PhyloChip hybridization uncovered an enormous bacterial diversity in the rhizosphere of different potato cultivars: many common and few cultivar-dependent taxa

The phylogenetic composition of bacterial communities in the rhizosphere of three potato cultivars grown at two distant field sites was analysed. Ribosomal gene fragments amplified from total community DNA were hybridized to PhyloChips. A total of 2432 operational taxonomic units (OTUs) were detected by the PhyloChips, of which 65% were found in the rhizosphere of all cultivars at both field sites. From all detected OTUs, 9% revealed a cultivar-dependent abundance at the one or the other field site and 4% at both sites. Differential abundance on the three cultivars was mainly observed for OTUs belonging to the Pseudomonadales, Actinomycetales and Enterobacteriales. More than 40% of OTUs belonging to Bradyrhizobiales, Sphingomonadales, Burkholderiales, Rhodocyclales, Xanthomonadales and Actinomycetales differed significantly in their abundance between the sites. A sequence analysis of six 16S rRNA gene clone libraries corresponded well with the taxonomic community structure evidenced by the PhyloChip hybridization. Most ribotypes matched OTUs detected by the PhyloChip. Those OTUs that responded to the potato cultivar at both field sites might be of interest in view of cultivar-specific effects on bacterial biocontrol strains and pathogens.     

Efficacy,Safety, and Dose of Pafuramidine,a New Oral Drug for Treatment of First Stage Sleeping Sickness,in a Phase 2a Clinical Study and Phase 2b Randomized Clinical Studies

Background

Sleeping sickness (human African trypanosomiasis [HAT]) is caused by protozoan parasites and characterized by a chronic progressive course, which may last up to several years before death. We conducted two Phase 2 studies to determine the efficacy and safety of oral pafuramidine in African patients with first stage HAT.

Methods

The Phase 2a study was an open-label, non-controlled, proof-of-concept study where 32 patients were treated with 100 mg of pafuramidine orally twice a day (BID) for 5 days at two trypanosomiasis reference centers (Angola and the Democratic Republic of the Congo [DRC]) between August 2001 and November 2004. The Phase 2b study compared pafuramidine in 41 patients versus standard pentamidine therapy in 40 patients. The Phase 2b study was open-label, parallel-group, controlled, randomized, and conducted at two sites in the DRC between April 2003 and February 2007. The Phase 2b study was then amended to add an open-label sequence (Phase 2b-2), where 30 patients received pafuramidine for 10 days. The primary efficacy endpoint was parasitologic cure at 24 hours (Phase 2a) or 3 months (Phase 2b) after treatment completion. The primary safety outcome was the rate of occurrence of World Health Organization Toxicity Scale Grade 3 or higher adverse events. All subjects provided written informed consent.

Findings/Conclusion

Pafuramidine for the treatment of first stage HAT was comparable in efficacy to pentamidine after 10 days of dosing. The cure rates 3 months post-treatment were 79% in the 5-day pafuramidine, 100% in the 7-day pentamidine, and 93% in the 10-day pafuramidine groups. In Phase 2b, the percentage of patients with at least 1 treatment-emergent adverse event was notably higher after pentamidine treatment (93%) than pafuramidine treatment for 5 days (25%) and 10 days (57%). These results support continuation of the development program for pafuramidine into Phase 3.     

Novel sulfenamides and sulfonamides based on pyridazinone and pyridazine scaffolds as CB1 receptor ligand antagonists

A series of sulfenamide and sulfonamide derivatives was synthesized and evaluated for the affinity at CB₁ and CB₂ receptors. The N-bornyl-S-(5,6-di-p-tolylpyridazin-3-yl)-sulfenamide, compound 11, displayed good affinity and high selectivity for CB₁ receptors (K_i values of 44.6?nM for CB₁ receptors and >40?μM for CB₂ receptors, respectively). The N-isopinocampheyl-sulfenamide 12 and its sulfonamide analogue 22 showed similar selectivity for CB₁ receptors with K_i values of 75.5 and 73.2?nM, respectively. These novel compounds behave as antagonists/inverse agonists at CB₁ receptor in the [³⁵S]-GTPγS binding assays, and none showed adequate predictive blood–brain barrier permeation, exhibiting low estimated LD₅₀. However, testing compound 12 in a supraspinal analgesic test (hot-plate) revealed that it was as effective as the classic CB₁ receptor antagonist rimonabant, in reversing the analgesic effect of a cannabinoid agonist.     

