High frequency of submicroscopic chromosomal imbalances in patients with syndromic craniosynostosis detected by a combined approach of microsatellite segregation analysis, multiplex ligation-dependent probe amplification and array-based comparative genome hybridisation

We present the first comprehensive study, to our knowledge, on genomic chromosomal analysis in syndromic craniosynostosis. In total, 45 patients with craniosynostotic disorders were screened with a variety of methods including conventional karyotype, microsatellite segregation analysis, subtelomeric multiplex ligation-dependent probe amplification) and whole-genome array-based comparative genome hybridisation. Causative abnormalities were present in 42.2% (19/45) of the samples, and 27.8% (10/36) of the patients with normal conventional karyotype carried submicroscopic imbalances. Our results include a wide variety of imbalances and point to novel chromosomal regions associated with craniosynostosis. The high incidence of pure duplications or trisomies suggests that these are important mechanisms in craniosynostosis, particularly in cases involving the metopic suture.

Screening of autosomal gene deletions in patients with hypogonadotropic hypogonadism using multiplex ligation-dependent probe amplification: detection of a hemizygosis for the fibroblast growth factor receptor 1

P>Objective Congenital hypogonadotropic hypogonadism with anosmia (Kallmann syndrome) or with normal sense of smell is a heterogeneous genetic disorder caused by defects in the synthesis, secretion and action of gonadotrophin-releasing hormone (GnRH). Mutations involving autosomal genes have been identified in approximately 30% of all cases of hypogonadotropic hypogonadism. However, most studies that screened patients with hypogonadotropic hypogonadism for gene mutations did not include gene dosage methodologies. Therefore, it remains to be determined whether patients without detected point mutation carried a heterozygous deletion of one or more exons. Measurements We used the multiplex ligation-dependent probe amplification (MLPA) assay to evaluate the potential contribution of heterozygous deletions of FGFR1, GnRH1, GnRHR, GPR54 and NELF genes in the aetiology of GnRH deficiency. Patients We studied a mutation-negative cohort of 135 patients, 80 with Kallmann syndrome and 55 with normosmic hypogonadotropic hypogonadism. Results One large heterozygous deletion involving all FGFR1 exons was identified in a female patient with sporadic normosmic hypogonadotropic hypogonadism and mild dimorphisms as ogival palate and cavus foot. FGFR1 hemizygosity was confirmed by gene dosage with comparative multiplex and real-time PCRs. Conclusions FGFR1 or other autosomal gene deletion is a possible but very rare event and does not account for a significant number of sporadic or inherited cases of isolated GnRH deficiency.

Identification of genetic alterations by multiple ligation-dependent probe amplification (MLPA) analysis in oligodendrogliomas

Concurrent deletion at 1p/19q is a common signature of oligodendrogliomas, and it may, be identified in low-grade tumours (grade II) suggesting it represents an early event in the development of these brain neoplasms. Additional non-random changes primarily involve CDKN2A, PTEN and EGFR. Identification of all of these genetic changes has become an additional parameter in the evaluation of the clinical patients` prognosis, including good response to conventional chemotherapy. Multiple ligation-dependent probe amplification (MLPA) analysis is a new methodology that allows an easy identification of the oligodendrogliomas` abnormalities in a single step. No need of the respective constitutional DNA from each patient is another advantage of this method. We used MLPA kits P088 and P105 to determine the molecular characteristics of a series of 40 oligodendrogliomas. Deletions at I p and 19q were identified in 45% and 65% of cases, respectively. Alterations of EGFR, CDKN2A, ERBB2, PTEN and TP53 were also identified in variable frequencies among 7% to 35% of tumours. These findings demonstrate that MLPA is a reliable technique to the detection of molecular genetic changes in oligodendrogliomas.

