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Biomedical subjects

C Bailly

Publications and source records attributed to C Bailly.

At least 217 records · Page 12Linked to original sources

Selective expression of PNA-binding glycoconjugates by invasive human melanomas: a new marker of metastatic potential.

Alterations of cell-surface glycoconjugates have been associated with invasiveness and metastatic capacity in a number of experimental and human tumors (bladder and colon cancer). We have recently shown that human melanoma cells from variants selected for high metastatic potential in an animal model bind the lectin peanut agglutinin (PNA), and that human melanoma cell populations enriched for PNA binding cells generated a higher frequency of metastases when xenografted into immune suppressed neonatal rats. We have therefore sought cells binding PNA in biopsied human melanocytic tumors and compared frequencies of PNA binding by cells from benign nevi, early and late primary melanomas, and metastatic melanomas. Sections of conventionally processed tissues were deparaffinised and exposed to biotinylated PNA; PNA fixation was revealed by the avidine/peroxidase/AEC technique. In 51 specimens tested, PNA appears to react electively with invasive tumors, since only one of the 7 early primary melanomas (Clark I-II) reacted while 13/23 late primary melanomas (Clark III-V), and 4/21 melanoma metastases were reactive. In addition, only 1/17 benign nevi bound PNA. In primary tumors, the reactive cells were exclusively invasive tumors cells in the dermis. PNA reactive material was observed in the cytoplasm and plasma membrane of reactive cells. Hence, alterations in composition and cellular localisation of glycoconjugates detectable by lectin histochemistry in melanoma cells may be markers of metastatic potential that may be applicable on an individual patient basis.

Animals↗

Cyclic guanosine monophosphate is the mediator of platelet-activating factor inhibition on transport by the mouse kidney thick ascending limb.

Since we have previously shown a direct inhibitory effect of platelet-activating factor (PAF) on Cl reabsorption in the medullary thick ascending limb of Henle's loop (TAL), the aim of this study was to extend this effect to the whole TAL and to further investigate the signaling pathway involved. In microperfused cortical TALs, PAF significantly decreased Cl reabsorption by 50.3 +/- 6.5%. On the one hand, this effect was not modified in the presence of staurosporine and was not mimicked by phorbol ester; chelating cytosolic Ca by BAPTA/AM failed to suppress the inhibitory effect of PAF on Cl reabsorption; moreover, no significant increase in intracellular Ca concentration could be observed in the presence of PAF on isolated tubules. On the other hand, 8-bromo cyclic GMP mimicked the PAF effect on Cl reabsorption and prevented a further effect of this agent; the PAF effect was significantly reduced by H-8, a cyclic GMP-dependent protein kinase inhibitor; in medullary TALs, PAF significantly increased by twofold cyclic GMP content, an effect inhibited by the PAF antagonist BN 50730, whereas PAF did not significantly modify cAMP content in basal or stimulated conditions. Finally, inhibition of nitric oxide production by NAME or NMMA failed to prevent the effect of PAF on Cl reabsorption. It is concluded that the PAF-induced inhibition of Cl reabsorption in the TAL was mediated by cyclic GMP, likely independent of a nitric oxide synthesis.

Amino Acid Oxidoreductases↗

Psoralen--lexitropsin hybrids: DNA sequence selectivity of photoinduced cross-linking from MPE footprinting and exonuclease III stop assay, and mode of binding from electric linear dichroism.

