In vivo determination of RNA structure-function relationships: analysis of the 790 loop in ribosomal RNA

The 790 loop is a conserved hairpin located between positions 786 and 796 of Escherichia coli 16 S rRNA that is required for ribosome function. Using a novel genetic approach, all positions in the loop were simultaneously mutated and functional mutant sequences were selected in vivo. This "instant evolution" experiment revealed that approximately 190 of the 262,144 possible mutant sequences were functional. Analysis of functional mutant sequences allowed discrimination between nucleotides directly involved in protein synthesis and those involved primarily in loop structure. Among the functional mutant sequences, positions 789 and 791 were invariant and extensive covariation was observed among the nucleotides at the base of the loop at positions 787, 788, 794 and 795. NMR and thermodynamic analyses of model 790 hairpins in vitro revealed weak pairing interactions between positions 787 and 795 and between positions 788 and 794 consistent with the in vivo mutational analysis. Functional analysis of site-directed mutants containing all possible nucleotide combinations at positions 787 and 795 in vivo showed that stable base-pairs at these positions prevent subunit association.     

Hypertrophic pulmonary osteoarthropathy and its occurrence with pulmonary metastases from renal carcinoma

Studies concerning the ability of an inhaled beta2-agonist to produce dose-related bronchodilatation are conflicting. In five asthmatic, five bronchitic, and five normal subjects, specific airway conductance (Gaw/VL), flow-volume curves, and single-breath nitrogen washout were recorded after noncummulative inhalation of 0.65 mg, 1.30 mg, 1.95 mg, and, in some subjects, 2.60 mg of metaproterenol sulfate. Bronchodilatation appeared to be dose-related and was best assessed by using Gaw/VL; in all but one subject with chronic bronchitis, there was a significant linear relationship between log dose and precent change in Gaw/VL. Measurements of flow rate could demonstrate significant log dose-responses in eight subjects, maximum midexpiratory flow being the most sensitive measurement of flow rate, followed in order by forced expiratory volume in one second, flow at 50 percent of forced vital capacity, peak expiratory flow rate, and flow at 75 percent of forced vital capacity. No log dose-respone curve could be observed by using the single-breath nitrogen-washout technique. This demonstration of significant log dose-responses to inhaled metaproterenol is consistent with the response to drugs acting upon receptors and suggests that patients may benefit from increasing doses of bronchodilators.     

Proteinase activity in the isolates of Trichomonas vaginalis according to their pathogenicity

Ten axenic isolates of Trichomonas vaginalis were subcutaneously injected to the BALB/c mice in order to assess their pathogenicity by means of so-called "mouse assay" method. All the isolates revealed neutral and acid proteinase activities both in their lysates and in culture media, but the specific activities of both proteinases in the severely pathogenic group were significantly higher than the mildly pathogenic group (p < 0.05). In the SDS-PAGE system in which the electrophoretic gels contained 0.4% gelatin as the substrate, five different banding patterns of trichomonal proteinases were detected, and the patterns were closely related with the pathogenicity of the isolates of T. vaginalis. All five bands might be regarded as cysteine proteinases group in the inhibitor assays. The cytotoxicity of the lysates of T. vaginalis to the target Chinese hamster ovarian (CHO) cell line was also significantly different according to the pathogenicity of the isolates, and generally lower in the lysates treated with cysteine proteinase inhibitors than in the control lysates. In summarizing the results, it might be considered that the proteinases of T. vaginalis showing five electrophoretic banding patterns are closely related with the pathogenicity and cytotoxicity of the isolates of T. vaginalis.     