Proposal for a Trigonometric Method to Evaluate the Abduction Angle of the Lower Limbs in Neonates

It is difficult to precisely measure articular arc movement in newborns using a goniometer. This article proposes an objective method based on trigonometry for the evaluation of lower limb abduction. With the newborn aligned in the dorsal decubitus position, 2 points are marked at the level of the medial malleolus, one on the sagittal line and the other at the end of the abduction. Using the right-sided line between these 2 points and a line from the medial malleolus to the reference point at the anterior superior iliac spine or umbilical scar, an isosceles triangle is drawn, and half of the inferential abduction angle is obtained by calculating the sine. Twenty healthy full-term newborns comprise the study cohort. Intersubject and intrasubject variability among the abduction angle values (mean [SD], 37 degrees [4]degrees) is low. This method is advantageous because the measurement is precise and because the sine can be used without approximation.

FURTHER INSIGHTS TOWARD VITAMIN C DETERMINATION AND STABILITY. PROPOSAL OF A NEW QUANTIFICATION METHOD

This work aimed at an evaluation of the classical iodine method for quantification of vitamin C (L-ascorbic acid) in fruit juices, as well as at a search into the stability of this so popular vitamin under different conditions of pH, temperature and light exposition, in addition to a proposal of a new quantification method. Our results point to the persistent reversibility of the blue color of the starch-triiodide complex at the end point when using the classical iodine titration, and the overestimation of the true vitamin concentration in fruit juices. A new quantification method is proposed in order to overcome this problem. Surprising conclusions were obtained regarding the controversial stability of L-ascorbic acid toward atmospheric oxygen, at low pH, even in fruit juice and at room temperature, showing that the major problem concerned with aging of fruit juices is proliferation of microorganisms rather than expontaneous oxidation of L-ascorbic acid.

Evaluation of three different adhesive systems using a bacterial method to develop secondary caries in vitro

Purpose: To assess the effects of three different dental adhesive systems on the formation of secondary root caries, in vitro, with a standardized interfacial gap in a filled cavity model. Methods: 40 sound human molars were selected and randomly assigned to four experimental groups: Clearfil SE Bond (CSEB), Xeno III (X-III), Scotchbond Multi-Purpose Plus (SBMP) and negative control (NC) without an adhesive system. After the standardized Class V cavity preparations on the buccal and lingual surfaces, restorations were placed with resin composite (Filtek Z250) using a standardized interfacial gap, using a 3 x 2 mm piece of 50 mu m metal matrix. The teeth were sterilized with gamma irradiation and exposed to a cariogenic challenge using a bacterial system with Streptococcus mutans. Depth and extension of wall lesions formed and the depth of outer lesions were measured by software coupled with light microscopy. Results: For wall lesion extension the ANOVA test showed differences between groups except between X-HI and SBMP (P= 0.294). The Tukey`s test of confidence intervals indicated smaller values for the CSEB group than for the others. For wall lesion depth the CSEB group also presented the smallest mean values of wall lesion depth when compared to the others (P< 0.0001) for all comparisons using Tukey`s test. Regarding outer lesion depth, all adhesives showed statistically similar behavior. SEM evaluation of the morphologic appearance of caries lesions confirmed the statistical results showing small caries lesion development for cavities restored with CSEB adhesive system, which may suggest that this adhesive system interdiffusion zone promoted a good interaction with subjacent dentin protecting the dental tissues from recurrent caries. (Am J Dent 2010;23:93-97).

