Evaluation of protective and restoring effects of a mixture of silanols on photoaging. Use of a device allowing the quantification of contractile strengths of human fibroblasts after UVA irradiation.

Chronic sun exposure and especially UVA wavelengths are responsible for long-term clinical skin changes such as photoageing and photocancers. The objectives of the present study were to analyse the contractile activity of fibroblasts irradiated with several doses of UVA and to evaluate the preventive, protective and restoring effects of a mixture of monomethylsilanetriol mannuronate and dimethylsilanediol salicylate. The forces generated by fibroblasts in tense collagen lattices were quantified using Glasbox device before and after UVA irradiation and the addition of a mixture of monomethylsilanetriol mannuronate and dimethylsilanediol salicylate. The production of collagen was also evaluated before and after irradiation and with and without the presence of a mixture of monomethylsilanetriol mannuronate and dimethylsilanediol salicylate. A dose of 3 J cm(-2) of UVA showed more than 50% of mortality in fibroblast population after 48 h and significant decreases in contractile forces developed by irradiated fibroblasts and collagen I production. One percentage of a mixture of monomethylsilanetriol mannuronate and dimethylsilanediol salicylate protected fibroblasts from UVA irradiation and made it possible to restore their capacity to the same level as fibroblasts that were not irradiated. It also tended to restore the capacity to synthesize collagen I. These results show that the use of the new device Glasbox makes it possible to evaluate a possible preventive and repairing effect of a cosmetic functional active on photoageing.

Carbachol and oxytocin stimulate the generation of inositol phosphates in the guinea pig myometrium.

In the guinea pig myometrium prelabelled with myo-[2-3H]inositol, carbachol and oxytocin enhanced a concentration-dependent and rapid release of IP3 which preceded that of IP2 and IP1. The specific receptor-mediated phospholipase C activation degrading PIP2 to IP3 did not require the presence of extracellular Ca2+. The ionophore A23187 as well as K+ depolarization failed to increase inositol phosphate accumulation. It is proposed that IP3 could have a role in the contraction of uterine smooth muscle elicited by the activation of muscarinic as well as of oxytocin receptors.

Characterization of G proteins in rat myometrium. A differential modulation of Gi2 alpha and Gi3 alpha during gestation.

Myometrial membranes, obtained from estrogen-dominated (day 0) rat uteri, were immunoblotted with antiserum (SG1), which recognizes the alpha subunits of both Gi1 and Gi2, with antiserum (LE2) specific for Gi2 alpha, and with I3B antiserum, specific for Gi3 alpha. The data revealed the absence of detectable levels of Gi1 alpha and the simultaneous presence of Gi2 alpha and Gi3 alpha as Gi subunits in rat myometrium. The expression of Gi proteins during gestation (days 0, 12, 21) was studied with the above antibodies. No qualitative change in the nature of Gi alpha species was observed during gestation: Gi1 alpha remained undetectable, Gi2 alpha and Gi3 alpha were both present on days 12 and 21. Of significance was the increase (160%) in the amount of Gi2 alpha at midgestation (day 12) compared to days 0 and 21. A different pattern was observed with Gi3 alpha, which decreased with advancing gestation (day 0 greater than 12 greater than 21). Immunodetection of beta subunits of G proteins indicated the presence of a 35/36 kDa doublet on days 0, 12 and 21, with an increase at midgestation. The simultaneous increase in Gi2 alpha and beta subunits may provide an explanation for the previously demonstrated alteration in adenylate cyclase stimulability detected at midgestation.

Identification of both Gi2 and a novel, immunologically distinct, form of Go in rat myometrial membranes.

Immunoblotting of rat myometrial membranes with an antiserum (SG1) which recognizes the alpha-subunits of both Gi1 and Gi2 indicated the presence of detectable levels of an apparently single form of some 40 kDa. A second antiserum (LE2) specific for Gi2 also recognized this protein, confirming its identity. Immunoblotting of the myometrial membranes with a series of antipeptide (OC1, IM1, ON1) and polyclonal (RV3) antisera, all of which recognize rat brain Go, produced a more complex pattern. Antisera OC1 and ON1 recognized a single polypeptide which essentially comigrated with rat brain Go. In contrast, antisera RV3 and IM1 did not recognize the myometrial protein. These data provide evidence for the presence of Gi2 and of a novel G-protein, related immunologically to Go, in rat myometrial membranes.

Differential role of microtubules in the control of prostaglandin E2 and beta-adrenergic stimulation of cyclic AMP accumulation in the rat myometrium.

