Diagnostic and Treatment Challenges of Paroxysmal Nocturnal Hemoglobinuria in Uganda

Introduction. Paroxysmal nocturnal hemoglobinuria (PNH) is a very rare disorder of the hematopoietic stem cells which is often underdiagnosed. Its incidence is about 5 cases per million inhabitants in a year, and currently, there are only 1610 patients in the International PNH Registry. In this report, we describe a case of PNH in southwestern Uganda. Case Presentation. A 34-year-old man, subsistence farmer, with a history of multiple prior presentations with anemia, jaundice, and dark-colored urine requiring blood transfusions presented to us again in July 2018 with a week’s history of palpitations, dizziness, and dark-colored urine. Investigations done suggested a direct antiglobulin test- (DAT-) negative hemolytic anemia, and subsequently, flow cytometry showed a large clone of PNH. He received many blood transfusions and hematinics on several occasions during the course of his admissions. Conclusions. Our report showed diagnostic and treatment challenges of PNH in health resource-limited setting.

Individual based genetic analyses support asexual hydrochory dispersal in Zostera noltei

Dispersal beyond the local patch in clonal plants was typically thought to result from sexual reproduction via seed dispersal. However, evidence for the separation, transport by water, and re-establishment of asexual propagules (asexual hydrochory) is mounting suggesting other important means of dispersal in aquatic plants. Using an unprecedented sampling size and microsatellite genetic identification, we describe the distribution of seagrass clones along tens of km within a coastal lagoon in Southern Portugal. Our spatially explicit individual-based sampling design covered 84 km2 and collected 3 185 Zostera noltei ramets from 803 sites. We estimated clone age, assuming rhizome elongation as the only mechanism of clone spread, and contrasted it with paleo-oceanographic sea level change. We also studied the association between a source of disturbance and the location of large clones. A total of 16 clones were sampled more than 10 times and the most abundant one was sampled 59 times. The largest distance between two samples from the same clone was 26.4 km and a total of 58 and 10 clones were sampled across more than 2 and 10 km, respectively. The number of extremely large clone sizes, and their old ages when assuming the rhizome elongation as the single causal mechanism, suggests other processes are behind the span of these clones. We discuss how the dispersal of vegetative fragments in a stepping-stone manner might have produced this pattern. We found higher probabilities to sample large clones away from the lagoon inlet, considered a source of disturbance. This study corroborates previous experiments on the success of transport and re-establishment of asexual fragments and supports the hypothesis that asexual hydrochory is responsible for the extent of these clones.

Multiple Approaches To Enhance the Cultivability of Bacteria Associated with the Marine SpongeHaliclona(gellius) sp.

ABSTRACTThree methods were examined to cultivate bacteria associated with the marine spongeHaliclona(gellius) sp.: agar plate cultures, liquid cultures, and floating filter cultures. A variety of oligotrophic media were employed, including media with aqueous and organic sponge extracts, bacterial signal molecules, and siderophores. More than 3,900 isolates were analyzed, and 205 operational taxonomic units (OTUs) were identified. Media containing low concentrations of mucin or a mixture of peptone and starch were most successful for the isolation of diversity, while the commonly used marine broth did not result in a high diversity among isolates. The addition of antibiotics generally led to a reduced diversity on plates but yielded different bacteria than other media. In addition, diversity patterns of isolates from agar plates, liquid cultures, and floating filters were significantly different. Almost 89% of all isolates wereAlphaproteobacteria; however, members of phyla that are less commonly encountered in cultivation studies, such asPlanctomycetes,Verrucomicrobia, andDeltaproteobacteria, were isolated as well. The sponge-associated bacteria were categorized into three different groups. The first group represented OTUs that were also obtained in a clone library from previously analyzed sponge tissue (group 1). Furthermore, we distinguished OTUs that were obtained from sponge tissue (in a previous study) but not from sponge isolates (group 2), and there were also OTUs that were not obtained from sponge tissue but were obtained from sponge isolates (group 3). The 17 OTUs categorized into group 1 represented 10 to 14% of all bacterial OTUs that were present in a large clone library previously generated fromHaliclona(gellius) sp. sponge tissue, which is higher than previously reported cultivability scores for sponge-associated bacteria. Six of these 17 OTUs were not obtained from agar plates, which underlines that the use of multiple cultivation methods is worthwhile to increase the diversity of the cultivable microorganisms from sponges.

Low Taxon Richness of Bacterioplankton in High-Altitude Lakes of the Eastern Tibetan Plateau, with a Predominance of Bacteroidetes and Synechococcus spp.

ABSTRACT Plankton samples were collected from six remote freshwater and saline lakes located at altitudes of 3,204 to 4,718 m and 1,000 km apart within an area of ca. 1 million km2 on the eastern Tibetan Plateau to comparatively assess how environmental factors influence the diversity of bacterial communities in high-altitude lakes. The composition of the bacterioplankton was investigated by analysis of large clone libraries of 16S rRNA genes. Comparison of bacterioplankton diversities estimated for the six Tibetan lakes with reference data previously published for lakes located at lower altitudes indicated relatively low taxon richness in the Tibetan lakes. The estimated average taxon richness in the four Tibetan freshwater lakes was only one-fifth of the average taxon richness estimated for seven low-altitude reference lakes. This cannot be explained by low coverage of communities in the Tibetan lakes by the established libraries or by differences in habitat size. Furthermore, a comparison of the taxonomic compositions of bacterioplankton across the six Tibetan lakes revealed low overlap between their community compositions. About 70.9% of the operational taxonomic units (99% similarity) were specific to single lakes, and a relatively high percentage (11%) of sequences were <95% similar to publicly deposited sequences of cultured or uncultured bacteria. This beta diversity was explained by differences in salinity between lakes rather than by distance effects. Another characteristic of the investigated lakes was the predominance of Cyanobacteria (Synechococcus) and Bacteroidetes. These features of bacterioplankton diversity may reflect specific adaptation of various lineages to the environmental conditions in these high-altitude lakes.

