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Biomedical subjects

P Nicolas

Publications and source records attributed to P Nicolas.

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

Structure of genes for dermaseptins B, antimicrobial peptides from frog skin. Exon 1-encoded prepropeptide is conserved in genes for peptides of highly different structures and activities.

We cloned the genes of two members of the dermaseptin family, broad-spectrum antimicrobial peptides isolated from the skin of the arboreal frog Phyllomedusa bicolor. The dermaseptin gene Drg2 has a 2-exon coding structure interrupted by a small 137-bp intron, wherein exon 1 encoded a 22-residue hydrophobic signal peptide and the first three amino acids of the acidic propiece; exon 2 contained the 18 additional acidic residues of the propiece plus a typical prohormone processing signal Lys-Arg and a 32-residue dermaseptin progenitor sequence. The dermaseptin genes Drg2 and Drg1g2 have conserved sequences at both untranslated ends and in the first and second coding exons. In contrast, Drg1g2 comprises a third coding exon for a short version of the acidic propiece and a second dermaseptin progenitor sequence. Structural conservation between the two genes suggests that Drg1g2 arose recently from an ancestral Drg2-like gene through amplification of part of the second coding exon and 3'-untranslated region. Analysis of the cDNAs coding precursors for several frog skin peptides of highly different structures and activities demonstrates that the signal peptides and part of the acidic propieces are encoded by conserved nucleotides encompassed by the first coding exon of the dermaseptin genes. The organization of the genes that belong to this family, with the signal peptide and the progenitor sequence on separate exons, permits strikingly different peptides to be directed into the secretory pathway. The recruitment of such a homologous 'secretory' exon by otherwise non-homologous genes may have been an early event in the evolution of amphibian.

Amino Acid Sequence↗
PubMed1997

Pulsed-field gel electrophoresis analysis of clonal relationships among Neisseria meningitidis A strains from different outbreaks.

Pulsed-field gel electrophoresis was used to analyze the DNA of 67 Neisseria meningitidis A strains from five African outbreaks occurring between 1988 and 1996. Endonuclease Bg/II was used to cut chromosomal DNA, generating 19 analyzable fragments. This technique allowed comparison of clonal relationships between outbreak strains and revealed that the same pulsotype was responsible for the outbreaks. This pulsotype is closely related to strain B54, subgroup III reference strain, Finland, 1975, showing only two fragment differences.

Africa↗
PubMed1997

Typing Tuber ectomycorrhizae by polymerase chain amplification of the internal transcribed spacer of rDNA and the sequence characterized amplified region markers.

Identification of some economically important Tuber species using classical morphological characteristics is sometimes difficult. We report here the molecular characterization of a species coming from China, Tuber indicum, mistaken with Tuber melanosporum species. Using restriction analysis of the amplified internal transcribed spacer (ITS) of rDNA, ITS sequence analysis, and sequence characterized amplified region markers, with DNA from fruit bodies or mycorrhizae, genetic variation was found between these two species, allowing to differentiate and characterize them.

Ascomycota↗
PubMed1997

Thyrotropin-releasing hormone immunoreactivity in rat adrenal tissue is localized in mast cells.

Pro-thyrotropin-releasing hormone (pro-TRH) has been shown to be present throughout the central nervous system and in several peripheral tissues. In adrenals, TRH immunoreactivity has been reported but not characterized. We show here that two rat pro-TRH-derived peptides, TRH and prepro-TRH[160-169] (Ps4), were detected in extracts of rat adrenal glands by enzyme immunoassay. Endogenous TRH and Ps4 were purified by gel exclusion chromatography and reverse-phase HPLC. Structural identification of each peptide was achieved by chromatographic comparison with synthetic standards. By using the indirect immunofluorescence technique, TRH-immunoreactive cell bodies were found rather widely scattered outside the adrenal, in the brown adipose tissue in which the gland is embedded. These immunofluorescent cells have the typical appearance of mast cells and are metachromatic after histological staining with acidic Toluidine Blue. Our findings suggest that pro-TRH-derived peptides exist in rat mast cells.

