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    ABSTRACT: Real-time visualization of collagen is important in studies on tissue formation and remodeling in the research fields of developmental biology and tissue engineering. Our group has previously reported on a fluorescent probe for the specific imaging of collagen in live tissue in situ, consisting of the native collagen binding protein CNA35 labeled with fluorescent dye Oregon Green 488 (CNA35-OG488). The CNA35-OG488 probe has become widely used for collagen imaging. To allow for the use of CNA35-based probes in a broader range of applications, we here present a toolbox of six genetically-encoded collagen probes which are fusions of CNA35 to fluorescent proteins that span the visible spectrum: mTurquoise2, EGFP, mAmetrine, LSSmOrange, tdTomato and mCherry. While CNA35-OG488 requires a chemical conjugation step for labeling with the fluorescent dye, these protein-based probes can be easily produced in high yields by expression in E. coli and purified in one step using Ni2+-affinity chromatography. The probes all bind specifically to collagen, both in vitro and in porcine pericardial tissue. Some first applications of the probes are shown in multicolor imaging of engineered tissue and two-photon imaging of collagen in human skin. The fully-genetic encoding of the new probes makes them easily accessible to all scientists interested in collagen formation and remodeling.
    PLoS ONE 12/2014; 9(12):e114983. DOI:10.1371/journal.pone.0114983
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    ABSTRACT: p38α is a significant target for drug designing against cancer. The overproduction of p38α MAPK promotes tumorigenesis in head and neck squamous cell carcinoma (HNSCC). The ATP binding and an allosteric site referred as DFG are the key sites of the p38α mitogen activated protein kinase (MAPK) exploited for the design of inhibitors. This study demonstrated design of peptide inhibitor on the basis of allosteric site using Glide molecular docking software and the biochemical analysis of the best modeled peptide. The best fitted tetrapeptide (FWCS) in the allosteric site inhibited the pure recombinant and serum p38α of HNSCC patients by 74 and 72%, respectively. The potency of the peptide was demonstrated by its IC50 (4.6 nM) and KD (3.41×10-10 M) values, determined by ELISA and by surface plasmon resonance (SPR) technology, respectively. The cell viability of oral cancer i.e. KB cell line was reduced in dose dependent manner by 60 and 97% by the treatment of peptide and the IC50 was 600 and 210 µM after 24 and 72 h incubation, respectively. Our result provides an insight for the development of a proficient small peptide as a promising anticancer agent targeting DFG site of p38α kinase.
    PLoS ONE 07/2014; 9(7):e101525. DOI:10.1371/journal.pone.0101525
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    ABSTRACT: Aim The aim of the present study was to measure the accuracy and reproducibility of probe forces in simulated assessments of periodontal pocket depth. The study included experienced and inexperienced examiners and used manual and pressure-sensitive probes. Materials and Methods Sixty-one participants were divided into seven groups and asked to probe selected anterior and posterior sites with three different probes (Williams 14W, Chapple UB-CF-15, and Vivacare TPS probes). The model was positioned on a digital electronic balance to measure force, which was recorded initially and after 15 minutes. Probe preferences were recorded. Accuracy was measured by comparing to a standardized 25 g force, and reproducibility was calculated for all duplicate measurements. Results The Vivacare probe produced the most accurate and most reproducible forces, whereas the Williams probe produced the least accurate and least reproducible forces. Probe forces were lighter at anterior sites compared to posterior sites at baseline. Probe forces were reduced at both sites after 15 minutes compared to baseline. Conclusions Vivacare TPS periodontal probes are more accurate and reproducible than Chapple and Williams probes. Many clinicians in this study preferred the Chapple probe.
    Saudi Dental Journal 04/2014; 26(2). DOI:10.1016/j.sdentj.2014.02.001
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    ABSTRACT: Treatment of oral mucositis (OM) is challenging. In order to develop and test useful treatment approaches, the development of reliable, reproducible and simpler methods than are currently available for assessment of OM is important. A Patient-Reported Oral Mucositis Symptom (PROMS) scale was assessed in patients with head and neck cancer to determine if the patient-reported OM experience, as determined by using the PROMS scale, correlate with OM assessed by clinician-based scoring tools. Fifty patients with head and neck cancer and undergoing radiotherapy consented to participate. They were examined before cancer treatment and twice weekly during 6-7 weeks of therapy and once 4-6 weeks after therapy. Signs of OM were evaluated using the 3 clinician-based scoring tools; NCI-CTCAE v.3, the OMAS criteria and the Total VAS-OMAS. The participants' OM experiences were recorded using PROMS-questionnaires consisting of 10 questions on a visual analogue scale. Spearman rank correlation test were applied between the PROMS scale values and the clinician-determined scores. Repeated measures mixed linear models were applied to appraise the strengths of correlation at the different time points throughout the observation period. Thirty-three participants completed all stages of the study. The participant experience of OM using the PROMS scale demonstrates good correlations (Spearman's Rho 0.65-0.78, p<0.001) with the clinician-determined scores on the group level over all time points and poor to good correlations (Spearman's Rho -0.12-0.70, p<0.001) on the group level at different time points during and after therapy. When mouth opening was problematic, i.e. during the 6th and 7th week after commencing cancer treatment, the Spearman's Rho varied between 0.19 and 0.70 (p<0.001). Patient experience of OM, as reported by the PROMS scale may be a feasible substitute for clinical assessment in situations where patients cannot endure oral examinations.
