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Biomedical subjects

S Vincent

Publications and source records attributed to S Vincent.

At least 91 records · Page 5Linked to original sources

Characterization of late response genes sequentially expressed during renewed growth of fibroblastic cells.

Many proto-oncogenes are rapidly and transiently activated during the early stages of the cellular transition from a resting G0 state to the DNA synthesis (S) phase. To get better understanding of the gene complexity involved at later stages, we isolated, by cDNA cloning, and identified 17 genes that are activated sequentially during the period of time from proto-oncogene expression to the onset of DNA synthesis in the hamster CCL39 fibroblastic cell line. When protein synthesis is inhibited, induced expression of these genes is unaffected for 10 of them, enhanced for four, in a fashion similar to the immediate-early response genes, and inhibited for three, as observed for delayed early-response genes. In addition to rhoG, a new member of the ras homolog gene family (Vincent et al., 1992), cDNA sequencing indicated that six of them correspond to cytoskeletal proteins (alpha-tubulin, vascular alpha-actin and skeletal gamma-actin), extracellular matrix protein (thrombospondin), secreted protease (plasminogen activator inhibitor-1) and energy-linked transporter (mitochondrial proton/phosphate symporter). This overall survey shows that numerous differentially regulated gene activations are associated with the cell cycle progression, and suggests that proteins involved in cellular reshaping participate actively in the control of cellular growth.

Actins↗

Biocontrol efficacy of Gerris (A) spinolae, Laccotrephes griseus and Gambusia affinis on larval mosquitoes.

Predation experiments using Gerris (A) spinolae, Laccotrephes griseus and Gambusia affinis were conducted against IV stage culicine larvae with varying prey densities. Ranking of individual predatory efficiency showed the sequence: large Gambusia > medium Gambusia > small Gambusia > female Laccotrephes > male Laccotrephes > Gerris. Predation under coexistence reveals the significance of predatory efficiency of different predator combinations with reference to prey density and exposure period.

Animals↗

Investigations on the role of hemopexin and albumin in plasma clearance and tissue distribution of Sn-protoporphyrin.

The mechanism of the clearance of circulating tin-protoporphyrin (Sn-PP), a competitive inhibitor of heme oxygenase in the degradation of heme to bilirubin, is unknown. Two serum proteins, albumin and hemopexin, which are instrumental in the delivery of iron-protoporphyrin (heme) to the liver, also bind metalloporphyrins with high affinity and may aid in targeting their tissue distribution. After intravenous injection of 1 mumol Sn-PP/kg, the serum concentration of hemopexin decreased in human subjects, rats, and rabbits within 24 hours to a similar extent (30% to 50%). This finding suggested that hemopexin may have a role in the tissue distribution of Sn-PP. However, when rats were injected with Sn-PP in saline solution or complexed with albumin, more Sn-PP was taken up by the liver and testes than when Sn-PP was complexed with hemopexin. These results indicate that hemopexin does not preferentially target Sn-PP to the liver and may not be the preferred vehicle for clearance of circulating Sn-PP.

Albumins↗

Vancomycin resistance in Enterococcus gallinarum.

The vancomycin resistance expressed by several strains of Enterococcus gallinarum was studied. Resistance was expressed constitutively, as demonstrated by analysis of growth and inhibition of peptidoglycan synthesis. E. gallinarum strains were moderately resistant to vancomycin (MIC, 16 micrograms/ml) but were as susceptible as vancomycin-susceptible enterococci to the glycopeptides, teicoplanin, A35512B, A47934, A4103A, and A41030E and the glycopeptide actaplanins A1, B2, and C1. Vancomycin resistance in E. gallinarum was inhibited by beta-lactam antibiotics at concentrations that saturated penicillin-binding protein 6 (PBP 6), as demonstrated by binding competition experiments. Spontaneous mutants (frequency, 10(-8)) were two- to fourfold more resistant to beta-lactam inhibition of vancomycin resistance than the parent strain. PBP binding competition experiments suggested that PBP 6 in the mutants bound less cefotaxime, while binding of penicillin and cefoxitin was unaffected. Both a bioassay method and high-performance liquid chromatography showed that E. gallinarum membranes have enzymatic activity which modifies a model pentapeptide yielding a product that is thought to be a tetrapeptide. This activity could be a D,D-carboxypeptidase. In both the parent E. gallinarum strain and its derivatives that were resistant to the synergistic drug combination, the activity was inhibited by beta-lactams at concentrations which correlated with those that inhibit vancomycin resistance and those that saturate PBP 6. These results suggest the possibility that PBP 6 may be involved in the vancomycin resistance of E. gallinarum and that the putative D,D-carboxypeptidase activity seen in E. gallinarum membranes may be attributable to PBP 6.