Adipose Tissue, Brown↗
PubMed1997

Nongenomic effect of triiodothyronine on cell surface beta-adrenoceptors in cultured embryonic cardiac myocytes.

We studied the time course of cell surface beta-adrenoceptors (BAR) in cardiomyocytes in response to a single triiodothyronine (T3) (10(-8) M) stimulation. An early first increase of BAR density was observed within 2 hr (+ 10% versus control cells, P < 0.05), and a plateau was maintained for 17-20 hr. This effect was followed by a much greater, late increase of BAR density, starting around 22 hr and lasting until 48 hr post T3 addition (+40% versus control cells; p < 0.05). Since reverse T3 studied in the same conditions had no effect in this system, we concluded that this T3 effect was specific. We hypothesized that the early response might be nongenomic because the early effect of T3 was still observed in the presence of cycloheximide (2 x 10(-5) M) whereas the late increase was totally suppressed by the drug. The early response to T3 required intact microtubules, since colchicine (2 x 10(-5) M) was able to block the increase in the cell surface BAR number, but it did not involve a change in BAR distribution between external and internal sites, as the external to total BAR ratio remained stable. The measurement of the rate of BAR disappearance from the cell surface allowed us to hypothesize that T3 induced a modification of the turnover in cell surface BAR.

Animals↗
PubMed1996

Covalent structure, synthesis, and structure-function studies of mesentericin Y 105(37), a defensive peptide from gram-positive bacteria Leuconostoc mesenteroides.

A 37-residue cationic antimicrobial peptide named mesentericin Y 105(37) was purified to homogeneity from cell-free culture supernatant of the Gram-positive bacterium Leuconostoc mesenteroides. The complete amino acid sequence of the peptide, KYYGNGVHCTKSGCSVNWGEAASAGIHRLANGGNGFW, has been established by automated Edman degradation, mass spectrometry, and solid phase synthesis. Mesentericin Y 105(37) contains a single intramolecular disulfide bond that forms a 6-membered ring within the molecule. Mesentericin Y 105(37) was synthesized by the solid phase method. The synthetic replicate was shown to be indistinguishable from the natural peptide with respect to electrophoretic and chromatographic properties, mass spectrometry analysis, automated amino acid sequence determination, and antimicrobial properties. At nanomolar concentrations, synthetic mesentericin Y 105(37) is active against Gram+ bacteria in the genera Lactobacillus and Carnobacterium. Most interestingly, the peptide is inhibitory to the growth of the food-borne pathogen Listeria. CD spectra of mesentericin Y 105(37) in low polarity medium, which mimic the lipophilicity of the membrane of target organisms, indicated 30-40% alpha-helical conformation, and predictions of secondary structure suggested that the peptide can be configured as an amphipathic helix spanning over residues 17-31. To reveal the molecular basis of the specificity of mesentericin Y 105(37) targetting and mode of action, NH2- or COOH-terminally truncated analogs together with point-substituted analogs were synthesized and evaluated for their ability to inhibit the growth of Listeria ivanovii. In sharp contrast with broad spectrum alpha-helical antimicrobial peptides from vertebrate animals, which can be shortened to 14-18 residues without deleterious effect on potency, molecular elements responsible for anti-Listeria activity of mesentericin Y 105(37) are to be traced at once to the NH2-terminal tripeptide KYY, the disulfide bridge, the putative alpha-helical domain 17-31, and the COOH-terminal tryptophan residue of the molecule. It is proposed that the amphipathic helical domain of the peptide interacts with lipid bilayers, leading subsequently to alteration of the membrane functions, whereas residues 1-14 form part of a recognition structure for a membrane-bound receptor, which may be critical for peptide targetting. Because mesentericin Y 105(37) is easy to synthesize at low cost, it may represent a useful and tractable tool as a starting point for the design of more potent analogs that may be of potential applicability in foods preservation.