Chemical compositions and source identification of PM₂.₅ aerosols for estimation of a diesel source surrogate

Exposure to traffic-related pollution during childhood has been associated with asthma exacerbation, and asthma incidence. The objective of the Cincinnati Childhood Allergy and Air Pollution Study (CCAAPS) is to determine if the development of allergic and respiratory disease is associated with exposure to diesel engine exhaust particles. A detailed receptor model analyses was undertaken by applying positive matrix factorization (PMF) and UNMIX receptor models to two PM_2.5 data sets: one consisting of two carbon fractions and the other of eight temperature-resolved carbon fractions. Based on the source profiles resolved from the analyses, markers of traffic-related air pollution were estimated: the elemental carbon attributed to traffic (ECAT) and elemental carbon attributed to diesel vehicle emission (ECAD).Application of UNMIX to the two data sets generated four source factors: combustion related sulfate, traffic, metal processing and soil/crustal. The PMF application generated six source factors derived from analyzing two carbon fractions and seven factors from temperature-resolved eight carbon fractions. The source factors (with source contribution estimates by mass concentrations in parentheses) are: combustion sulfate (46.8%), vegetative burning (15.8%), secondary sulfate (12.9%), diesel vehicle emission (10.9%), metal processing (7.5%), gasoline vehicle emission (5.6%) and soil/crustal (0.7%). Diesel and gasoline vehicle emission sources were separated using eight temperature-resolved organic and elemental carbon fractions. Application of PMF to both datasets also differentiated the sulfate rich source from the vegetative burning source, which are combined in a single factor by UNMIX modeling. Calculated ECAT and ECAD values at different locations indicated that traffic source impacts depend on factors such as traffic volumes, meteorological parameters, and the mode of vehicle operation apart from the proximity of the sites to highways. The difference in ECAT and ECAD, however, was less than one standard deviation. Thus, a cost benefit consideration should be used when deciding on the benefits of an eight or two carbon approach.     

Polarity of axoplasmic microtubules in the olfactory nerve of the frog

Pieces of olfactory nerve of the bullfrog were extracted in a tubulin assembly buffer medium containing detergents. With incubation at 37 degrees C in such medium containing soluble tubulin, ribbons of protofilaments are formed on the surfaces of microtubules, with the ribbons curving in a clockwise or counterclockwise direction. The direction of hooking reflects the polarity of the microtubule. In nerve pieces oriented such that cross sections could be viewed toward the perikarya of the axons, over 90% of the ribbons on microtubules showed a clockwise orientation. When observers were looking toward the axonal terminals, most ribbons on microtubules showed a counterclockwise direction. In single axons in which ribbons appeared on all the contained microtubules, the ribbons showed a single directionality. The evidence suggests that microtubules in axons have a single polarity, probably reflecting their assembly from the perikarya outward through the axoplasm. If bidirectional transport is assumed in these axons, it is not reflected by the polarity of their microtubules, which may mean that the directionality of transport is provided by components other than microtubules.     

Trends in mortality of childhood-onset insulin-dependent diabetes mellitus in Leicestershire: 1940-1991

The relative risk of death by calendar date of diagnosis was investigated in a population-based incident cohort of 845 (463 males:382 females) IDDM diagnosed in Leicestershire before the age of 17 years between 1940 and 1989. The mortality status of 844 (99.9%) patients was determined as of the 31 December 1991, representing 14,346 person-years of risk. Trends in relative risk of death were investigated using Cox proportional hazards modelling for within cohort comparisons and age/sex and calendar time adjusted standardized mortality ratios (SMR) using generalized linear modelling for external comparisons. Median age at diagnosis was 10 years (range 3 months to 16 years); median duration of diabetes 15 years (range 1-51 years). Forty-four patients had died (5.2%; median age at death 31 years, range 11-51 years). A further four patients died at presentation (within 24 h) from ketoacidosis and are excluded from all analyses. Calendar date of diagnosis was found to be an important predictor of mortality. Adjusting for attained age there was evidence of a decline in relative risk of death with calendar date of diagnosis of 3.4% (95% CI, 0.005-6.9%) per annum, equivalent to a 32% fall per decade (95% CI, 5-51%), or 84% (95% CI, 21-97) from 1940 to 1989. The data are consistent with a large fall in mortality between the 1940s and 1950s representing over 50% of the total reduction in mortality between 1940 and 1991. Neither sex nor age at diagnosis were significant predictors of mortality. Over the study period 1940-89 the SMR (male and female combined) fell from 981 (541-1556) to 238 (60-953) relative to the general population. This population-based study shows that the prognosis for Type 1 (insulin-dependent) diabetes mellitus has improved markedly over the period 1940-1991.