High correlation between isoenzyme classification and kinetoplast DNA variability in Trypanosoma cruzi

By means of 14 restriction enzymes, we have studied the kinetoplast DNA polymorphism in 21 Trypanosoma cruzi isolates previously classified into 19 different genotypes based on the analysis of 15 isozyme loci. We have found a high correlation (p less than 0.001) between the proportion of restriction bands (fragments) common to any two given isolates and the corresponding genetic identities calculated from the isozyme data. This shows that the two classifications (kDNA and isozymes) corrobate one another and strongly suggests that the two types of variability are correlated with time (molecular clocks). The phylogenic classifications so obtained can be used as rational bases for medical and epidemiological studies. Although they are correlated, the two types of analysis are complementary as they do not yield identical results. Like the isozyme genetic distances and genetic identities, the values obtained for the proportion of common restriction bands (fragments) exhibit a continuum. This seems to confirm that natural T. cruzi populations exhibit a wide range of genotypes rather than a few well-differentiated clusters of strains.     

Site-directed mutagenesis of dicarboxylic acid residues of the penicillin-binding module of the Escherichia coli penicillin-binding protein 3

Abstract The glutamic acid E396, aspartic acid D409 and glutamic acid E411 residues of the Escherichia coli penicillin-binding protein 3 were each converted into an alanine residue. As deduced from penicillin-binding and complementation experiments, none of these dicarboxylic acid residues is involved in the mechanism of acylation by penicillin and none of them is essential for the in vivo functioning of the PBP. The mutation E396, however, causes an increased thermolability of the protein.     

Historical relationships of the Mexican cloud forests: a preliminary vicariance model applying Parsimony Analysis of Endemicity to vascular plant taxa

Mexican cloud forests, situated between 600 and 3000 m of elevation, exhibit a remarkable high biotic diversity. They follow a fragmented pattern, similar to that of an archipelago, that makes them suitable to vicariance modelling. A Parsimony Analysis of Endemicity (PAE) was applied to the presence/absence of 1267 species of vascular plants (gymnosperms, angiosperms, and pteridophytes) from twenty-four patches of Mexican cloud forests, in order to postulate a preliminary hypothesis of relationships. The single cladogram obtained grouped the twenty-four cloud forests into five clades. These results indicate that the Sierra Madre Oriental, Sierra Madre del Sur, and Serranías Meridionales floristic provinces do not represent natural units. A preliminary vicariance model is presented to explain the sequence of fragmentation of the Mexican cloud forests.     

Primer design for the cloning of immunoglobulin heavy-chain leader-variable regions from mouse hybridoma cells using the PCR

To facilitate the rapid cloning and sequencing of rearranged murine heavy-chain variable regions, we have designed a set of universal primers using conserved sequences of leader (signal peptide), framework one and constant regions of the immunoglobulin heavy-chain genes. RNA was extracted from the mouse hybridoma cells secreting monoclonal antibodies: IOR-T3 (anti-CD3), C6 (anti-P1 of N. meningitidis B385), IOR-T1 (anti-CD6), CB-CEA.1 (anti-carcinoembryonic antigen), CB-Fib.1 (anti-human fibrin) and CB-Hep.2 (anti-hepatitis B surface antigen). First-strand cDNA was synthesized and amplified using PCR. The primers successfully amplified correct size fragments from cDNA prepared from all hybridomas. These methods will facilitate the cloning and sequencing of mouse immunoglobulin variable regions.     

Isospora manchacensis n. sp., an intranuclear coccidian from the Louisiana ground skink, Scincella lateralis (Say, 1823) (Lacertilia: Scincidae)

Isospora manchacensis n. sp. is described from ground skinks, Scincella lateralis (Say, 1823) from Louisiana. Overall prevalence at 6 sites near Lake Ponchartrain was 43.1% (59/137) and ranged from 8% (1/13) to as high as 60% (6/10). Endogenous stages develop inside the nuclei of epithelial cells in the small intestine. Infected hypertrophic nuclei migrate from the basal lamina of the host cell to the luminal striated border. Oocysts in freshly passed fecal pellets usually contain a single contracted sporont that divides to form 2 sporoblasts. These undergo a brief pyramid stage followed by sporulation within 45-50 hr. Sporulated oocysts have a single-layered wall and measure 25.0 X 22.6 (20.0-28.9 X 18.6-26.0) micron. The lemon-shaped sporocysts measure 12.8 X 10.2 (11.1-15.2 X 9.0-11.0) micron and contain a Steida body, a spherical to oval substeida body, and a dispersed, granular sporocyst residuum. Prepatent periods in skinks fed 700 and 1,400 oocysts ranged from 24 to 32 days. Experimentally infected skinks produced large numbers of oocysts continuously during the 3-4 wk they were monitored after the onset of patency, but exhibited no signs of disease. Experimental doses of 200 oocysts failed to produce infections in skinks monitored for as long as 7 wk.     

Boolean Models: Maximum Likelihood Estimation from Circular Clumps

This paper deals with the problem of making inferences on the maximum radius and the intensity of the Poisson point process associated to a Boolean Model of circular primary grains with uniformly distributed random radii. The only sample information used is observed radii of circular clumps (DUPAC, 1980). The behaviour of maximum likelihood estimation has been evaluated by means of Monte Carlo methods.