Promotion of functional heterotrimeric type I collagen via transfection in osteogenesis imperfecta fibroblasts

Abstract only availableOsteogenesis imperfecta (OI) is a heritable disorder due to mutations in type I collagen. Normal type I collagen forms a heterotrimeric protein comprised of two pro1(I) chains and one pro2(I) chain [1(I)22(I)]. The osteogenesis imperfecta murine (oim) model mouse contains a single nucleotide deletion in the pro2(I) gene (COL1A2) resulting in non-functional pro2(I) chains and production of homotrimeric type I collagen containing three pro1(I) collagen chains, [1(I)3], resulting in small body size, increased bone fragility and altered bone mineralization. The overall goal of this study is to correct the oim defect by introducing normal COL1A2 genes into oim cells. Oim dermal fibroblasts were transfected with a series of COL1A2 gene constructs containing the full-length murine pro2(I) collagen cDNA driven by various lengths of the murine COL1A2 promoter (1.5kb, 3.0kb, and 6.0kb) along with a COL1A2 enhancer. These DNA constructs were cotransfected with pcDNA3 containing a neomycin resistance gene, which allows for selection of stably transfected cell lines. Various assays have been developed to monitor proa2(I) collagen expression at the DNA, RNA and protein levels. A PCR assay was used to confirm genomic incorporation of transgenic COL1A2 gene constructs and an RT-PCR assay used to confirm expression of normal pro2(I) collagen mRNA. Denaturing urea-SDS polyacrylamide gel electrophoresis along with Western blotting analyses using anti-murine 1(I) and 2(I) collagen antibodies were used to confirm normal pro2(I) collagen expression at the protein level as well as its incorporation into normal heterotrimeric type I collagen. All the necessary tools have been established for evaluating the efficacy of transfection. Currently, the first series of stably transfected oim cell lines are being expanded for analyses as described above. Although this study is aimed at 'fixing' the oim mutation via gene therapy, valuable data will also be collected regarding the effectiveness of the variable length promoter regions in enhancing the expression of the normal COL1A2 gene.Life Sciences Undergraduate Research Opportunity Progra

Second order superintegrable systems in conformally flat spaces. IV. The classical 3D Stäckel transform and 3D classification theory

This article is one of a series that lays the groundwork for a structure and classification theory of second order superintegrable systems, both classical and quantum, in conformally flat spaces. In the first part of the article we study the Stäckel transform (or coupling constant metamorphosis) as an invertible mapping between classical superintegrable systems on different three-dimensional spaces. We show first that all superintegrable systems with nondegenerate potentials are multiseparable and then that each such system on any conformally flat space is Stäckel equivalent to a system on a constant curvature space. In the second part of the article we classify all the superintegrable systems that admit separation in generic coordinates. We find that there are eight families of these systems

Families of classical subgroup separable superintegrable systems

We describe a method for determining a complete set of integrals for a classical Hamiltonian that separates in orthogonal subgroup coordinates. As examples, we use it to determine complete sets of integrals, polynomial in the momenta, for some families of generalized oscillator and Kepler-Coulomb systems, hence demonstrating their superintegrability. The latter generalizes recent results of Verrier and Evans, and Rodriguez, Tempesta and Winternitz. Another example is given of a superintegrable system on a non-conformally flat space.Comment: 9 page

Lagrangian Formalism for nonlinear second-order Riccati Systems: one-dimensional Integrability and two-dimensional Superintegrability

The existence of a Lagrangian description for the second-order Riccati equation is analyzed and the results are applied to the study of two different nonlinear systems both related with the generalized Riccati equation. The Lagrangians are nonnatural and the forces are not derivable from a potential. The constant value $E$ of a preserved energy function can be used as an appropriate parameter for characterizing the behaviour of the solutions of these two systems. In the second part the existence of two--dimensional versions endowed with superintegrability is proved. The explicit expressions of the additional integrals are obtained in both cases. Finally it is proved that the orbits of the second system, that represents a nonlinear oscillator, can be considered as nonlinear Lissajous figuresComment: 25 pages, 7 figure

Can i take your subdomain? Exploring same-site attacks in the modern web

Related-domain attackers control a sibling domain of their target web application, e.g., as the result of a subdomain takeover. Despite their additional power over traditional web attackers, related-domain attackers received only limited attention from the research community. In this paper we define and quantify for the first time the threats that related-domain attackers pose to web application security. In particular, we first clarify the capabilities that related-domain attackers can acquire through different attack vectors, showing that different instances of the related-domain attacker concept are worth attention. We then study how these capabilities can be abused to compromise web application security by focusing on different angles, including cookies, CSP, CORS, postMessage, and domain relaxation. By building on this framework, we report on a large-scale security measurement on the top 50k domains from the Tranco list that led to the discovery of vulnerabilities in 887 sites, where we quantified the threats posed by related-domain attackers to popular web applications

Minimally invasive (flapless) crown lengthening by erbium: YAG laser in aesthetic zone

