Transpupillary collagen photocrosslinking for targeted modulation of ocular biomechanics.

The central vision-threatening event in glaucoma is dysfunction and loss of retinal ganglion cells (RGCs), thought to be promoted by local tissue deformations. Here, we sought to reduce tissue deformation near the optic nerve head by selectively stiffening the peripapillary sclera, i.e. the scleral region immediately adjacent to the optic nerve head. Previous scleral stiffening studies to treat glaucoma or myopia have used either pan-scleral stiffening (not regionally selective) or regionally selective stiffening with limited access to the posterior globe. We present a method for selectively stiffening the peripapillary sclera using a transpupillary annular light beam to activate methylene blue administered by retrobulbar injection. Unlike prior approaches to photocrosslinking in the eye, this approach avoids the damaging effects of ultraviolet light by employing red light. This targeted photocrosslinking approach successfully stiffened the peripapillary sclera at 6 weeks post-treatment, as measured by whole globe inflation testing. Specifically, strain was reduced by 47% when comparing treated vs. untreated sclera within the same eye (n = 7, p=0.0064) and by 54% when comparing the peripapillary sclera of treated vs. untreated eyes (n = 7, p<0.0001). Post-treatment characterization of RGCs (optic nerve axon counts/density, and grading), retinal function (electroretinography), and retinal histology revealed that photocrosslinking was associated with some ocular toxicity. We conclude that a transpupillary photocrosslinking approach enables selective scleral stiffening targeted to the peripapillary region that may be useful in future treatments of glaucoma.

Factors controlling sediment and nutrient fluxes in a small microtidal salt marsh within the Venice Lagoon.

Coastal salt marshes are among the Earth's most productive ecosystems and provide a number of ecosystem services. Water quality regulation by storing, transforming and releasing nutrients, organic matter and suspended sediment is recognized as one of the most important functions of salt marshes. The self-purification capacity of intertidal ecosystems contributes to mitigating nutrient inputs, acting as a buffer zone between watersheds and coastal waters, especially in relation to climate change and the increasing frequency of impulsive extreme events. Understanding sediment and nutrient fluxes and assessing the mechanisms that control them are valuable for the preservation and future restoration of salt marshes and for enhancing their effectiveness in providing water quality regulation services. To investigate the main driving factors in microtidal environments, changes in suspended sediment and nutrient concentrations were measured during several tidal cycles in a small microtidal reconstructed salt marsh in the Venice Lagoon, which was selected as the experimental site. The tidal amplitude, nutrients and total suspended solid concentrations were measured during 6 tidal cycles between September 2011 and October 2013. The water discharge was derived by forcing the hydrodynamic MIKE-DHI numerical model with the measured tidal levels. Fluxes were assessed by coupling field data with the calculated discharges. The salt marsh filtering function was related to the inflow matter concentrations and tidal amplitude. When high suspended solid and nutrient concentrations enter the salt marsh, accumulation processes prevail on release. In contrast, in the case of low concentrations and high tidal excursion, the salt marsh functions as a nutrient and sediment source. During all campaigns, the nitrogen removal function was more effective within the intertidal vegetated areas. Sediment release was the dominant process in the outermost creek, whereas sedimentation prevailed in the inner area of the salt marsh because of the attenuation of hydrodynamic forcing during tide propagation.

The impact of prolapse mesh on vaginal smooth muscle structure and function.

