Evaluation of endothelium-protective effects of drugs in experimental models of endothelial damage

Endothelium-protective properties of pharmacological agents may be assessed by using different experimental models of endothelial dysfunction or injury. The model of endothelial dysfunction induced by vessel perfusion with polymorphonuclear leukocytes (PMN) was used for evaluation of pentoxifylline (PTX) effects on vasoconstrictor responses to noradrenaline (NA) in the rabbit renal artery. Addition of PMN into the perfusion solution significantly increased the responses to NA at all doses. PTX administration (10(-5) mol x l(-1)) significantly diminished the constrictor responses to NA in vessels perfused with PMN+PTX when compared to the responses in PMN-perfused vessels (at dose 0.1 microg: 32.25 vs. 14.25, at dose 1 microg: 51 vs. 27.75 (p<0.01), at dose 10 microg 74.25 vs. 39.75 (p<0.05), all values expressed as median of perfusion pressure in mm Hg). The model of endothelial damage induced by repeated NA administration in 5 doses (10-50 microg of NA) was used for evaluation of the endothelium-protective effect of sulodexide (SLX). It was found that SLX (120 U/l) significantly decreased the number of desquamated endothelial cells (EC) compared to the control group (controls: 131.4+/-20.1 EC, +SLX: 83.3+/-13.8 EC, p<0.01). These results confirmed the favorable endothelium-protective effects of pentoxifylline and sulodexide in the two experimental models.     

Bryophytes of Kédougou (Eastern Senegal), with a key to the Fissidens of Senegal

The bryophyte flora of the Sahelian region of sub-Saharan Africa is characterized by a low species richness, but is also extremely poorly documented. We present here the results of a floristic survey in Kedougou (eastern Senegal), which yielded 22 species. Two liverworts and 15 moss species are new to the country, so that the bryophyte flora of Senegal now includes a total of 4 liverwort and 34 moss species. An identification key is provided for Fissidens, which, with eight species, is the richest moss genus of the country.     

The Population Genomics of Trans-Specific Inversion Polymorphisms in Anopheles gambiae

