Mutación CCDC103: una variante rara como causa de discinesia ciliar primaria / CCDC103 mutation: a rare variant as cause of primary ciliary dyskinesia

Introducción. La discinesia ciliar primaria (DCP) es un conjunto de enfermedades caracterizadas por una disfunción de las estructuras ciliadas del epitelio bronquial y gonadal. Puede deberse a que los cilios sean inmóviles, tengan un movimiento ineficaz o, menos frecuentemente, a la ausencia total de los mismos. Como consecuencia de esta disfunción se producen las manifestaciones clínicas: bronquiectasias, sinusitis crónica y, en ocasiones, esterilidad en los varones. Se trata de la segunda causa más frecuente de enfermedad congénita respiratoria, por detrás de la fibrosis quística, con una herencia autosómica recesiva. Actualmente se están intentando identificar y definir los genes y mutaciones causantes de la enfermedad. Paciente. Se presenta un caso de DCP asociada a situs inversus en el que el diagnóstico se estableció por estudio genético. Presentamos el caso por la singularidad de la sospecha diagnóstica desde el periodo neonatal, por haberse realizado el diagnóstico a los 2 años, 9 meses (antes que la edad media habitual) y por la escasa prevalencia de la variante encontrada en la población europea. Resultados. Se identifica en este caso, en homozigosis, una variante: c.461A>C (p.H154P) en el gen CCDC103 (NM-213607.2), rara, ya que se encuentra en un 0,1% de la población europea (AU)

Introduction. Primary ciliary dyskinesia (DCP) englobes a set of diseases in which the microscopic cells in the respiratory and gonadal system do not function normally. This dysfunction is produced by immotile cilia, abnormal movement or, less frequent, absence of cilia at all. This ciliary dysfunction produces the different clinic manifestations, such as: chronic sinusitis, bronchiectasis and sometimes infertility in men. It usually follows autosomal recessive genetic inheritance. The genes responsible of this disorder are actually being identified. Patient. We present a case of DCP asociated to situs inversus, in wich the diagnosis was established with genetic study. This is a singular case, because the diagnosis was done sooner than the medium age (2 years and 9 months old), it was also suspected in the neonatal period and the low prevalence of this variant. Results. Exome sequencing identified a homozygous missense variant in CCDC103 (C.461A>C, p.H154P). This is a rare variant which only occurs in approximately 0.1% of european population (AU)

Does shaking increase the pressure inside a bottle of champagne?

Colas, beers and sparkling wines are all concentrated solutions of carbon dioxide in aqueous solvents. Any such carbonated liquid is ordinarily conditioned inside a closed bottle or a metal can as a liquid-gas 2-phase system. At thermodynamic equilibrium, the partial pressure of carbon-dioxide in the gas phase and its concentration in the liquid are proportional (Henry's law). In practical conditions and use (transport, opening of the container, exterior temperature change, etc.), Henry's equilibrium can be perturbed. The goal of this paper is to describe and understand how the system responds to such perturbations and evolves towards a new equilibrium state. Formally, we investigate the dynamics around Henry's equilibrium of a closed system, through dedicated experiments and modeling. We focus on the response to a sudden pressure change and to mechanical shaking (the latter point inspired the article's title). Observations are rationalized through basic considerations including molecular diffusion, bubble dynamics (based on Epstein-Plesset theory) and chemi-convective hydrodynamic instabilities.

Reduced capacitative calcium entry in the mesenteric vascular bed of bile duct-ligated rats.

