The mouse formin (Fmn) gene: abundant circular RNA transcripts and gene-targeted deletion analysis.

BACKGROUND: Mutations in the mouse formin (Fmn) gene result in limb deformities and incompletely penetrant renal aplasia. A molecular genetic approach was taken to characterize novel circular RNAs from the Fmn gene and to understand the developmental effects of gene-targeted mutations. MATERIALS AND METHODS: RT-PCR and ribonuclease protection analyses were done to characterize the circular RNA transcripts arising from the Fmn gene. Two lines of mice with gene-targeted deletions of specific Fmn exons, namely exon 4 or exon 5, were generated and analyzed. RESULTS: In our analysis of formin cDNAs, we discovered a class of transcripts in which the exon order is reversed such that downstream exons are joined to the acceptor end of a specific exon that lies 5' to them in the genome. RT-PCR and ribonuclease protection analyses indicate that these transcripts are circular and are the major transcripts arising from this locus in adult brain and kidney. To gain insight into the biological function of these transcripts, we have systematically deleted the relevant exons using gene-targeted homologous recombination. The resulting mice fail to produce circular transcripts, but appear to produce normal amounts of the linear RNA isoforms from this locus. While these deficient mice have normal limbs, they display variably penetrant renal aplasia characteristic of other mutant formin alleles. CONCLUSIONS: Our results demonstrate novel circular transcripts arising from the Fmn gene. Moreover, their high levels of expression suggest that they are not products of aberrant splicing events, but instead, may play important biological roles. Mice with gene-targeted deletions of Fmn exons 4 or 5 lack these circular transcripts and have an incompletely penetrant renal agenesis phenotype. While the biologic function of circular Fmn RNA transcripts is not entirely known, our work suggests their possible involvement in kidney development.

Deaths in tuberculosis patients in British Columbia, 1980-1984.

Records of all 1884 newly notified tuberculosis cases, over the 5-year period 1980-1984 in British Columbia, Canada, were reviewed and 201 deaths were identified, including 48 diagnosed only after death, and 153 who died while on treatment; 56 of unrelated causes, 67 in whom tuberculosis was a contributing cause, and 30 in whom it was the principal cause. Significant predictors of death while on treatment (specific to tuberculosis) were the extent of disease, history of previous disease and sputum smear-positive for acid-fast organisms. Significant predictors of failure of diagnosis, in patients who died, were the presence of disseminated disease and the absence of a history of previous disease. The presenting features were not different in those dying, in whom the diagnosis was made before, as compared with after, death. The most frequent mode of death due to tuberculosis was respiratory failure, followed by multiple organ system failure and haemoptysis. The case fatality rate was low (1.6%) and did not change over 10 years. One-half of patients whose death was due to tuberculosis were diagnosed only after death and this had not changed over 10 years. We conclude that death due to tuberculosis is uncommon in patients while on treatment and that the main reason for death due to tuberculosis is that some patients are not diagnosed, and therefore not treated, before they die of the disease.

Tuberculosis in British Columbia among immigrants from five Asian countries, 1982-85.

The influence of immigration from six selected Asian countries--Japan, Korea, Philippines, India, China and Hong Kong--on the incidence of tuberculosis in British Columbia has been examined. During the period 1982-1985 the average annual incidence of bacillary tuberculosis in these immigrants was more than six times as great as the overall British Columbia rate and contributed a quarter of the cases of active bacillary tuberculosis in this province whereas the immigrants from these countries represented only 3.7% of the total population. The clinical patterns of active tuberculosis by birthplace were analysed. A high proportion of cases of lymphadenitis was seen among all immigrants from Asia, particularly those from the Philippines. Both primary and secondary drug resistance was substantially higher than in Canadian-born patients. The frequency of primary drug resistance was higher among patients aged less than 40 years than those aged 40 or more.

Inner-city survey for tuberculosis: evaluation of diagnostic methods.

A total of 1,271 persons living in a socially and economically depressed, inner-city area of Vancouver, British Columbia, voluntarily attended a tuberculosis case-finding campaign. Chest x-ray, on-the-spot specimen of sputum, and tuberculin skin test were offered at the time of the first attendance. All 3 diagnostic methods were found to be well accepted, with 93% of the participants having an x-ray, over 95% producing a sputum specimen, and almost 95% having a tuberculin test (a quarter of these did not, however, report for reading of the test). Eight cases of bacteriologically confirmed pulmonary tuberculosis were found: 6 suspected on x-ray (the remaining 2 films were abnormal but not diagnostic of tuberculosis), and 6 being positive on smear and/or culture of the initial on-the-spot sputum specimen. Examination of a second specimen of sputum diagnosed all of the 8 active cases identified by the survey. These results suggest that, in this particular setting, a chest radiogram taken by a transportable chest x-ray apparatus or examination of 2 sputum specimens might be equally successful at detecting all cases of active pulmonary tuberculosis within the time required for sputum culture. Examination of the sputum smear immediately identifies all the more infectious cases of pulmonary tuberculosis. The prevalence rate of 629 per 100,000 among those presenting themselves to this campaign illustrates the high-yield which might be achieved by active case-finding projects in known high-incidence segments of a generally low-incidence population.

Immunological study of proteins and mucosubstance in saline soluble human ocular mucus.

Proteins and mucosubstance of the saline extract of human ocular mucus were studied by immunological analysis. A minor study was made with human tears for comparison. Immunoelectrophoresis of proteins from these two sources consistently revealed similar characteristic gel patterns. Proteins were found as the major constituents of both samples. However, more mucosubstance was present in the saline extract of human ocular mucus than in tears. Seventeen proteins were identified in the mucus extract. Albumin, IgA, and lactoferrin appeared to be the three major proteins, while lysozyme, lactoferrin, tear prealbumin, and ocular mucoisolate were tear and ocular mucus specific. Although saline soluble mucoisolate is complex in structure, it seemed to resist electrical dissociation, producing only one major precipitation line along with a line of IgA during immunoelectrophoresis. The ocular mucoisolate accounted for about 12% of the saline extractable proteins of human ocular mucus.

Isolation and geographic distribution of Mycobacterium other than M. tuberculosis in British Columbia, 1972-81.

It has been suggested that the incidence of infection with mycobacteria other than typical tubercle (MOTT) bacilli is increasing. Laboratory and epidemiologic information relating to MOTT infection in British Columbia between 1972 and 1981 was analysed. Patient records for 1960-81 were also analysed. Of the 313 661 laboratory specimens 13 474 yielded Mycobacterium tuberculosis isolates and 3172, MOTT isolates. Over the 10 years the number of M. tuberculosis isolates declined, whereas the absolute and relative numbers of MOTT isolates increased. Members of the highly drug-resistant MAIS complex (M. avium-intracellulare, M. scrofulaceum and M. simiae) accounted for 73.3% of the 1778 potentially pathogenic MOTT isolates. MAIS isolation rates varied geographically. Analysis of patient records revealed 217 MOTT infections, of which 152 (70%) were due to MAIS organisms. Further studies are needed to determine the source of MAIS organisms in order that the infection and the disease may be more clearly understood.