Changes of Body Mass Index in Relation to Mortality: Results of a Cohort of 42,099 Adults

Background

High Body-Mass-Index (BMI) is associated with increased all-cause mortality, but little is known about the effect of short- and long-term BMI change on mortality. The aim of the study was to determine how long-term weight change affects mortality.

Methods and findings

Within a population-based prospective cohort of 42,099 Austrian men and women (mean age 43 years) with at least three BMI measurements we investigated the relationship of BMI at baseline and two subsequent BMI change intervals of five years each with all-cause mortality using Cox proportional Hazard models. During median follow-up of 12 years 4,119 deaths were identified. The lowest mortalities were found in persons with normal weight or overweight at baseline and stable BMI over 10 years. Weight gain (≥0.10 kg/m²/year) during the first five years was associated with increased mortality in overweight and obese people. For weight gain during both time intervals mortality risk remained significantly increased only in overweight (Hazard Ratio (HR): 1.39 (95% confidence interval: 1.01; 1.92)) and obese women (1.85 (95% confidence interval: 1.18; 2.89)). Weight loss (< −0.10 kg/m²/year) increased all-cause mortality in men and women consistently. BMI change over time assessed using accepted World Health Organisation BMI categories showed no increased mortality risk for people who remained in the normal or overweight category for all three measurements. In contrast, HRs for stable obese men and women were 1.57 (95% CI: 1.31; 1.87) and 1.46 (95% CI: 1.25; 1.71) respectively.

Conclusion

Our findings highlight the importance of weight stability and obesity avoidance in prevention strategy.     

Eocene Diversification of Crown Group Rails (Aves: Gruiformes: Rallidae)

Central to our understanding of the timing of bird evolution is debate about an apparent conflict between fossil and molecular data. A deep age for higher level taxa within Neoaves is evident from molecular analyses but much remains to be learned about the age of diversification in modern bird families and their evolutionary ecology. In order to better understand the timing and pattern of diversification within the family Rallidae we used a relaxed molecular clock, fossil calibrations, and complete mitochondrial genomes from a range of rallid species analysed in a Bayesian framework. The estimated time of origin of Rallidae is Eocene, about 40.5 Mya, with evidence of intrafamiliar diversification from the Late Eocene to the Miocene. This timing is older than previously suggested for crown group Rallidae, but fossil calibrations, extent of taxon sampling and substantial sequence data give it credence. We note that fossils of Eocene age tentatively assigned to Rallidae are consistent with our findings. Compared to available studies of other bird lineages, the rail clade is old and supports an inference of deep ancestry of ground-dwelling habits among Neoaves.     

Atrial granules in the dog heart after cardiac denervation

Summary Because the increase in sodium excretion during left atrial distension in conscious dogs is abolished after chronic cardiac denervation, we have investigated whether this is a result of the disappearance of specific atrial granules. Electron microscopy and light-microscopical and ultrastructural immunohistochemistry of canine atria show that atrial granules displaying immunoreactivity for cardiac hormones of the cardiodilatin/atrial natriuretic polypeptide (CDD/ANP) family are still present in denervated left and right atria, although reduced in quantity. It is concluded that the atrial-induced natriuresis is not only related to the existence of specific atrial granules. The functional link between atrial-induced natriuresis provoked by atrial distension and the release of atrial polypeptide hormones remains uncertain because the denervated heart can secrete CDD although the diuretic-natriuretic effect is altered.     