In this work, we analyzed the interaction of nitric oxide (NO) with some of the mechanisms known to regulate intracellular calcium levels in order to gain insight into the mechanisms responsible for the reduced vascular pressor response to vasoconstrictors observed in an experimental model of liver cirrhosis. Specifically, we hypothesized that the entry of calcium through capacitative channels is defective in this model. The experiments were performed with isolated, Krebs-perfused and de-endothelialized mesenteric arterial bed of rats with bile duct ligation (4 weeks) and their controls. Pretreatment with thapsigargin to inhibit calcium uptake into sarcoplasmic reticulum potentiated the pressor responses to methoxamine, but the response of the cirrhotic vessels was significantly lower than that of the controls. Under the same conditions, perfusion of the mesenteries with zero calcium-Krebs resulted in lower pressor responses to methoxamine, especially in the mesenteries of the bile duct-ligated rats. To specifically analyze the entry of calcium through store-operated calcium channels, the pressor response to the addition of calcium was studied in mesenteries perfused with zero calcium-Krebs and in the presence of thapsigargin. Again, the response of the cirrhotic mesenteric beds was significantly lower than that of the control vessels. Under all these experimental conditions, the differences between control and cirrhotic responses were abolished by pretreatment with the NO synthesis inhibitor N(w)-nitro-L-arginine (NNA). These results indicate that, in the mesenteric bed of bile duct-ligated rats, an excess of nitric oxide interferes with the release of calcium from thapsigargin-sensitive internal stores and also reduces the capacitative entry of calcium into vascular muscular cells induced by the depletion of calcium from internal stores. This mechanism may have an important role in the reduced pressor response observed in the mesenteric vascular bed in cirrhosis.

Role of vascular nitric oxide in experimental liver cirrhosis.

One of the most important features of liver cirrhosis is the splanchnic and systemic arterial vasodilation, related to an increase in vascular capacity and an active vasodilation. This arterial vasodilation seems to be the consequence of the excessive generation of vasodilating substances, which also contributes to a lower than normal pressor response to circulating nervous or humoral substances. The following review analyzes the mechanisms responsible for the vascular hyporesponse to vasoconstrictors observed in the experimental models of liver cirrhosis. It has become increasingly clear that, among the great variety of substances studied, nitric oxide (NO) seems to be one of the main contributors to this vascular alteration, since elimination of the endothelium or inhibition of its synthesis corrects it. The mechanism by which NO interferes with the contractile apparatus in smooth muscle cells seems to be related to a direct effect on calcium entry from the extracellular space and release from the internal stores.

Electrically induced microflows probed by fluorescence correlation spectroscopy.

We report on the experimental characterisation of electrically induced flows at the micrometer scale through Fluorescence Correlation Spectroscopy (FCS) measurements. We stress the potential of FCS as a useful characterisation technique in microfluidics devices for transport properties cartography. The experimental results obtained in a model situation are in agreement with previous calculations (F. Nadal, F. Argoul, P. Kestener, B. Pouligny, C. Ybert, A. Ajdari, Eur. Phys. J. E 9, 387 (2002)) predicting the structure and electric-field dependency of the induced flow. Additionally, the present study evidences a complex behaviour of the probe nanobeads under electric field whose precise understanding might prove relevant for situations where nano-objects interact with an external electric field.

Interaction of nitric oxide with calcium in the mesenteric bed of bile duct-ligated rats.

We have analysed the interaction of NO with calcium in order to study the molecular mechanisms responsible for the vascular hyporesponse of liver cirrhosis. The experiments have been performed in the isolated and perfused mesenteric arterial bed of rats with bile duct ligation (BDL) and their controls. While perfusing the vessels with normal Krebs, methoxamine (MTX) or KCl produced a lower pressor response in the BDL mesenteries. The NO synthesis inhibitor N(w)-nitro-L-arginine (NNA) potentiated those responses and abolished the differences between groups. The administration of MTX under perfusion with zero calcium-Krebs, to analyse the intracellular release of calcium, also induced a lower response in the BDL mesenteries and NNA potentiated and normalized the response. To investigate calcium entry, the vessels were perfused with zero-calcium Krebs containing high potassium to open voltage-dependent calcium channels. Then, the addition of calcium (10(-1) - 3 x 10(-3) M) produced a lower pressor response in the BDL vessels, that was corrected by NNA. To study calcium entry through receptor-operated channels, the vessels were perfused with zero-calcium Krebs containing MTX. The addition of calcium elevated the perfusion pressure less in the BDL mesenteries than in the control and NNA potentiated the responses and eliminated the between groups differences. When calcium entry through both voltage- and receptor-operated channels was simultaneously analysed, similar results were obtained. In the mesenteric bed of bile duct ligated rats, an excess of nitric oxide affects vascular calcium regulation through an interaction with both calcium entry and intracellular calcium release.

