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Biomedical subjects

S Lemaire

Publications and source records attributed to S Lemaire.

At least 19 recordsLinked to original sources
PubMed2004

[Fluoroscopy guided L5-S1 transforaminal injection as a treatement for s1 radiculopathy].

OBJECTIVE: To assess the efficacy of fluoroscopy guided L5-S1 transforaminal steroid injections for the treatment of S1 radiculopathy secondary to intervertebral disk disorder. METHOD: 41 patients were included: prospective study (20 patients) and retrospective study (21 patients). All patients suffered from S1 radiculopathy secondary to nerve root compression by intervertebral disk material, as demonstrated by lumbar spine CT. The patients underwent 2 fluoroscopy guided L5-S1 transforaminal injections of steroid (Hydrocotancyl 125 mg), at 8 days interval. RESULTS: 60% of patients showed significant improvement of their painful radiculopathy at day 8 (n:41), 60-67% at day 30 (n:41) and 67% at day 90 (n:18). CONCLUSION: Fluoroscopy guided L5-S1 transforaminal injection showed good efficacy in the treatment of S1 radiculopathy.

Adrenal Cortex Hormones↗
PubMed2003

Target-controlled infusion of propofol and remifentanil in cardiac anaesthesia: influence of age on predicted effect-site concentrations.

BACKGROUND: Propofol-anaesthesia administrated via target-controlled infusion (TCI) has been proposed for cardiac surgery. Age-related changes in pharmacology explain why propofol dose requirement is reduced in elderly patients. However, the Marsh pharmacokinetic model incorporated in the Diprifusor propofol device does not take age into account as a covariable. In the absence of depth of anaesthesia monitoring, this limitation could cause adverse cardiovascular effects resulting from propofol overdose in older patients. We assessed the influence of age on effect-site propofol concentrations predicted by the Diprifusor and titrated to the bispectral index score (BIS) during cardiac anaesthesia. METHODS: Forty-five patients received propofol by Diprifusor and remifentanil by software including Minto model. Propofol and remifentanil effect-site concentrations were adapted to BIS (40-60) and haemodynamic profile, respectively. The influence of age on effect-site concentrations was assessed by dividing patients into two groups: young (<65 yr) and elderly (> or =65 yr). RESULTS: For a similar depth of anaesthesia, effect-site propofol concentrations were significantly lower in elderly patients at the different stages of cardiac surgery. The mean dose of propofol required to perform tracheal intubation was significantly lower in elderly patients. However, the overall doses of propofol were comparable in both groups. Neither effect-site remifentanil concentrations nor overall doses of remifentanil were significantly different between the two groups. CONCLUSIONS: In cardiac anaesthesia, target concentrations of propofol must be reduced in elderly patients. Although this probably contributes to improving intraoperative haemodynamic stability, the absence of decrease in overall dose requirement of propofol suggests that this adjustment is relatively moderate.

Adolescent↗
PubMed2002

Early evaluation of dressage ability in different breeds.

Dressage is one of the Olympic equestrian sports practiced in several countries using different horse breeds. Specific characteristics of the walk, trot and canter are required for dressage. It has been assumed that some of these traits could be selected for genetically and contribute to dressage performance. The purpose of this study was to compare the walk, trot and conformation characteristics in young horses of different breeds used for dressage. A total of 142 horses age 3 years were classified into 3 groups of breeds (German, French and Spanish saddle horses) and tested using the same procedure. The skeletal conformation measurements were made by image analysis. Gait variables of the walk and trot were measured by the accelerometric gait analysis system Equimetrix. Discriminant analysis could explain the variability between the groups by taking into account the walk (P<0.0003), trot (P<0.0001) and conformation variables (P<0.0001). Many gait and conformation variables were significantly different between the breeds. In summary, the German horses had gait characteristics more adapted for dressage competition, and the results of this group could be used as a reference for early evaluation in dressage. Purebred Spanish horses could be considered as a reference for collected gaits used for farm work and old academic dressage. The gait and conformation tests could be applied in a breeding or crossing plan to detect more accurately young horses with good dressage ability.

