Derivation of human induced pluripotent stem cell line EURACi004-A from skin fibroblasts of a patient with Arrhythmogenic Cardiomyopathy carrying the heterozygous PKP2 mutation c.2569_3018del50

Arrhythmogenic Cardiomyopathy (ACM) is an inherited cardiac disease characterized by arrhythmias and fibro-fatty replacement in the ventricular myocardium. Causative mutations are mainly reported in desmosomal genes, especially in plakophilin2 (PKP2). Here, using a virus-free reprogramming approach, we generated induced pluripotent stem cells (iPSCs) from skin fibroblasts of one ACM patient carrying the frameshift heterozygous PKP2 mutation c.2569_3018del50. The iPSC line (EURACi004-A) showed the typical morphology of pluripotent cells, possessed normal karyotype and exhibited pluripotency markers and trilineage differentiation potential, including cardiomyogenic capability. Thus, this line can represent a human in vitro model to study the molecular basis of ACM

Exploring digenic inheritance in arrhythmogenic cardiomyopathy

Background: Arrhythmogenic cardiomyopathy (ACM) is an inherited genetic disorder, characterized by the substitution of heart muscle with fibro-fatty tissue and severe ventricular arrhythmias, often leading to heart failure and sudden cardiac death. ACM is considered a monogenic disorder, but the low penetrance of mutations identified in patients suggests the involvement of additional genetic or environmental factors. Methods: We used whole exome sequencing to investigate digenic inheritance in two ACM families where previous diagnostic tests have revealed a PKP2 mutation in all affected and some healthy individuals. In family members with PKP2 mutations we determined all genes that harbor variants in affected but not in healthy carriers or vice versa. We computationally prioritized the most likely candidates, focusing on known ACM genes and genes related to PKP2 through protein interactions, functional relationships, or shared biological processes. Results: We identified four candidate genes in family 1, namely DAG1, DAB2IP, CTBP2 and TCF25, and eleven candidate genes in family 2. The most promising gene in the second family is TTN, a gene previously associated with ACM, in which the affected individual harbors two rare deleterious-predicted missense variants, one of which is located in the protein's only serine kinase domain. Conclusions: In this study we report genes that might act as digenic players in ACM pathogenesis, on the basis of co-segregation with PKP2 mutations. Validation in larger cohorts is still required to prove the utility of this model

Effect Of 4-(n,n-dimethylamino) Phenethyl Alcohol On Degree Of Conversion And Cytotoxicity Of Photo - Polymerized Cq - Based Resin Composites