Electrically induced flows in the vicinity of a dielectric stripe on a conducting plane.

We report a theoretical and experimental study of the hydrodynamic flow induced by an a.c. electric field in the vicinity of a dielectric stripe deposited on a conducting plate. In the theoretical part, we model the stripe as a small change of the surface capacitance of the plate, and a perturbative approach is used to perform the calculations. This approach predicts an outwards rectified electro-osmotic slip along the surface that generates two steady counter-rotating rolls, the size of which decreases with the frequency. In the experimental section, we use tracers to determine the structure of the flow and investigate its dependence on the frequency and the amplitude of the applied voltage. The structure and amplitude of the observed flow compares satisfactorily with the theoretical analysis. This could guide the design of surface-controlled flows and help to understand the collective behavior of colloids near electrodes.

Mechanisms of the increased pressor response to vasopressors in the mesenteric bed of nitric oxide-deficient hypertensive rats.

In the present study we analyzed mesenteric vascular reactivity of chronic nitric oxide (NO)-deficient hypertensive rats (NW-nitro-L-Arginine Methyl Ester, L-NAME, 50 mg/kg/day, oral, 3 weeks). Perfusion pressure changes in response to cumulative additions of methoxamine and KCl were significantly increased in the mesenteric vessels of the L-NAME-treated as compared with vessels of the controls. Verapamil reduced the responses to methoxamine, but those of the hypertensive rats were still enhanced. In contrast, responses to KCl were almost completely abolished by verapamil. In mesenteric vessels perfused with zero calcium and high-potassium Krebs, pressor responses to the re-addition of calcium were also significantly enhanced in the hypertensive rats compared to the controls. Vasodilator responses to acetylcholine in KCl-preconstricted vessels, while still significant, were reduced in the L-NAME-treated rats. In this case, acute inhibition of NO blocked the vasodilator responses to acetylcholine and abolished the differences between the two groups. In methoxamine-preconstricted vessels and in the presence of acute inhibition of NO and prostaglandins, vasodilator responses to acetylcholine were significantly greater in the hypertensive vessels than in controls. In conclusion, the mesenteric vessels of L-NAME hypertensive rats show an enhanced response to vasopressors which is related to calcium entry. These data also reveal the existence of an enhanced role of a NO and prostaglandin-independent vasodilator factor, probably endothelium-derived hyperpolarizing factor that may play a compensatory role in the deficiency of NO.

Mesenteric hyporesponsiveness in cirrhotic rats with ascites: role of cGMP and K+ channels.

The mechanisms that mediate hyporesponsiveness to vasoconstrictors in liver cirrhosis are not completely established. In the present study we have explored the role of NO and potassium channels by studying the pressor response to methoxamine in rats with carbon tetrachloride-induced cirrhosis with ascites. Experiments were performed in the isolated and perfused mesenteric arterial bed of control rats and of cirrhotic rats with ascites. Pressor responses to methoxamine, an alpha-adrenergic agonist, were analysed under basal conditions, after inhibition of guanylate cyclase with Methylene Blue (MB; 10 microM), after inhibition of NO synthesis with N(G)-nitro-L-arginine (L-NNA; 100 microM) and after blockade of potassium channels with tetraethylammonium (TEA; 3 mM). Compared with those from controls, preparations from cirrhotic rats showed a lower pressor response to methoxamine (maximum: controls, 114.4+/-6.8 mmHg; cirrhotic rats, 74.7+/-7.3 mmHg). Pretreatment with MB or L-NNA increased the responses in both groups, but without correcting the lower than normal response of the cirrhotic rats. Pretreatment with TEA alone did not modify the responses as compared with the untreated groups. Pretreatment with TEA plus MB or TEA plus L-NNA also potentiated the responses, and the responses of the cirrhotic animals were greater than those of the groups treated with MB or L-NNA alone. However, no treatment completely normalized the lower response of the mesenteries from cirrhotic animals, suggesting that factors other than NO and potassium channels also participate, although to a lesser degree, in the lower pressor response of the mesenteric arterial bed of animals with cirrhosis. These results confirm that NO and potassium channels are important mediators of the lower vascular pressor response of the mesenteric bed of cirrhotic rats with ascites. This effect seems to be mediated by the NO-dependent formation of cGMP and by the NO-dependent and -independent activation of potassium channels.