Animals↗
PubMed2002

The effect of reins on kinetic variables of locomotion.

The equestrian literature and reports of experienced riders indicated that attitude of a horse's head and neck is related to the equine locomotion characteristics. The purpose of this study was to quantify the effects of 3 types of reins on locomotion at walk and trot. Eight saddle horses were tested in an automatic walker without reins, with rubber bands, Chambons and Back lift, using the same procedure. The rubber bands closed the head-neck angle, whereas the Chambons and the Back lift opened it. The gait characteristics were measured with the Equimetrix gait analysis system. Two accelerometric devices were fixed on the sternum and sacrum to measure the kinetic characteristics of the fore- and hindlimbs at walk and trot. Analysis of variance was used to test the effects of reins on the fore- and hindlimbs. All 3 types of reins significantly (P<0.05) modified walk and trot, with increased forelimb propulsion at the trot (P<0.05). Chambons increased the dorsoventral activity of the hindlimbs at the trot and hindlimb propulsion at the walk (P<0.05). Back lift increased forelimb dorsoventral activity both at the trot and walk (P<0.05). It was concluded that the Chambons increased the activity of the hindlimbs while Back lift increased that of the forelimbs. The use of reins modified the head placement and the fore- and hindlimbs activity. The specific effect of each type of rein can be used for training, rehabilitation or management of locomotor's injury.

Acceleration↗
PubMed2002

Quality of life in multiple sclerosis: influence of interferon-beta1 a (Avonex) treatment.

BACKGROUND: Numerous data argue for initiating treatment with interferon-beta (IFN-beta) at an early stage in multiple sderosis (MS). The consequences of its use may negatively influence the MS patient's quality of life (QoL). OBJECTIVE: To evaluate the QoL of MS patients before and after a one-year period of treatment with IFN-beta1a (Avonex). PATIENTS AND METHODS: QoL was assessed using the SF-36 in 121 relapsing-remitting MS patients. We compared QoL before and after treatment and with data from a normal population. We also studied the possible influence of disease progression on the SF-36 scores. RESULTS: One hundred six patients completed the study (87%). Compared to a normal population, patients were, at baseline, worse off for all QoL scales, varying from a minimum decrease of 0.73 SD in mental health, to a maximum decrease of 1.55 SD in general health. After treatment, we found no significant changes in any of the QoL scores, except for physical function, where we noted a slight but significant decrease (p = 0.03). Furthermore, there was no significant change either in the physical component summary (PCS) or mental component summary (MCS). The 'reported health transition' item was significantly improved compared to baseline (p = 0.001). At indusion, significant correlations were found between EDSS scores and scores of physical function (p < 0.001), role - physical (p < 0.01), general health and social function (both p < 0.01), and with the PCS (p < 0.01). Patients with dinical relapses and/or disability progression had a more significant decease in physical function (p < 0.05) and also in social function (p < 0.05). CONCLUSION: The QoL, assessed by the SF-36 scale, is correlated with disability in MS. IFN-beta1a treatment (Avonex) has no negative effect on MS patient's QoL.

Adjuvants, Immunologic↗
PubMed2001

Ceramide generation occurring during 7beta-hydroxycholesterol- and 7-ketocholesterol-induced apoptosis is caspase independent and is not required to trigger cell death.