The properties of certain hybrids 3 and 5 bearing a photoactivatable psoralen group attached to DNA sequence recognizing lexitropsin carriers have been examined. The hybrids bind to poly(dA-dT) with Kapp of 2.8 and 0.9 x 10(7) M-1, i.e. greater than or equal to that of netropsin (Kapp = 1.0 x 10(7) M-1), indicating that the psoralen moiety may contribute to binding in the case of 5. Photoinduced cross-linking of DNA by 3 and 5, while efficient, is less so than that of individual psoralens and reaches a maximum at a ligand to DNA base pair ratio (r) of 0.2. Complementary strand methidium-propyl-EDTA (MPE).Fe(II) footprinting demonstrated that, in the dark, the sequence preferential recognition of hybrids 3 and 5 is dominated by the lexitropsin moiety. Examination of 360 nm photoinduced DNA cross-linking by the hybrids 3 and 5 was carried out using an exonuclease III stop assay. This revealed that > 95% of the DNA remained double stranded, indicating that 3 and 5 generate primarily biadducts at AT-rich sequences. This assay also located individual monoadduct sites, some of which are remote from the dominant cross-linked sites. When the samples were exposed to 254 nm UV light before loading onto the gel to reverse the photoproducts, the pattern of the exonuclease III stop bands was not altered significantly compared with the experiment without 254 nm irradiation. It is concluded that these termination sites include both mono- and biadducts. Electric linear dichroism examination of the DNA complexes of hybrids 3 and 5 (without light activation) provides evidence that the lexitropsin portion binds in the minor groove, while the psoralen portion intercalates in a suitably located site for subsequent photoinduced cross-linking.

Antineoplastic Agents↗

Sequence-selective binding to DNA of cis- and trans- butamidine analogues of the anti-Pneumocystis carinii pneumonia drug pentamidine.

Footprinting experiments using both DNase I and methidium propyl-EDTA.Fe(II) have been used to investigate the sequence selectivity in binding to DNA of pentamidine and four butamidine analogues active against the Pneumocystis carinii pathogen, which afflicts patients with acquired immunodeficiency syndrome. In common with pentamidine, the butamidine drugs, which contain cis- or trans-1,4-but-2-ene linkers and either bis(amidine) or bis(imidazolidine) terminal groups, bind selectively to DNA sequences composed of at least 4 consecutive A.T base pairs. None of the drugs tolerates the presence of a G.C base pair within the binding site. Consistently in the DNase I and methidium propyl-EDTA.Fe(II) footprinting experiments, the cis-isomers produce stronger footprints than do the trans-isomers, despite their similar hydrogen-bonding potentialities. The present experimental data support the view that the conformation of the drug plays a determining role in the binding reaction. Starting from the known structure of a pentamidine-oligonucleotide complex, it is possible to rationalize the different capacities of the cis- and trans-butamidine analogues to recognize defined DNA sequences in terms of the radius of curvature of the molecule and the distance between the positively charged terminal groups. Together, these features constitute critical factors favoring (cis-conformation) or hampering (trans-conformation) the fitting of the drugs into the minor groove of DNA. In terms of structure-activity relationships, the AT-specific recognition of DNA by this series of butamidine derivatives cannot be directly correlated with their potencies against Pneumocystis carinii pneumonia.

Base Sequence↗

[Desmoplastic tumors with multiple differentiation. A new entity. Six cases].

Desmoplastic tumours with divergent differentiation are principally located in the abdomen and develop locally or regionally. They occur in adolescents or young adults and are characterized at histology by a proliferation of undifferentiated small cells surrounded by a dense stroma. Only immuno-histo-chemistry provides the diagnosis. Since their chemosensitivity is rare and often partial the outcome is usually lethal. Six new cases of this recently described entity are presented here. The authors are in favour of a multidisciplinary and aggressive management, combining intensive polychemotherapy, extensive surgical exerisis and total abdominal radiotherapy.

Abdominal Neoplasms↗

The different binding modes of Hoechst 33258 to DNA studied by electric linear dichroism.

The binding mode of the bisbenzimidazole derivative Hoechst 33258 to a series of DNAs and polynucleotides has been investigated by electric linear dichroism. Positive reduced dichroisms were measured for the poly(dA-dT).poly(dA-dT)- and poly(dA).poly(dT)-Hoechst complexes in agreement with a deep penetration of the drug into the minor groove. Similarly, the drug displays positive reduced dichroism in the presence of the DNAs from calf thymus, Clostridium perfringens and Coliphage T4. Conversely, negative reduced dichroisms were obtained when Hoechst 33258 was bound to poly(dG-dC).poly(dG-dC), poly(dA-dC).poly(dG-dT) and poly(dG).poly(dC) as well as with the GC-rich DNA from Micrococcus lysodeikticus indicating that in this case minor groove binding cannot occur. Substitution of guanosines for inosines induces a reversal of the reduced dichroism from negative to positive. Therefore, as anticipated it is the 2-amino group of guanines protruding in this groove which prevents Hoechst 33258 from getting access to the minor groove of GC sequences. The ELD data obtained with the GC-rich biopolymers are consistent with an intercalative binding. Competition experiments performed with the intercalating drug proflavine lend credence to the involvement of an intercalative binding rather than to an external or major groove binding of Hoechst 33258 at GC sequences.