Negative regulation of mitogen-activated protein kinase activation by integrin alphaIIbbeta3 in platelets.

Activation of the mitogen-activated protein (MAP) kinase pathway in nucleated cells is dependent on both growth factor receptors and integrins engaged in cell adhesion. Human platelets are an interesting model for studying cell adhesion and the involvement of integrin engagement on extracellular signal-regulated kinase (ERK) activation, independently from the nuclear-DNA signal pathway. Maximal phosphorylation and activity of ERK2 occurred late during thrombin-induced platelet aggregation (90 s and later), an alphaIIbbeta3 integrin-dependent event. Surprisingly, alphaIIbbeta3 inhibition by the RGDS ligand peptide, or (Fab')2 fragments of the AP-2 monoclonal antibody, resulted in a 2-fold enhancement in ERK2 phosphorylation and activity. A similar 2-fold enhancement of ERK2 activation was observed in thrombasthenic platelets which are defective in alphaIIbbeta3 and do not aggregate. This suggests that ERK2 activation in thrombin-induced platelet aggregation is dependent on thrombin rather than on alphaIIbbeta3 and is down-regulated by alphaIIbbeta3 engaged in ligand (fibrinogen) binding and/or aggregation. Finally, in the absence of stirring which allows fibrinogen binding to alphaIIbbeta3 but prevents aggregation, ERK2 was again overactivated. This overactivation appears to be consecutive to inhibition of aggregation itself and to alphaIIbbeta3 ligand binding. We conclude that in platelets, alphaIIbbeta3 engaged in aggregation down-regulates thrombin-induced ERK2 activation. To our knowledge, this is the first report of a down-regulation of the MAP kinase pathway by integrin engagement.

[Delivery after 2 previous cesarean sections. A series of 41 uterine trials]. / Accouchement des utérus bicicatriciels. Une série de 41 épreuves utérines.

It has been possible to consider how delivery should be carried out in view of the progress that has been made handling scarred uteruses. Over 21 months 41 tests of uterine function have been authorized in our department out of 67 cases where there were two scars in the uterus (67%). 26 patients delivered vaginally (63.4%) and 5 had Caesarean sections because of failure of the test of the scar. When the relationship between the fetus and the pelvis was satisfactory, the fact that the cervix was not ripe and the presenting part was not engaged, did not prevent carrying out a trial of scar in 78% of cases. It is important to assess conditions continuously during labour and this assessment should include fetal heart monitoring, internal tokometry, fetal pH assessment and ultrasound of the scar at the onset of labour. Oxytocics had to be used in 96.2% of cases because there was at the outset marked dynamic dystocia. Epidural anaesthesia was used in 90.2% of cases. A full obstetric team must be present throughout the whole labour so that the conduct of the labour can be observed, and if necessary corrected quickly if anything in its progress is becoming abnormal. It has become reasonable to carry out tests of uterine scars even after two scars have been made in the uterus because of the absence of any maternal or fetal complications in this series or in the literature.

[Indications for operative hysteroscopy. A series of 418 interventions]. / Les indications de l'hystéroscopie opératoire. A propos d'une série de 418 interventions.