The aim of this study was to evaluate the degree of conversion (DC) and the cytotoxicity of photo-cured experimental resin composites containing 4-(N,N-dimethylamino) phenethyl alcohol (DMPOH) combined to the camphorquinone (CQ) compared with ethylamine benzoate (EDAB). The resin composites were mechanically blended using 35 wt% of an organic matrix and 65 wt% of filler loading. To this matrix was added 0.2 wt% of CQ and 0.2 wt% of one of the reducing agents tested. 5x1 mm samples (n=5) were previously submitted to DC measurement and then pre-immersed in complete culture medium without 10% (v/v) bovine serum for 1 h or 24 h at 37 °C in a humidifier incubator with 5% CO2and 95% humidity to evaluate the cytotoxic effects of experimental resin composites using the MTT assay on immortalized human keratinocytes cells. As a result of absence of normal distribution, the statistical analysis was performed using the nonparametric Kruskal-Wallis to evaluate the cytotoxicity and one-way analysis of variance to evaluate the DC. For multiple comparisons, cytotoxicity statistical analyses were submitted to Student-Newman-Keuls and DC analysis to Tukey’s HSD post-hoc test (α=0.05). No significant differences were found between the DC of DMPOH (49.9%) and EDAB (50.7%). 1 h outcomes showed no significant difference of the cell viability between EDAB (99.26%), DMPOH (94.85%) and the control group (100%). After 24 h no significant difference were found between EDAB (48.44%) and DMPOH (38.06%), but significant difference was found compared with the control group (p>0.05). DMPOH presented similar DC and cytotoxicity compared with EDAB when associated with CQ.256538542Furuse, A.Y., Mondelli, J., Watts, D.C., Network structures of Bis-GMA/ TEGDMA resins differ in DC, shrinkage-strain, hardness and optical properties as a function of reducing agent (2011) Dent Mat, 27, pp. 497-506Mundim, F., Pires-de-Souza, F., Garcia, L., Consani, S., Colour stability, opacity and cross-link density of composites submitted to accelerated artificial aging (2010) Eur J Prosthodont Rest Dent, 18, pp. 89-93Neumann, M.G., Miranda, W.G., Jr., Schmitt, C.C., Rueggeberg, F.A., Correa, I.C., Molar extinction coefficients and the photon absorption efficiency of dental photoinitiators and light curing units (2005) J Dent, 33, pp. 525-532Alvim, H.H., Alecio, A.C., Vasconcellos, W.A., Furlan, M., Oliveira, J.E., Saad, R.C., Analysis of camphorquinone in composite resins as a function of shade (2007) Dent Mat, 23, pp. 1245-1249Ruyter, I., Nilner, K., Moller, B., Color stability of dental resin materials for crown and bridge veneers (1987) Dent Mat, 3, pp. 246-251Heydecke, G., Zhang, F., Razzoog, M., In vitro color stability of double-layer veneers after accelerated aging (2001) J Prosthet Dent, 85, pp. 551-557Stansbury, J.W., Curing dental resins and composites by photopolymerization (2000) J Esthet Dent, 12, pp. 300-308Darvell, B., (2002) Materials science for dentistry, , 7th ed. Hong Kong: BW DarvelSilami, F.D.J., Mundim, F.M., Garcia, L.F.R., Sinhoreti, M.A.C., Pires-De-Souza, C.P., Colour stability of experimental composites containing different photoinitiators (2012) J Dent, 41, pp. e62-e66Albuquerque, P.P.A., Moreira, A.L., Moraes, R.R., Cavalcante, L.M., Schneider, F.J., Colour stability, conversion, water sorption and solubility of dental composites formulated with different photoinitiator systems (2012) J Dent, 41, pp. e67-e72Ely, C., Schneider, L.F.J., Ogliari, F.A., Schmitt, C.C., Correa, I.C., Lima, J.S., Polymerization kinetics and reactivity of alternative initiator systems for use in light-activated dental resins (2012) Dent Mat, 28, pp. 1199-1206Brandt, W.C., Tomaselli, L.O., Correr-Sobrinho, L., Sinhoreti, A.C., Can phenylpropanedione influence Knoop hardness, rate of polymerization and bond strength of resin composite restorations? (2011) J Dent, 39, pp. 438-447Schroeder, W.F., Vallo, C.I., Effect of different photoinitiator systems on conversion profiles of a model unfilled light-cured resin (2007) Dent Mat, 23, pp. 1313-1321Bowen, R.L., Argentar, H., Amine accelerators for methacrylate resin systems (1971) J Dent Res, 50, pp. 923-928Braden, M., Causton, B.E., Clarke, R.L., Diffusion in water in composite filling materials (1976) J Dent Res, 55, pp. 730-732Atai, M., Watts, D.C., A new kinetic model for the photopolymerization shrinkage-strain of dental composites and resin-monomers (2006) Dent Mat, 22, pp. 785-791Cook, W.D., Factors affecting the depth of cure of UV-polymerized composites (1980) J Dent Res, 59, pp. 800-808Lee, Y.K., Powers, J.M., Influence of background color on the color changes of resin composites after accelerated aging (2007) Am J Dent, 20, pp. 27-30Ota, M., Ando, S., Endo, H., Ogura, Y., Miyazaki, M., Hosoya, Y., Influence of refractive index on optical parameters of experimental resin composites (2012) Acta Odontol Scand, 70, pp. 362-367Fugita, K., Nishiyama, N., Nemoto, K., Okada, T., Ikemi, T., Effect of base monomer’s refractive index on curing depth and polymerization conversion of photo-cured resin composites (2005) Dent Mat J, 24, pp. 403-408Vargas, M.A., Cobb, D.S., Schmit, J.L., Polymerization of composite resins: Argon laser vs conventional light (1998) Oper Dent, 23, pp. 87-93Watts, D.C., Cash, A.J., Analysis of optical transmission by 400–500 nm visible light into aesthetic dental biomaterials (1994) J Dent, 22, pp. 112-117Porto, I.C.C.M., Andrade, A.K.M., Guenes, G.M.T., Ribeiro, A.I.A.M., Braz, R., Castro, M.B., In vitro potential cytotoxicity of an adhesive system to alveolar macrophages (2009) Braz Dent J, 20, pp. 195-200Ratanasathien, S., Wataha, J.C., Hanks, C.T., Dennison, J.B., Cytotoxic interactive effects of dentin bonding components on mouse fibroblasts (1995) J Dent Res, 74, pp. 1602-1606Ferracane, J.L., Elution of leachable components from composites (1994) J Oral Rehabil, 21, pp. 441-45