Effect of processing induced particle alignment on the fracture toughness and fracture behavior of multiphase dental ceramics

Objective. To investigate the processing induced particle alignment on fracture behavior of four multiphase dental ceramics (one porcelain, two glass-ceramics and a glass-infiltrated-alumina composite). Methods. Disks (empty set12mm x 1.1 mm-thick) and bars (3 mm x 4 mm x 20 mm) of each material were processed according to manufacturer instructions, machined and polished. Fracture toughness (K(IC)) was determined by the indentation strength method using 3-point bending and biaxial flexure fixtures for the fracture of bars and disks, respectively. Microstructural and fractographic analyses were performed with scanning electron microscopy, energy dispersive spectroscopy and X-ray diffraction. Results. The isotropic microstructure of the porcelain and the leucite-based glass-ceramic resulted in similar fracture toughness values regardless of the specimen geometry. On the other hand, materials containing second-phase particles with high aspect ratio (lithium disilicate glass-ceramic and glass-infiltrated-alumina composite) showed lower fracture toughness for disk specimens compared to bars. For the lithium disilicate glass-ceramic disks, it was demonstrated that the occurrence of particle alignment during the heat-pressing procedure resulted in an unfavorable pattern that created weak microstructural paths during the biaxial test. For the glass-infiltrated-alumina composite, the microstructural analysis showed that the large alumina platelets tended to align their large surfaces perpendicularly to the direction of particle deposition during slip casting of green preforms. Significance. The fracture toughness of dental ceramics with anisotropic microstructure should be determined by means of biaxial testing, since it results in lower values. (C) 2009 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Influence of Preflaring on the Accuracy of Length Determination With Four Electronic Apex Locators

Introduction: The aim of this study was to compare the influence of preflaring on the accuracy of 4 electronic apex locators (EALs): Root ZX, Elements Diagnostic Unit and Apex Locator, Mini Apex Locator, and Apex DSP. Methods: Forty extracted teeth were preflared by using S1 and SX ProTaper instruments. The working length was established by reducing 1 mm from the total length (TL). The ability of the EALs to detect precise (-1 mm from TL) and acceptable (-1+/-0.5 mm from TL) measurements in unflared and preflared canals was determined. Results: The precise and acceptable (P/A) readings in unflared canals for Root ZX, Elements Diagnostic Unit and Apex Locator, Mini Apex and Apex DSP were 50%/97.5%, 47.5%/95%, 50%/97.5%, and 45%/67.5%, respectively. For preflared canals, the readings were 75%/97.5%, 55%/95%, 75%/97.5%, and 60%/87.5%, respectively. For precise criteria, the preflared procedure increased the percentage of accurate electronic readings for the Root ZX and the Mini Apex Locator (P < .05). For acceptable criteria, no differences were found among Root ZX, Elements Diagnostic Unit and Apex Locator, and Mini Apex Locator (P > .05). Fisher test indicated the lower accuracy for Apex DSP (P < .05). Conclusions: The Root ZX and the Mini Apex Locator devices increased significantly the precision to determine the real working length after the preflaring procedure. All the EALs showed an acceptable determination of the working length between the ranges of+/-0.5mm except for the Apex DSP device, which had the lowest accuracy. (J Endod 2009;35:1300-1302)

Effect of curing protocol on the polymerization of dual-cured resin cements

Objectives. The purpose of this study was to evaluate how curing protocol affects the extent of polymerization of dual-cured resin cements. Methods. Four commercial resin cements were used (DuoLink, Panavia F 2.0, Variolink II and Enforce). The extent of polymerization of the resin cements cured under different conditions was measured using a (1)H Stray-Field MRI method, which also enabled to probe molecular mobility in the kHz frequency range. Results. Resin cements show well distinct behaviours concerning chemical cure. Immediate photo-activation appears to be the best choice for higher filler loaded resin cements (Panavia F 2.0 and Variolink). A photo-activation delay (5 min) did not induce any significant difference in the extent of polymerization of all cements. Significance. The extent of polymerization of dual-cured resin cements considerably changed among products under various curing protocols. Clinicians should optimize the materials choice taking into account the curing characteristics of the cements. (C) 2010 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

The relationship between interimplant distances and vascularization of the interimplant bone