Biological activities of oxysterols seem tightly regulated. Therefore, the ability to induce cell death of structurally related oxysterols, such as those oxidized at C7(7alpha-, 7beta-hydroxycholesterol, and 7-ketocholesterol), was investigated on U937 cells at different times of treatment in a concentration range of 5-80 microg/ml. Whereas all oxysterols accumulate inside the cells, strong inhibition of cell growth and increased permeability to propidium iodide were observed only with 7beta-hydroxycholesterol and 7-ketocholesterol, which trigger an apoptotic process characterized by the occurrence of cells with fragmented and/or condensed nuclei, and by various cellular dysfunctions: loss of mitochondrial transmembrane potential, cytosolic release of cytochrome c, activation of caspase-9 and -3 with subsequent enhanced activity of caspase-3, degradation of poly(ADP-ribose) polymerase, and increased accumulation of cellular C16 : 0 and C24 : 1 ceramide species. This ceramide generation is not attributed to caspase activation since inhibition of 7beta-hydroxycholesterol- and 7-ketocholesterol-induced apoptosis by Z-VAD-fmk (100 microM), a broad spectrum caspase inhibitor, did not reduce C16 : 0 and C24 : 1 ceramide species accumulation. Conversely, when U937 cells were treated with 7beta-hydroxycholesterol and 7-ketocholesterol in the presence of fumonisin B1 (100 microM), a specific inhibitor of ceramide synthase, C16 : 0 and C24 : 1 ceramide species production was completely abrogated whereas apoptosis was not prevented. Noteworthy, 7alpha-hydroxycholesterol induced only a slight inhibition of cell growth. Collectively, these results are consistent with the notion that the alpha or beta hydroxyl radical position of oxysterols oxidized at C7 plays a key role in the induction of the apoptotic process. In addition, our findings demonstrate that 7beta-hydroxycholesterol- and 7-ketocholesterol-induced apoptosis involve the mitochondrial signal transduction pathway and they suggest that C16 : 0 and C24 : 1 ceramide species generated through ceramide synthase play a minor role in the commitment of 7beta-hydroxycholesterol- and 7-ketocholesterol-induced cell death.

Amino Acid Chloromethyl Ketones↗
PubMed2000

Phorbol ester treatment of HL 60 leukemia cells results in increase of beta-(1 --> 4)-galactosyltransferase.

We previously showed that HL 60 leukemia cells exhibit various changes in their cellular glycans after phorbol 12-myristate 13-acetate (PMA) treatment. These changes could originate largely from changes in one or several glycosyltransferases. In this report, we show using enzymatic measures, fluorescence microscopy, immunoblotting and Northern blot that beta-(1 --> 4)-galactosyltransferase I (GalT I) activity was higher (> x 2) in PMA-treated compared with untreated HL 60 cells. Immunoblotting showed an increased intensity of the GalT I band at 49 kDa and Northern blot a weak increase of the GalT I transcript band, after PMA treatment. Moreover, Northern blot performed after actinomycin-D treatment of the cells, which inhibits transcription, suggests that the observed increase of GalT I expression could originate, in part, from increase of the stability of GalT I transcripts.

Blotting, Northern↗
PubMed2000

Interactions of dynorphin A-(1-13) and nociceptin with cardiac D2 binding sites: inhibition of ischemia-evoked release of noradrenaline from synaptosomal-mitochondrial fractions.