OBJECTIVE: To evaluate the impact of prolapse meshes on vaginal smooth muscle structure (VaSM) and function, and to evaluate these outcomes in the context of the mechanical and textile properties of the mesh. DESIGN: Three months following the implantation of three polypropylene prolapse meshes with distinct textile and mechanical properties, mesh tissue explants were evaluated for smooth muscle contraction, innervation, receptor function, and innervation density. SETTING: Magee-Womens Research Institute at the University of Pittsburgh. POPULATION: Thirty-four parous rhesus macaques of similar age, parity, and pelvic organ prolapse quantification (POP-Q) scores. METHODS: Macaques were implanted with mesh via sacrocolpopexy. The impact of Gynemesh(™)  PS (Ethicon; n = 7), Restorelle(®) (Coloplast; n = 7), UltraPro(™) parallel and UltraPro(™) perpendicular (Ethicon; n = 6 and 7, respectively) were compared with sham-operated controls (n = 7). Outcomes were analysed by Kruskal-Wallis ANOVA, Mann-Whitney U-tests and multiple regression analysis (P < 0.05). MEAN OUTCOME MEASURES: Vaginal tissue explants were evaluated for the maximum contractile force generated following muscle, nerve, and receptor stimulation, and for peripheral nerve density. RESULTS: Muscle myofibre, nerve, and receptor-mediated contractions were negatively affected by mesh only in the grafted region (P < 0.001, P = 0.002, and P = 0.008, respectively), whereas cholinergic and adrenergic nerve densities were affected in the grafted (P = 0.090 and P = 0.008, respectively) and non-grafted (P = 0.009 and P = 0.005, respectively) regions. The impact varied by mesh property, as mesh stiffness was a significant predictor of the negative affect on muscle function and nerve density (P < 0.001 and P = 0.013, respectively), whereas mesh and weight was a predictor of receptor function (P < 0.001). CONCLUSIONS: Mesh has an overall negative impact on VaSM, and the effects are a function of mesh properties, most notably, mesh stiffness. TWEETABLE ABSTRACT: Prolapse mesh affects vaginal smooth muscle.

The protection of individuals affected with Specific Learning Disorders in the Italian Legislation.

Specific Learning Disorders (SLDs) affect specific abilities in individuals with an otherwise normal academic development. Among Italian School population, their reported prevalence is between 2.5% and 3.5%. Dysfunctions at the base of these disorders interfere with the normal acquisition process of reading, writing and/or mathematical abilities, leading to various degrees of adjustment difficulties in the affected individuals. The aim of this study was to assess the support that Italian Government offers to its citizens affected with SLDs, with a particular focus on assistance during the school-age years, particularly through the introduction of the Law 170/2010 and successive guidelines, supplementing the existing regulations to offer more efficient means and legal instruments aimed at achieving earlier diagnoses.

Penetrating Ocular Trauma with Retained Intraocular Foreign Body: Management, Follow-up and Medico-legal Evaluation.

Ocular trauma is the leading cause of acquired monocular blindness, accounting for 1.97-6% of such cases. Particularly, penetrating ocular injuries are among the most common eye injuries with this kind of outcome. Early diagnosis and prompt management are crucial to avoid complications, and the especially dreaded enucleation. In this article, the authors describe the clinical management, and evaluate the visual and anatomical results obtained in a case of ocular injury with retained intraocular foreign body (IOFB) in a 20-year old female patient. The course of treatment involved a combination of penetrating keratoplasty with a temporary keratoprosthesis, phacoemulsification with intraocular lens implantation and pars plana vitrectomy. At three years from the initial injury, the patient was able to count fingers at 30 centimetres and anatomical restitutio ad integrum of the globe had been achieved.

Sleep disorders and driving licence: the current Italian legislation and medico-legal issues.

Road traffic accidents (RTA) are a serious issue in all industrialized countries and have dramatic social and healthcare-related implications. Fatigue (sleepiness) is the principal identifiable and preventable cause of road traffic accidents. Obstructive sleep apnoea syndrome (OSAS) and narcolepsy are two of the leading causes of excessive daytime sleepiness. In this article, the authors analyze the current Italian legislation regarding driving licence issuance and fitness to drive, in order to evaluate the potential implications of sleep disorders, particularly OSAS and narcolepsy. In European Legislation and in Italy, OSAS and narcolepsy are not included among the illnesses or invalidating conditions that limit the fitness to drive for driving licence issuance purposes. In fact, they are not included in the Annex III of the European Council Directive 91/439/EEC of the 29th of July 1991 on driving licences. Some Countries of the European Union (Belgium, France, Finland, Great Britain, the Netherlands, Spain and Sweden) had implemented the 91/439/EEC Directive with national restrictions on driving licence issuance policies in case of OSAS and narcolepsy. Given the well-established scientific evidence available, in Italy, the lack of legislation regulating the assessment of the psychophysical requisites for the issuance and renewal of driving licences of individuals affected by sleep disorders seems extremely worrying. Furthermore, the current lack of legal obligation in Italy for healthcare facilities to disclose such diagnoses to the organs responsible for issuing driving licences (such as the Motorizzazione Civile - the Department of motor vehicles) remains the subject of heated debate.

