IRF6 is a mediator of Notch pro-differentiation and tumour suppressive function in keratinocytes.

While the pro-differentiation and tumour suppressive functions of Notch signalling in keratinocytes are well established, the underlying mechanisms remain poorly understood. We report here that interferon regulatory factor 6 (IRF6), an IRF family member with an essential role in epidermal development, is induced in differentiation through a Notch-dependent mechanism and is a primary Notch target in keratinocytes and keratinocyte-derived SCC cells. Increased IRF6 expression contributes to the impact of Notch activation on growth/differentiation-related genes, while it is not required for induction of 'canonical' Notch targets like p21(WAF1/Cip1), Hes1 and Hey1. Down-modulation of IRF6 counteracts differentiation of primary human keratinocytes in vitro and in vivo, promoting ras-induced tumour formation. The clinical relevance of these findings is illustrated by the strikingly opposite pattern of expression of Notch1 and IRF6 versus epidermal growth factor receptor in a cohort of clinical SCCs, as a function of their grade of differentiation. Thus, IRF6 is a primary Notch target in keratinocytes, which contributes to the role of this pathway in differentiation and tumour suppression.

IRF6 is a mediator of Notch pro-differentiation and tumour suppressive function in keratinocytes.

While the pro-differentiation and tumour suppressive functions of Notch signalling in keratinocytes are well established, the underlying mechanisms remain poorly understood. We report here that interferon regulatory factor 6 (IRF6), an IRF family member with an essential role in epidermal development, is induced in differentiation through a Notch-dependent mechanism and is a primary Notch target in keratinocytes and keratinocyte-derived SCC cells. Increased IRF6 expression contributes to the impact of Notch activation on growth/differentiation-related genes, while it is not required for induction of 'canonical' Notch targets like p21(WAF1/Cip1), Hes1 and Hey1. Down-modulation of IRF6 counteracts differentiation of primary human keratinocytes in vitro and in vivo, promoting ras-induced tumour formation. The clinical relevance of these findings is illustrated by the strikingly opposite pattern of expression of Notch1 and IRF6 versus epidermal growth factor receptor in a cohort of clinical SCCs, as a function of their grade of differentiation. Thus, IRF6 is a primary Notch target in keratinocytes, which contributes to the role of this pathway in differentiation and tumour suppression.

Missale ad usum ecclesiae Parisiensis .