Anaesthetic Efficacy Of Bupivacaine 2-hydroxypropyl-β-cyclodextrin For Dental Anaesthesia After Inferior Alveolar Nerve Block In Rats

Bupivacaine is a long-acting local anaesthetic that is widely used in medicine and dentistry. The duration and intensity of its sensory blockade in animal models is increased by its inclusion in complexes with cyclodextrins. The aim of the present study was to evaluate the anaesthetic efficacy of bupivacaine 2-hydroxypropyl-β-cyclodextrin (HPβCD) inclusion complex for dental anaesthesia after inferior alveolar nerve block in rats. Thirty rats were each given an injection close to the mandibular foramen of 0.2 ml of one of the following formulations: 0.5% bupivacaine alone; 0.5% bupivacaine with 1:200,000 epinephrine; and 0.5% bupivacaine-HPβCD inclusion complex (bupivacaine-HPβCD). The other sides were used as controls, with either 0.9% saline or anaesthetic-free HPβCD solution being injected. The onset, success, and duration of pulpal anaesthesia were assessed by electrical stimulation ("pulp tester") on inferior molars. Results were analysed using ANOVA (Tukey), log rank, and chi square tests (α = 5%). There were no differences among the formulations in onset of anaesthesia (p = 0.59) or between the bupivacaine plus epinephrine and bupivacaine plus HPβCD in duration of anaesthesia, but bupivacaine plus epinephrine gave significantly higher values than bupivacaine alone (p = 0.007). Bupivacaine plus epinephrine was a better anaesthetic than bupivacaine alone (p = 0.02), while Bupi-HPβCD gave intermediate results, and therefore did not differ significantly from the other 2 groups (p = 0.18 with bupivacaine alone; and p = 0.44 with bupivacaine plus epinephrine). The bupivacaine-HPβCD complex showed similar anaesthetic properties to those of bupivacaine with epinephrine. © 2014 The British Association of Oral and Maxillofacial Surgeons.525452457Laskin, J.L., Wallace, W.R., Deleo, B., Use of bupivacaine hydrochloride in oral surgery - A clinical study (1977) J Oral Surg, 35, pp. 25-29Moore, P.A., Bupivacaine: A long-lasting local anesthetic for dentistry (1984) Oral Surg Oral Med Oral Pathol, 58, pp. 369-374Volpato, M.C., Ranali, J., Ramacciato, J.C., Anesthetic efficacy of bupivacaine solutions in inferior alveolar nerve block (2005) Anesth Prog, 52, pp. 132-135Fernandez, C., Reader, A., Beck, M., A prospective, randomized, double-blind comparison of bupivacaine and lidocaine for inferior alveolar nerve blocks (2005) J Endod, 31, pp. 499-503Branco, F.P., Ranali, J., Ambrosano, G.M., A double-blind comparison of 0.5% bupivacaine with 1:200,000 epinephrine and 0.5% levobupivacaine with 1:200,000 epinephrine for the inferior alveolar nerve block (2006) Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 101, pp. 442-447De Paula, E., Cereda, C., Tofoli, G., Drug delivery systems for local anesthetics (2010) Recent Pat Drug Deliv Formul, 4, pp. 23-34Weiniger, C.F., Golovanevski, L., Domb, A.J., Extended release formulations for local anaesthetic agents (2012) Anaesthesia, 67, pp. 906-916De Paula, E., Cereda, C.M., Fraceto, L.F., Micro and nanosystems for delivering local anesthetics (2012) Expert Opin Drug Deliv, 9, pp. 1505-1524Welliver, M., McDonough, J., Anesthetic related advances with cyclodextrins (2007) ScientificWorld J, 7, pp. 364-371Pinto, L.M., Fraceto, L.F., Santana, M.H., Physico-chemical characterization of benzocaine-beta-cyclodextrin inclusion complexes (2005) J Pharm Biomed Anal, 39, pp. 956-963Fréville, J.C., Dollo, G., Le Corre, P., Controlled systemic absorption and increased anesthetic effect of bupivacaine following epidural administration of bupivacaine-hydroxypropyl-beta- cyclodextrin complex (1996) Pharm Res, 13, pp. 1576-1580Araujo, D.R., Braga Ade, F., Moraes, C.M., Complexation of 50% enantiomeric