Differentiation of ruminant transmissible spongiform encephalopathy isolate types, including bovine spongiform encephalopathy and CH1641 scrapie

With increased awareness of the diversity of transmissible spongiform encephalopathy (TSE) strains in the ruminant population, comes an appreciation of the need for improved methods of differential diagnosis. Exposure to bovine spongiform encephalopathy (BSE) has been associated with the human TSE, variant Creutzfeldt–Jakob disease, emphasizing the necessity in distinguishing low-risk TSE types from BSE. TSE type discrimination in ruminants such as cattle, sheep, goats and deer, requires the application of several prion protein (PrP)-specific antibodies in parallel immunochemical tests on brain homogenates or tissue sections from infected animals. This study uses in a single incubation step, three PrP-specific antibodies and fluorescent Alexa dye-labelled anti-mouse Fabs on a Western blot. The usual amount of brain tissue needed is 0.5 mg. This multiplex application of antibodies directed towards three different PrP epitopes enabled differential diagnosis of all established main features of classical scrapie, BSE and Nor98-like scrapie in sheep and goats, as well as the currently known BSE types C, H and L in cattle. Moreover, due to an antibody-dependent dual PrP-banding pattern, for the first time CH1641 scrapie of sheep can be reliably discriminated from the other TSE isolate types in sheep

Generalizations of Sturmian sequences associated with NN-continued fraction algorithms

Given a positive integer $N$ and $x$ irrational between zero and one, an $N$-continued fraction expansion of $x$ is defined analogously to the classical continued fraction expansion, but with the numerators being all equal to $N$. Inspired by Sturmian sequences, we introduce the $N$-continued fraction sequences $\omega(x,N)$ and $\hat{\omega}(x,N)$, which are related to the $N$-continued fraction expansion of $x$. They are infinite words over a two letter alphabet obtained as the limit of a directive sequence of certain substitutions, hence they are $S$-adic sequences. When $N=1$, we are in the case of the classical continued fraction algorithm, and obtain the well-known Sturmian sequences. We show that $\omega(x,N)$ and $\hat{\omega}(x,N)$ are $C$-balanced for some explicit values of $C$ and compute their factor complexity function. We also obtain uniform word frequencies and deduce unique ergodicity of the associated subshifts. Finally, we provide a Farey-like map for $N$-continued fraction expansions, which provides an additive version of $N$-continued fractions, for which we prove ergodicity and give the invariant measure explicitly.Comment: 23 pages, 2 figure

The transcription factor GATA6 is essential for early extraembryonic development

The gene coding for the murine transcription factor GATA6 was inactivated by insertion of a beta-galactosidase marker gene. The analysis of heterozygote GATA6/lacZ mice shows two inductions of GATA6 expression early in development. It is first expressed at the blastocyst stage in part of the inner mass and in the trophectoderm. The second wave of expression is in parietal endoderm (Reichert's membrane) and the mesoderm and endoderm that form the heart and gut. Inactivation leads to a lethality shortly after implantation (5.5 days postcoitum). Chimeric experiments show this to be caused by an indirect effect on the epiblast due to a defect in an extraembryonic tissue

Variability in disease phenotypes within a single PRNP genotype suggests the existence of multiple natural sheep scarpie strains within Europe

Variability of pathological phenotypes within classical sheep scrapie cases has been reported for some time, but in many instances it has been attributed to differences in the PRNP genotype of the host. To address this issue we have examined by immunohistochemistry (IHC) and Western blotting (WB) for the disease-associated form of the prion protein (PrPd), the brains of 23 sheep from five European countries, all of which were of the same ARQ/ARQ genotype. As a result of IHC examinations, sheep were distributed into five groups with different phenotypes and the groups were the same regardless of the scoring method used, ‘long’ or ‘short’ PrPd profiling. The groups made did not respond to the geographical origin of the cases and did not correlate with the vacuolar lesion profiles, which showed a high individual variability. Discriminatory IHC and WB methods coincided to detect a ‘CH1641-like’ case but otherwise correlated poorly in the classification of disease phenotypes. No other polymorphisms of the PRNP gene were found that could account for the pathological differences, except perhaps for a sheep from Spain with a mutation at codon 103 and a unique pathological phenotype. Preliminary evidence indicates that those different IHC phenotypes correlate with distinct biological properties on bioassay, suggesting that they are indicative of strain diversity. We therefore conclude that natural scrapie strains exist and that they can be revealed by detailed pathological examinations, which can be harmonized between laboratories to produce comparable results

Direct evidence of the failure of electric-dipole approximation in second-harmonic generation from a chiral polymer film

Second-harmonic generation from Langmuir-Blodgett films of a polythiophene is strongly influenced by the chirality of the polymer. The polarization dependence of the process cannot be explained in the elec.-dipole approxn. Evidence of contributions beyond elec. dipoles is obtained directly from individual second-harmonic signal