    PLoS ONE 03/2014; 9(3):e91733. DOI:10.1371/journal.pone.0091733
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    ABSTRACT: SrO and SrF2 are widely used to replace CaO and CaF2 in ionomer glasses to produce radiopaque glass ionomer cements (GIC). The purpose of this study was to evaluate the effects of this substitution on release of ions from GIC as well as its effect on esthetics (translucency) and radiopacity. Cements were produced from ionomer glasses with varying content of Sr, Ca and F. The cements were stored in dilute acetic acid (pH 4.0) for up to 7 days at 37°C. Thereafter, the cements were removed and the solution was tested for F(-), Sr(2+), Ca(2+), and Al(3+) release. Radiopacity and translucency were measured according to BS EN ISO 9917-1:2003. Ion release was linear to t(1/2) suggesting that this is a diffusion controlled mechanism rather than dissolution. The fluoride release from the cements is enhanced where some or all calcium is replaced by strontium. Radiopacity shows a strong linear correlation with Sr content. All cements were more opaque than the C0.70 0.55 standard but less opaque than the C0.70 0.90 standard which is the limit for the ISO requirement for acceptance. This study shows that the replacement of calcium by strontium in a glass ionomer glass produces the expected increase in radiopacity of the cement without adverse effects on visual properties of the cement. The fluoride release from the cements is enhanced where some or all calcium is replaced by strontium.
    Dental materials: official publication of the Academy of Dental Materials 01/2014; DOI:10.1016/j.dental.2013.12.003
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    ABSTRACT: The potential of antimicrobial nanoparticles to control oral infections is reviewed. Such particles can be classified as having a size no greater than 100 nm and are produced using traditional or more novel techniques. Exploitation of the toxic properties of nanoparticles to bacteria, fungi and viruses, in particular metals and metal oxides, as well as their incorporation into polymeric materials have increased markedly over the past decade. The potential of nanoparticles to control the formation of biofilms within the oral cavity, as a function of their biocidal, anti-adhesive and delivery capabilities, is now receiving close attention. Latest insights into the application of nanoparticles within this field, including their use in photodynamic therapy, will be reviewed. Possible approaches to alter biocompatibility and desired function will also be covered.
    International journal of antimicrobial agents 11/2013; 43(2). DOI:10.1016/j.ijantimicag.2013.11.002
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    ABSTRACT: Crohn's disease presents in the orofacial region with a characteristic lip swelling that can progress to facial disfigurement.1 ,2 To date, assessments of these changes are based on clinical observation together with patient satisfaction reports and semiquantitative methods. Although useful, the development of an analytical approach that permits objective, reproducible and quantitative evaluation of morphological changes is required. We report the use of facial biometrics to objectively quantify changes associated with orofacial Crohn's disease in a 28-year-old woman.3 The patient was referred by the gastroenterologist for treatment of repeated upper lip swelling. In this patient a baseline facial scan was acquired when minimal lip swelling was present (figure 1A). Two months later she represented with markedly increased perioral swelling and a second scan was captured (figure 1B). Utilising coordinate metrology, figure 1A,B were identically positioned and registered. This allowed areas of swelling to be visualised (figure 1b) and a defined patient-specific template (highlighted in figure 1b–e) to be generated. An initial intralesional triamcinolone injection (40 mg/mL) resulted in some resolution of the swelling; a volume reduction of 6540.8 mm3 (figure 1c) postregistration of the third scan (figure 1C) against the second was noted. Four months after the third scan, the upper lip swelling recurred (figure 1D), with the right side more severely affected (figure 1d). This informed the precise position (* in figure 1D) for guided infiltration of intralesional triamcinolone (40 mg/mL). Two weeks post-treatment, registration of figure 1D,E revealed a volume reduction of 9656.4 mm3. At this point she reported considerable satisfaction with the outcome of treatment.