Cf. notice du ms. par Leroquais, Sacramentaires, II, 180-183 n° 366. Les incipit des proses ont été relevés dans la table des incipit. F. 1-6v Calendrier de Paris : 3 janv., en rouge, «Genovefe virg. IX lc.» (1); 22 avr., «Inventio corporis s. Dyonisii sociorumque ejus. Oportune virg. semid.» (2v); 28 mai, en rouge, «Germani ep. Parisiensis semid. Carauni mart. memoria» (3); 25juin, en violet, «Translatio s. Eligii ep.» (3v); en violet, «In prima dominica hujus mensis [augusti] fit d. de cruce» [réception à Notre-Dame d'un fragment de la vraie Croix] (4v); 9 oct., en rouge, «Dyonisii sociorumque ejus duplex» (5v); 28 oct., «Germani ep. Translatio s. Genovefe virg. mart. memoria» (5v); 3 nov. «Marcelli ep. Parisiensis dupl.»; 13 nov., «Gendulphi ep. et conf. [Parisiensis] dupl.»; 26 nov., en violet, «Genovefe virg. de miraculo [ardentium] IX lc.» (6); 4 déc., «Susceptio capillorum b. Marie et capitis b. Dyonisii in ecclesia Parisiensi» (6v). Le calendrier ne contient pas la fête de s. Louis au 25 août (établie en 1298) ni au 17 mai la translation du chef de s. Louis à la Sainte-Chapelle (en 1306); — Ajout du XIVe s. au 30 avr., «Eutropii mart. et conf.» [culte établi à Paris en 1296] (2v); cf. Leroquais, Bréviaires, I, CXII-CXIII, Tableau chronologique des fêtes parisiennes. — Au début de chaque mois, vers sur les jours égyptiaques (éd. Hennig, Traditio, XI (1955), 84 III), sauf celui de janvier (1) qui correspond au premier vers de la série éd. par Riese (Anthol. lat., I2 (1906), 680a); — À la fin de février, vers sur l'année bissextile : «Byssextum sexte martis tenuere kalende...»; cf. ms. Latin 3162, f. 102 (1v); — À la fin de mars, vers sur la date de Pâques : «Post martis nonas ubi sit nova luna requiras...» (2); — Notations astrologiques et de comput, passim. F. 7-353v Temporal, avec parties chantées notées. Incomplet du début par la perte du premier f., il commence dans la prose du premier dimanche de l'Avent : «... [eterna indefici]ens mundi vita...» (7). À remarquer : «dominica IIIa [in Adventu] more ecclesie Parisiensis» (24v); — Cérémonie des Cendres avec rubriques liturgiques (62v-66v); — Dimanche des Rameaux, «congregatis processionibus conventualibus in ecclesia B. Marie, capiceriis portantibus capsam et tribus clericis in albis paratis tres textus, exitur de ecclesia nichil cantando et sic eundum est ad ecclesiam Sancte Genovefe de Monte...» (147); cf. Leroquais, 181; — Dans l'évangile des Rameaux, le Christ est désigné par la lettre L, le narrateur par la lettre C et les autres par la lettre S (153-158); dans les autres évangiles de la Passion (162-186 passim), ces lettres suscrites ont été effacées et remplacées à la mine de plomb de façon sporadique, le Christ étant alors désigné par une croix; — «Oremus pro papa nostro N...» (186v); «... pro christianissimo rege nostro N...» (187); — Parmi les litanies, «s.Stephane... s. Dyonisi...» (191v)..., « s. Lucane... s. Justine... s. Gendulphe... s. Germane...» (199)..., « s. Genovefa...» (201v). — «Incipit ordo misse. Sacerdos primo induat se rocheto dicens : Actiones nostras...» (203v-206); pour les rubriques liturgiques, cf. Leroquais, 181; — Préfaces notées : «Incipiunt prefationes...» (206-210); — Canon de la messe (210-213); pour les rubriques liturgiques, cf. Leroquais, 182; — «In die Re[sur]rectionis» (214); — «In die sancto [Pentecostes]» (262v); — [De sancta Trinitate] (280v); — «Dominica XXVa» (348v); — «In dedicatione ecclesie» (350-353v). La fête du Saint-Sacrement ne figure pas. F. 354-496v Sanctoral, avec parties chantées notées. À noter parmi les saints parisiens : 3 janv., «S. Genovefe virg.» (373v); 22 avr., «Inventio corporis s. Dyonisii, Rustici et Eleuterii» (398v); — 28 mai, «S. Germani ep. Parisiensis» (407v); — 10 juin, «S. Landerici Parisiensis ep.» (410); — 26 juill., «In translatione s. Marcelli» (432); — 11août, «De receptione s. corone...» (443v); —, 4 oct., «Auree virg. [abb. Parisiis]» (480); 9 oct., «In die [s. Dyonisii]», avec octave (481v); — 28oct., «In translatione s. Genovefe» (487); — 3nov., «S. Marcelli Parisiensis ep.», avec octave (489v); — 26 nov., «S. Genovefe de miraculo ardentium» (495); — «De s. Gendulpho. Prosa» (495). F. 497-541 Commun des saints, avec pièces chantées notées. F. 541-559 Messes votives, sans parties notées à l'exception de la messe des morts (551v-556). Aux ff. 546v-547, en marge de la messe «pro amico», deux additions d'une écriture cursive de la fin du XIVe s. précisent : «ducem nostrum et duxissem ejusque prolem». — Prières diverses (556-559). F. 559-563 Rituel de mariage. «Incipit ordo ad sponsam benedicendam. Cum venerint ante valvas ecclesie sponsus et sponsa, accinctus sacerdos alba et stola... auxilium et argentum super scutum positum benedicat dicens : Manda Deus... Tunc aspergatur aqua benedicta et thurificetur et sponsus et sponsa; quo facto sacerdos dicat : Bones genz nos avons faiz les bans III foiz de ces II genz et encore les faison nous, que se il i a nul ne nule qui sache enpeschement par quoi l'un ne puisse avoir l'autre par loi de mariage, si le die. Et re[spon]dent assistentes : Nous ni savons se bien non. Quo audito, accipiat sacerdos manum dexteram sponse et ponat in dextera manu sponsi et dicat... nominandos eos : Vos Marie et vos Jeham vous prometez, fianciez et jurez l'un à l'autre à garder la foi et la loiauté du mariage... Tunc sacerdos tradat anulum sponso et sponsus autem per manum sacerdotis primo in police sponse... dicens... : Marie de cest annel t'espous et de mon cors t'ennor et de douaire qui est devisiez entre mes amis et les tiens. In nomine... Secundo in indice dicens... Tercio in medio dicens : Et Spiritus sancti. Amen... Postea sacerdos extensa manu super illos dicat orationes... Tunc sacerdos centus [sic] sponsum per manum dexteram et sponsam et introducat eos in ecclesiam...» (559-559bis); — Messe de mariage, comportant deux préfaces notées : «Deinde celebratur missa...» (559bis-562v); pour les rubriques liturgiques, cf. Leroquais, 183. — «Missa celebrata, recedant sponsus et sponsa et, ipsis stantibus ante hostium domus illorum presentibus pane et vinum [sic], faciat sacerdos benedictionem super panem dicens : Benedic Domine creaturam... Tunc sponsus mordet in pane, postea sponsa. Item benedictio super vinum... Tunc sponsus bibat, postea sponsa. Quo facto introducit eos sacerdos per manum in domum... Item in sero benedictio thalami... Tunc turificet thalamum, postea sponsum et sponsam sedentes vel jacentes in lecto suo benedicat dicens : Benedic Domine adulescentulos...» (562v-563) (éd. avec variantes par E. Martène, De antiquis Ecclesiae ritibus, II, 374-376, d'après le ms. Latin 859A); même ordo dans le ms. NAL 2649, f. 333-336. F. 563-565v Exorcisme de l'eau et du sel (563-564v). — «Pro Terra sancta» (564v-565). — «Pro rege nostro» (565). — «Benedictio pere et baculi peregrinorum» (565-565v). — «Benedictio crucis... Benedictio novorum fructuum... Benedictio panis» (565v).