Over a period of 38 months, from October 1, 1987 to December 31, 1990, 418 surgical hysteroscopies were carried out following systematic pre-operative hysteroscopy. Two-hundred and eighty-two patients presenting with a benign intercavitary lesion were treated by transcervical endo-uterine resection (EUR). The efficacy of the surgical method was assessed from the control of hemorrhagic phenomena, the most frequent sign. 89.7 percent of the patients who presented with menometrorrhagia, had become asymptomatic from the first cycle after EUR and 83.5 percent remained asymptomatic after a mean follow-up period of 28 months, 71 endometrectomies were carried out by EUR during this period, after careful hysteroscopic and histological evaluation of the endometrium. The results after one year of cycles were satisfactory in 82 percent of patients. Twenty-two patients presenting with a uterine septum were treated by endoscopic hysteroplasty. In most cases, the authors used the tip of the Charriere 21 resector. The follow-up period exceeds 1 year in 15 patients, 11 of whom became pregnant, giving birth to 9 live infants, 43 cases of synechia were treated by hysteroscopy in the context of Ashermann's syndrome. The functional results were good in 90 percent of patients, with the restoration of normal cycles. Only limited results were obtained in fertility in cases of muscular or fibrous synechia. The risks linked to surgical hysteroscopy are analysed. They appear to be acceptable, on condition that rigorous methods are used. The quality of sequelae and the short hospitalization make operative hysteroscopy preferable in the selected indications.

[Conservative treatment of cancer of the ovary]. / Le traitement conservateur du cancer de l'ovaire.

The authors assess the potential of conservative treatment in a series of cases of ovarian cancer. Surgical evaluation is essential, in order to confirm that the conditions required for conservative treatment are present. The tumor must be a stage IA tumor, with a low histoprognostic score and small in size. Conservative treatment must be proposed in cases of tumors of potential malignancy and tumors of the sexual cords. It is debateable in epithelial tumors. The question of the necessity of further surgery to remove the genital tract after the desired pregnancies may be debateable in cases of epithelial tumors.

[Hysteroscopic endometrial reduction. Analysis of 85 cases]. / La réduction endométriale hystéroscopique. Analyse de 85 observations.

The authors report the results of their experience of hysteroscopic endometrial reduction through a retrospective series of 85 patients who underwent surgery between October 1988 and February 1991. The indication consisted of hemorrhage (menorrhagia and metrorrhagia) of endometrial origin in women with a group mean age of 46 years who had no intention of becoming pregnant in the future. The pre-treatment assessment was based on diagnostic hysteroscopy and was intended to eliminate neoplasm of the endometrium or its precursors. The endometrial reduction procedure always preserved an isthmic ring of endometrial tissue. No peroperative complication was reported. The mean duration of surgery was about 39 minutes. With a follow-up period of more than one year, bleeding had disappeared in 78% of cases. The authors conclude that the technic of endometrial resection in the treatment of benign lesions of the endometrium, on condition that the possibility of any disorder calling for laparotomy is ruled out and that there is regular postoperative follow-up.

[Effects of preinduction by mifepristone on therapeutic abortions of the second and third trimesters induced by intravenous sulprostone]. / Effets d'une préinduction par la mifépristone sur les interruptions médicales des deuxième et troisième trimestres de grossesse réalisées par sulprostone par voie intraveineuse.

The authors investigate the effects of preinduction by administering 600 mg of mifepristone (RU486) per os before medical termination of pregnancy during the third to ninth months of pregnancy by intravenous sulprostone (Nalador). The study included a population of 35 patients treated with RU486 and then Nalador versus a population of 38 patients treated with Nalador only. The characteristics of the two groups did not show any statistically significant differences (medical, surgical and obstetric difference, indication for termination). The gestational age was lower in the RU486-Nalador group (group mean age : 20.9 weeks of amenorrhea versus 23.3, p less than 0.01). The group receiving RU486 significantly demonstrated the following : reduced time for induction of labour to modifications of the cervix (7.2 h versus 10.9 h, p less than 0.05), a higher number of rapid expulsions (within 10 h in 37.1% versus 15.7%, p less than 0.05) and a reduction of side effects (45.7% versus 71.05%, p less than 0.05). The other parameters analysed (duration of termination, total dose of prostaglandin) appeared to be better with RU486, but the values were not significantly different.