Mechanical biocompatibility of prosthetic meshes: a comprehensive protocol for mechanical characterization.

This study is aimed at a comprehensive and extensive characterization of the mechanical biocompatibility of mesh prostheses. A robust and simple experimental protocol and a set of parameters is proposed, addressing stiffness under uniaxial and biaxial loading conditions, anisotropy, influence of prior deformation history, local mismatch of deformation mechanisms, and changes in mechanical properties when embedded in a homogeneous matrix. These parameters can form the basis for comparison of different mesh types and for evaluation of their mechanical biocompatibility. Measurements were performed on nine mesh types used for hernia and/or pelvic repair, including heavier as well as lighter implants and covering a wide range of mechanical responses. A total of 93 experiments were performed and all parameters are represented for each mesh in a single diagram to facilitate the quantitative assessment of implant characteristics and their comparison. The mechanics of implants is a critical factor determining clinical performance which should be accounted for in mesh selection and for development of future implants.

The p85 regulatory subunit of PI3K mediates cAMP-PKA and insulin biological effects on MCF-7 cell growth and motility.

Recent studies have shown that hyperinsulinemia may increase the cancer risk. Moreover, many tumors demonstrate an increased activation of IR signaling pathways. Phosphatidylinositol 3-kinase (PI3K) is necessary for insulin action. In epithelial cells, which do not express GLUT4 and gluconeogenic enzymes, insulin-mediated PI3K activation regulates cell survival, growth, and motility. Although the involvement of the regulatory subunit of PI3K (p85α (PI3K)) in insulin signal transduction has been extensively studied, the function of its N-terminus remains elusive. It has been identified as a serine (S83) in the p85α (PI3K) that is phosphorylated by protein kinase A (PKA). To determine the molecular mechanism linking PKA to insulin-mediated PI3K activation, we used p85α (PI3K) mutated forms to prevent phosphorylation (p85A) or to mimic the phosphorylated residue (p85D). We demonstrated that phosphorylation of p85α (PI3K)S83 modulates the formation of the p85α (PI3K)/IRS-1 complex and its subcellular localization influencing the kinetics of the insulin signaling both on MAPK-ERK and AKT pathways. Furthermore, the p85α (PI3K)S83 phosphorylation plays a central role in the control of insulin-mediated cell proliferation, cell migration, and adhesion. This study highlights the p85α (PI3K)S83 role as a key regulator of cell proliferation and motility induced by insulin in MCF-7 cells breast cancer model.

The role of E-cadherin down-regulation in oral cancer: CDH1 gene expression and epigenetic blockage.

BACKGROUND: The prognosis of the oral squamous cell carcinoma (OSCC) patients remains very poor, mainly due to their high propensity to invade and metastasize. E-cadherin reduced expression occurs in the primary step of oral tumour progression and gene methylation is a mode by which the expression of this protein is regulated in cancers. In this perspective, we investigated E-cadherin gene (CDH1) promoter methylation status in OSCC and its correlation with Ecadherin protein expression, clinicopathological characteristics and patient outcome. METHODS: Histologically proven OSCC and paired normal mucosa were analyzed for CDH1 promoter methylation status and E-cadherin protein expression by methylation-specific polymerase chain reaction and immunohistochemistry. Colocalization of E-cadherin with epidermal growth factor (EGF) receptor (EGFR) was evidenced by confocal microscopy and by immunoprecipitation analyses. RESULTS: This study indicated E-cadherin protein down-regulation in OSCC associated with protein delocalization from membrane to cytoplasm. Low E-cadherin expression correlated to aggressive, poorly differentiated, high grade carcinomas and low patient survival. Moreover, protein down-regulation appeared to be due to E-cadherin mRNA downregulation and CDH1 promoter hypermethylation. In an in vitro model of OSCC the treatment with EGF caused internalization and co-localization of E-cadherin with EGFR and the addition of demethylating agents increased E-cadherin expression. CONCLUSION: Low E-Cadherin expression is a negative prognostic factor of OSCC and is likely due to the hypermethylation of CDH1 promoter. The delocalization of E-cadherin from membrane to cytoplasm could be also due to the increased expression of EGFR in OSCC and the consequent increase of E-cadherin co-internalization with EGFR.

