Comparison of different methods for thin section EM analysis of Mycobacterium smegmatis

Bacteria are generally difficult specimens to prepare for conventional resin section electron microscopy and mycobacteria, with their thick and complex cell envelope layers being especially prone to artefacts. Here we made a systematic comparison of different methods for preparing Mycobacterium smegmatis for thin section electron microscopy analysis. These methods were: (1) conventional preparation by fixatives and epoxy resins at ambient temperature. (2) Tokuyasu cryo-section of chemically fixed bacteria. (3) rapid freezing followed by freeze substitution and embedding in epoxy resin at room temperature or (4) combined with Lowicryl HM20 embedding and ultraviolet (UV) polymerization at low temperature and (5) CEMOVIS, or cryo electron microscopy of vitreous sections. The best preservation of bacteria was obtained with the cryo electron microscopy of vitreous sections method, as expected, especially with respect to the preservation of the cell envelope and lipid bodies. By comparison with cryo electron microscopy of vitreous sections both the conventional and Tokuyasu methods produced different, undesirable artefacts. The two different types of freeze-substitution protocols showed variable preservation of the cell envelope but gave acceptable preservation of the cytoplasm, but not lipid bodies, and bacterial DNA. In conclusion although cryo electron microscopy of vitreous sections must be considered the 'gold standard' among sectioning methods for electron microscopy, because it avoids solvents and stains, the use of optimally prepared freeze substitution also offers some advantages for ultrastructural analysis of bacteria.

Ground reaction force profiles after partial and pancarpal arthrodesis in dogs

OBJECTIVES: To evaluate and compare long-term functional outcome after partial carpal arthrodesis and pancarpal arthrodesis in dogs using kinetic gait analysis. METHODS: Fourteen dogs with 19 partial carpal or pancarpal arthrodeses were retrospectively examined and underwent force-plate gait analysis. Mean times since surgery were 29.4 and 24.4 months for pancarpal and partial carpal arthrodesis respectively. Vertical and braking-propulsive ground reaction force profiles were compared between treatment groups, and to those of normal dogs (control group) using Kruskal-Wallis one-way analysis of variance. RESULTS: With the exception of time to vertical peak that occurred earlier in dogs with pancarpal than in dogs with partial carpal arthrodesis (p <0.01), there was no difference between the two treatment groups. Several parameters differed significantly between operated and healthy dogs (p <0.01): vertical impulses were significantly lower in both treatment groups, braking forces and impulses were also reduced after both techniques. Propulsive forces and impulses were only reduced in dogs with pancarpal arthrodesis. When comparing gait parameters of sound limbs of unilateral operated dogs to those of control dogs, braking forces and impulses (p <0.01; p <0.05) were significantly higher in the sound legs of unilateral operated dogs. CLINICAL SIGNIFICANCE: Long-term outcome after partial carpal and pancarpal arthrodesis is good and comparable to each other. Propulsive action may be altered more in dogs with pancarpal arthrodesis.

Reconstruction of patient-specific 3D bone surface from 2D calibrated fluoroscopic images and point distribution model

Reconstruction of patient-specific 3D bone surface from 2D calibrated fluoroscopic images and a point distribution model is discussed. We present a 2D/3D reconstruction scheme combining statistical extrapolation and regularized shape deformation with an iterative image-to-model correspondence establishing algorithm, and show its application to reconstruct the surface of proximal femur. The image-to-model correspondence is established using a non-rigid 2D point matching process, which iteratively uses a symmetric injective nearest-neighbor mapping operator and 2D thin-plate splines based deformation to find a fraction of best matched 2D point pairs between features detected from the fluoroscopic images and those extracted from the 3D model. The obtained 2D point pairs are then used to set up a set of 3D point pairs such that we turn a 2D/3D reconstruction problem to a 3D/3D one. We designed and conducted experiments on 11 cadaveric femurs to validate the present reconstruction scheme. An average mean reconstruction error of 1.2 mm was found when two fluoroscopic images were used for each bone. It decreased to 1.0 mm when three fluoroscopic images were used.

Basistests Suisse Sport Test Konzept: Validierung einer sportmotorischen Basistestbatterie für den Schul- und Nachwuchssport