The effect of dynorphin A (Dyn A)-related peptides and nociceptin on the binding of the D2 receptor antagonist, [(3)H]raclopride, was examined in membrane preparations of rat heart. Non-linear regression saturation binding analysis of [(3)H]raclopride binding revealed the presence of a single high-affinity binding site with a K(d)of 4.1 n M and a B(max)of 220 fmol/mg protein. The D2 stereospecificity of [(3)H]raclopride binding was demonstrated by competition experiments using two enantiomer pairs of antagonists. (+)-Butaclamol (IC(50): 8.0 n M) and (-)-sulpiride (IC(50): 112.3 n M) were 27 000 and 24 times more potent than (-)-butaclamol (IC(50): >100 microm) and (+)-sulpiride (IC(50): 2666 n M), respectively. Nociceptin and Dyn A-(1-13) were also potent inhibitors of the binding of [3H]raclopride with shallow inhibition curves that fitted best with two sites model. Their order of potency on the low affinity site [alpha -Neo-endorphin>nociceptin>Dyn A-(2-13)>Dyn A-(1-13)>Dyn B>Dyn A-(6-10)] correlated well with their ability to inhibit the binding of [3H]nociceptin (r=0.82). The indirect nature of the inhibitory effects of the peptides on the D2 receptor was demonstrated by their inability to inhibit [(3)H]raclopride binding to a membrane preparation (Sf9 cells transfected with the human D2(long)receptor) that does not contain the ORL(1)receptor and the lack of effect of raclopride (0.1 n M-10 microm) on both [(3)H]nociceptin and [(3)H]Dyn A-(1-13) binding. Isolated cardiac mitochondrial-synaptosomal fractions submitted to ischemic conditions (1 m M iodoacetate +2 m M NaCN, 5 min at 37 degrees C) released 10.9% of their content in preloaded [(3)H]noradrenaline ([(3)H]NA). Dyn A-(1-13) (10 microm), nociceptin (10 microm) and the selective D2 receptor agonist, quinpirole (10 microm) were potent blockers of the release of [(3)H]NA evoked by the ischemic conditions. The inhibitory effect of Dyn A-(1-13), nociceptin and quinpirole were antagonized by the selective D2 receptor antagonist, raclopride (10 microm); whereas naloxone, at a concentration (1 microm) known to affect the ORL(1)receptor, blocked the effects of the peptides but not those of quinpirole. The results demonstrate the presence of D2 receptors in rat heart and suggest that Dyn A-(1-13) and nociceptin modulate ischemia-induced NA release by a mechanism that involves the participation of both ORL(1)and D2 receptors.

Animals↗
PubMed2000

Non-opioid antinociceptive effects of supraspinal histogranin and related peptides: possible involvement of central dopamine D(2) receptor.

The antinociceptive effects of intracerebroventricular (ICV) administration of histogranin (HN) and related peptides were assessed in the mouse writhing and tail-flick assays. In the writhing test, the peptides displayed dose-dependent analgesic effects with an AD(50) of 23.9 nmol/mouse for HN and the following order for other peptides: HN-(7-15)<histone H4-(86-100) approximately HN approximately HN-(7-10)<[Ser(1)]HN<osteogenic growth peptide (OGP) approximately HN-(1-10). HN-(6-9) and HN-(8-10) did not show any significant analgesic activity at 50 nmol/mouse. The importance of the C- and N-terminal amino acids in the analgesic activity of the peptides was demonstrated by the prolonged effects of HN and [Ser(1)]HN ( approximately 30 min) compared with those of HN fragments (HN-(7-15), HN-(1-10) and HN-(7-10): 5-10 min). The analgesic activity of [Ser(1)]HN (50 nmol/mouse) was not affected by the coadministration of opioid (naloxone, 1 nmol/mouse), NMDA (CPP, 0.3 and MK-801, 0.3 nmol/mouse) and D(1) (SCH-23390, 0.5 nmol/mouse) receptor antagonists, but it was significantly antagonized by the coinjection of the D(2) receptor antagonist raclopride (0.5 nmol/mouse). In the mouse tail-flick assay, HN and related peptides (50 nmol/mouse) also showed significant analgesic activity (15-35% MPE). The analgesic effect of [Ser(1)]HN was dose-dependent and, at 75 nmol/mouse, lasted for up to 45 min, and was partially blocked by the coadministration of raclopride (1 nmol/mouse), but not naloxone (2 nmol/mouse). In the mouse rotarod assay, relative high doses (75-100 nmol/mouse) of HN and related peptides did not significantly affect motor coordination. These results indicate that supraspinal administration of HN and related peptides induce significant non-opioid analgesic effects devoid of motor activity by a mechanism that involves the participation of central dopamine D(2) receptors.

Amino Acid Sequence↗
PubMed1999

The internal Cys-207 of sorghum leaf NADP-malate dehydrogenase can form mixed disulphides with thioredoxin.

