Rapid activation and down-regulation of protein kinase C alpha in 12-O-tetradecanoylphorbol-13-acetate -induced differentiation of human rhabdomyosarcoma cells

Human rhabdomyosarcoma RD cells express the myogenic regulatory factors MyoD and myogenin but differentiate spontaneously very poorly. Prolonged treatment of RD cells with the protein kinase C (PKC) activator 12-O-tetradecanoylphorbol-13-acetate (TPA) induces growth arrest and myogenic differentiation as shown by the accumulation of alpha-actin and myosin light and heavy chains, without affecting the expression of MyoD and myogenin. In this study, we show that short-term phorbol ester treatment of the cultures is sufficient to trigger myogenic differentiation but not growth arrest. Furthermore, PKC inhibitors, such as staurosporine or calphostin C, prevent TPA-induced differentiation but not cell growth arrest. These data suggest that the two events are mediated by different pathways; a possible interpretation is that the activation of one or more PKC isoforms mediates the induction of differentiation, whereas the down-regulation of the same or different isoforms mediates the growth arrest. To address the mechanism whereby TPA affects cell growth and differentiation in RD cells, we first analyzed PKC isoenzyme distribution. We found that RD cells express the alpha, beta 1, gamma, and sigma PKC isoenzymes. Only the alpha isoform is exclusively found in the soluble fraction, but it translocates to the membrane fraction within 5 min of TPA treatment and is completely down-regulated after 6 h. The other isoenzymes are found associated to both the soluble and the particulate fractions and are down-regulated after long-term TPA treatment. By immunofluorescence analysis, we show that the PKC alpha down-regulation is specific for those cells that respond to TPA by activating the muscle phenotype. We propose that TPA-induced differentiation in RD cells is mediated by the transient activation of PKC alpha, which activates some of the intracellular events that are necessary for MyoD and myogenin transacting activity and for the induction of terminal differentiation of RD cells. By contrast, the constitutively active beta 1 and sigma are responsible for the maintenance of cell growth, and their down-regulation is responsible for long-term TPA-induced cell growth arrest

Reconstructive periodontal therapy with simultaneous ridge augmentation. A clinical and histological case series report

Treatment of intrabony periodontal defects with a combination of a natural bone mineral (NBM) and guided tissue regeneration (GTR) has been shown to promote periodontal regeneration in intrabony defects. In certain clinical situations, the teeth presenting intrabony defects are located at close vicinity of the resorbed alveolar ridge. In these particular cases, it is of clinical interest to simultaneously reconstruct both the intrabony periodontal defect and the resorbed alveolar ridge, thus allowing insertion of endosseous dental implants. The aim of the present study was to present the clinical and histological results obtained with a new surgical technique designed to simultaneously reconstruct the intrabony defect and the adjacently located resorbed alveolar ridge. Eight patients with chronic advanced periodontitis displaying intrabony defects located in the close vicinity of resorbed alveolar ridges were consecutively enrolled in the study. After local anesthesia, mucoperiosteal flaps were raised, the granulation tissue removed, and the roots meticulously scaled and planed. A subepithelial connective tissue graft was harvested from the palate and sutured to the oral flap. The intrabony defect and the adjacent alveolar ridge were filled with a NBM and subsequently covered with a bioresorbable collagen membrane (GTR). At 11–20 months (mean, 13.9 ± 3.9 months) after surgery, implants were placed, core biopsies retrieved, and histologically evaluated. Mean pocket depth reduction measured 3.8 ± 1.7 mm and mean clinical attachment level gain 4.3 ± 2.2 mm, respectively. Reentry revealed in all cases a complete fill of the intrabony component and a mean additional vertical hard tissue gain of 1.8 ± 1.8 mm. The histologic evaluation indicated that most NBM particles were surrounded by bone. Mean new bone and mean graft area measured 17.8 ± 2.8% and 32.1 ± 8.3%, respectively. Within their limits, the present findings indicate that the described surgical approach may be successfully used in certain clinical cases to simultaneously treat intrabony defects and to reconstruct the resorbed alveolar ridge

Calcium Sulfate and Platelet-Rich Plasma make a novel osteoinductive biomaterial for bone regeneration