Anti-Bacterial Agents↗

Growth-regulated expression of rhoG, a new member of the ras homolog gene family.

Cellular transition from the resting state to DNA synthesis involves master switches genes encoding transcriptional factors (e.g., fos, jun, and egr genes), whose targets remain to be fully characterized. To isolate coding sequences specifically accumulated in late G1, a differential screening was performed on a cDNA library prepared from hamster lung fibroblasts stimulated for 5 h with serum. One of the positive clones which displayed a sevenfold induction, turned out to code for a protein sharing homology to Ras-like products. Cloning and sequence analysis of the human homolog revealed that this putative new small GTPase, referred to as rhoG, is more closely related to the rac, CDC42, and TC10 members of the rho (ras homolog) gene family and might have diverged very early during evolution. rhoG mRNA accumulates in proportion to the mitogenic strength of various purified growth factors used for the stimulation, as a consequence of transcriptional activation. G1-specific RNA accumulation is impaired upon addition of antimitogenic cyclic AMP and is enhanced when protein synthesis is inhibited, mainly as a result of RNA stabilization. rhoG mRNA expression is observed in a wide variety of human organs but reaches a particularly high level in lung and placental tissues.

Amino Acid Sequence↗

Lytic effect of two fluoroquinolones, ofloxacin and pefloxacin, on Escherichia coli W7 and its consequences on peptidoglycan composition.

Examination of biochemical changes in Escherichia coli W7 after exposure to ofloxacin or pefloxacin revealed distinct concentration-dependent responses. At levels close to the MIC, extensive filamentation was followed by a lytic event, which involved an active protein synthesis. This lysis was correlated with changes in the peptidoglycan composition, particularly a decrease in the average glycan chain length, involving the action of the autolysines. At higher concentrations, no lysis occurred and the growth was totally inhibited as well as the protein synthesis. The peptidoglycan composition exhibited an increase in the average glycan chain length, suggesting an apparent reduced activity of the lytic transglycosylase. These results show that exposure to low concentrations of quinolones leads to the induction of lysis and peptidoglycan modifications which might contribute to the bactericidal effects of quinolones.

Bacterial Proteins↗

Vancomycin susceptibility and identification of motile enterococci.

Thirty-seven clinical isolates of Enterococcus gallinarum and Enterococcus casseliflavus and three type or reference strains of the species were studied with respect to vancomycin susceptibility and key identification characteristics. With the exception of one clinical isolate of E. casseliflavus (MIC, 4 micrograms/ml), MICs of vancomycin were 8 to 32 micrograms/ml. The type strain of E. gallinarum, NCDO 2313, and five of the clinical isolates had similar penicillin-binding protein profiles and shared 90 to 100% DNA homology. Two isolates, identified as E. gallinarum by conventional tests, were shown to be non-pigment-producing E. casseliflavus on the basis of penicillin-binding protein profile and DNA homology. The type and reference strains of E. casseliflavus, ATCC 25788 and ATCC 25789, were nonmotile in our experiments. However, both shared 65 to 100% DNA homology with each other and with five clinical isolates of E. casseliflavus. These data suggest that the MICs of vancomycin observed for strains of E. gallinarum and E. casseliflavus are higher than those usually associated with other enterococci and may be a common property of these species. Additionally, pigment production and motility may occasionally be misleading criteria for definitive identification of these organisms.

Bacterial Proteins↗

In-series compliance of gastrocnemius muscle in cat step cycle: do spindles signal origin-to-insertion length?