Physical fitness can be evaluated in competitive and school sports with different field tests under different conditions and goals. To produce valid results, a field test must be practical and reach high standards of test criteria (objectivity, reliability, validity). The purpose of this study was to investigate the test criteria and the practicability of a group of field tests called «SUISSE Sport Test Konzept Basis Feldtestbatterie». For 20-m sprint, ventral trunk muscle test, standing long jump, 2-kg medicine ball shot, obstacle course and cooper-test, test quality and practicability were evaluated. 221 children and adolescents from competitive sports and different school levels took part in the study. According to school level, they were divided into 3 groups (P: 7–11.5 y, S1: 11.6–15.5 y, S2: 15.6–21.8 y). Objectivity was tested for time or distance measurement in all tests as well as for error rating in obstacle test. For reliability measurement, 162 subjects performed the field tests twice within a few weeks. For validity results of standing long jump were compared with counter movement jump performance on a force plate. Correlation analysis was performed and level of significance was set for p < 0.05. For accuracy standard error was calculated. All tests achieved sufficient to excellent objectiv - ity with correlation-coefficient (r) lying between 0.85 and 0.99. Reliability was very good (r = 0.84–0.97). In cooper- and trunk test, reliability was higher for athletes than for pupils (trunk test: r = 0.95 vs. r = 0.62, cooper-test: r = 0.90 vs. r = 0.78). In those tests the reliability decreases with increasing age (cooper-test: P: r = 0.84, S1: r = 0.69, S2: r = 0.52; trunk-test: P: r = 0.69, S1: r = 0.71; S2: r = 0.39). Validity for standing long jump was good (r = 0.75–0.86). The standard error of the mean was between 4–8%, with the exception for cooper-test (athletes: 6%, pupils: 11%) and trunk test (athletes: 14%, pupils: 46%). The results show that the evaluated group of field tests is a practicable, objective and reliable tool to determine physical skills in young athletes as well as in a scholar setting over a broad age range. Most of the tests achieved the test criteria with the grades good to excellent. The lower coefficient of reliability for cooper- and trunk test by the pupils could be explained by motivational problems in this setting. For up to 20 subjects, a tester can accomplish the tests within 3 h. Finally, age-dependent grades were elaborated

The Locking Compression Paediatric Hip Plate: technical guide and critical analysis

Osteotomies of the proximal femur and stable fixation of displaced femoral neck fractures are demanding operations. An LCP Paediatric Hip Plate was developed to make these operations safer and less demanding. The article focuses on the surgical technique and critically analyses the device.

Investigating interocclusal perception in tactile teeth sensibility using symmetric and asymmetric analysis

The purpose of this clinical trial was to determine the active tactile sensibility of natural teeth and to obtain a statistical analysis method fitting a psychometric function through the observed data points. On 68 complete dentulous test persons (34 males, 34 females, mean age 45.9 ± 16.1 years), one pair of healthy natural teeth each was tested: n = 24 anterior teeth and n = 44 posterior teeth. The computer-assisted, randomized measurement was done by having the subjects bite on thin copper foils of different thickness (5-200 µm) inserted between the teeth. The threshold of active tactile sensibility was defined by the 50% value of correct answers. Additionally, the gradient of the sensibility curve and the support area (90-10% value) as a description of the shape of the sensibility curve were calculated. For modeling the sensibility curve, symmetric and asymmetric functions were used. The mean sensibility threshold was 14.2 ± 12.1 µm. The older the subject, the higher the tactile threshold (r = 0.42, p = 0.0006). The support area was 41.8 ± 43.3 µm. The higher the 50% threshold, the smaller the gradient of the curve and the larger the support area. The curves showing the active tactile sensibility of natural teeth demonstrate a tendency towards asymmetry, so that the active tactile sensibility of natural teeth can mathematically best be described by using the asymmetric Weibull function.

Morphology of the nasopalatine canal and dental implant surgery: a radiographic analysis of 100 consecutive patients using limited cone-beam computed tomography

To analyze the dimensions and anatomic characteristics of the nasopalatine canal and the corresponding buccal bone plate of the alveolar process, using limited cone-beam computed tomography (CBCT) imaging.

Nanoscale switching characteristics of nearly tetragonal BiFeO3 thin films

We have investigated the nanoscale switching properties of strain-engineered BiFeO(3) thin films deposited on LaAlO(3) substrates using a combination of scanning probe techniques. Polarized Raman spectral analysis indicates that the nearly tetragonal films have monoclinic (Cc) rather than P4mm tetragonal symmetry. Through local switching-spectroscopy measurements and piezoresponse force microscopy, we provide clear evidence of ferroelectric switching of the tetragonal phase, but the polarization direction, and therefore its switching, deviates strongly from the expected (001) tetragonal axis. We also demonstrate a large and reversible, electrically driven structural phase transition from the tetragonal to the rhombohedral polymorph in this material, which is promising for a plethora of applications.