A possible modulatory role of microtubules was investigated for the beta-adrenergic and prostaglandin E2 (PGE2)-induced cyclic AMP accumulation in the estrogen-treated rat myometrium. Colchicine, vinblastine and nocodazole, three different antitubulin drugs, enhanced cyclic AMP accumulation caused by PGE2. The effect of inhibitors was dose-(0.1-5 microM) and time-dependent, increased maximal responses without changing EC50 for PGE2, did not occur with trimethyl-colchicinic acid, which does not bind to tubulin, and was totally prevented in tissues pretreated with taxol, an agent that enhances polymerization and stabilization of microtubules. Concomitantly, colchicine reduced the rate and extent of PGE2-induced refractoriness in terms of cyclic AMP. In contrast, the antitubulin drugs failed to affect the rise in cyclic AMP evoked by isoproterenol and cholera toxin but enhanced the response to prostacyclin (PGI2), which is assumed to share common receptors with PGE2. Colchicine and vinblastine also failed to alter the ability of the beta-adrenergic agonist to provoke a cyclic AMP refractory state. Stimulations induced by all effectors were totally insensitive to cytochalasin B. The data suggest a relation between the constraints associated with the microtubules and/or membrane tubulin of the myometrium and the efficacy of PGE2 and PGI2 (but not the beta-adrenergic agonist or cholera toxin) to interact with the cyclic AMP forming system.

Modulation of cyclic AMP content of the rat myometrium: desensitization to isoproterenol, PGE2 and prostacyclin.

Exposure of the oestrogen-dominated rat myometrium to either isoproterenol or PGE2 resulted in a rapid but transient accumulation of cyclic AMP, with a progressive loss of responsiveness to the corresponding agonist. Induction of refractoriness was a time- and dose-related phenomenon. In the earliest time, desensitization was agonist-specific but was followed, with continued exposure, by a cross desensitization between isoproterenol and PGE2 and vice versa. Differential time courses for development and reversal of specific and heterologous refractoriness indicate at least 2 different processes for the 2 phenomena, the non-specific type being possibly mediated by cyclic AMP. Exposure to isoproterenol or PGE2 also caused an attenuated cyclic AMP response to prostacyclin (PGI2). Kinetics for PGE2-induced desensitization to PGI2 were comparable to that of an agonist-specific refractoriness, indicating that PGE2 and PGI2 may share common receptor sites. PGF2 alpha, PGD2 and 6-keto PGF1 alpha, which contract the myometrium but are ineffective on adenylate-cyclase activity, did not promote cyclic AMP refractoriness to PGE2, PGI2 or isoproterenol. Isoproterenol also caused refractoriness to its own relaxing activity, whereas PGE2 did not affect isoproterenol-induced relaxation despite a marked attenuation of the beta-adrenergic response to cyclic AMP. These results provide further evidence for the non-exclusive role of cyclic AMP in mediating uterine relaxation.

Cyclic AMP binding to intracellular receptor proteins in rat myometrium. Effect of epinephrine and prostaglandin E1.

In estrogen-pretreated rat myometrium, the relaxing effect exerted by theophylline or epinephrine has been correlated with their ability to raise cyclic AMP levels (Vesin and Harbon, 1974). The present study demonstrates that such a correlation can be quantitatively extended to the degree of saturation of intracellular cyclic AMP receptors. The rise in cyclic AMP induced by theophylline and/or epinephrine in intact myometrial strips was accompanied by a decrease in the ability of the corresponding extracts to bind exogenous 3H-labeled cyclic AMP. Total intracellular cyclic AMP binding sites were not modified and averaged a value of 0.22 muM. Accurate estimation of intracellular receptor-cyclic AMP complex has been correlated with the corresponding level of cyclic AMP in the tissue, the apparent intracellular Kd for cyclic AMP has been evaluated at 450 nm. Stimulation of myometrial strips with prostaglandin E1 (PGE1) which has been shown previously to induce contractions, although elevating cyclic AMP levels, was accompanied by a parallel increase in the saturation of the endogenous receptor, in an identical manner to that found with epinephrine or theophylline. The postulated hypothesis for a compartmentalization of cyclic AMP, or an interference of PGE1 with the intracellular cyclic AMP binding equilibrium has not been verified. The cyclic AMP system cannot be considered as the exclusive mechanism regulating uterine relaxation.

[Olfactory esthesioneuroma. Clinical, histological and therapeutic aspects; apropos of 6 cases]. / Les esthésioneuromes olfactifs. Aspects cliniques, histologiques, et thérapeutiques; à propos de six observations.