The role of the internal Cys-207 of sorghum NADP-malate dehydrogenase (NADP-MDH) in the activation of the enzyme has been investigated through the examination of the ability of this residue to form mixed disulphides with thioredoxin mutated at either of its two active-site cysteines. The h-type Chlamydomonas thioredoxin was used, because it has no additional cysteines in the primary sequence besides the active-site cysteines. Both thioredoxin mutants proved equally efficient in forming mixed disulphides with an NADP-MDH devoid of its N-terminal bridge either by truncation, or by mutation of its N-terminal cysteines. They were poorly efficient with the more compact WT oxidised NADP-MDH. Upon mutation of Cys-207, no mixed disulphide could be formed, showing that this cysteine is the only one, among the four internal cysteines, which can form mixed disulphides with thioredoxin. These experiments confirm that the opening of the N-terminal disulphide loosens the interaction between subunits, making Cys-207, located at the dimer contact area, more accessible.

Animals↗
PubMed1999

Hepatic acute phase induction of murine beta-galactoside alpha 2,6 sialyltransferase (ST6Gal I) is IL-6 dependent and mediated by elevation of exon H-containing class of transcripts.

Hepatic expression of CMP-NeuAc:Gal beta 1,4GlcNAc alpha 2,6-sialyltransferase (ST6Gal I) is induced as part of the acute phase response in mammals by mechanisms that remain poorly understood. Previous work suggests that murine liver ST6Gal I mRNA contains an additional and novel region that is not found on ST6Gal I mRNA from human HepG2 hepatoma cells and from rat liver. This novel region, residing 5' of the common Exon I sequence, is encoded by a discrete upstream exon, Exon H. Here we provide evidence that the Exon H-containing transcript is the murine counterpart of the human and rat ST6Gal I mRNAs transcribed from the hepatic-specific promoter, P1. Exon H-containing ST6Gal I mRNA is expressed in all three mice strains examined: balb/c, C57B46, and 129Sv. Furthermore, murine RNA tissue survey indicates that presence of Exon H-containing transcripts is restricted to the liver. When mice are subjected to subcutaneous injection of turpentine to elicit the hepatic acute phase response, greater than 4-fold elevation in liver ST6Gal I mRNA was observed. Consistent with the view that Exon H-containing transcripts is regulated by the murine P1 promoter, 5'-RACE analysis indicates that the majority of these transcripts contains the Exon H sequence. This is consistent with the view that Exon H-containing transcripts are regulated by the murine P1 region. To assess the mechanism of ST6Gal I response in the hepatic acute phase reaction, mice harboring lesions in both alleles of the IL-6 gene were examined. IL-6(-/-) animals expressed normal levels of ST6Gal I mRNA in liver, with Exon H-containing transcripts remaining the predominant mRNA isoform. However, hepatic ST6Gal I is not elevated upon turpentine injection in the IL-6(-/-) animals. These results indicate that ST6Gal I induction in mouse liver during the acute phase reaction is mediated predominantly by the IL-6 pathway, and results in the induction of the Exon H-containing class of ST6Gal I mRNA that is specific to the liver.

Acute-Phase Reaction↗
PubMed1999

Heavy-metal regulation of thioredoxin gene expression in chlamydomonas reinhardtii

Heavy metals are highly toxic compounds for cells. In this report we demonstrate that the expression of Chlamydomonas reinhardtii thioredoxins (TRX) m and h is induced by heavy metals. Upon exposure of the cells to Cd and Hg, a strong accumulation of both messengers was observed. Western-blot experiments revealed that among these two TRXs, only TRX h polypeptides accumulated in response to the toxic cations. A biochemical analysis indicated that heavy metals inhibit TRX activity, presumably by binding at the level of their active site. Sequence analysis of the C. reinhardtii TRX h promoter revealed the presence of cis-acting elements related to cadmium induction. The origins and purposes of this regulation are discussed. Our data suggest, for the first time to our knowledge, a possible implication of TRXs in defense mechanisms against heavy metals.