In the malaria mosquito Anopheles gambiae polymorphic chromosomal inversions may play an important role in adaptation to environmental variation. Recently, we used microarray-based divergence mapping combined with targeted resequencing to map nucleotide differentiation between alternative arrangements of the 2La inversion. Here, we applied the same technique to four different polymorphic inversions on the 2R chromosome of An. gambiae. Surprisingly, divergence was much lower between alternative arrangements for all 2R inversions when compared to the 2La inversion. For one of the rearrangements, 2Ru, we successfully mapped a very small region (∼100 kb) of elevated divergence. For the other three rearrangements, we did not identify any regions of significantly high divergence, despite ample independent evidence from natural populations of geographic clines and seasonal cycling, and stable heterotic polymorphisms in laboratory populations. If these inversions are the targets of selection as hypothesized, we suggest that divergence between rearrangements may have escaped detection due to retained ancestral polymorphism in the case of the youngest 2R rearrangements and to extensive gene flux in the older 2R inversion systems that segregate in both An. gambiae and its sibling species An. arabiensis.MORE than 70 years ago Dobzhansky and Sturtevant (1938) first discovered polymorphic inversion arrangements carried by various Drosophila pseudoobscura populations. After observing correlations between environmental conditions and inversion frequencies, Dobzhansky proposed that inversions are under strong selection due to their role in promoting local adaptation to the heterogeneous conditions a species encounters both spatially and temporally (Dobzhansky 1944, 1948; Powell 1997). More recent studies have implicated chromosomal inversions in the adaptation of a diversity of eukaryotes including humans (Coluzzi et al. 1979; Feder et al. 2003; Hoffmann et al. 2004; Stefansson et al. 2005). Long known to be common in dipteran insects, more recent HapMap data suggest that polymorphic inversions may be numerous in human populations and by extension other mammals (Bansal et al. 2007). Given their potential importance in facilitating adaptation, surprisingly little is known about the mechanism(s) or the genes responsible for maintaining inversion polymorphisms in natural populations.Gene exchange between inverted and standard arrangements, although reduced, can still occur through gene flux: the action of gene conversion and multiple crossovers in inversion heterozygotes (heterokaryotypes) (Chovnick 1973; Navarro et al. 1997; Schaeffer and Anderson 2005). Over time allelic variation unrelated to ecological adaptation should become homogenized between arrangements, while alleles which are under divergent selection pressures should remain in linkage disequilibrium with each other and with the inversion itself, leading to heightened differentiation between standard and inverted arrangements at and near the target loci. In principle, this process allows the identification of specific loci involved in adaptive divergence (Schaeffer et al. 2003; Schaeffer and Anderson 2005; Storz 2005). Consistent with this model, previous low-resolution studies of Drosophila inversions revealed heterogeneous patterns of nucleotide diversity relative to divergence, as well as the interspersion of regions of high and low genetic association potentially due to the interaction of selection and gene flux (Schaeffer et al. 2003; Kennington et al. 2006; but see Munte et al. 2005). The application of high-resolution tools flowing from completely sequenced genomes will facilitate the mapping of genes that are the targets of divergent natural selection within gene arrangements.Although Drosophila has been the favored model, the African malaria vector Anopheles gambiae sensu stricto also provides an excellent system for studying the maintenance of inversion polymorphisms, not only within a species but across speciation events of different ages in the An. gambiae sibling species complex. The nominal species An. gambiae s.s. (hereafter, An. gambiae) is synanthropic: almost exclusively biting humans, resting indoors, and exploiting anthropogenic larval habitats (Coluzzi 1999). This close association with humans, vital to making An. gambiae one of the most proficient vectors of malaria, is likely to have been facilitated by chromosomal inversions thought to confer adaptive benefits in heterogeneous climatic and ecological settings in Africa. Seven common polymorphic inversions exist on the second chromosome. Six of these are located on the right arm (2R): j, b, c, u, d, and k, while 2La is the only inversion on the left arm (Coluzzi et al. 2002). Facilitated by the sequenced reference genome (Holt et al. 2002), some of the breakpoints for these polymorphic inversions have been localized to small genomic regions (Sharakhov et al. 2006; Coulibaly et al. 2007; Sangare 2007). Most of these inversions appear to be the targets of strong selection. Five of the inversions (2La and 2Rb, -c, -d, and -u) are nonrandomly associated with degree of aridity; each cycles seasonally with rainfall, and all except 2Ru form stable geographic clines in frequency from mesic forest to xeric regions bordering the Sahara (Coluzzi et al. 1979; Toure et al. 1994, 1998; Powell et al. 1999). Inversion 2Rj is not clinal, but its distribution in Mali is consistent with adaptation to novel rockpool niches (Coluzzi et al. 1985; Manoukis et al. 2008).In the An. gambiae species complex, inversion polymorphisms can be maintained across the boundaries of emerging and even full species. An. gambiae and its sibling An. arabiensis, strictly sympatric throughout most of their extensive ranges in sub-Saharan Africa, differ by multiple fixed chromosomal rearrangements on the X but share three chromosome 2 inversions: 2La, fixed in An. arabiensis and polymorphic in An. gambiae; and 2Rb and -c, polymorphic in both species (Coluzzi et al. 1979, 2002). Moreover, these same inversions and all other common An. gambiae inversions with the exception of 2Rj are shared and polymorphic in two lineages apparently undergoing ecological speciation within An. gambiae—the assortatively mating M and S molecular forms (della Torre et al. 2002, 2005). Inversion frequencies are correlated with climatic and ecological conditions in parallel in both lineages (Costantini et al. 2009; Simard et al. 2009). Unlike the full species, the M and S incipient species are not distinguished by any fixed inversion differences. Indeed, genomewide divergence mapping between the M and S forms revealed that significant differentiation was confined to two small low-recombination regions adjacent to the centromeres of 2L and X which are distant from any inversions (Turner et al. 2005). Thus, in distinction to models of speciation invoking inversions as facilitating the persistence of hybridizing species (Noor et al. 2001; Rieseberg 2001; Ortiz-Barrientos et al. 2002; Navarro and Barton 2003), the An. gambiae data suggest that chromosome 2 inversions are not directly responsible for reproductive isolation. Instead, the same chromosome 2 inversion polymorphisms appear to confer similar ecological benefits, within and across species boundaries. A long-term research goal is to identify the mechanisms and the genes controlling these processes.Previously we conducted the first high-density genomic scan of divergence across a chromosomal inversion (2La) in An. gambiae (White et al. 2007). By hybridizing genomic DNA from S form mosquitoes homokaryotypic for alternate gene arrangements on chromosome 2L (2La or 2L+^a) to oligonucleotide microarrays we were able to measure divergence across the 22-Mb inversion at nearly 14,000 markers. Differentiation in the rearranged region was significantly higher than in collinear portions of chromosome 2L. Between breakpoints the pattern of differentiation was heterogeneous: two genomic clusters of significantly higher divergence were identified near but not adjacent to the breakpoints. Directed resequencing within the S form confirmed these results and suggested that both clusters contained genes targeted by selection. Observed levels of linkage disequilibrium between the 2La breakpoints and markers in the clusters are highly unlikely under a neutral scenario, in light of known recombination rates and plausible estimates of the age of the inversion.The present study characterizes the patterns of genetic variation in polymorphic rearrangements on the opposite (right) arm of chromosome 2: 2Rj, -b, -c, and -u. With the goal of identifying candidate genes maintaining these inversions in natural populations, we applied microarray-based divergence mapping to measure differentiation between alternative 2R arrangements. Because three of four inversions have taxonomic distributions that span incipient and/or completed speciation events, we validated the microarray findings by targeted sequencing in multiple taxa: sympatric Malian populations of An. gambiae M and S forms, and the sibling species An. arabiensis.     