Amino Acid Sequence↗
PubMed1996

The putative immunity protein of the gram-positive bacteria Leuconostoc mesenteroides is preferentially located in the cytoplasm compartment.

Immunity proteins are though to protect bacteriocin-producing bacterial strains against the bactericidal effects of their own bacteriocin. The immunity protein which protects the lactic acid bacterium Leuconostoc mesenteroides against mesentericin Y105(37) bacteriocin was detected and localized by immunofluorescence and electron microscopy, using antibodies directed against the C-terminal end of the predicted immunity protein. The antibodies recognized the immunity proteins of various strains of Leuconostoc, including Leuconostoc mesenteroides and Leuconostoc gelidum. This study demonstrated that immunity proteins produced by Leuconostoc mesenteroides accumulated in the cytoplasmic compartment of the bacteria. This is in contrast with other known immunity proteins, such as the colicin immunity proteins, which are integral membrane proteins possessing three to four transmembrane domains.

Amino Acid Sequence↗
PubMed1996

Employment implications of declining tobacco product sales for the regional economies of the United States.

OBJECTIVE: To determine whether declines in tobacco product sales significantly reduce employment in the United States, as the tobacco industry claims. DESIGN: Computer simulation of the economies of the Southeast Tobacco region and 8 nontobacco regions of the United States, with domestic tobacco expenditures eliminated or reduced and the equivalent spending redistributed, according to consumers' normal spending patterns. We compared these results with baseline forecasts of the regional economies that include normal tobacco expenditures. MAIN OUTCOME MEASURE: Number of jobs. RESULTS: Had there been no spending on tobacco products in the United States in 1993, the Southeast Tobacco region would have had 303 000 fewer jobs. Collectively, however, the 8 nontobacco regions would have gained enough employment to completely offset losses in the Southeast Tobacco region, with every nontobacco region gaining jobs. By the year 2000, the absence of tobacco spending would mean a loss of 222 000 jobs in the Southeast Tobacco region, but a gain of 355 000 throughout the rest of the country. In the more realistic scenario of doubling the downward trend in tobacco consumption, the Southeast Tobacco region would lose 6300 jobs in 1993 (0.03% of regional employment) and 36 600 jobs by 2000 (0.2%). The 8 nontobacco regions would gain 6400 jobs in 1993 and 56 300 jobs in 2000, with each of the nontobacco regions gaining employment in every year. CONCLUSIONS: Contrary to the tobacco industry's claims, reductions in spending on tobacco products will boost employment in every one of the 8 nontobacco regions and will not diminish employment in the Southeast Tobacco region by as much as the industry estimates. The primary concern about tobacco should be the enormity of its toll on health and not its impact on employment.

Commerce↗
PubMed1996

Broad spectrum antibiotic activity of the skin-PYY.

Neuropeptide Y (NPY) and polypeptide YY (PYY) are two ubiquitous neuropeptides, found in brain and intestines, respectively, where they exert important regulatory functions. In this study, a new member of the YY family recently isolated from amphibian skin, skin-PYY (SPYY), is reported to inhibit irreversibly the proliferation of a broad spectrum of pathogenic microorganisms. NPY and PYY are shown to be endowed with the same activity. Their potency is similar to that of other antibacterial peptides which have been shown to exert their function by disintegrating the bacterial membrane. These findings and the fact that the C-terminal alpha-helical domain SPYY14-36, which is highly conserved among family members, was responsible for killing microorganisms and for permeation of phospholipid vesicles, suggested that the antibiotic activity may emerge via a membrane permeation mechanism. These findings also raise the question whether NPY and PYY exert in vivo a similar function in mammals.

Amino Acid Sequence↗
PubMed1996

Folded conformations of the delta-selective opioid dermenkephalin with head-to-tail interactions. A simulated annealing study through NMR restraints.