A mutation causing pseudohypoaldosteronism type 1 identifies a conserved glycine that is involved in the gating of the epithelial sodium channel.

Pseudohypoaldosteronism type 1 (PHA-1) is an inherited disease characterized by severe neonatal salt-wasting and caused by mutations in subunits of the amiloride-sensitive epithelial sodium channel (ENaC). A missense mutation (G37S) of the human ENaC beta subunit that causes loss of ENaC function and PHA-1 replaces a glycine that is conserved in the N-terminus of all members of the ENaC gene family. We now report an investigation of the mechanism of channel inactivation by this mutation. Homologous mutations, introduced into alpha, beta or gamma subunits, all significantly reduce macroscopic sodium channel currents recorded in Xenopus laevis oocytes. Quantitative determination of the number of channel molecules present at the cell surface showed no significant differences in surface expression of mutant compared with wild-type channels. Single channel conductances and ion selectivities of the mutant channels were identical to that of wild-type. These results suggest that the decrease in macroscopic Na currents is due to a decrease in channel open probability (P(o)), suggesting that mutations of a conserved glycine in the N-terminus of ENaC subunits change ENaC channel gating, which would explain the disease pathophysiology. Single channel recordings of channels containing the mutant alpha subunit (alphaG95S) directly demonstrate a striking reduction in P(o). We propose that this mutation favors a gating mode characterized by short-open and long-closed times. We suggest that determination of the gating mode of ENaC is a key regulator of channel activity.