Animals↗

Preferential intercalation at AT sequences in DNA by lucanthone, hycanthone, and indazole analogs. A footprinting study.

DNAase I footprinting has been used to probe the DNA sequence selectivity of the antitumor intercalating agents lucanthone (1), hycanthone (2), 6-chlorolucanthone (7), and four indazole analogs (IA-3-IA-6). The latter have a benzothiopyranoindazole chromophore substituted with a diethylaminoethyl side chain identical to that attached to the thioxanthenone chromophore of compounds 1, 2, and 7. IA-3 and IA-5 are lucanthone analogs bearing a methyl group at position 4, whereas IA-4 and IA-6 are hycanthone analogs bearing a hydroxymethyl group. IA-3 and IA-4 have an additional chloro group at position 6. Studies employing the 160-bp tyrT DNA fragment as substrate to assay inhibition of DNAase I-mediated cleavage show that both lucanthone and hycanthone bind preferentially to AT sites. They discriminate against GC-rich sequences as well as short runs of a single base, which are often cut more readily in the presence of the drugs compared to the control. The indazole analogs exhibit more pronounced selectivity of binding to AT sequences and promote enhanced DNAase I cleavage both at GC-rich sequences and at homooligomeric runs of adenines or thymines. The results of further DNAase I cleavage inhibition assays, performed with three more restriction fragments having different base pair arrangements, are fully consistent with those obtained with the tyrT fragment. They reveal that the preferred binding sequences for lucanthone, hycanthone, and the indazole analogs are predominantly composed of alternating A and T residues.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗

A tandem repeat of the SPKK peptide motif induces psi-type DNA structures at alternating AT sequences.

The interaction between a tandem repeat of the SPKK peptide motif and calf thymus DNA or several polynucleotides has been investigated by circular dichroism. The octapeptide SPKKSPKK does not induce any important changes in the CD spectra of the polynucleotides poly(dG).poly(dC), poly(dG-dC).poly(dG-dC) and poly(dA).poly(dT) while the spectrum of calf thymus DNA is slightly modified. Binding of this basic peptide to the alternating copolymer poly(dA-dT).poly(dA-dT) results in a marked psi-type condensation in a manner similar to that induced by the entire C-terminal domain of histone H1.

Amino Acid Sequence↗

Sequence-selective binding of amiloride to DNA.

Nuclease footprinting techniques have been employed to investigate the interaction between the diuretic drug amiloride, a sodium channel blocker with potential therapeutic use in the treatment of cystic fibrosis, and three DNA fragments of defined sequence. Using either DNAse I or micrococcal nuclease as probes, an unusual pattern of sequence-selective recognition of DNA has been detected. Amiloride binds selectively to sites rich in adenine and thymine residues, frequently with an apparent preference for 5'-TpX-3' steps, and discriminates strongly against GC-rich sequences which are sometimes cut more readily in the presence of the drug compared to the control. A detailed comparison with the actions of known selective DNA-binding antibiotics and drugs reveals a unique pattern of binding sites, different from those of typical intercalators on the one hand and those of minor groove-binders on the other. Amiloride is believed to adopt a pH-dependent tricyclic hydrogen-bonded conformation in solution which allows it to intercalate into DNA; consistent with this belief, we find that the footprinting pattern largely disappears at pH values above the pKa. Preliminary studies with three amiloride analogues have indicated the importance of two functional groups in the recognition of DNA. The possible relevance of selective DNA binding to activity in vivo is considered.

Amiloride↗

Luminal and intracellular cGMP inhibit the mTAL reabsorptive capacity through different pathways.