Despite similar tripeptide N-termini, dermorphin (Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH2) and dermenkephalin (Tyr-D-Met-Phe-His-Leu-Met-Asp-NH2), naturally occuring opioid peptides from frog skin, exhibit high affinity but contrasting selectivity for the mu- and delta-opioid receptors, respectively. Structure-activity relationship studies have shown that the N-terminal tripeptide, Tyr-D-Xaa-Phe (where Xaa is either Ala or Met), is necessary for binding with both the mu- and delta-receptors while the nature and/or the conformation of the C-terminus His-Leu-Met-Asp-NH2 of dermenkephalin are responsible for addressing the peptide to the delta-receptor. In order to examine the conformational characteristics that are related to the selectivity of dermenkephalin towards the delta-receptor, 50 NOE restraints (10 between non-adjacent residues), and 7 dihedral angles, derived from a two-dimensional 1H-NMR study of dermenkephalin in dimethyl sulfoxide, were used in simulated annealing and energy minimization procedures. Twenty-four resulting conformers (60% of the generated structures) with no severe distance restraint violation were pooled into seven groups and three related families. These 24 conformers show close proximity between the two methionine residues, S-shaped structures, mean planes of N-terminal and C-terminal moieties almost at right angles to each other, a C-terminus region above the plane of the N-terminal region and g- as preferential orientation in the side chain of Phe. Aside these similarities, families of conformers differ by the preferential orientation in the side chain of Tyr (t or g-) and proximity between Tyr and Asp, or Tyr and the C-terminus. In contrast to previous models, practically no beta-turn structures exist for dermenkephalin, most of the NH hydrogen bonds participating to gamma-turns. The possible relationship between the conformational characteristics of dermenkephalin and the delta-opioid receptor selectivity is discussed.

Animals↗
PubMed1996

Identification and cellular localization of thyrotropin-releasing hormone-related peptides in rat testis.

Endogenous TRH-like products were analyzed in rat testis using a TRH enzyme immunoassay coupled to molecular sieve filtration and HPLC identification. Of the three immunoreactive peptides detected, the two major forms exhibited the same chromatographic properties as synthetic TRH and pGlu-Phe-Pro-NH2. These peptides accounted respectively for 33% and 54% of the total TRH immunoreactivity in the testis. In rat serum, HPLC analysis showed the presence of only one immunoreactive peak with a retention time similar to that of authentic TRH. Ethylene dimethanesulfonate treatment of adult rat was used to assess the effect of Leydig cell destruction on the TRH immunoreactivity content. The concentration of the three TRH immunoreactive peptides fell gradually after treatment and reached a minimum at day 21, where a marked decrease (98%) was observed. At day 41, the regeneration of Leydig cells was achieved, as shown by histochemistry and measurements of serum testosterone and testicular weight. However, no restoration of the TRH immunoreactive content was achieved, the three TRH-related peptides being only detectable in trace amount. On the other hand, no significant change in hypothalamic levels of TRH was observed at any treatment time, indicating that hypothalamic TRH biosynthesis was not influenced by testosterone. By using the indirect immunofluorescence technique, TRH immunoreactive cells were found in the interstitial space of the testis. These results suggest that Leydig cells are the only source of authentic TRH and TRH-like peptides in the rat testis. A paracrine, or autocrine role for these products is suggested.

Animals↗
PubMed1996

Effect of insulin concentration on bioavailability during nasal spray administration.

The bioavailability of rapid-acting insulin administered as a nasal spray was studied in 6 type 1 (insulin-dependent) diabetic patients. They received long-acting bovine insulin (Ultratardum 40 U/ml, Organon) as basal treatment at 8 a.m. Rapid-acting insulin was also administered at 8 a.m., then at noon and 6 p.m, subcutaneously on day 1 as a 100 U/ml solution and intranasally by aerosol spray as a 100 U/ml and 500 U/ml with 1% (w/v) 9 lauryl ether solution on day 2 and day 3 respectively. On days 2 and 3, the dose of insulin was at least nine times higher than the subcutaneous dose on day 1. Free and total plasma insulin concentrations were assayed after the noon insulin administration. The peaks of the free and total plasma insulin levels were reached earlier and the return to basal levels was obtained earlier after nasal insulin administration than after insulin injected subcutaneously. The bioavailability of nasal spray insulin versus subcutaneous insulin with a 100 U/ml insulin solution was similar to that with a 500 U/ml insulin solution: 5.14 +/- 0.38% (m +/- SEM) and 4.64 +/- 0.46% according to the total plasma insulin level. This study suggests that the bioavailability of nasal spray insulin is not increased by increasing insulin concentration in our experimental conditions.