Crown lengthening is a surgical procedure aimed at exposure of a larger tooth surface by gingivectomy alone or with cortical bone remodelling for aesthetic purposes in the anterior zone of the maxilla or for reconstruction of teeth affected by subgingival caries. We report two cases of crown lengthening in the anterior maxilla for aesthetic purposes by gingival and bone re-contouring performed by erbium-doped yttrium aluminium garnet (erbium:YAG) laser. As highlighted in this report, the erbium:YAG laser-assisted crown lengthening is less invasive and also leads to faster clinical outcomes in contrast to the conventional surgical technique by scalpel incision, flap elevation and osteoplastic

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR): A critical overview on the most promising applications of molecular scissors in oral medicine

The scientific community is continuously working to translate the novel biomedical techniques into effective medical treatments. CRISPR-Cas9 system (Clustered Regularly Interspaced Short Palindromic Repeats-9), commonly known as the “molecular scissor”, represents a recently developed biotechnology able to improve the quality and the efficacy of traditional treatments, related to several human diseases, such as chronic diseases, neurodegenerative pathologies and, interestingly, oral diseases. Of course, dental medicine has notably increased the use of biotechnologies to ensure modern and conservative approaches: in this landscape, the use of CRISPR-Cas9 system may speed and personalize the traditional therapies, ensuring a good predictability of clinical results. The aim of this critical overview is to provide evidence on CRISPR efficacy, taking into specific account its applications in oral medicine

A Case of Osteonecrosis of the Jaw in a Patient with Crohn's Disease Treated with Infliximab

Background: Medication-related osteonecrosis of the jaw (MRONJ) is a severe adverse drug reaction, occurring in patients undergoing treatments with antiresorptive or antiangiogenic agents, such as bisphosphonates, denosumab, or bevacizumab, for different oncologic and non-oncologic diseases. The aim of this study was to report a case of MRONJ in a patient taking infliximab, an anti-TNF-alpha antibody used to treat Crohn's disease, rheumatoid arthritis, ulcerative colitis, ankylosing spondylitis, psoriatic arthritis, and plaque psoriasis. Case Report: A 49-year-old female patient affected by Crohn's disease, who had been undergoing 250 mg intravenous infliximab every six weeks for 12 years, with no history of antiresorptive or antiangiogenic agent administration, came to our attention for post-surgical MRONJ, associated with a wide cutaneous necrotic area of her anterior mandible. Following antibiotic cycles, the patient underwent surgical treatment with wide bone resection and debridement of necrotic tissues; after prolonged follow-up (16 months), the patient completely healed without signs of recurrence. Conclusions: Prevention of MRONJ by dental check-up before and during treatments with antiresorptive treatments (bisphosphonates or denosumab) is a well-established procedure. Although further studies are required to confirm the role of infliximab in MRONJ, based on the results of this study, we propose that patients who are going to be treated with infliximab should also undergo dental check-up before starting therapy, to possibly avoid MRONJ onset

Gender-related time course of sleep disturbances and psychological symptoms during the COVID-19 lockdown: a longitudinal study on the Italian population

Italy was the first western hotspot of the COVID-19 pandemic. In order to contain the spread of the virus, the Italian Government imposed home confinement to the entire population for almost two months. The present study is the first large-scale longitudinal report of the sleep and mental health changes during the prolonged lockdown due to the COVID-19 outbreak. We focused on the gendered vulnerability in a sample of the Italian population since cross-sectional research identified women to be more at-risk than men during this unprecedented situation. A total of 2701 individuals (mean age ± standard deviation, 32.37 ± 11.62; range, 18–82) participated in a web-based longitudinal survey consisting of two measurements. Participants were first-time recruited on social networks and via telephone messages through a snowball sampling and tested during the third week of the lockdown period. Subsequently, a follow-up evaluation was carried out during the seventh week of restraining measures. The survey assessed sleep quality, insomnia and depression symptoms, perceived stress, and anxiety, using the following questionnaires: the Pittsburgh Sleep Quality Index, the Insomnia Severity Index, the Beck Depression Inventory-second edition, the 10-item Perceived Stress Scale, and the State-Anxiety Inventory. Female gender showed the worst condition for all the examined dimensions in both the assessments. Nevertheless, at the follow-up women reported a reduction in insomnia and depression severity symptoms, perceived stress, and anxiety. On the other hand, male participants showed a worsening of sleep quality, insomnia symptoms, and perceived stress. Consequently, the gender prevalence gap of clinical conditions such as insomnia and depression was largely reduced under lockdown. Our investigation pointed to a different time course of sleep and mental health between genders during the home confinement period. Women seemed to show greater long-term resilience during the lockdown. Meanwhile, the male gender emerges as the most vulnerable category to the extension of the restraining measures. Our results suggest that there is no “weaker gender” after a prolonged lockdown. Indeed, the Italian population transversely presented signs of psychological suffering and significant sleep disturbances after the protracted and stressful lockdown period due to the COVID-19 pandemic