Background Long-term success of the implant restorations is based upon the biology and vasculature of the bone surrounding the implants, especially for the bone between two implants. Purpose The aim of this study was to evaluate how loaded implants placed 2 or 3 mm apart influence bone vessel organization. Material and methods Six mongrel dogs were used for the study. The four mandibular premolars were extracted and 3 months later, four 4.5 x 10 mm implants were placed on each side of the mandible. The implants were placed so that two adjacent implants were 2 mm (group 1) or 3 mm (group 2) distant from each other. After 12 weeks, the implants were loaded with provisional prostheses, then metallic crowns were placed 4 weeks later. Both temporary and metallic restorations were made so that the distance between the contact point and the bone crest was 5 mm. The animals were sacrificed after 8 weeks. The hemi-mandibles were removed and prepared for analysis. The interimplant bone vasculature of the two groups was studied using scanning electron microscopic images fractal analysis. The fractal dimension (D(f)) was calculated using the box-counting method. Results The values of the D(f) for the blood vessels were significantly higher (P <.05) in the specimens of the group 2 (1.969 +/- 0.169) than the group 1 (1.556 +/- 0.246). Conclusion The presence of more blood vessels in the group 2 is another indication that 3 mm is a preferable distance for contiguous implants than the 2 mm distance. To cite this article:Traini T, Novaes AB, Piattelli A, Papalexiou V, Muglia VA. The relationship between interimplant distances and vascularization of the interimplant bone.Clin. Oral Impl. Res. 21, 2010; 822-829.doi: 10.1111/j.1600-0501.2010.01926.x.

Platyhelminth phylogenetics - a key to understanding parasitism?

The comparative method, the inference of biological processes from phylogenetic patterns, is founded on the reliability of the phylogenetic tree. In attempting to apply the comparative method to the understanding of the evolution of parasitism in the phylum Platyhelminthes, we have highlighted several points we consider to be of value along with many problems. We discuss four of these topics. Firstly, we view the group at a phylum level, in particular discussing the importance of establishing the sister taxon to the obligate parasite group, the Neodermata, for addressing such questions as the monophyly, parasitism or the endo or ectoparasitic nature of the early parasites. The variety of non-congruent phylogenetic trees presented so far, utilising either or both morphological and molecular data, gives rise to the suggestion that any evolutionary scenarios presented at this stage be treated as interesting hypotheses rather than well-supported theories. Our second point of discussion is the conflict between morphological and molecular estimates of monogenean evolution. The Monogenea presents several well-established morphological autapomorphies, such that morphology consistently estimates the group as monophyletic, whereas molecular sequence analyses indicate paraphyly, with different genes giving different topologies. We discuss the problem of reconciling gene and species trees. Thirdly, we use recent phylogenetic results on the tapeworms to interpret the evolution of strobilation, proglottization, segmentation and scolex structure. In relation to the latter, the results presented indicate that the higher cestodes are diphyletic, with one branch difossate and the other tetrafossate. Finally, we use a SSU rDNA phylogenetic tree of the Trematoda as a basis for the discussion of an aspect of the digenean life-cycle, namely the nature of the first intermediate host. Frequent episodes of host-switching, between gastropod and bivalve hosts or even into annelids, are indicated.

Continuous quantum measurement of two coupled quantum dots using a point contact: A quantum trajectory approach

We obtain the finite-temperature unconditional master equation of the density matrix for two coupled quantum dots (CQD's) when one dot is subjected to a measurement of its electron occupation number using a point contact (PC). To determine how the CQD system state depends on the actual current through the PC device, we use the so-called quantum trajectory method to derive the zero-temperature conditional master equation. We first treat the electron tunneling through the PC barrier as a classical stochastic point process (a quantum-jump model). Then we show explicitly that our results can be extended to the quantum-diffusive limit when the average electron tunneling rate is very large compared to the extra change of the tunneling rate due to the presence of the electron in the dot closer to the PC. We find that in both quantum-jump and quantum-diffusive cases, the conditional dynamics of the CQD system can be described by the stochastic Schrodinger equations for its conditioned state vector if and only if the information carried away from the CQD system by the PC reservoirs can be recovered by the perfect detection of the measurements.