Administration, Intranasal↗
PubMed1995

Separation of oviposition-stimulating peptides and myotropic factors from head extracts of Galleria mellonella L.: comparative effects of myotropic and non-myotropic factors on egg laying.

Methanolic extracts from heads of the wax moth, Galleria mellonella L. contain several factors that stimulate oviposition of virgin females in vivo and spontaneous contractions of the oviduct in vitro of the cricket, Gryllus bimaculatus. The myotropic and egg-laying activities behaved distinctly during all steps of purification, including fractionation on reverse-phase high-performance liquid chromatography. A novel neuropeptide was isolated from Galleria mellonella. The peptide does not increase the motility of isolated cricket oviducts. Among 13 other myotropic or non-myotropic factors studied, none were found to be as potent as the new peptide in stimulating egg-laying activity. The dose-response curves of myotropic and non-myotropic factors indicate that (i) proctolin, L-glutamate, octopamine, leucokinin-VI, leucopyrokinin, ecdysone and 20-hydroxy-ecdysone, or equivalent structures, might be involved in Galleria mellonella oviposition control, and (ii) that only the novel wax moth neuropeptide has the ability to trigger egg laying in that insect. The significance of these findings is discussed.

Amino Acid Sequence↗
PubMed1995

Ex vivo studies of polymorphonuclear neutrophils from patients with early-onset periodontitis (III). CR3 and LFA-1 expression by peripheral blood and gingival crevicular polymorphonuclear neutrophils.

In this study, we assessed the LFA-1 (CD18/CD11a) and CR3 (CD18/CD11b) expression on peripheral polymorphonuclear leukocytes (PB-PMN) and crevicular fluid polymorphonuclear leukocytes (CF-PMN), by subjects with a healthy periodontium (n = 7), gingivitis (n = 8), early-onset periodontitis (n = 17) and adult periodontitis (n = 8). Using flow cytometry analysis, the %s of CD18, CD11a and CD11b positive cells and the absolute numbers of fluorescent molecules were determined. No significant difference could be found among the 4 groups, for these 2 kinds of parameters, in PB-PMN or CF-PMN. However, a great difference could be noted between the results obtained from PB-PMN and those obtained from CF-PMN. The %s of positive CF-PMN were significantly lower than those of PB-PMN for the 3 sub-units (p < 0.001). The levels of CD18 and CD11b expressed by CF-PMN were higher than those expressed by PB-PMN and the difference was significant for CD11b (p < 0.001). On the contrary, the level of CD11a expressed on CF-PMN was significantly lower than that expressed by PB-PMN (p < 0.001). Hence, our current results show that early-onset periodontitis PMN can be quite normal and this fact is not surprising insofar as, in our study, these cells were perfectly functional and all the subjects were in good health. We concluded that the analysis of the leukocyte adhesion receptors expression on PB-PMN does not appear useful for helping to establish a differential diagnosis between the different forms of periodontitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Expression of the alpha-subunit of CR4 (CD11c) by peripheral blood and crevicular fluid neutrophils.

This work determined the levels of expression of CD11c by neutrophils (PMNs) collected from subjects with various periodontal conditions. The percentages of CD11c-positive crevicular fluid PMNs were significantly lower than those of peripheral blood PMNs, but the levels of CD11c expression were similar in PB-PMNs and CF-PMNs (P<0.001). On the other hand, no significant difference could be found between the groups, either for the percentages of CD11c-positive cells or for the CD11c expression levels.

Adult↗