Synthetic Statistical Approach Reveals a High Degree of Richness of Microbial Eukaryotes in an Anoxic Water Column

ABSTRACT Molecular surveys suggest that communities of microbial eukaryotes are remarkably rich, because even large clone libraries seem to capture only a minority of species. This provides a qualitative picture of protistan richness but does not measure its real extent either locally or globally. Statistical analysis can estimate a community's richness, but the specific methods used to date are not always well grounded in statistical theory. Here we study a large protistan molecular survey from an anoxic water column in the Cariaco Basin (Caribbean Sea). We group individual 18S rRNA gene sequences into operational taxonomic units (OTUs) using different cutoff values for sequence similarity (99 to 50%) and systematically apply parametric models and nonparametric estimators to the OTU frequency data to estimate the total protistan diversity. The parametric models provided statistically sound estimates of protistan richness, with biologically meaningful standard errors, maximal data usage, and extensive model diagnostics and were preferable to the available nonparametric tools. Our clone library exceeded 700 clones but still covered only a minority of species and less than half of the larger protistan clades. Our estimates of total protistan richness portray the target community as very rich at all OTU levels, with hundreds of different populations apparently co-occurring in the small (3-liter) volume of our sample, as well as dozens of clades of the highest taxonomic order. These estimates are among the first for microbial eukaryotes that are obtained using state-of-the-art statistical methods and can serve as benchmark numbers for the local diversity of protists.

Comparison of the beta-lactamase gene cluster in clonally distinct strains of Enterococcus faecalis.

Ten beta-lactamase-producing Enterococcus faecalis isolates were examined for the presence of the staphylococcal beta-lactamase repressor and antirepressor genes. Four isolates, previously shown to be unrelated to each other by pulsed-field gel electrophoresis analysis, were positive for both genes by PCR, although beta-lactamase production was not induced with methicillin. Six isolates, previously shown to be clonally related, were negative for both genes by PCR. The blaZ sequences of eight beta-lactamase-producing E. faecalis isolates were determined. Seven isolates from five distinct clones had sequences identical to that previously reported for E. faecalis HH22, regardless of whether the repressor or antirepressor was demonstrated by PCR. However, blaZ from one isolate differed from those of the other enterococci by 11 nucleotides; this isolate is part of the large clone, as defined by pulsed-field gel electrophoresis and multilocus enzyme analysis, that includes HH22. These findings suggest either that enterococci have acquired the bla gene cluster from more than one source or that the gene cluster has undergone considerable change since acquisition by this clone.

Variation within and among Blueberry Taxa in Flower Size and Shape

Variation was studied within and among five Vaccinium taxa for the flower parameters corolla length, corolla aperture diameter, stigma location relative to the apex of the corolla tube, position of the anthers relative to the stigma and to the apex of the corolla, and style length. The objective was to determine whether there was enough genetic variation to breed cultivars with flower shapes that might favor pollination by a wider range of bee species. The taxa studied were cultivated rabbiteye (V. ashei Reade), cultivated southern highbush (mainly V. corymbosum L. with up to 30% introgression from V. darrowi Camp), F1 V. ashei × V. constablaei A. Gray hybrids, V. darrowi, and V. elliottii Chapm. Vaccinium elliottii flowers differed from all others in having short styles that were not exserted from the corolla tube. Vaccinium elliottii was also unusual in that the end of the anther tube extended nearly to the stigmatic surface. Vaccinium ashei corollas were longer and had smaller apertures than those of southern highbush, possibly making them less suitable for honeybee (Apis) pollination. For corolla length and aperture diameter, F1 V. ashei × V. constablaei hybrids were similar to southern highbush, indicating that V. constablaei introgression could be used to breed hexaploid cultivars with shorter, more open flowers. Large clone-to-clone variation within taxa for each flower characteristic indicates much potential for changing the shape of the blueberry flower by breeding, if the shape that maximizes fruit set can be determined.

Design and analysis of chromosome physical mapping experiments

Mathematical and statistical aspects of constructing ordered-clone physical maps of chromosomes are reviewed. Three broad problems are addressed: analysis of fingerprint data to identify configurations of overlapping clones, prediction of the rate of progress of a mapping strategy and optimal design of pooling schemes for screening large clone libraries.

Interaction of Cottonwood Clones with Site and Planting Year

Height and diameter growth to age three and the number of first year branches were analyzed for 25 cottonwood clones grown in six plantations in central Mississippi. Plantations were on two contrasting sites and planted in three consecutive years. Results showed small clone × planting year interactions and large clone × site interactions for all parameters. In the lower Mississippi Valley, therefore, emphasis should be placed on testing over a range of sites rather than replicating over time.