Reconstructing Colonization Dynamics of the Human Parasite Schistosoma mansoni following Anthropogenic Environmental Changes in Northwest Senegal

BackgroundAnthropogenic environmental changes may lead to ecosystem destabilization and the unintentional colonization of new habitats by parasite populations. A remarkable example is the outbreak of intestinal schistosomiasis in Northwest Senegal following the construction of two dams in the ‘80s. While many studies have investigated the epidemiological, immunological and geographical patterns of Schistosoma mansoni infections in this region, little is known about its colonization history.Conclusions/SignificanceOur results show that S. mansoni parasites are very successful in colonizing new areas without significant loss of genetic diversity. Maintaining high levels of diversity guarantees the adaptive potential of these parasites to cope with selective pressures such as drug treatment, which might complicate efforts to control the disease.     

MicroRNA‐92b‐5p modulates melatonin‐mediated osteogenic differentiation of bone marrow mesenchymal stem cells by targeting ICAM‐1

Osteoporosis is closely associated with the dysfunction of bone metabolism, which is caused by the imbalance between new bone formation and bone resorption. Osteogenic differentiation plays a vital role in maintaining the balance of bone microenvironment. The present study investigated whether melatonin participated in the osteogenic commitment of bone marrow mesenchymal stem cells (BMSCs) and further explored its underlying mechanisms. Our data showed that melatonin exhibited the capacity of regulating osteogenic differentiation of BMSCs, which was blocked by its membrane receptor inhibitor luzindole. Further study demonstrated that the expression of miR‐92b‐5p was up‐regulated in BMSCs after administration of melatonin, and transfection of miR‐92b‐5p accelerated osteogenesis of BMSCs. In contrast, silence of miR‐92b‐5p inhibited the osteogenesis of BMSCs. The increase in osteoblast differentiation of BMSCs caused by melatonin was attenuated by miR‐92b‐5p AMO as well. Luciferase reporter assay, real‐time qPCR analysis and western blot analysis confirmed that miR‐92b‐5p was involved in osteogenesis by directly targeting intracellular adhesion molecule‐1 (ICAM‐1). Melatonin improved the expression of miR‐92b‐5p, which could regulate the differentiation of BMSCs into osteoblasts by targeting ICAM‐1. This study provided novel methods for treating osteoporosis.     