Since, in the presence of ANF, urinary cGMP was shown to be of glomerular origin, a possible paracrine effect of luminal cGMP on the medullary thick ascending limb (mTAL) function was investigated. Net chloride reabsorption (JCl) was determined on isolated microperfused tubules from mouse kidney. Addition of 10(-6) M cGMP to the lumen significantly decreased JCl by 46.5 +/- 4.6%. A concentration-dependent decrease of the transepithelial voltage was observed, with a 10(-8) M threshold. Added to the bath, ANF (10(-7) M) as well as urodilatin (6 x 10(-8) M) decreased JCl by 29.8 +/- 3.9% and 36.9 +/- 5.1%, respectively, an effect reproduced by 8-bromo cGMP and associated with a significant increase in tubular cGMP content. The inhibitory effect of ANF was similar whether or not cGMP was present in the lumen. Furthermore, increasing intracellular cGMP content by 8-bromo cGMP did not prevent a further effect of luminal cGMP. Finally, H-8, which blocked the effect of ANF, urodilatin, and 8-bromo cGMP, failed to abolish the luminal cGMP-induced decrease of JCl, suggesting that this effect did not require a cGMP-dependent protein kinase activation. It is concluded that luminal cGMP inhibits the reabsorptive function of the mTAL through a pathway different from the intracellular cGMP production.

Absorption↗

Man-fly contact in the Gambian trypanosomiasis focus of Nola-Bilolo (Central African Republic).

A study using bipyramid tetse fly traps in the Nola-Bilolo sleeping sickness focus (Central African Republic) reveals ecological and behavioural differences between two vectors, Glossina palpalis palpalis and G. fuscipes fuscipes. The latter species inhabits mainly open water sites and surrounding forest, whereas G. p. palpalis occurs mainly in coffe plantations near villages. Consequently, the man-fly contact differs considerably according to the species. The intensity of trypanosomiasis transmission, estimated by the probable distribution of cases, showed significant positive correlation with the density of the flies. Analysis of the fly blood meals in two villages show that, unlike G. g. palpalis, G. f. fuscipes feeds on men more than on pigs. Trypanosoma vivax infection was observed only in G. fuscipes fuscipes. The differences in occupation of the environment between the two vectors must be taken in account in trapping programmes which may modify this distribution.

Animals↗

[An epidemiological survey to discover the probable places of infection with sleeping sickness in the Central African Republic].

The sleeping sickness focus at Nola-Bilolo in the forest region was the subject of a survey of 142 patients in order to discover their probable places of infection. Seventy of these patients had been detected by active case-finding during a survey conducted in January-February 1991; the other 12 had been discovered by passive case-finding during 1990. The sample of actively detected patients is significantly younger (average 22 +/- 3 years) than the sample detected passively (30 +/- 3 years). This survey highlights the geographical heterogeneity of transmission in the focus. The places where infection is highest are M'Poyo, which with 18% of cases forms the epicentre of the focus and the main reservoir of infecting flies, followed by Bilolo (9%), Mékara (8%), Modigui-Kouna, Ziendi and Domissili (7%). The patients, 95% of whom belong to the M'Bimou tribe, all engage in agriculture; the main activities are coffee-growing for men and the steeping of casava in the river for women. These two activities determine the main places and times of human-tsetse contact. The information obtained by listing the cases detected passively from medical registers is inadequate for locating the places of transmission. The information gathered during active case-finding campaigns by the mobile teams offers a better approach. The method used here, which is recommended, consists of matching the replies given by the patients about their points of contact with the tsetse flies, not just with their places of residence but also with their successive movements and their places of work. The rigour demanded by this kind of approach can only be ensured by a specific survey. This survey also made it possible to demonstrate substantial movements from village to village, which appear to be characteristic of this focus. The results indicate that poorly targeted vector control is likely to be doomed to failure. The study makes it possible to specify priority areas for vector control by trapping and to avoid a "blind" control strategy whereby the whole focus is swamped with traps. The latter strategy is not financially feasible for many countries of tropical Africa.