excess (S75-R25) bupivacaine with cyclodextrins and spinal block anesthesia in rats (2006) Rev Bras Anestesiol, 56, pp. 495-506. , (in Portuguese)Araújo, D.R., Fraceto, L.F., Braga Ade, F., Drug-delivery systems for racemic bupivacaine (S50-R50) and bupivacaine enantiomeric mixture (S75-R25): Cyclodextrins complexation effects on sciatic nerve blockade in mice (2005) Rev Bras Anestesiol, 55, pp. 316-328. , (in Portuguese)De Araujo, D.R., Tsuneda, S.S., Cereda, C.M., Development and pharmacological evaluation of ropivacaine-2- hydroxypropyl-beta-cyclodextrin inclusion complex (2008) Eur J Pharm Sci, 33, pp. 60-71Franco De Lima, R.A., De Jesus, M.B., Cereda, C.M., Improvement of tetracaine antinociceptive effect by inclusion in cyclodextrins (2012) J Drug Target, 20, pp. 85-96Moraes, C.M., Abrami, P., De Paula, E., Study of the interaction between S(-) bupivacaine and 2-hydroxypropyl-beta-cyclodextrin (2007) Int J Pharm, 331, pp. 99-106Weisleder, R., Yamauchi, S., Caplan, D.J., The validity of pulp testing: A clinical study (2009) J Am Dent Assoc, 140, pp. 1013-1017Dreven, L.J., Reader, A., Beck, M., An evaluation of an electric pulp tester as a measure of analgesia in human vital teeth (1987) J Endod, 13, pp. 233-238Marenduzzo, A., Abbondanza, P., Technic of root anesthesia in the inferior alveolar process in the albino rat (1969) Arch Stomatol (Napoli), 10, pp. 151-156. , (in Italian)Naftel, J.P., Richards, L.P., Pan, M., Course and composition of the nerves that supply the mandibular teeth of the rat (1999) Anat Rec, 256, pp. 433-447Malamed, S., (2004) Handbook of Local Anesthesia, , 5th ed St. Louis MosbyDe Jong, D.J., (1994) Local Anesthesics, , 2nd ed. Charles C Thomas SpringfieldDollo, G., Thompson, D.O., Le Corre, P., Inclusion complexation of amide-typed local anesthetics with β-cyclodextrin and its derivatives, III. Biopharmaceutics of bupivacaine-SBE7-β-CD complex following percutaneous sciatic nerve administration in rabbits (1998) Int J Pharm, 164, pp. 11-19Karashima, K., Taniguchi, M., Nakamura, T., Prolongation of intrathecal and sciatic nerve blocks using a complex of levobupivacaine with maltosyl-beta-cyclodextrin in rats (2007) Anesth Analg, 104, pp. 1121-1128Cereda, C.M., Tofoli, G.R., Maturana, L.G., Local neurotoxicity and myotoxicity evaluation of cyclodextrin complexes of bupivacaine and ropivacaine (2012) Anesth Analg, 115, pp. 1234-124

Restless legs syndrome: Epidemiological and clinicogenetic study in a South tyrolean population isolate.

Genetic contributions to restless legs syndrome (RLS) have been consistently recognized from population and family studies. To determine the clinical and genetic features of RLS in a population isolate and explore linkage to three previously described susceptibility loci on chromosomes 12q, 14q, and 9p, respectively, an isolated population in the South Tyrolean Alps was identified and 530 adults participated in the study. Using a two-step strategy, 47 patients with idiopathic RLS were ascertained. The prevalence in the population was 8.9%. Twenty-eight patients (59.6%) had at least one affected first-degree relative and were classified as hereditary cases. In a single extended pedigree, linkage to known RLS loci was investigated specifying autosomal dominant and recessive models; parametric and nonparametric multipoint linkage scores were computed. None of the calculated linkage scores was suggestive of linkage between RLS and any of the three investigated loci. This study was conducted in a population isolate providing for a homogeneous genetic and environmental background. The absence of a suggestive linkage signal at the three known RLS susceptibility loci is indicative of further locus heterogeneity of this frequent disorder and encourages further studies to unveil the genetic causes of RLS