The reliability of immunohistochemistry as a prescreening method for the diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC) - Results of an international collaborative study

Hereditary nonpolyposis colorectal cancer syndrome (HNPCC) is an autosomal dominant condition accounting for 2–5% of all colorectal carcinomas as well as a small subset of endometrial, upper urinary tract and other gastrointestinal cancers. An assay to detect the underlying defect in HNPCC, inactivation of a DNA mismatch repair enzyme, would be useful in identifying HNPCC probands. Monoclonal antibodies against hMLH1 and hMSH2, two DNA mismatch repair proteins which account for most HNPCC cancers, are commercially available. This study sought to investigate the potential utility of these antibodies in determining the expression status of these proteins in paraffin-embedded formalin-fixed tissue and to identify key technical protocol components associated with successful staining. A set of 20 colorectal carcinoma cases of known hMLH1 and hMSH2 mutation and expression status underwent immunoperoxidase staining at multiple institutions, each of which used their own technical protocol. Staining for hMSH2 was successful in most laboratories while staining for hMLH1 proved problematic in multiple labs. However, a significant minority of laboratories demonstrated excellent results including high discriminatory power with both monoclonal antibodies. These laboratories appropriately identified hMLH1 or hMSH2 inactivation with high sensitivity and specificity. The key protocol point associated with successful staining was an antigen retrieval step involving heat treatment and either EDTA or citrate buffer. This study demonstrates the potential utility of immunohistochemistry in detecting HNPCC probands and identifies key technical components for successful staining.

Minimum acceptable standards for digital compression of a fetal ultrasound video-clip

If the Internet could be used as a method of transmitting ultrasound images taken in the field quickly and effectively, it would bring tertiary consultation to even extremely remote centres. The aim of the study was to evaluate the maximum degree of compression of fetal ultrasound video-recordings that would not compromise signal quality. A digital fetal ultrasound videorecording of 90 s was produced, resulting in a file size of 512 MByte. The file was compressed to 2, 5 and 10 MByte. The recordings were viewed by a panel of four experienced observers who were blinded to the compression ratio used. Using a simple seven-point scoring system, the observers rated the quality of the clip on 17 items. The maximum compression ratio that was considered clinically acceptable was found to be 1:50-1:100. This produced final file sizes of 5-10 MByte, corresponding to a screen size of 320 x 240 pixels, running at 15 frames/s. This study expands the possibilities for providing tertiary perinatal services to the wider community.

A pilot study of the manual force levels required to produce manipulation induced hypoalgesia

Objective. A pilot investigation of the influence of different force levels on a treatment technique's hypoalgesic effect. Design. Randomised single blind repeated measures. Background. Optimisation of such biomechanical treatment variables as the point of force application, direction of force application and the level of applied manual force is classically regarded as the basis of best practice manipulative therapy. Manipulative therapy is frequently used to alleviate pain, a treatment effect that is often studied directly in the neurophysiological, paradigm and seldom in biomechanical research. The relationship between the level of force applied by a technique (e.g. biomechanics) and its hypoalgesic effect was the focus of this study. Method. The experiment involved the application of a lateral glide mobilisation with movement treatment technique to the symptomatic elbow of six subjects with lateral epicondylalgia. Four different levels of force, which were measured with a flexible pressure-sensing mat, were randomly applied while the subject performed a pain free grip strength test. Results. Standardised manual force data varied from 0.76 to 4.54 N/cm, lower-upper limits 95 Cl, respectively. Pain free grip strength expressed as a percentage change from pre-treatment values was significantly greater with manual forces beyond 1.9 N/cm (P = 0.014). Conclusions. This study, albeit a pilot, provides preliminary evidence that in terms of the hypoalgesic effect of a mobilisation with movement treatment technique, there may be an optimal level of applied manual force.