On the basis of 6 cases, the authors review the clinical, histological and therapeutic aspects of olfactory esthesioneuromas. These rare tumours, showing varying rates and degrees of progression from one patient to another, generally have a severe prognosis. Diagnosis is based upon precise histological criteria which may be clarified by electromicroscopic data. In difficult cases it may be useful to seek the aid of immuno-histochemical techniques in order to demonstrate the presence in tumour cells of specific neuronal enolase and the labelling of such cells by anti-protein S-100 antibodies. The treatment of choice would appear to be radio-surgical completed by chemotherapy similar to that used in neuroblastomas.

[Value of a scapular flap in the treatment of axillary hidradenitis. Apropos of a clinical case]. / Intérêt du lambeau scapulaire dans le traitement des hidrosadénites axillaires. A propos d'un cas clinique.

A severe case of axillary hidradenitis suppurativa, treated bilaterally by large excision followed by reconstruction with a pedicled scapular flap is presented. This reliable and thin fasciocutaneous flap provides good cover of large axillary defects with minimal donor site consequences. Its advantage is discussed versus the other surgical procedures used in this pathology for axillary reconstruction.

["Chinese hat" flap]. / Le lambeau "en chapeau chinois".

A design of a new type of advancement flap is described and called "chinese hat" flap. It uses the properties of the traditional advancement flap combined with rotation at the two distal edges, that overlap the height of the defect like two claws. This allows direct suture for larger defects with a prominent dog ear that become level with time. Two cases illustrate the procedure.

[Giant sacrococcygeal teratoma in adults]. / Tératome sacro-coccygien géant de l'adulte.

Sacro-coccygeal teratomas (SCT) are rare congenital tumors (1/40,000 births) but the commonest neoplasms in the newborn, generally discovered at birth; they are well known by pediatric surgeons who operate on them during the first days of life because of the high risk of malignant transformation when the diagnosis is delayed after several months of life. Exceptionally these tumors are observed in the adult without being diagnosed during infancy. The case of a 67 year old man is reported; the tumor presented as a solid mass complicated by necrosis and ulceration. The operation involved en bloc resection of the tumor and its sacro-coccygeal bony attaches. The tumor was found to be benign on histological examination. The main problems concerning diagnosis are discussed: possible association with meningocele and intra-pelvic extensions; these two factors must be defined preoperatively as they may change the approach to treatment. TSC can only be treated surgically, and the classical inverted V incision performed with the patient in the prone position is discussed versus other procedures designed a midline sacral scar, including Z plasty. Radical resection of the tumor with its pedunculated last two sacral and coccygeal vertebrae must be performed to avoid recurrence that may be malignant even if the primary lesion was benign.

Heterologous regulations of cAMP responses in pregnant rat myometrium. Evolution from a stimulatory to an inhibitory prostaglandin E2 and prostacyclin effect.

The regulation of cAMP generation in rat myometrium during gestation was investigated comparatively to the stimulations evoked in the estrogen-dominated myometrium. At the onset of gestation, there was a progressive attenuation in both tissue cAMP accumulation and adenylate cyclase activation in response to isoproterenol and prostaglandins (PGs) (PGE2 and prostacyclin, PGI2), as well as to cholera toxin and to forskolin. Minimal responses were observed at midgestation (day 12). The decline in isoproterenol-mediated cAMP accumulation could not be ascribed to alterations in either beta-adrenergic receptor density or coupling properties. Treatment of day 12 myometrium with islet-activating protein (IAP), pertussis toxin, caused a reversal of the attenuated beta-adrenergic--as well as cholera toxin--responses, suggesting that the inhibitory regulatory protein, Gi, was involved. However IAP treatment did not improve the PGE2-mediated stimulatory effect. In membranes from day 12 myometrium, the amount of [32P]NAD incorporated by IAP into the Mr = 41,000 peptides (alpha i, subunit of Gi was markedly increased compared to membranes from day 0 tissue. There was also a consistent decrease (25%) of the label incorporated by cholera toxin into the Mr = 52,000 (major) and 45,000/53,000 (minor) peptides (alpha s, subunit of Gs). The advanced stages of gestation were associated with a progressive restoration of cAMP responses to isoproterenol, cholera toxin, and forskolin with a full responsiveness before parturition (day 22). By contrast, the inability of PGE2 to generate any active Gs (stimulatory regulatory protein)-C complex persisted and coincided with the development of PGE2-induced inhibition of cAMP generation due to isoproterenol. The inhibitory effect, which was similarly evoked by PGE2 as well as by PGI2 and PGF2 alpha, was prevented by IAP. The data suggest that alterations of the stimulatory Gs-mediated pathway, which culminated at midgestation, is due at least in part to an increase in the abundance of Gi relative to Gs. Additional alterations operate at the level of the prostaglandin receptor(s) with a conversion from a stimulatory (Gs-mediated) cAMP response in the estrogen-dominated myometrium to an inhibitory (Gi-mediated) effect, fully expressed in the final stage of gestation.