Design-based stereology to quantify structural properties of artificial and natural snow using thin sections

The quantification of the structural properties of snow is traditionally based on model-based stereology. Model-based stereology requires assumptions about the shape of the investigated structure. Here, we show how the density, specific surface area, and grain boundary area can be measured using a design-based method, where no assumptions about structural properties are necessary. The stereological results were also compared to X-ray tomography to control the accuracy of the method. The specific surface area calculated with the stereological method was 19.8 ± 12.3% smaller than with X-ray tomography. For the density, the stereological method gave results that were 11.7 ± 12.1% larger than X-ray tomography. The statistical analysis of the estimates confirmed that the stereological method and the sampling used are accurate. This stereological method was successfully tested on artificially produced ice beads but also on several snow types. Combining stereology and polarisation microscopy provides a good estimate of grain boundary areas in ice beads and in natural snow, with some limitatio

Coronary plaque composition of culprit/target lesions according to the clinical presentation: a virtual histology intravascular ultrasound analysis

AIMS: To evaluate the plaque composition obtained by virtual histology (VH) IVUS according to the clinical presentation and to compare those data to previously published histopathology data. METHODS AND RESULTS: VH was performed on 95 de novo significant lesions (>75% stenosis) in 85 patients [28 acute coronary syndrome (ACS) patients, 30 lesions; 57 stable angina pectoris (SAP) patients, 65 lesions]. There were a higher prevalence of positive remodelling (47 vs. 22%, P=0.013), thrombus (20 vs. 1.5%, P=0.0037), and echo-lucent area (23.3 vs. 7.7%, P=0.047) in ACS patients. At the minimal lumen site, fibrous plaque area was significantly larger in ACS lesions than in SAP lesions (66.0+/-10.7 vs. 61.4+/-8.9%, P=0.034), whereas necrotic core and dense calcium plaque area were smaller in ACS lesions (Necrotic core: 6.8+/-6.0 vs. 11.0+/-8.3%, P=0.02; Dense calcium: 2.6+/-3.0 vs. 4.9+/-5.8%, P=0.03). No differences in rate of thin cap fibroatheroma, thick fibrotheroma, or for the presence of multiple necrotic core layers were observed between both groups. CONCLUSION: Plaque composition obtained by VH-IVUS shows less necrotic core and more fibrous tissue in ACS compared to SAP lesions, which is in contradiction with previously published histopathologic data.

Late postoperative opacification of hydrogel intraocular lenses: analysis of 13 explanted lenses

PURPOSE: We report the clinical, morphological, and ultrastructural findings of 13 consecutively explanted opacified Hydroview(R) (hydrogel) intraocular lenses (IOLs). Our purpose was to provide a comprehensive account on the possible factors involved in late postoperative opacification of these IOLs. PATIENTS AND METHODS: Thirteen consecutive opacified hydrogel IOLs (Hydroview H 60 M, Bausch ; Lomb) were explanted due to the significant visual impairment they caused. The IOLs underwent macroscopical examination, transmission electron microscopy (TEM), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), and electrophoresis for protein detection. Three unused control Hydroview IOLs served for comparison. RESULTS: Macroscopical examination showed a diffuse or localized grey-whitish opacification within the IOL optic. TEM confirmed the presence of lesions inside the optic in all the explanted IOLs and revealed 3 patterns of deep deposits: a) diffuse, thick, granular, electron-dense ones; b) small, thin, lattice-like ones, with prominent electron-lucent areas; and c) elongated electron-dense formations surrounded by electron-lucent halos. SEM showed surface deposits on four IOLs. EDS revealed oxygen and carbon in all IOLs and documented calcium, phosphorus, silicon and/or iron in the deposits. Two of the patients with iron in their IOLs had eye surgery prior to their phacoemulsification. Iron correlated well with the second TEM pattern of deep lesions, whereas calcium with the third TEM pattern. No protein bands were detected on electrophoresis. Control lenses did not show any ultrastructural or chemical abnormality. CONCLUSIONS: The present study supports the presence of chemical alterations inside the polymer of the optic in late postoperative opacification of Hydroview IOLs. This opacification does not follow a unique pathway but may present under different ultrastructular patterns depending on the responsible factors. Mechanical stress during surgery may initiate a sequence of events where ions such as calcium, phosphorus, silicon, and/or iron, participate in a biochemical cascade that leads to gradual alteration of the polymer network. Intraocular inflammation due to previous operation may be a factor inducing opacification through increase of iron-binding capacity in the aqueous humour. Calcification accounts only partially for the opacification noted in this type of IOL.