Adolescent↗

[Giant cell fibroblastoma. Apropos of 2 cases].

Two cases of giant cell fibroblastoma (GCF) are reported. One presented as cervical tumor in a 11 year old child and the second localized in axillar region of 14 year old boy. Histologically both showed a typical distinctive appearance of this entity when focal fusocellular cells arranged in a storiform pattern was also constated in one. The immunohistochemical and ultrastructural study ruled out a vascular origin. The clinical and pathological findings suggest that CGF represent a juvenile form of dermatofibrosarcoma protuberans.

Adolescent↗

Binding to DNA, cellular uptake and biological activity of a distamycin-ellipticine hybrid molecule.

A hybrid molecule which conjugates the minor groove binding agent distamycin and an ellipticine derivative was synthesized and evaluated for cytostatic and cytotoxic activities against L1210 leukaemia cells in vitro. The binding of the hybrid molecule, named 'Distel', to a range of natural DNAs and synthetic polynucleotides with different base pair arrangements was studied by electric linear dichroism. The interaction with DNA simultaneously implicates binding of the distamycin part in the minor groove and intercalation of the ellipticine chromophore. The drug binds to DNA without any apparent preference for AT or GC polynucleotides, and can accommodate both homopolymeric and co-polymeric sequences as a binding site. However, the geometry of the drug-DNA complex varies depending on the targeted sequence. The lower activity of the hybrid as compared to the ellipticine derivative cannot be explained in terms of DNA binding. Taking advantage of the fluorescence of the pyridocarbazole chromophore, fluorescence microscopy was used to map cellular uptake of the hybrid molecule compared to the ellipticine derivative. Both the conjugate and the ellipticine derivative preferentially accumulate in the nuclei of HeLa cells rather than in the cytoplasm. Nuclei of ellipticine derivative-treated cells appear markedly more fluorescent than those of cells treated with the hybrid, which seems to be preferentially located in the nucleoli. Therefore, we consider the possibility that the difference in cytotoxicity between the two ellipticine-containing drugs is due to different intranuclear concentrations of these two compounds.

Animals↗

[Superinfection of ovarian dermoid cyst. Apropos of a case].

One case of a rare complication of dermoid ovarian cyst is reported: infection. Sonographic examination was not helpful, mainly due to a great quantity of gas within the mass. However radiography of the abdomen with the patient upright and CT showed a gas-fluid level, teeth and presence of fat. These findings permitted diagnosis of a complicated ovarian dermoid cyst before surgery.

Adult↗

Nevi, other than dysplastic and Spitz nevi.

Cutaneous nevi are common lesions that develop by proliferation of melanocyte-derived cells. The majority develop as junction nevi from melanocytes at the epidermo-dermal junction. Cells from this proliferation pass into the underlying dermis forming compound nevi. Later junctional melanocytic activity ceases, leaving an intradermal nevus. A minority of nevi, mainly blue nevi, arise from intradermal melanocytes. Histological variants of melanocytic nevi exist and can be the source of difficult diagnostic problems. Nevi are important as clinical and histological simulators of cutaneous melanoma, as precursor lesions for melanoma (although the actual chance of malignant transformation of an individual nevus is low) and as cosmetic problems (mainly large congenital nevi). Cutaneous nevi are to be separated clinically and histologically from melanomas that are comprised of nevocyte-like cells (minimal deviation melanoma).

Adult↗

The 2-amino group of guanine is absolutely required for specific binding of the anti-cancer antibiotic echinomycin to DNA.

The 2-amino group of guanine is believed to be a critical determinant of potential DNA binding sites for echinomycin and related quinoxaline antibiotics. In order to probe its importance directly we have studied the interaction between echinomycin and DNA species in which guanine N(2) is deleted by virtue of substitution of inosine for guanosine residues. The polymerase chain reaction was used to prepare inosine-substituted DNA. Binding of echinomycin, assessed by DNAse I footprinting, was practically abolished by incorporation of inosine into one or both strands of DNA. We conclude that both the purines in the preferred CpG binding site need to bear a 2-amino group to interact with echinomycin.

Base Sequence↗