Pharmacological evidence for distinct muscarinic receptor subtypes coupled to the inhibition of adenylate cyclase and to the increased generation of inositol phosphates in the guinea pig myometrium.

In the guinea pig myometrium, muscarinic receptor activation leads to contraction and elicits two biochemical responses viz. an increased formation of inositol phosphates (via a guanine nucleotide regulatory protein, distinct from the stimulatory and inhibitory G proteins of the adenylate cyclase system and a decreased synthesis of cyclic AMP involving inhibitory G protein activation. We now describe two major differences in the effects of muscarinic agonists. First, the greater potency of carbachol in inhibiting cyclic AMP formation (EC50 = 8 nM) than in stimulating the accumulation of inositol phosphates and tension (EC50 = 15 and 2 microM, respectively). Second, carbachol, oxotremorine and pilocarpine were equally effective in eliciting cyclic AMP inhibition but the order of potency for inositol phosphate formation was carbachol greater than oxotremorine and pilocarpine was without effect. The partial agonists, pilocarpine and oxotremorine, inhibited carbachol-mediated inositol phosphate formation. Pirenzepine, selective for muscarinic M1 receptor subtype, displayed a low affinity for antagonizing cyclic AMP inhibition, inositol phosphate generation and tension due to carbachol (Ki = 286, 92 and 110 nM, respectively). AF-DX116 (11-[( 2-[(diethylamino)methyl]-1- piperidinyl]acetyl)-5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepine- 6-one), selective for cardiac M2 receptors blocked cyclic AMP inhibition with high affinity (Ki = 1.14 nM) while it antagonized inositol phosphate formation with low affinity (Ki = 346 nM). Both high (Ki = 1 nM) and low (Ki = 100 nM) affinities were displayed by AF-DX116 in antagonizing contractions due to carbachol (24 and 76% inhibition, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Fluoroaluminates mimic muscarinic- and oxytocin-receptor-mediated generation of inositol phosphates and contraction in the intact guinea-pig myometrium. Role for a pertussis/cholera-toxin-insensitive G protein.

1. In the intact guinea-pig myometrium, carbachol and oxytocin stimulated a specific receptor-mediated phospholipase C activation, catalysing the breakdown of PtdIns(4,5)P2 with the sequential generation of InsP3, InsP2 and InsP. Stimulation of muscarinic receptors also triggered an inhibition of cyclic AMP accumulation caused by prostacyclin. 2. NaF plus AlCl3 mimicked the effects of carbachol and oxytocin by inducing, in a dose-dependent manner, the generation of all three inositol phosphates as well as uterine contractions. AlCl3 enhanced the fluoride effect, supporting the concept that A1F4- was the active species. Under similar conditions, fluoroaluminates activated the guanine nucleotide regulatory protein Gi, reproducing the inhibitory effect of carbachol on cyclic AMP concentrations. 3. Both carbachol- and oxytocin-mediated increases in inositol phosphates, as well as contractions, were insensitive to pertussis toxin, under conditions where the expression of Gi was totally prevented. Cholera toxin, which activates Gs and enhances cyclic AMP accumulation, failed to affect basal or oxytocin-evoked inositol phosphate generation, but induced a slight, though consistent, attenuation of the muscarinic inositol phosphate response, which was similarly evoked by forskolin. 4. The data provide evidence that, in the myometrium, (a) a G protein mediates the generation of inositol phosphates and the Ca2+-dependent contractile event, (b) the relevant G protein that most probably couples muscarinic and oxytocin receptors to phospholipase C is different from Gi and Gs, the proteins that couple receptors to adenylate cyclase, and (c) cyclic AMP does not seem to control the phosphoinositide cycle, but rather exerts a negative regulation at the muscarinic-receptor level.

Pervanadate mediated an increased generation of inositol phosphates and tension in rat myometrium. Activation and phosphorylation of phospholipase C-gamma 1.