[Triplet pregnancies in France. Results of a retrospective, multicenter study of two years (1987-1988). Proposals for optimal management]. / Les grossesses triples en France. Résultats d'une enquête rétrospective multicentrique sur deux ans (1987-1988). Propositions pour une prise en charge optimale.

The explosion of procedures for medically assisted parenthood (MAP) has resulted in a previously little known type of pregnancy: triple pregnancies. In order to assess the current obstetrical and pediatric situation, the authors have carried out a major retrospective, multicenter survey in France concerning the triple pregnancies from 1987 to 1988: 156 case histories have been collected. The finds are compared with those reported in the French and international literature, showing that in France, three-quarters of such pregnancies result from MAP, mainly due to ovulation-inducing agents. Hospitalization is prolonged (averaging 27 days), early (24 WA) and imposed by complications (in 8 out of 10 cases), prophylactic hospitalization being rarely prescribed. The main complications encountered are late miscarriages (1.9%), in-utero death (6.41%), dysgravidia (16.6%), hydramnios (5.12%) and serious cardio-pulmonary complications related to the use of beta-mimetics. A Cesarian is performed in only 87 percent of cases. The mean birth weight was 1,776 g. Neonatal mortality is on the decline, but still equivalent to 80.3 per thousand. Hypotrophy is common (27%). Birth is nearly always premature (99.3%), but very early prematurity (28-32 WA) was reduced. The approach suggested for optimum management is based on the personal experience of the authors, the findings of their survey and of the international literature. It is based on the prevention of prematurity and a fundamentally multidisciplinary approach.

[Treatment of urinary stress incontinence by a modified Pereyra procedure:Chrub's operation]. / Traitement de l'incontinence urinaire d'effort par un procédé de Pereyra modifié: l'intervention de Chrub.

The authors report the utility of suspending the cervix by means of the pubo-urethrax ligaments anchored to Cooper's ligaments in treating exercise-related urinary incontinence. A prospective series of 40 female patients gave 97.5% of successful outcomes with regard to the incontinence. The use of the pubo-urethrax ligaments and the absence of subsphincterian dissection confers major urodynamic advantages, particularly with regard to closing pressure. These anatomical and physiological arguments appear to give the procedure an advantage over suspensions using the vagina wall, whether these make use of the upper or lower route. Anchoring to Cooper's ligaments should ensure long-term stability of the outcome.

[The results of urethro-cervicolysis after surgical over-correction of stress urinary incontinence. Apropos of 10 cases]. / Les résultats des urétro-cervicolyses après hypercorrection dans la chirurgie de l'incontinence urinaire d'effort. A propos de dix observations.

Between 1985 and 1988, 10 female patients between the ages of 35 and 76 years, suffering from dysuria after surgical correction of urinary stress incontinence (7 sub-urethral bands, 3 colposuspensions) were observed. The abnormality of the course of the urethra compressed anteriorly against the symphysis pubis, observed on clinical examination and on intravenous pyelography as well as urodynamic investigations allowed this dysuria to be attributed to a cervico-urethral obstruction resulting from over correction during the initial surgical operation. The 10 patients were treated by urethro-cervicolysis (9 out of 10) associated with repeated colposuspension in 3 cases. The overall results were disappointing as although 8 of the 10 patients were improved at 3 month, only 4 out of 9 were still improved at 6 months. These poor results raise the question of the value of combining urethro-cervicolysis with colposuspension and possible VY plasty of the bladder neck.