    Case Reports 11/2013; 2013(nov20_1). DOI:10.1136/bcr-2013-010460
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    ABSTRACT: To identify the timing of significant arch dimensional increases during orthodontic alignment involving round and rectangular nickel-titanium (NiTi) wires and rectangular stainless steel (SS). A secondary aim was to compare the timing of changes occurring with conventional and self-ligating fixed appliance systems. Additional data from a multicenter randomized trial initially involving 96 patients, aged 16 years and above, were analyzed. The main outcome measures were the magnitude and timing of maxillary intercanine, interpremolar, and intermolar dimensions. Each participant underwent alignment with a standard Damon (Ormco, Orange, Calif) wire sequence for a minimum of 34 weeks. Blinding of clinicians and patients was not possible; however, outcome assessors and data analysts were kept blind to the appliance type during data analysis. Complete data were obtained from 71 subjects. Significant arch dimensional changes were observed relatively early in treatment. In particular, changes in maxillary inter-first and second premolar dimensions occurred after alignment with an 0.014" NiTi wire (P< 0.05). No statistical differences in transverse dimensions were found between rectangular NiTi and working SS wires for each transverse dimension (P> 0.05). Bracket type had no significant effect on the timing of the transverse dimensional changes. Arch dimensional changes were found to occur relatively early in treatment, irrespective of the appliance type. Nickel-Titanium wires may have a more profound effect on transverse dimensions than previously believed.
    Journal of dentistry 11/2013; 42(1). DOI:10.1016/j.jdent.2013.11.010
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    ABSTRACT: The complex biological, physicochemical process of human dental enamel formation begins in utero and for most teeth takes several years to complete. Lost enamel tissue cannot regenerate, therefore a better understanding of the spatial and temporal progression of mineralization of this tissue is needed in order to design improved in vivo mineral growth processes for regenerative dentistry and allow the possibility to grow a synthetic whole or partial tooth. Human dental enamel samples across a range of developmental stages available through archaeological collections have been used to explore the spatial and temporal progression of enamel biomineralization. Position sensitive synchrotron X-ray diffraction was used to quantify spatial and temporal variations in crystallite organization, lattice parameters and crystallite thickness at three different stages in enamel maturation. In addition X-ray microtomography was used to study mineral content distributions. An inverse correlation was found between the spatial variation in mineral content and the distribution of crystallite organization and thickness as a function of time during enamel maturation. Combined X-ray microtomography and synchrotron X-ray diffraction results show that as enamel matures the mineral content increases and the mineral density distribution becomes more homogeneous. Starting concurrently but proceeding at a slower rate, the enamel crystallites become more oriented and larger; and the crystallite organization becomes spatially more complex and heterogeneous. During the mineralization of human dental enamel, the rate of mineral formation and mineral organization are not identical. Whilst the processes start simultaneously, full mineral content is achieved earlier, and crystallite organization is slower and continues for longer. These findings provide detailed insights into mineral development in human dental enamel which can inform synthetic biomimetic approaches for the benefit of clinical dentistry.
    Archives of oral biology 11/2013; 58(11):1726-1734. DOI:10.1016/j.archoralbio.2013.08.012
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    ABSTRACT: The prevalence of waterpipe tobacco smoking (WPTS) is increasing worldwide. The aims of this study were (a) to evaluate the psychometric properties of the Lebanon Waterpipe Dependence Scale (LWDS-11) and (b) to assess, estimate, and identify factors associated with waterpipe tobacco dependence symptoms among U.K. male adult resident waterpipe tobacco smokers. A total of 180 waterpipe tobacco smokers were recruited during random visits to 7 outlets serving waterpipe tobacco. Data were collected via face-to-face interviews using the WPTS module of the Global Adult Tobacco Survey and the LWDS-11. Descriptive statistics, exploratory psychometric, univariate, and Poisson regression analysis were employed. Participants' M/SD age was 29.46/±9.41 years, Arabic ethnicity accounted for 58.3%, and 53.9% had completed above secondary education. Psychometric analyses for the LWDS-10, after removing 1 item from the generic scale, revealed Cronbach's alpha coefficient = 0.74. About 47% of the sample demonstrated waterpipe tobacco dependence. Being Arab (p = .040, OR = 2.63, 95% CI = 1.05-6.62), smoking waterpipe daily in the past (p = .003, OR = 2.13, 95% CI = 1.30-3.49), and an increase in length in the last session of WPTS (p = .044, OR = 1.15, 95% CI = 1.00-1.32) were identified as risk factors for waterpipe tobacco dependence. This study demonstrated the potential of LWDS-11 and revealed 2 domains, positive and negative reinforcement and physiological dependence. Results indicated that waterpipe tobacco smokers in the United Kingdom demonstrated tobacco dependence symptoms, which were associated with socio-behavioral factors. Generalizability of these findings and their implications in public health are yet to be investigated.
    Nicotine & Tobacco Research 10/2013; 16(3). DOI:10.1093/ntr/ntt148
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