Stimulation of [3H]inositol-labeled rat myometrial strips with pervanadate, formed by mixing orthovanadate and H2O2, induced a dose-dependent accumulation of [3H]inositol phosphates. Orthovanadate or H2O2 added alone had no effect. Pretreatment of myometrium with two tyrosine kinase inhibitors, namely genistein and tyrphostin 47 (at 100 microM), reduced pervanadate-stimulated inositol phosphate formation by 50%. Pervanadate induced a time-sequential formation of inositol phosphates in the order inositol trisphosphate, inositol bisphosphate, and inositol monophosphate. The inhibitory effect of genistein was observed at the level of the three inositol phosphates. Pervanadate induced contraction of the myometrium; the response was dose-dependent. H2O2 or orthovanadate was without effect. Pervanadate-mediated contraction was inhibited (50%) by genistein and tyrphostin 47 (100 microM). Western blot analysis, using anti-phosphotyrosine antibodies, revealed that phosphorylated proteins were present in detergent extracts from pervanadate-stimulated myometrium. Tyrosine phosphorylation was reduced by a preincubation with 100 microM genistein or tyrphostin 47. Phospholipase C-gamma1 was immunodetected in myometrial extracts and was identified as one of the substrates subject to tyrosine phosphorylation following pervanadate treatment. The results demonstrate that, in myometrium, protein tyrosine kinase/phosphatase activities controlled both phosphorylation and activation of phospholipase C-gamma1, contributing to the modulation of the generation of inositol phosphates and tension.

[SIN-1 interactions with the generation of cyclic nucleotides and arachidonate oxide metabolites in the uterine muscle]. / Interactions du SIN-1 avec la génération des nucléotides cycliques et des métabolites oxydés de l'arachidonate dans le muscle utérin.

In the uterine smooth muscle, SIN-1 stimulated cGMP accumulation independently of the presence of Ca2+ and activated the soluble form of guanylate-cyclase through mechanisms apparently similar to those involved in the stimulations evoked by NO-containing compounds. These activations appear different from those induced by hydroperoxy-unsaturated fatty acids and which contribute to the carbachol-mediated cGMP accumulation. SIN-1 did not influence the rise in cAMP of the biosynthesis of PG1(2) and 12-HETE due to exogenous arachidonic acid. By contrast, SIN-1 markedly inhibited the increased synthesis of PG1(2) induced by the ionophore A23187 which was due to a prior, Ca2+-dependent, liberation of endogenous arachidonic acid. The data suggests an interference of SIN-1 with the generation and/or the expression of the Ca2+ signal.

[Morbidity of iliac bone grafts. A study apropos of 100 consecutive cases]. / Morbidité des prises de greffes osseuses iliaques. Etude à propos de 100 cas consécutifs.

A study of the real morbidity after iliac bone graft harvesting was conducted on a homogenous series of 100 consecutive cases. Functional and aesthetic consequences were evaluated in relation to the immediate post-operative course and the long-term follow-up and in relation to the indication, the technique used and the amount of bone removed. Considering the small number of sequelae observed, autogenous iliac bone graft remains the best material for craniomaxillofacial reconstruction. The main disadvantage consists of the resorptions noted; which raises the possibility of using other types of bone grafts or bio-materials in some indications.

Carbachol-induced desensitization of PLC-beta pathway in rat myometrium: downregulation of Gqalpha/G11alpha.

In the estrogen-treated rat myometrium, carbachol increased the generation of inositol phosphates by stimulating the muscarinic receptor-Gq/G11-phospholipase C-beta3 (PLC-beta3) cascade. Exposure to carbachol resulted in a rapid and specific (homologous) attenuation of the subsequent muscarinic responses in terms of inositol phosphate production, PLC-beta3 translocation to membrane, and contraction. Refractoriness was accompanied by a reduction of membrane muscarinic binding sites and an uncoupled state of residual receptors. Protein kinase C (PKC) altered the functionality of muscarinic receptors and contributed to the initial period of desensitization. A delayed phase of the muscarinic refractoriness was PKC independent and was associated with a downregulation of Gqalpha/G11alpha. Atropine failed to induce desensitization as well as Gqalpha/G11alpha downregulation, indicating that both events involve active occupancy of the receptor. Prolonged exposure to AlF-4 reduced subsequent AlF-4 as well as carbachol-mediated inositol phosphate responses and similarly induced downregulation of Gqalpha/G11alpha. Data suggest that a decrease in the level of Gqalpha/G11alpha is subsequent to its activation and may account for heterologous desensitization.