Deterioration in biomechanical properties of the vagina following implantation of a high-stiffness prolapse mesh.

OBJECTIVE: To define the impact of prolapse mesh on the biomechanical properties of the vagina by comparing the prototype Gynemesh PS (Ethicon) to two new-generation lower stiffness meshes, SmartMesh (Coloplast) and UltraPro (Ethicon). DESIGN: A study employing a nonhuman primate model. SETTING: University of Pittsburgh, PA, USA. POPULATION: Forty-five parous rhesus macaques. METHODS: Meshes were implanted via sacrocolpopexy after hysterectomy and compared with sham. Because its stiffness is highly directional, UltraPro was implanted in two directions: UltraPro Perpendicular (less stiff) and UltraPro Parallel (more stiff), with the indicated direction referring to the position of the blue orientation lines relative to the longitudinal axis of the vagina. The mesh-vaginal complex (MVC) was excised in toto after 3 months. MAIN OUTCOME MEASURES: Active mechanical properties were quantified as the contractile force generated in the presence of 120 mmol/l KCl. Passive mechanical properties (a tissue's ability to resist an applied force) were measured using a multiaxial protocol. RESULTS: Vaginal contractility decreased by 80% following implantation with the Gynemesh PS (P = 0.001), 48% after SmartMesh (P = 0.001), 68% after UltraPro Parallel (P = 0.001) and was highly variable after UltraPro Perpendicular (P = 0.16). The tissue contribution to the passive mechanical behaviour of the MVC was drastically reduced for Gynemesh PS (P = 0.003), but not for SmartMesh (P = 0.9) or UltraPro independent of the direction of implantation (P = 0.68 and P = 0.66, respectively). CONCLUSIONS: Deterioration of the mechanical properties of the vagina was highest following implantation with the stiffest mesh, Gynemesh PS. Such a decrease associated with implantation of a device of increased stiffness is consistent with findings from other systems employing prostheses for support.

Vaginal degeneration following implantation of synthetic mesh with increased stiffness.

OBJECTIVE: To compare the impact of the prototype prolapse mesh Gynemesh PS with that of two new-generation lower stiffness meshes, UltraPro and SmartMesh, on vaginal morphology and structural composition. DESIGN: A mechanistic study employing a nonhuman primate model. SETTING: Magee-Womens Research Institute at the University of Pittsburgh. POPULATION: Parous rhesus macaques, with similar age, weight, parity and Pelvic Organ Prolapse-Questionnaire scores. METHODS: Following Institutional Animal Care Use Committee approval, 50 rhesus macaques were implanted with Gynemesh PS (n = 12), UltraPro with its blue line perpendicular to the longitudinal axis of vagina (n = 10), UltraPro with its blue line parallel to the longitudinal axis of vagina (n = 8) or SmartMesh (n = 8) via sacrocolpopexy following hysterectomy. Sham-operated animals (n = 12) served as controls. MAIN OUTCOME MEASURES: The mesh-vagina complex was removed after 12 weeks and analysed for histomorphology, in situ cell apoptosis, total collagen, elastin, glycosaminoglycan content and total collagenase activity. Appropriate statistics and correlation analyses were performed accordingly. RESULTS: Relative to sham and the two lower stiffness meshes, Gynemesh PS had the greatest negative impact on vaginal histomorphology and composition. Compared with sham, implantation with Gynemesh PS caused substantial thinning of the smooth muscle layer (1557 ± 499 µm versus 866 ± 210 µm, P = 0.02), increased apoptosis particularly in the area of the mesh fibres (P = 0.01), decreased collagen and elastin content (20%, P = 0.03 and 43%, P = 0.02, respectively) and increased total collagenase activity (135%, P = 0.01). Glycosaminoglycan, a marker of tissue injury, was highest with Gynemesh PS compared with sham and other meshes (P = 0.01). CONCLUSION: Mesh implantation with the stiffer mesh Gynemesh PS induced a maladaptive remodelling response consistent with vaginal degeneration.

Graft-related complications and biaxial tensiometry following experimental vaginal implantation of flat mesh of variable dimensions.