Journal Article↗
PubMed1999

Interaction of thioredoxins with target proteins: role of particular structural elements and electrostatic properties of thioredoxins in their interplay with 2-oxoacid dehydrogenase complexes.

The thioredoxin action upon the 2-oxoacid dehydrogenase complexes is investigated by using different thioredoxins, both wild-type and mutated. The attacking cysteine residue of thioredoxin is established to be essential for the thioredoxin-dependent activation of the complexes. Mutation of the buried cysteine residue to serine is not crucial for the activation, but prevents inhibition of the complexes, exhibited by the Clamydomonas reinhardtii thioredoxin m disulfide. Site-directed mutagenesis of D26, W31, F/W12, and Y/A70 (the Escherichia coli thioredoxin numbering is employed for all the thioredoxins studied) indicates that both the active site and remote residues of thioredoxin are involved in its interplay with the 2-oxoacid dehydrogenase complexes. Sequences of 11 thioredoxin species tested biochemically are aligned. The thioredoxin residues at the contact between the alpha3/3(10) and alpha1 helices, the length of the alpha1 helix and the charges in the alpha2-beta3 and beta4-beta5 linkers are found to correlate with the protein influence on the 2-oxoacid dehydrogenase complexes (the secondary structural elements of thioredoxin are defined according to Eklund H et al., 1991, Proteins 11:13-28). The distribution of the charges on the surface of the thioredoxin molecules is analyzed. The analysis reveals the species specific polarization of the thioredoxin active site surroundings, which corresponds to the efficiency of the thioredoxin interplay with the 2-oxoacid dehydrogenase systems. The most effective mitochondrial thioredoxin is characterized by the strongest polarization of this area and the highest value of the electrostatic dipole vector of the molecule. Not only the magnitude, but also the orientation of the dipole vector show correlation with the thioredoxin action. The dipole direction is found to be significantly influenced by the charges of the residues 13/14, 51, and 83/85, which distinguish the activating and inhibiting thioredoxin disulfides.

3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)↗
PubMed1999

Pharmacological manipulation of Ins(1,4,5)P3 signaling mimics preconditioning in rabbit heart.

Recent evidence revealed biphasic alterations in myocardial concentrations of the second messenger inositol (1,4,5)-trisphosphate [Ins(1,4,5)P3] with ischemic preconditioning (PC), i.e., increase during brief PC ischemia and decrease early during sustained test occlusion. Our aim was to determine whether an agonist and an antagonist of Ins(1,4,5)P(3) signaling (D-myo-inositol-1,4,5-trisphosphate hexasodium salt [D-myo-Ins(1,4, 5)P3] and 2-aminoethoxydiphenyl borate (2-APB), respectively), given such that they mimic this biphasic profile, would mimic infarct size reduction with PC. To test this concept, isolated, buffer-perfused rabbit hearts received no intervention (control), ischemic PC, D-myo-Ins(1,4,5)P3, D-myo-Ins(1,4,5)P(3) + PC, 2-APB, or 2-APB + PC. All hearts then underwent 30-min coronary occlusion and 2 h reflow, and infarct size was delineated by tetrazolium staining. In addition, the effects of D-myo-Ins(1,4,5)P3 and 2-APB on Ins(1,4,5)P3 signaling were evaluated in isolated fura 2-loaded rat cardiomyocytes. Mean infarct size was reduced with PC and in all D-myo-Ins(1,4,5)P3- and 2-APB-treated groups versus control (59 and 42-55%, respectively, vs. 80% of myocardium at risk, P < 0.05). Thus pharmacological manipulation of Ins(1,4,5)P3 signaling mimics the cardioprotection achieved with ischemic PC in rabbit heart.

Animals↗
PubMed1998

Different patterns of IL-1beta secretion, adhesion molecule expression and apoptosis induction in human endothelial cells treated with 7alpha-, 7beta-hydroxycholesterol, or 7-ketocholesterol.

