Two-laboratory collaborative study on identification of mycobacteria: molecular versus phenotypic methods

Previous studies have indicated that the conventional tests used for the identification of mycobacteria may (i) frequently result in erroneous identification and (ii) underestimate the diversity within the genus Mycobacterium. To address this issue in a more systematic fashion, a study comparing phenotypic and molecular methods for the identification of mycobacteria was initiated. Focus was given to isolates which were difficult to identify to species level and which yielded inconclusive results by conventional tests performed under day-to-day routine laboratory conditions. Traditional methods included growth rate, colonial morphology, pigmentation, biochemical profiles, and gas-liquid chromatography of short-chain fatty acids. Molecular identification was done by PCR-mediated partial sequence analysis of the gene encoding the 16S rRNA. A total of 34 isolates was included in this study; 13 of the isolates corresponded to established species, and 21 isolates corresponded to previously uncharacterized taxa. For five isolates, phenotypic and molecular analyses gave identical results. For five isolates, minor discrepancies were present; four isolates remained unidentified after biochemical testing. For 20 isolates, major discrepancies between traditional and molecular typing methods were observed. Retrospective analysis of the data revealed that the discrepant results were without exception due to erroneous biochemical test results or interpretations. In particular, phenotypic identification schemes were compromised with regard to the recognition of previously undescribed taxa. We conclude that molecular typing by 16S rRNA sequence determination is not only more rapid (12 to 36 h versus 4 to 8 weeks) but also more accurate than traditional typing.     

SOLUBILITY OF CO2 AND H2S IN A HINDERED AMINE SOLUTION

The solubility of H₂S and CO₂ in aqueous solutions of the sterically hindered amine, 2-amino-2-methyl-1-propanol (AMP), was determined at 40 and 100°C. Partial pressures of C)₂ ranged from approximately 2 to 6000 kPa and of H₂S from 2 to 2200 kPa. The solubility results were compared with previously reported acid gas solubilities in aqueous monoethanolamine (MEA) solutions.     

The selectivity of porous catalysts: parallel reactions

The influence of intraparticle mass diffusion on the overall and relative rates of the parallel reactions A → B and A → C is examined for the case of an isothermal catalyst particle with no external concentration or temperature gradients. Calculations for three pairs of reaction orders: (0,1), (0,2) and (1,2), show that internal concentration gradients can cause the yield of the product formed in the higher-order reaction to decrease by as much as 80 percent. Extension of the analysis to other reaction orders is discussed. Criteria for close approach to various asymptotic limits are presented, together with an asymptotic expression which allows the maximum effect of pore diffusion on selectivity to be estimated for any pair of reaction orders.     

Assessing the reliability of ordered categorical scales using kappa-type statistics

Methods for the analysis of reliability of ordered categorical scales are discussed, focussing on the limitation of the single summary-weighted kappa coefficients. A symmetric matrix of kappa-type coefficients is suggested as an alternative. The method is proposed as being suitable for ordinal scale where there is no underlying continuum. Their application is illustrated using two data sets from reliability studies. If, instead, distances between categories can be specified, a weighted mean of the matrix terms can be used as a summary measure. This is equal to a weighted kappa coefficient with squared weights, provided distances between adjacent categories are equal. When a study design corresponds to a one-way random effects model, estimates of precision of kappa-type coefficient, including the coefficients described here, can be obtained using the delta-method, bootstrap resampling by subjects or jack-knifing by subjects. In the case of interobserver reliability studies, where there may be systematic differences between observers, the investigator may wish to generalise to a population of observers and subjects. In this case, jack-knifing by observer and subject is suggested. Empirical comparisons are made between standard error estimates based on the delta-method, on jack-knifing by subjects and a two-way jack-knife by subjects and observers. The results suggest that standard errors based on the delta-method or jack-knifing by subject alone may be overly precise.     

A dislocation model for the two critical stress intensities required for threshold fatigue crack propagation

We have examined the variations, with decreasing load ratio, of threshold peak and cyclic stress intensities required for fatigue crack growth in stage I (mainly mode II loading) using a simple model simulating dislocation motion near a crack tip. In this model the crack grows by dislocations running into the crack during loading and unloading phases. Initially we have studied the behaviour of a crack with a dislocation source relatively far away from the crack tip. Crack propagation rates showed a Paris regime at high ΔK, and an abrupt threshold value ΔK_th below which no crack growth occurred. The variation with load ratio of the peak (K_th) and cyclic (ΔK_th) stress intensities at the fatigue threshold showed that two different processes controlled the behaviour. At high load ratios dislocations are generated readily during loading and the threshold is controlled by the need for sufficient unloading to allow dislocations to run back into the crack, so that the criterion ΔK ≥ ΔK∗ results. At negative load ratios it is the generation of dislocations during the loading phase that controls the threshold condition, since once generated, the large unloading and reversed loading easily forces dislocations back to the crack. Under these conditions the threshold criterion becomes K_max ≥ K∗.     

Chromium distribution between liquid iron and molten basic slags

An equilibrium study was made of the distribution of chromium and oxygen between liquid iron, containing less than 1 pct Cr, and simple slags of the CaO(MgO)-SiO₂-FeO-Cr₂O₃ type in the temperature range 1526° to 1734°C The effects of slag oxidation, temperature, and basicity were observed.