OBJECTIVE: To compare the occurrence of graft-related complications (GRCs) and biomechanical properties of meshes implanted vaginally and abdominally. DESIGN: In vivo animal experiment. SETTING: Centre for Surgical Technologies, Medical Faculty, KU Leuven, Belgium. POPULATION: Twenty adult parous Texel ewes. METHODS: Sheep were implanted with Gynemesh M, a 28-g/m² polypropylene mesh reinforced with polyglecaprone fibres, under general anaesthesia. Dissection into the rectovaginal septum was performed to accommodate a flat 50 × 50 mm (n = 10) or 35 × 35 mm (n = 10) mesh, which was sutured to the underlying tissues. A 50 × 50 mm mesh was laid over a primarily sutured, full-thickness, 40-mm longitudinal abdominal wall incision. Sacrifice was at 60 days (n = 10) or 90 days (n = 5). MAIN OUTCOME MEASURES: The occurrence of exposure, the degree of contraction and examination of the biomechanical properties of explants with a minimum radius of 32 mm via biaxial tensiometry. RESULTS: Insertion of a 50 × 50 mm mesh led to exposures in 30% (3/10) of cases, and the average contraction rate was 52 ± 14%. In the 35 × 35 mm implants, there were no exposures, and the contraction rate was 25 ± 26.3%. Vaginal explants with no GRCs and of sufficient size had biomechanical properties that were comparable with those of abdominal explants. CONCLUSION: Vaginal mesh insertion is associated with GRCs, such as exposure and contraction. Although other factors probably play a role, this study illustrates that mesh size may also induce these complications. In a vaginal surgery model, clinically occurring GRCs can be reproduced. In addition, biomechanics of uncomplicated vaginal explants are comparable with those measured on abdominal explants.

Cytokines and VEGF induction in orthodontic movement in animal models.

Orthodontics is a branch of dentistry that aims at the resolution of dental malocclusions. The specialist carries out the treatment using intraoral or extraoral orthodontic appliances that require forces of a given load level to obtain a tooth movement in a certain direction in dental arches. Orthodontic tooth movement is dependent on efficient remodeling of periodontal ligament and alveolar bone, correlated with several biological and mechanical responses of the tissues surrounding the teeth. A periodontal ligament placed under pressure will result in bone resorption whereas a periodontal ligament under tension results in bone formation. In the primary stage of the application of orthodontic forces, an acute inflammation occurs in periodontium. Several proinflammatory cytokines are produced by immune-competent cells migrating by means of dilated capillaries. In this paper we summarize, also through the utilization of animal models, the role of some of these molecules, namely, interleukin-1ß and vascular endothelial growth factor, that are some proliferation markers of osteoclasts and osteoblasts, and the macrophage colony stimulating factor.

Prognostic significance of N-Cadherin expression in oral squamous cell carcinoma.

BACKGROUND: N-Cadherin (CDH2) is a calcium-dependent adhesion protein, whose de novo expression, re-expression, up-regulation and down-regulation in human tumors has been demonstrated. The aim of the present work was to define the prognostic role of N-Cadherin in a large series of oral squamous cell carcinomas (OSCCs). MATERIALS AND METHODS: A total of 94 selected OSCCs were quantitatively and qualitatively analyzed by immunohistochemistry for N-Cadherin. The association between protein expression and clinico-pathological parameters was assessed by statistical analysis. RESULTS: In neoplastic tissue, N-Cadherin levels were more evident than in normal peritumoral epithelium (p<0.05). Protein staining was mainly detected in the neoplastic cells, and only focal nuclear positivity was observed. Expression of cytoplasmic N-Cadherin correlated significantly with poor histological differentiation (p<0.05). Furthermore, we have observed significant a statistical trend for stage and a correlation with worst patient outcome, also confirmed by Kaplan-Meier estimates. CONCLUSION: Our work has underlined the key-role of N-Cadherin in oral carcinogenesis and in the prognostic stratification of patients.

Treatment of obesity in three rural primary care practices.

The experience of three rural primary care practices in treating obesity is described. Treatment alternatives utilized include behavior modification with a balanced deficit diet, use of the protein-sparing modified fast in a group setting, and use of this technique in conjunction with behavior modification on an individual basis. The results of these three case studies are evaluated by an index approach as well as the traditional mean pounds lost. Although treatment of obesity is often avoided because of reported low success rates, this study demonstrates that a family physician has more success than previously reported, with one of the sites showing maintenance of weight loss in 84 percent of a selected group of its patients.