Among oxysterols oxidized at C7 (7alpha-, 7beta-hydroxycholesterol, and 7-ketocholesterol), 7beta-hydroxycholesterol and 7-ketocholesterol involved in the cytotoxicity of oxidized low density lipoproteins (LDL) are potent inducers of apoptosis. Here, we asked whether all oxysterols oxidized at C7 were able to trigger apoptosis, to stimulate interleukin (IL)-Ibeta and/or tumor necrosis factor (TNF)-alpha secretion, and to enhance adhesion molecule expression (intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin) on human umbilical venous endothelial cells (HUVECs). Only 7beta-hydroxycholesterol and 7-ketocholesterol were potent inducers of apoptosis and of IL-1beta secretion. TNF-alpha secretion was never detected. Depending on the oxysterol considered, various levels of ICAM-1, VCAM-1 and E-selectin expression were observed. So, oxysterols oxidized at C7 differently injure and activate HUVECs, and the alpha- or beta-hydroxyl radical position plays a key role in apoptosis and IL-1beta secretion.

Apoptosis↗
PubMed1998

The single mutation Trp35-->Ala in the 35-40 redox site of Chlamydomonas reinhardtii thioredoxin h affects its biochemical activity and the pH dependence of C36-C39 1H-13C NMR.

The role of the invariant Trp residue at the redox site of thioredoxins was investigated by site-directed mutagenesis of a Chlamydomonas reinhardtii thioredoxin h. Though being still redox active with NADPH-thioredoxin reductase and chemical substrates [dithiothreitol and 5,5'-dithio-bis(2-nitrobenzoic acid)] the Trp35-->Ala-mutated protein completely lost the capacity to activate the thiol-regulated NADPH-dependent malate dehydrogenase. However, it was able to activate a mutant malate dehydrogenase where only the most exposed disulfide was retained. The pH dependence of the redox-site Cys beta 1H/13C-NMR frequencies of the wild-type and mutated proteins, in both the reduced and oxidised states, were compared over the pH range 5.8-10. The mutation does not affect the conserved buried Asp30, which titrates with a pKa of 7.5 in the oxidised proteins in agreement with previous studies. However, for the reduced forms of the proteins, the pH dependence of resonances of both Cys was strongly affected by the mutation. In the case of the wild-type thioredoxin, two apparent pKa values were found around 7.0 and 9.5 and could be assigned to the titration of Cys36 and Cys39 thiol, respectively, similar to the case of Escherichia coli thioredoxin. For the mutated thioredoxin a single pKa was found around 8.3. This result can be interpreted as a single pKa of either Cys36 or Cys39 or both. While the mutation clearly affects ionisations, the measured redox potentials of the active-site Cys pair are not significantly affected by the Trp35-->Ala mutation. Possible roles of an aromatic side chain on the reactivity of the catalytic Cys residues in thioredoxins are proposed.

Animals↗
PubMed1998

Poly A-containing histone H4 mRNA variant (H4-v. 1): isolation and sequence determination from bovine adrenal medulla.

A histone H4 cDNA variant (H4-v.1) was cloned from a bovine adrenal medullary phage library using PCR as a method of detection. The isolated clones contained a short 5' untranslated region (UTR) followed by the histone H4 coding region and a long atypical 3'UTR. The 3'UTR comprised the palindromic and purine-rich sequences typical of cell-cycle dependent histone mRNAs, and a 1.1 kb extension downstream of the palindromic sequence ending with a poly(A) track typical of cell-cycle independent histone mRNAs. Northern blot and RT-PCR analyses indicate that the transcript is fully expressed in bovine adrenal medulla. Thus, bovine histone H4-v.1 mRNA represents the first example of a histone H4 transcript that contains both 3'UTR characteristics of cell-cycle dependent and cell-cycle independent histone mRNAs.

Adrenal Medulla↗