BACKGROUND: With the present study we introduce a novel and simple biomaterial able to induce regeneration of bone. We theorized that nourishing a bone defect with calcium and with a large amount of activated platelets may initiate a series of biological processes that culminate in bone regeneration. Thus, we engineered CS-Platelet, a biomaterial based on the combination of Calcium Sulfate and Platelet-Rich Plasma in which Calcium Sulfate also acts as an activator of the platelets, therefore avoiding the need to activate the platelets with an agonist. METHODS: First, we tested CS-Platelet in heterotopic (muscle) and orthotopic (bone) bone regeneration bioassays. We then utilized CS-Platelet in a variety of dental and craniofacial clinical cases, where regeneration of bone was needed. RESULTS: The heterotopic bioassay showed formation of bone within the muscular tissue at the site of the implantation of CS-Platelet. Results of a quantitative orthotopic bioassay based on the rat calvaria critical size defect showed that only CS-Platelet and recombinant human BMP2 were able to induce a significant regeneration of bone. A non-human primate orthotopic bioassay also showed that CS-Platelet is completely resorbable. In all human clinical cases where CS-Platelet was used, a complete bone repair was achieved. CONCLUSION: This study showed that CS-Platelet is a novel biomaterial able to induce formation of bone in heterotopic and orthotopic sites, in orthotopic critical size bone defects, and in various clinical situations. The discovery of CS-Platelet may represent a cost-effective breakthrough in bone regenerative therapy and an alternative or an adjuvant to the current treatments

Genetic-Background Modulation of Core and Variable Autistic-Like Symptoms in Fmr1 Knock-Out Mice

International audienc

Spatially Controlled Single Photon Emitters in hBN-Capped WS2 Domes

Monolayers (MLs) of transition-metal dichalcogenides host efficient single-photon emitters (SPEs) usually associated to the presence of nanoscale mechanical deformations or strain. Large-scale spatial control of strain would enhance the scalability of such SPEs and allow for their incorporation into photonic structures. Here, the formation of regular arrays of strained hydrogen-filled one-layer-thick micro-domes obtained by H-ion irradiation and lithography-based approaches is reported. Typically, the H-2 liquefaction for temperatures T<32 K causes the disappearance of the domes preventing their use as potential SPEs. Here, it is shown that the dome deflation can be overcome by hBN heterostructuring, that is by depositing thin hBN flakes on the domes. This leads to the preservation of the dome structure at all temperatures, as found by micro-Raman and micro-photoluminescence (mu-PL) studies. Eventually, spatially controlled hBN-capped WS2 domes show the appearance, at 5 K, of intense emission lines originating from localized excitons, which are shown to behave as quantum emitters here. The electronic properties of the emitters are addressed by time-resolved mu-PL yielding time decays of 1-10 ns, and by magneto-mu-PL measurements. The latter provide an exciton magnetic moment a factor of two larger than the value observed in planar strain-free MLs

National Italian Guidelines for caries prevention in 0 to 12 years-old children

Aim Oral and dental health improved tremendously over the last fifty years in Italy but still prevalence of dental caries in children remains a significant clinical problem. This report describes the National Italian Guidelines for caries prevention. Methodology A panel of experts coordinated by the Italian Society of Paediatric Dentistry (SIOI) planned to elaborate the national Italian guidelines for caries prevention in children. The structure of the guidelines has been planned to follow the principles of modern caries treatment and management as well as science based dentistry. The main procedure was based on a hierarchic evaluation of literature. Conclusion The guidelines are planned for dentist working in primary dental care, however, they are also designed to be of interest for other care professionals such as paediatricians, gynecologists, pharmacists and general medical practitioners and also for parents and/or guardians of the children

National Italian Guidelines for caries prevention in 0 to 12 years-old children.

AIM: Oral and dental health improved tremendously over the last fifty years in Italy but still prevalence of dental caries in children remains a significant clinical problem. This report describes the National Italian Guidelines for caries prevention. METHODOLOGY: A panel of experts coordinated by the Italian Society of Paediatric Dentistry (SIOI) planned to elaborate the national Italian guidelines for caries prevention in children. The structure of the guidelines has been planned to follow the principles of modern caries treatment and management as well as science based dentistry. The main procedure was based on a hierarchic evaluation of literature. CONCLUSION: The guidelines are planned for dentist working in primary dental care, however, they are also designed to be of interest for other care professionals such as paediatricians, gynecologists, pharmacists and general medical practitioners and also for parents and/or guardians of the children