Cartilage thickness at the posterior medial femoral condyle is increased in femorotibial knee osteoarthritis: a cross-sectional CT arthrography study (Part 2).

OBJECTIVE: To evaluate the thickness of cartilage at the posterior aspect of the medial and lateral condyle in Osteoarthritis (OA) knees compared to non-OA knees using computed tomography arthrography (CTA). DESIGN: 535 consecutive knee CTAs (mean patient age = 48.7 ± 16.0; 286 males), were retrospectively analyzed. Knees were radiographically classified into OA or non-OA knees according to a modified Kellgren/Lawrence (K/L) grading scheme. Cartilage thickness at the posterior aspect of the medial and lateral femoral condyles was measured on sagittal reformations, and compared between matched OA and non-OA knees in the whole sample population and in subgroups defined by gender and age. RESULTS: The cartilage of the posterior aspect of medial condyle was statistically significantly thicker in OA knees (2.43 mm (95% confidence interval (CI) = 2.36, 2.51)) compared to non-OA knees (2.13 mm (95%CI = 2.02, 2.17)) in the entire sample population (P < 0.001), as well as for all subgroups of patients over 40 years old (all P ≤ 0.01), except for females above 60 years old (P = 0.07). Increase in cartilage thickness at the posterior aspect of the medial condyle was associated with increasing K/L grade in the entire sample population, as well as for males and females separately (regression coefficient = 0.10-0.12, all P < 0.001). For the lateral condyle, there was no statistically significant association between cartilage thickness and OA (either presence of OA or K/L grade). CONCLUSIONS: Cartilage thickness at the non-weight-bearing posterior aspect of the medial condyle, but not of the lateral condyle, was increased in OA knees compared to non-OA knees. Furthermore, cartilage thickness at the posterior aspect of the medial condyle increased with increasing K/L grade.

Improved smoking cessation in smokers given ultrasound photographs of their own atherosclerotic plaques.

BACKGROUND: We examined whether making smokers aware that they had developed peripheral atherosclerosis would improve smoking cessation. METHODS: Smokers selected from the general population were randomly allocated to undergo high-resolution B-mode ultrasonography of their carotid and femoral arteries. All smokers received quit-smoking counseling. Smokers with > or =1 atherosclerotic plaque were given two photographs of a plaque with a relevant explanation. Quit rates were assessed by telephone 6 months later. RESULTS: Seventy-nine smokers did not undergo ultrasonography (A). Among the 74 smokers submitted to ultrasonography, 20 had no plaque (B) and 54 had > or =1 plaque (C). Quit rates were, respectively, 6.3, 5.0, and 22.2% in groups A, B, and C. Quit rates were higher in smokers submitted to ultrasonography (B + C vs A; P = 0.031) and in those receiving photographs (C vs A + B; P = 0.003). Smoking cessation was independently associated with intervention C (OR = 6.2; 95% CI = 1.8-21) and a white-collar job but not with age or gender. CONCLUSIONS: Providing smokers with photographs demonstrating atherosclerosis on their own person was an effective adjunct to physician's advice to quit smoking. Since ultrasonography is used increasingly often in clinical practice for cardiovascular risk stratification, this can provide an additional opportunity and means to deter smokers from smoking.

Colon-specific deletion of epithelial sodium channel causes sodium loss and aldosterone resistance.