Diffusion tensor imaging of two unrelated Chinese men with hereditary spastic paraplegia associated with thin corpus callosum

Hereditary spastic paraplegia (HSP) associated with thin corpus callosum is a rare autosomal recessive neurodegenerative disorder characterized by an abnormally thin corpus callosum, normal motor development, slowly progressive spastic paraparesis and cognitive deterioration. To investigate and localize abnormalities in the brains of two Chinese patients with HSP-TCC, with mutations in the spatacsin gene. Diffusion tensor imaging (DTI) was used to determine the mean diffusion (MD) and fractional anisotropy (FA) in the brains of the patients in comparison to 20 healthy subjects. Voxel-based analysis (VBA) of both the diffusion and anisotropy values were performed using statistical parametric mapping (SPM). Significant changes with MD increase and FA reduction were found in the already known lesions including the corpus callosum, cerebellum and thalamus. In addition, changes were also found in regions that appear to be normal in conventional MRI, such as the brain stem, internal capsule, cingulum and subcortical white matter including superior longitudinal fascicle and inferior longitudinal fascicle. Neither increase in FA nor reduction in MD was detected in the brain. Our study provides clear in vivo MR imaging evidence of a more widespread brain involvement of HSP-TCC. MD is more sensitive than FA in detecting lesions in thalamus and subcortical white matter, suggesting that MD may be a better marker of the disease progression.

Loss of alpha10beta1 integrin expression leads to moderate dysfunction of growth plate chondrocytes

Integrin alpha10beta1 is a collagen-binding integrin expressed on chondrocytes. In order to unravel the role of the alpha10 integrin during development, we generated mice carrying a constitutive deletion of the alpha10 integrin gene. The mutant mice had a normal lifespan and were fertile but developed a growth retardation of the long bones. Analysis of the skeleton revealed defects in the growth plate after birth characterized by a disturbed columnar arrangement of chondrocytes, abnormal chondrocyte shape and reduced chondrocyte proliferation. Electron microscopy of growth plates from newborn mice revealed an increased number of apoptotic chondrocytes and reduced density of the collagen fibrillar network compared to these structures in control mice. These results demonstrate that integrin alpha10beta1 plays a specific role in growth plate morphogenesis and function.

Ridge alterations post-extraction in the esthetic zone: a 3D analysis with CBCT

Dimensional alterations of the facial bone wall following tooth extractions in the esthetic zone have a profound effect on treatment outcomes. This prospective study in 39 patients is the first to investigate three-dimensional (3D) alterations of facial bone in the esthetic zone during the initial 8 wks following flapless tooth extraction. A novel 3D analysis was carried out, based on 2 consecutive cone beam computed tomographies (CBCTs). A risk zone for significant bone resorption was identified in central areas, whereas proximal areas yielded only minor changes. Correlation analysis identified a facial bone wall thickness of ≤ 1 mm as a critical factor associated with the extent of bone resorption. Thin-wall phenotypes displayed pronounced vertical bone resorption, with a median bone loss of 7.5 mm, as compared with thick-wall phenotypes, which decreased by only 1.1 mm. For the first time, 3D analysis has allowed for documentation of dimensional alterations of the facial bone wall in the esthetic zone of humans following extraction. It also characterized a risk zone prone to pronounced bone resorption in thin-wall phenotypes. Vertical bone loss was 3.5 times more severe than findings reported in the existing literature.

Mid-term results in relation to age and analysis of predictive factors after fixation of acetabular fractures using the modified Stoppa approach.

INTRODUCTION Data concerning outcome after management of acetabular fractures by anterior approaches with focus on age and fractures associated with roof impaction, central dislocation and/or quadrilateral plate displacement are rare. METHODS Between October 2005 and April 2009 a series of 59 patients (mean age 57 years, range 13-91) with fractures involving the anterior column was treated using the modified Stoppa approach alone or for reduction of displaced iliac wing or low anterior column fractures in combination with the 1st window of the ilioinguinal approach or the modified Smith-Petersen approach, respectively. Surgical data, accuracy of reduction, clinical and radiographic outcome at mid-term and the need for endoprosthetic replacement in the postoperative course (defined as failure) were assessed; uni- and multivariate regression analysis were performed to identify independent predictive factors (e.g. age, nonanatomical reduction, acetabular roof impaction, central dislocation, quadrilateral plate displacement) for a failure. Outcome was assessed for all patients in general and in accordance to age in particular; patients were subdivided into two groups according to their age (group "<60yrs", group "≥60yrs"). RESULTS Forty-three of 59 patients (mean age 54yrs, 13-89) were available for evaluation. Of these, anatomic reduction was achieved in 72% of cases. Nonanatomical reduction was identified as being the only multivariate predictor for subsequent total hip replacement (Adjusted Hazard Ratio 23.5; p<0.01). A statistically significant higher rate of nonanatomical reduction was observed in the presence of acetabular roof impaction (p=0.01). In 16% of all patients, total hip replacement was performed and in 69% of patients with preserved hips the clinical results were excellent or good at a mean follow up of 35±10 months (range: 24-55). No statistical significant differences were observed between both groups. CONCLUSION Nonanatomical reconstruction of the articular surfaces is at risk for failure of joint-preserving management of acetabular fractures through an isolated or combined modified Stoppa approach resulting in total joint replacement at mid-term. In the elderly, joint-preserving surgery is worth considering as promising clinical and radiographic results might be obtained at mid-term.