Steuerung und potentielle Evolutionsgeschwindigkeit des obligaten Teilzieherverhaltens: Ergebnisse eines Zweiweg-Selektionsexperiments mit der M?nchsgrasmücke (Sylvia atricapilla)

Zusammenfassung 1. In Südfrankreich lebt eine obligat teilziehende Population der Mönchsgrasmücke (Sylvia atricapilla) mit rund drei Vierteln Zugvögeln und einem Viertel Standvögeln. Das Teilzugverhalten dieser Population wird in beträchtlichem Umfang von genetischen Faktoren gesteuert (Berthold &Querner 1982). Der starke Selektionserfolg eines Zweiweg-Selektionsexperiments bis zur F₁-Generation ließ eine hohe potentielle Evolutionsgeschwindigkeit des obligaten Teilzieherverhaltens vermuten (2). Wir haben das Evolutionspotential in einem Langzeit-Selektionsversuch experimentell untersucht.2. Wir zogen insgesamt 267 Individuen der freilebenden Mönchsgrasmückenpopulation in fünf Gruppen von Hand auf, und alle fünf Stichproben zeigten entsprechende Verhältnisse von Ziehern und Nichtziehern. Wir konnten das Langzeit-Zweiweg-Selektionsexperiment bis zu einer F₆-Generation durchführen und konnten dabei insgesamt 455 Mönchsgrasmücken in Volieren züchten.3. Das Zweiweg-Selektionsexperiment zeigt, daß die untersuchte teilziehende Mönchsgrasmückenpopulation bereits nach drei Generationen eine ausschließlich ziehende und nach vier bis sechs Generationen eine fast nicht mehr ziehende Population werden kann.4. Das Zweiweg-Selektionsexperiment zeigt starken Selektionserfolg und hohe Heritabilitätswerte (ca. 0,6–1), die deutlich höher liegen als bisher für das Verhalten von (meist domestizierten) Wirbeltieren angegeben. Demnach sind genetische Faktoren für die Steuerung dieses obligaten Teilzugs sehr bedeutsam und wahrscheinlich dominant über Umwelteinflüsse.5. Die Versuchsergebnisse lassen im Falle von schnellen Umweltveränderungen und der Wirkung von starken Selektionsfaktoren ein enorm großes Evolutionspotential des obligaten Teilzieherverhaltens annehmen und damit allgemein eine schnelle Mikroevolution der Ortsbewegung von Vögeln. Sollten die von Meteorologen prophezeiten Klimaänderungen in Bälde eintreten (Erwärmung aufgrund von Verunreinigungen der Atmosphäre durch den Menschen, Treibhauseffekt), dann könnten viele obligate Teilzieher auf genetischer Basis rasch zu Standvögeln werden und wenig ausgeprägte Zieher rasch zu Teilziehern. Unter diesem allgemeinen Anwachsen von Standvögeln könnten ausgeprägte Zugvögel mehr und mehr zu leiden haben.6. Eine genetische Kontrolle obligaten Teilzugs scheint weit verbreitet zu sein, und sie stellt wohl auch für alle Individuen den zuverlässigsten Steuerungsmechanismus dar. Beim fakultativen Teilzug hingegen dürften Umweltfaktoren mehr unmittelbar steuernd wirken.

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Control and evolutionary potential of obligate partial migration: results of a two-way selective breeding experiment with the Blackcap (Sylvia atricapilla)