Aldosterone promotes electrogenic sodium reabsorption through the amiloride-sensitive epithelial sodium channel (ENaC). Here, we investigated the importance of ENaC and its positive regulator channel-activating protease 1 (CAP1/Prss8) in colon. Mice lacking the αENaC subunit in colonic superficial cells (Scnn1a(KO)) were viable, without fetal or perinatal lethality. Control mice fed a regular or low-salt diet had a significantly higher amiloride-sensitive rectal potential difference (∆PDamil) than control mice fed a high-salt diet. In Scnn1a(KO) mice, however, this salt restriction-induced increase in ∆PDamil did not occur, and the circadian rhythm of ∆PDamil was blunted. Plasma and urinary sodium and potassium did not change with regular or high-salt diets or potassium loading in control or Scnn1a(KO) mice. However, Scnn1a(KO) mice fed a low-salt diet lost significant amounts of sodium in their feces and exhibited high plasma aldosterone and increased urinary sodium retention. Mice lacking the CAP1/Prss8 in colonic superficial cells (Prss8(KO)) were viable, without fetal or perinatal lethality. Compared with controls, Prss8(KO) mice fed regular or low-salt diets exhibited significantly reduced ∆PDamil in the afternoon, but the circadian rhythm was maintained. Prss8(KO) mice fed a low-salt diet also exhibited sodium loss through feces and higher plasma aldosterone levels. Thus, we identified CAP1/Prss8 as an in vivo regulator of ENaC in colon. We conclude that, under salt restriction, activation of the renin-angiotensin-aldosterone system in the kidney compensated for the absence of ENaC in colonic surface epithelium, leading to colon-specific pseudohypoaldosteronism type 1 with mineralocorticoid resistance without evidence of impaired potassium balance.

The inositol Inpp5k 5-phosphatase affects osmoregulation through the vasopressin-aquaporin 2 pathway in the collecting system.

Inositol Inpp5k (or Pps, SKIP) is a member of the inositol polyphosphate 5-phosphatases family with a poorly characterized function in vivo. In this study, we explored the function of this inositol 5-phosphatase in mice and cells overexpressing the 42-kDa mouse Inpp5k protein. Inpp5k transgenic mice present defects in water metabolism characterized by a reduced plasma osmolality at baseline, a delayed urinary water excretion following a water load, and an increased acute response to vasopressin. These defects are associated with the expression of the Inpp5k transgene in renal collecting ducts and with alterations in the arginine vasopressin/aquaporin-2 signalling pathway in this tubular segment. Analysis in a mouse collecting duct mCCD cell line revealed that Inpp5k overexpression leads to increased expression of the arginine vasopressin receptor type 2 and increased cAMP response to arginine vasopressin, providing a basis for increased aquaporin-2 expression and plasma membrane localization with increased osmotically induced water transport. Altogether, our results indicate that Inpp5k 5-phosphatase is important for the control of the arginine vasopressin/aquaporin-2 signalling pathway and water transport in kidney collecting ducts.

HIV-1 Subtype Is an Independent Predictor of Reverse Transcriptase Mutation K65R in HIV-1 Patients Treated with Combination Antiretroviral Therapy Including Tenofovir.

Subtype-dependent selection of HIV-1 reverse transcriptase resistance mutation K65R was previously observed in cell culture and small clinical investigations. We compared K65R prevalence across subtypes A, B, C, F, G, and CRF02_AG separately in a cohort of 3,076 patients on combination therapy including tenofovir. K65R selection was significantly higher in HIV-1 subtype C. This could not be explained by clinical and demographic factors in multivariate analysis, suggesting subtype sequence-specific K65R pathways.

Effects of thyromimetic drugs on aldosterone-dependent sodium transport in the toad bladder.

Aldosterone increases transepithelial Na+ transport in the urinary bladder of Bufo marinus. The response is characterized by 3 distinct phases: 1) a lag period of about 60 min, ii) an initial phase (early response) of about 2 hr during which Na+ transport increases rapidly and transepithelial electrical resistance falls, and iii) a late phase (late response) of about 4 to 6 hr during which Na+ transport still increases significantly but with very little change in resistance. Triiodothyronine (T3, 6 nM) added either 2 or 18 hr before aldosterone selectively antagonizes the late response. T3 per se (up to 6 nM) has no effect on base-line Na+ transport. The antagonist activity of T3 is only apparent after a latent period of about 6 to 8 hr. It is not rapidly reversible after a 4-hr washout of the hormone. The effects appear to be selective for thyromimetic drugs since reverse T3 (rT3) is inactive and isopropyldiiodothyronine (isoT2) is more active than T3. The relative activity of these analogs corresponds to their relative affinity for T3 nuclear binding sites which we have previously described. Our data suggest that T3 might control the expression of aldosterone by regulating gene expression, e.g. by the induction of specific proteins, which in turn will inhibit the late mineralocorticoid response, without interaction with the early response.