Satellite DNA from the Y chromosome of the malaria vector Anopheles gambiae

Satellite DNA is an enigmatic component of genomic DNA with unclear function that has been regarded as "junk." Yet, persistence of these tandem highly repetitive sequences in heterochromatic regions of most eukaryotic chromosomes attests to their importance in the genome. We explored the Anopheles gambiae genome for the presence of satellite repeats and identified 12 novel satellite DNA families. Certain families were found in close juxtaposition within the genome. Six satellites, falling into two evolutionarily linked groups, were investigated in detail. Four of them were experimentally confirmed to be linked to the Y chromosome, whereas their relatives occupy centromeric regions of either the X chromosome or the autosomes. A complex evolutionary pattern was revealed among the AgY477-like satellites, suggesting their rapid turnover in the A. gambiae complex and, potentially, recombination between sex chromosomes. The substitution pattern suggested rolling circle replication as an array expansion mechanism in the Y-linked 53-bp satellite families. Despite residing in different portions of the genome, the 53-bp satellites share the same monomer lengths, apparently maintained by molecular drive or structural constraints. Potential functional centromeric DNA structures, consisting of twofold dyad symmetries flanked by a common sequence motif, have been identified in both satellite groups.     

Laboratory and Field Evaluation of the Partec CyFlow MiniPOC for Absolute and Relative CD4 T-Cell Enumeration

BackgroundA new CD4 point-of-care instrument, the CyFlow miniPOC, which provides absolute and percentage CD4 T-cells, used for screening and monitoring of HIV-infected patients in resource-limited settings, was introduced recently. We assessed the performance of this novel instrument in a reference laboratory and in a field setting in Senegal.MethodologyA total of 321 blood samples were obtained from 297 adults and 24 children, all HIV-patients attending university hospitals in Dakar, or health centers in Ziguinchor. Samples were analyzed in parallel on CyFlow miniPOC, FACSCount CD4 and FACSCalibur to assess CyFlow miniPOC precision and accuracy.ResultsAt the reference lab, CyFlow miniPOC, compared to FACSCalibur, showed an absolute mean bias of -12.6 cells/mm³ and a corresponding relative mean bias of -2.3% for absolute CD4 counts. For CD4 percentages, the absolute mean bias was -0.1%. Compared to FACSCount CD4, the absolute and relative mean biases were -31.2 cells/mm³ and -4.7%, respectively, for CD4 counts, whereas the absolute mean bias for CD4 percentages was 1.3%. The CyFlow miniPOC was able to classify HIV-patients eligible for ART with a sensitivity of ≥ 95% at the different ART-initiation thresholds (200, 350 and 500 CD4 cells/mm3). In the field lab, the room temperature ranged from 30 to 35°C during the working hours. At those temperatures, the CyFlow miniPOC, compared to FACSCount CD4, had an absolute and relative mean bias of 7.6 cells/mm³ and 2.8%, respectively, for absolute CD4 counts, and an absolute mean bias of 0.4% for CD4 percentages. The CyFlow miniPOC showed sensitivity equal or greater than 94%.ConclusionThe CyFlow miniPOC showed high agreement with FACSCalibur and FACSCount CD4. The CyFlow miniPOC provides both reliable absolute CD4 counts and CD4 percentages even under the field conditions, and is suitable for monitoring HIV-infected patients in resource-limited settings.