Handling of spurious sequences affects the outcome of high-throughput 16S rRNA gene amplicon profiling

16S rRNA gene amplicon sequencing is a popular approach for studying microbiomes. However, some basic concepts have still not been investigated comprehensively. We studied the occurrence of spurious sequences using defined microbial communities based on data either from the literature or generated in three sequencing facilities and analyzed via both operational taxonomic units (OTUs) and amplicon sequence variants (ASVs) approaches. OTU clustering and singleton removal, a commonly used approach, delivered approximately 50% (mock communities) to 80% (gnotobiotic mice) spurious taxa. The fraction of spurious taxa was generally lower based on ASV analysis, but varied depending on the gene region targeted and the barcoding system used. A relative abundance of 0.25% was found as an effective threshold below which the analysis of spurious taxa can be prevented to a large extent in both OTU- and ASV-based analysis approaches. Using this cutoff improved the reproducibility of analysis, i.e., variation in richness estimates was reduced by 38% compared with singleton filtering using six human fecal samples across seven sequencing runs. Beta-diversity analysis of human fecal communities was markedly affected by both the filtering strategy and the type of phylogenetic distances used for comparison, highlighting the importance of carefully analyzing data before drawing conclusions on microbiome changes. In summary, handling of artifact sequences during bioinformatic processing of 16S rRNA gene amplicon data requires careful attention to avoid the generation of misleading findings. We propose the concept of effective richness to facilitate the comparison of alpha-diversity across studies

Practical assessment on the run – iPads as an effective mobile and paperless tool in physical education and teaching

This paper investigates the use of iPads in the assessment of predominantly second year Bachelor of Education (Primary/Early Childhood) pre-service teachers undertaking a physical education and health unit. Within this unit, practical assessment tasks are graded by tutors in a variety of indoor and outdoor settings. The main barriers for the lecturer or tutor for effective assessment in these contexts include limited time to assess and the provision of explicit feedback for large numbers of students, complex assessment procedures, overwhelming record-keeping and assessing students without distracting from the performance being presented. The purpose of this pilot study was to investigate whether incorporating mobile technologies such as iPads to access online rubrics within the Blackboard environment would enhance and simplify the assessment process. Results from the findings indicate that using iPads to access online rubrics was successful in streamlining the assessment process because it provided pre-service teachers with immediate and explicit feedback. In addition, tutors experienced a reduction in the amount of time required for the same workload by allowing quicker forms of feedback via the iPad dictation function. These outcomes have future implications and potential for mobile paperless assessment in other disciplines such as health, environmental science and engineering

Electrochemical Oxidation Assessment and Interaction of 2-aminoethanol and N, N-diethylethanamine Propagation in Acidic Medium

Electro�oxidation and inhibitor performance of copper specimens in 1 M hydrochloric acid solu� tion was investigated at room temperature by linear potentiodynamic polarization and gravimetric method in the presence of 2�aminoethanol (A) and N, N�diethylethanamine (D) as an inorganic inhibitor. The effect of the inhibitory concentration on the corrosion behavior of copper was studied over 288 hrs at 298°K. The inhibitory efficiency rise up to 96% for single induced and 98% for synergistic behavior. The adsorption mechanism characteristic was supported by SEM/EDX analysis and adsorption isotherm. From all indica� tion, the inhibitive efficiency of these compounds majorly depends on their molecular structure and concen� tration. The blocking effects of the surface interface were also explained on the basis of the inhibitor active action. 2�aminoethanol and N, N�diethylethanamine inhibits copper in 1 M HCl by strictly affecting both the anodic and cathodic sites. Portion of the surface covered calculated was also found to follow Langmuir adsorption isotherm

Respiratory mechanics in mice: strain and sex specific differences.

To assess the contribution of genetic background to respiratory mechanics, we developed a ventilator unit to measure lung function parameters in the mouse. We studied two commonly used inbred mice strains originating from Mus musculus domesticus (C57BL/6 and C3HeB/FeJ) and a third strain derived from Mus musculus molossinus [Japanese fancy mouse 1 (JF1)]. The ventilator allows for accurate performance of the different breathing manoeuvres required for measuring in- and expiratory reserve capacity, quasi-static and dynamic compliance, and airway resistance. In combination with a mass spectrometer for monitoring gas concentrations, single-breath manoeuvres were performed and He-expirograms obtained, from which dead space volume and slope of phase III were determined. From each strain and each sex, 10, 2-month old animals were studied immediately after being killed by an intraperitoneal overdose of xylazine and ketamine. C3HeB/FeJ and C57BL/6 exhibited comparable lung volumes. In male C3HeB/FeJ mice, e.g. vital capacity (VC) was 1072 +/- 79 microL, inspiratory reserve capacity 782 +/- 88 microL, and dead space volume at total lung inflation 216 +/- 18 microL. Lung volumes of JF1 were significantly lower (e.g. VC 611 +/- 53 microL, P < 0.01) even when normalized to body weight. In all three strains, specific lung volumes were significantly higher in females than in males, possibly explained by a higher oxygen demand during pregnancy and lactation, both of which fill most of their life times. Static compliance in C3HeB/FeJ was 64.3 +/- 5.4 microL cmH2O-1. It was smaller in C57BL/6 and JF1 mice, even when related to the lung volume. Analysis of the degree of genetic vs. non-genetic components of the phenotypic variation revealed that at least 80% of the total variation of lung volumes and static compliance in the mixed population is attributable to genetic differences between individuals. These differences will be verified in further studies by segregation and genetic linkage analysis

Preparation and characterization of atomically clean, stoichlometric surfaces of AIN(0001)

In situ exposure of the. (0001) surface of AlN thin films to flowing ammonia at 1120 degreesC and 10(-4) Torr removes oxygen/hydroxide and hydrocarbon species below the detectable limits of x-ray photoelectron spectroscopy and decreases the Al/N ratio from 1.3 to 1.0. The positions of the Al 2p and the N 1s core level peaks acquired from the cleaned surfaces were 75.0 +/- 0.1 eV and 398.2 +/- 0.1 eV, respectively, which were similar to the values determined for the as-loaded samples. The cleaning process left unchanged the (1 X 1) low energy electron diffraction pattern, the step-and-terrace microstructure, and the root mean square roughness values observed for the surfaces of the as-loaded samples; i.e., the surface structure and microstructure were not changed by the high-temperature exposure to ammonia at low pressures. Vacuum annealing under 10(-7) Torr at 1175 degreesC for 15 min removed all detectable hydrocarbons; however, it did not remove the oxygen/hydroxide species. (C) 2005 American Vacuum Society.Author has checked copyrightAD 09/01/201