A novel vasopressin-induced transcript promotes MAP kinase activation and ENaC downregulation.

In the principal cell of the renal collecting duct, vasopressin regulates the expression of a gene network responsible for sodium and water reabsorption through the regulation of the water channel and the epithelial sodium channel (ENaC). We have recently identified a novel vasopressin-induced transcript (VIT32) that encodes for a 142 amino acid vasopressin-induced protein (VIP32), which has no homology with any protein of known function. The Xenopus oocyte expression system revealed two functions: (i) when injected alone, VIT32 cRNA rapidly induces oocyte meiotic maturation through the activation of the maturation promoting factor, the amphibian homolog of the universal M phase trigger Cdc2/cyclin; and (ii) when co-injected with the ENaC, VIT32 cRNA selectively downregulates channel activity, but not channel cell surface expression. In the kidney principal cell, VIP32 may be involved in the downregulation of transepithelial sodium transport observed within a few hours after vasopressin treatment. VIP32 belongs to a novel gene family ubiquitously expressed in oocyte and somatic cells that may be involved in G to M transition and cell cycling.

Trabecular bone score: a noninvasive analytical method based upon the DXA image.

The trabecular bone score (TBS) is a gray-level textural metric that can be extracted from the two-dimensional lumbar spine dual-energy X-ray absorptiometry (DXA) image. TBS is related to bone microarchitecture and provides skeletal information that is not captured from the standard bone mineral density (BMD) measurement. Based on experimental variograms of the projected DXA image, TBS has the potential to discern differences between DXA scans that show similar BMD measurements. An elevated TBS value correlates with better skeletal microstructure; a low TBS value correlates with weaker skeletal microstructure. Lumbar spine TBS has been evaluated in cross-sectional and longitudinal studies. The following conclusions are based upon publications reviewed in this article: 1) TBS gives lower values in postmenopausal women and in men with previous fragility fractures than their nonfractured counterparts; 2) TBS is complementary to data available by lumbar spine DXA measurements; 3) TBS results are lower in women who have sustained a fragility fracture but in whom DXA does not indicate osteoporosis or even osteopenia; 4) TBS predicts fracture risk as well as lumbar spine BMD measurements in postmenopausal women; 5) efficacious therapies for osteoporosis differ in the extent to which they influence the TBS; 6) TBS is associated with fracture risk in individuals with conditions related to reduced bone mass or bone quality. Based on these data, lumbar spine TBS holds promise as an emerging technology that could well become a valuable clinical tool in the diagnosis of osteoporosis and in fracture risk assessment.

Elytra colour polymorphism and randomness of matings in Chauliognathus fallax Germar 1824 from southern Brazil (Coleoptera, Cantharidae)

Beetles of the species Chauliognathus fallax Germar 1824 are polymorphic for elytron colouration with six morphs distinguished on the basis of black pigmentation on a yellow background. We investigated samples of C. fallax taken in eight consecutive weeks aiming to determine the frequency of the morphs which were grouped, for statistical analysis, into three classes according to elytra pigmentation as well as the possible occurrence of assortative mating. Our results showed a variation in the frequency of the classes throughout the season, both in males and females, with the maximum frequency of each class at the fourth and fifth week. The three classes (A, B, C) showed the same pattern of variation, and class B was always the more frequent. To test randomness of matings two approaches were taken: in the first, we compared the frequency of each class in copulating and non-copulating insects. In the second, the frequency of each class in the whole sample was taken as the probability of occurrence of the respective class; then, using the criterion of the probability of independent events we calculated the expected proportion of copulating insects for each pair of events. Both methods gave non-significant differences, suggesting that the matings were random.