids reflected dietary fatty acid composition. 4. The consumption of heated sunflower and linseed oils reduced alpha-tocopherol status, altered fatty acid composition of muscle lipids and increased susceptibility of muscle to lipid oxidation. 5. Supplementation of diets containing heated oils with alpha-tocopheryl acetate resulted in some alleviation of these effects. 6. The results indicate that caution should be exercised in the use of thermally oxidised oils in poultry diets if undesirable changes in composition and oxidative stability of carcase lipids are to be avoided.

Animals↗