Severe tracheal and bronchial collapse in adults with type II mucopolysaccharidosis

Background: Mucopolysaccharidosis type II (MPSII) patients frequently suffer from dyspnoea caused by restrictive airway disease due to skeletal abnormalities as well as glycosaminoglycans (GAG) accumulation at different levels of the airway, including the trachea. In this study we describe the extent of the tracheal and bronchial narrowing, the changes in airway diameter during respiration and the effects of these obstructions on respiratory function in adult MPSII patients. Methods: Five adult MPSII patients (mean age 40 years) were included. Pulmonary function tests and in- and expiratory chest CT scans were obtained. Cross-sectional areas of trachea and main bronchi were measured at end-inspiration and -expiration and percentage collapse was calculated. Results: There was diffuse narrowing of the entire intra-thoracic trachea and main bronchi and severe expiratory collapse of the trachea in all patients. At 1 cm above the aortic arch the median % collapse of the trachea was 68 (range 60 to 77 %), at the level of the aortic arch 64 (range 21-93 %), for the main bronchi this was 58 (range 26-66 %) on the left and 44 (range 9-76 %) on the right side. The pulmonary function tests showed that this airway collapse results in obstructive airway disease in all patients, which was severe (forced expiratory volume <50 % of predicted) in four out of five patients. Conclusion: In adult MPS II patients, central airways diameters are strikingly reduced and upon expiration there is extensive collapse of the trachea and main bronchi. This central airways obstruction explains the severe respiratory symptoms in MPSII patients

To Improve Your Surgical Drilling Skills, Make Use of Your Index Fingers

BACKGROUND: Surgery has greatly benefited from various technologic advancements over the past decades. Surgery remains, however, mostly manual labor performed by well-trained surgeons. Little research has focused on improving osseous drilling techniques. The objective of this study was to compare the accuracy and precision of different orthopaedic drilling techniques involving the use of both index fingers. QUESTIONS/PURPOSES: (1) Does the shooting grip technique and aiming at the contralateral index finger improve accuracy and precision in drilling? (2) Is the effect of drilling technique on accuracy and precision affected by the experience level of the performer? METHODS: This study included 36 participants from two Dutch training hospitals who were subdivided into three groups (N = 12 per group) based on their surgical experience (that is, no experience, residents, and surgeons). The participants had no further experience with drilling outside the hospital nor were there other potential confounding variables that could influence the test outcomes. Participants were instructed to drill toward a target exit point on a synthetic bone model. There were four conditions: (1) clenched grip without aiming; (2) shooting grip without aiming; (3) clenched grip with aiming at the contralateral index finger; and (4) shooting grip aiming at the contralateral index finger. Participants were only used to a clenched grip without aiming in clinical practice. Each participant had to drill five times per technique per test, and the test was repeated after 4 weeks. Accuracy was defined as the systematic error of all measurements and was calculated as the mean of the five distances between the five exit points and the target exit point, whereas precision was defined as the random error of all measurements and calculated as the SD of those five distances. Accuracy and precision were analyzed using mixed-design analyses of variance. RESULTS: Accuracy was highest when using a clenched grip with aiming at the index finger (mean 4.0 mm, SD 1.1) compared with a clenched grip without aiming (mean 5.0 mm, SD 1.2, p = 0.004) and a shooting grip without aiming (mean 4.9 mm, SD 1.4, p = 0.015). The shooting grip with aiming at the index finger (mean 4.1 mm, SD 1.2) was also more accurate than a clenched grip without aiming (p = 0.006) and a shooting grip without aiming (p = 0.014). Shooting grip with aiming at the opposite index finger (median 2.0 mm, interquartile range [IQR] 1.2) showed the best precision and outperformed a clenched grip without aiming (median 2.9 mm, IQR 1.1, p = 0.016), but was not different than the shooting grip without aiming (median 2.2 mm, IQR 1.4) or the clenched grip with aiming (median 2.4 mm, IQR 1.3). The accuracy of surgeons (mean 4.1 mm, SD 1.1) was higher than the inexperienced group (mean 5.0 mm, SD 1.1, p = 0.012). The same applied for precision (median 2.2 mm, IQR 1.0 versus median 2.8 mm, IQR 1.4, p = 0.008). CONCLUSIONS: A shooting grip combined with aiming toward the index finger of the opposite hand had better accuracy and precision compared with a clenched grip alone. Based on this study, experience does matter, because the orthopaedic surgeons outperformed the less experienced participants. Based on our study, we advise surgeons to aim at the index finger of the opposite hand when possible and to align the ipsilateral index finger to the drill bit. LEVEL OF EVIDENCE: Level II, therapeutic study