1. It has been claimed that stretch in the non-contractile (extramysial) portion of muscles is substantial, and may produce large discrepancies between the origin-to-insertion muscle length and the internal length variations 'seen' by muscle spindle endings. 2. In eight pentobarbitone-anaesthetized cats, we estimated stretch in the extramysial portion of medial gastrocnemius (MG) muscle with a method similar to the spindle null technique. 3. Length variations of MG previously monitored in a normal step cycle were reproduced with a computer-controlled length servo. The responses of test MG spindle endings were monitored in dorsal root filaments. Distributed stimulation of ventral root filaments, rate-modulated by the step-cycle EMG envelope, served to reproduce step-cycle forces. The filaments were selected so as to have no fusimotor action on the test spindle. 4. Spindle responses in active cycles were compared with those in passive cycles (stretch, but no distributed stimulation). In some cases concomitant tonic fusimotor stimulation was used to maintain spindle responsiveness throughout the cycle, both in active and passive trials. Generally, small discrepancies in spindle firing were seen. The passive trials were now repeated, with iterative adjustments of the length function, until the response matched the spindle firing profile in the active trial. The spindle 'saw' the same internal length change in the final passive trial as in the active trial. Any difference between the corresponding length profiles was attributed to extramysial displacement. 5. Extramysial displacement estimated in this was was maximal at short mean muscle lengths, reaching about 0.5 mm in a typical step cycle (force rising from 0 to 10 N). At longer mean muscle lengths where muscle force rose from say 2 to 12 N in the cycle, extramysial displacement was in the range 0.2-0.4 mm. 6. Except at very short lengths, the displacement was probably mainly tendinous. On this assumption, our results suggested that the stiffness of the MG tendinous compartment was force related, and about double that of cat soleus muscle at any given force. Calculations indicated that though the stretch was small, the MG tendon would store and release enough strain energy per cycle to contribute significantly to the E3 phase of the step cycle. The discrepancies in spindle firing were generally quite subtle, so we reject the claim that extramysial stretch poses a serious difficulty for inferences about fusimotion from chronic spindle afferent recordings.

Action Potentials↗

Ensemble proprioceptive activity in the cat step cycle: towards a representative look-up chart.

Analysis of the control of movement in tasks such as stepping is severely restricted by the lack of quantitative data on the ensemble activity of afferents in the numerous muscles involved. We have started to build up a quantitative "look-up-chart" of the ensemble afferent and efferent profiles in the cat step cycle. To this end, we have developed software which allows us to digitize afferent firing, muscle length and electromyogram (EMG) activity, and to align segments for averaging by choosing one or more reference points in the step cycle. The ensemble firing of triceps surae Ia afferents showed lower than expected mean and peak rates, whereas triceps group II and Ib afferents were more active than predicted. There were small but significant transients in Ia firing at foot-off and touch-down which could not be explained in terms of origin-to-insertion length lone. They were most likely caused by propagated mechanical transients or tendon compliance effects giving rise to small differences between the origin-to-insertion length and the intramuscular length "seen" by spindles. Net ensemble Ia rates, based on previous estimates of spindle populations, probably exceed 25 kilo-impulses/second (ki.p.s.) in some muscles. Inputs as large as this are likely to contribute significantly to reflex control.

Animals↗

What influences doctors' prescribing? Sore throats revisited.

An audit of two practices in 1987 revealed a wide range of antibiotic prescribing for acute sore throat among the general practitioners. The data were presented at a postgraduate meeting and recommendations were made for a practice policy on antibiotic prescribing. The results of studies that looked at the objectives of treatment were included at that meeting. This paper presents a re-evaluation of the same doctors' antibiotic prescribing one-year later. Changes had occurred in the range and costs of drugs chosen, but individual doctors' prescribing rates remained broadly similar, in other words it was easier to influence what, but not whether, a doctor prescribes for this clinical condition. The existence of a prescribing 'threshold' within the individual doctor is supported.

Anti-Bacterial Agents↗

Pulsatile nature of luteinizing hormone release is maintained during luteinizing hormone-releasing hormone stimulation in normal male rats.