Summary 1. In southern France breeds an obligate partially migratory population of the Blackcap with about three quarters of migrants and one quarter of residents. Partial migration of this population is considerably controlled by genetic factors (Berthold &Querner 1982). The large selection response of a two-way selective breeding experiment up to F₁-generations suggested a high evolutionary potential of obligate partial migration (2). We have this evolutionary potential experimentally investigated in a long-term selective breeding study.2. We raised by hand a total of 267 individuals of the free-living Blackcap population in five groups, and all five samples showed corresponding ratios of migrants and nonmigrants. We were able to conduct the long-term selective breeding experiment up to a F₆-generation, and thereby a total of 455 Blackcaps could be bred in aviaries.3. The two-way selective breeding experiment shows, that the investigated partially migratory Blackcap population can become completely migratory in just three generations and almost exclusively sedentary in just four to six generations.4. The two-way selective breeding experiment yielded large selection responses and high heritability values (about 0.6–1) which are distinctly higher than those previously reported for behavioural traits of (mostly domesticated) vertebrates. Hence, genetic factors are very important for the control of this obligate partial migration and are presumedly dominant over environmental influences.5. In case of rapid environmental changes and the effect of strong selection factors the experimental results suggest a strikingly high evolutionary potential of obligate partial migration and thus in general a rapid microevolution of bird movements. If climatic changes would occur in the near future as they are predicted by meteorologists (rise in temperature due to man-made pollution of the atmosphere, greenhouse-effect) then many obligate partial migrants could rapidly become sedentary on a genetic basis and many less typical migrants could then considerably suffer from the general increase of residents.6. A genetic control of obligate partial migration appears to be widespread and it also appears to be the most reliable control mechanism for all individuals of such a partially migratory population. Facultative partial migration, however, appears to be controlled more directly by environmental factors.

     

Maintenance of Leukemia-Initiating Cells Is Regulated by the CDK Inhibitor Inca1

Functional differences between healthy progenitor and cancer initiating cells may provide unique opportunities for targeted therapy approaches. Hematopoietic stem cells are tightly controlled by a network of CDK inhibitors that govern proliferation and prevent stem cell exhaustion. Loss of Inca1 led to an increased number of short-term hematopoietic stem cells in older mice, but Inca1 seems largely dispensable for normal hematopoiesis. On the other hand, Inca1-deficiency enhanced cell cycling upon cytotoxic stress and accelerated bone marrow exhaustion. Moreover, AML1-ETO9a-induced proliferation was not sustained in Inca1-deficient cells in vivo. As a consequence, leukemia induction and leukemia maintenance were severely impaired in Inca1^−/− bone marrow cells. The re-initiation of leukemia was also significantly inhibited in absence of Inca1^−/− in MLL—AF9- and c-myc/BCL2-positive leukemia mouse models. These findings indicate distinct functional properties of Inca1 in normal hematopoietic cells compared to leukemia initiating cells. Such functional differences might be used to design specific therapy approaches in leukemia.     

First molecular epidemiological study of cutaneous leishmaniasis in Libya

Background

Cutaneous leishmaniasis (CL) is a major public health problem in Libya. The objective of this study was to investigate, for the first time, epidemiological features of CL outbreaks in Libya including molecular identification of parasites, the geographical distribution of cases and possible scenarios of parasite transmission.

Methodology/Principal Findings

We studied 450 patients that came from 49 areas distributed in 12 districts in north-west Libya. The patients'' ages ranged from 9 months to 87 years (median age 25 years); 54% of the cases were males. Skin scrapings spotted on glass slides were collected for molecular identification of causative agent. The ribosomal internal transcribed spacer 1 (ITS1) was amplified and subsequently characterized by restriction fragment length polymorphism (RFLP) analysis. In total, 195 samples were successfully identified of which 148 (75.9%) were Leishmania major, and 47 (24.1%) Leishmania tropica. CL cases infected with L. major were found in all CL areas whereas L. tropica cases came mainly from Al Jabal Al Gharbi (46.4%), Misrata (17.8%) and Tarhuna districts (10.7%). A trend of seasonality was noticed for the infections with L. major which showed a clear peak between November and January, but was less pronounced for infections by L. tropica.

Conclusion

The first molecular study on CL in Libya revealed that the disease is caused by L. major and L. tropica and the epidemiological patterns in the different foci were the same as in other Mediterranean foci of CL.