The plasma LH concentration is believed to be reasonably steady in normal male rats. We found that LH is released in a regular pulsatile fashion. The overall mean concentration of plasma LH in normal male rats was 46.6 +/- 4.4 (mean +/- SEM) ng/ml. The normal male rats showed periodic LH pulses: the mean pulse amplitude was 144.4 +/- 25.5 ng/ml and the inter-peak interval was 22.5 +/- 2.0 min. Each pulse lasted 9.7 +/- 0.8 min. When LH-RH (1 microgram/kg) was injected as a bolus, the peak concentration was attained in 10-30 min reaching a peak concentration of 279.4 +/- 39.6 ng/ml. Distinct pulsatile bursts of plasma LH were discernible during the period of elevated plasma LH concentration. When a higher dose of LH-RH (5 micrograms/kg) was administered, the LH concentration slowly increased to a peak concentration of 400.2 +/- 38.7 ng/ml in 20-40 min. The pulsatile nature of the LH concentration was recognizable with distinct bursts. We have observed that: (a) normal male rats release LH in a pulsatile fashion with an approximate 20-min inter-peak interval; (b) mean LH pulses last less than 10 min, and (c) the LH pulses are visible even with elevated LH and LH-RH concentrations in the general circulation.

Animals↗

Distribution of anionic sites on the capillary endothelium in an experimental brain tumor model.

The distribution of anionic domains on the capillary endothelium of experimental brain tumors was determined using cationic ferritin (CF) in order to ascertain whether the pattern of these domains is different from that on normal cerebral capillaries. Tumors were induced by stereotaxic injection of cultured neoplastic glial cells, A15A5, into the caudate nucleus of Sprague-Dawley rats. Following a 14-21 day growth period tumors appeared as vascularized, sharply circumscribed masses which caused compression of the surrounding brain tissue. Anionic domains were distributed in a patchy and irregular pattern on the luminal plasma membrane of the endothelia of blood vessels in the tumors. Some variability in this pattern was observed infrequently in limited regions of the tumor where there was either a continuous layer of CF or an absence of CF binding. Plasmalemmal vesicles, coated vesicles, coated pits, multivesicular bodies, and some junctional complexes showed varying degrees of labeling with the probe. Capillaries in the tumor periphery and normal cerebral vessels showed a uniform distribution of anionic groups. These results indicate that there is an altered surface charge on the endothelial luminal plasma membrane of blood vessels in brain tumors. A correlation may exist between the altered surface charge and the degree to which the blood-brain barrier is impaired in these vessels.

Animals↗

Involvement of penicillin-binding protein 2 with other penicillin-binding proteins in lysis of Escherichia coli by some beta-lactam antibiotics alone and in synergistic lytic effect of amdinocillin (mecillinam).

Compared with cefotaxime, ceftazidime, moxalactam, and aztreonam, ceftriaxone produced the best lytic and bactericidal effects when each was added at about 10 times the MIC to Escherichia coli W7. When each of these antibiotics was added at its MIC, only bacteriostasis occurred, but the simultaneous addition of amdinocillin (mecillinam) was synergistic in causing rapid lysis and bactericidal effects. Induction of lysis of two E. coli mutants containing either a thermosensitive penicillin-binding protein (PBP) 2 or 3 by relatively PBP 3-specific (aztreonam) and PBP 2-specific (amdinocillin) antibiotics indicated that inhibition of only PBPs 2 and 3 can cause lysis. Examination of the interactions of cefotaxime, aztreonam, and cefsulodin, with or without amdinocillin, with their targets suggested that other combinations of PBPs could be involved in the onset of lysis. However, inhibition of both PBPs 2 and 3 may explain the better lysis-inducing activity of ceftriaxone (which binds well to both of these PBPs), as well as the synergistic effect of amdinocillin when added together with low concentrations of other beta-lactam antibiotics that interact with PBP 3.

Acyltransferases↗

Distribution of substance P in brain and periphery and its possible role as a co-transmitter.

Substance P is widely distributed in the nervous system. In brain and spinal cord it may act as a transmitter, for example at the central branches of primary sensory neurons. It may also be released from the sensory nerve endings and is thought to be involved in antidromic vasodilatation and in synaptic transmission in autonomic ganglia. In some central neurons substance P is stored together with 5-hydroxytryptamine and thyrotropin-releasing hormone. These neurons project to the ventral horn of the spinal cord, amongst other places. In another system substance P coexists with a cholecystokinin-like peptide. These neurons are localized in the periaqueductal central grey matter and also project to the spinal cord. Finally, injection of a substance P antagonist into the ventral mesencephalon causes marked morphological changes in neurons that contain dopamine, substance P and gamma-aminobutyric acid (GABA).

Animals↗