Mobility of a restriction-modification system revealed by its genetic contexts in three hosts

The flow of genes among prokaryotes plays a fundamental role in shaping bacterial evolution, and restriction-modification systems can modulate this flow. However, relatively little is known about the distribution and movement of restriction-modification systems themselves. We have isolated and characterized the genes for restriction-modification systems from two species of Salmonella, S. enterica serovar Paratyphi A and S. enterica serovar Bareilly. Both systems are closely related to the PvuII restriction-modification system and share its target specificity. In the case of S. enterica serovar Paratyphi A, the restriction endonuclease is inactive, apparently due to a mutation in the subunit interface region. Unlike the chromosomally located Salmonella systems, the PvuII system is plasmid borne. We have completed the sequence characterization of the PvuII plasmid pPvu1, originally from Proteus vulgaris, making this the first completely sequenced plasmid from the genus Proteus. Despite the pronounced similarity of the three restriction-modification systems, the flanking sequences in Proteus and Salmonella are completely different. The SptAI and SbaI genes lie between an equivalent pair of bacteriophage P4-related open reading frames, one of which is a putative integrase gene, while the PvuII genes are adjacent to a mob operon and a XerCD recombination (cer) site.     

Discovery and development of a novel class of phenoxyacetyl amides as highly potent TRPM8 agonists for use as cooling agents

The paper presents the activity trends for a novel series of phenoxyacetyl amides as human TRPM8 receptor agonists. This series encompasses in vitro activity values ranging from the micromolar to the picomolar levels. Sensory evaluation of these molecules highlights their relevance as cooling agents for oral applications. The positive outcome of the complete evaluation of N-(1H-pyrazol-3-yl)-N-(thiophen-2-ylmethyl)-2-(p-tolyloxy)acetamide resulted in its approval for Generally Recognized As Safe (GRAS) status by the Flavor & Extract Manufacturer Association (FEMA) as FEMA 4809.     

Viscosity as an additional physico-chemical parameter for studying chromatin structure

The viscosity values of chromatin in higher order structures, which range from 0.1 to 0.4 dl/g, are considerably lower than those of isolated DNA. These low values are consistent with other physico-chemical parameters, such as sedimentation and diffusion coefficients. When deducing molecular mass and compact shape of chromatin molecules in solvents of nearly physiological ionic strength, all these parameters are in general agreement. A decrease in ionic strength increases viscosity and decreases s-value. Both effects are consistent with a chromatin model postulating a very compact quaternary structure which unravels in low ionic environment to an unfolded but not completely extended tertiary structure.     

Effect of Storage at Low Temperatures on Development and Survival of Postparasitic Juveniles of Romanomermis culicivorax (Nematoda: Mermithidae)

Development of postparasites and adult females of Romanomermis culicivorax Ross and Smith, 1976 may be retarded by keeping cultures at 10 or 15 C for 15 wk. Storage at 5 C resulted in high mortality of postparasites. A storage temperature of 10 C is suitable if development of postparasites, young adults, and gravid females is to be greatly retarded. None of the females were gravid when postparasites (1-6 days old) were kept at 10 C for 15 wk. Development resumed after the cultures were returned to 26 C and no significant mortality occurred. A storage temperature of 15 C is suitable if development is to be only moderately retarded. After a 15-wk storage period, only a few weeks were required to return the cultures to full production status.     

Synthesis and biological evaluation of novel 4-benzylpiperazine ligands for sigma-1 receptor imaging

We report the synthesis and evaluation of 4-benzylpiperazine ligands (BP-CH(3), BP-F, BP-Br, BP-I, and BP-NO(2)) as potential σ(1) receptor ligands. The X-ray crystal structure of BP-Br, which crystallized with monoclinic space group P2(1)/c, has been determined. In vitro competition binding assays showed that all the five ligands exhibit low nanomolar affinity for σ(1) receptors (K(i)=0.43-0.91nM) and high subtype selectivity (σ(2) receptor: K(i)=40-61nM; K(i)σ(2)/K(i)σ(1)=52-94). [(125)I]BP-I (1-(1,3-benzodioxol-5-ylmethyl)-4-(4-iodobenzyl)piperazine) was prepared in 53±10% isolated radiochemical yield, with radiochemical purity of >99% by HPLC analysis after purification, via iododestannylation of the corresponding tributyltin precursor. The logD value of [(125)I]BP-I was found to be 2.98±0.17, which is within the range expected to give high brain uptake. Biodistribution studies in mice demonstrated relatively high concentration of radiolabeled substances in organs known to contain σ(1) receptors, including the brain, lung, kidney, heart, and spleen. Administration of haloperidol 5min prior to injection of [(125)I]BP-I significantly reduced the concentration of radioactivity in the above-mentioned organs. The accumulation of radiolabeled substance in the thyroid was quite low suggesting that [(125)I]BP-I is relatively stable to in vivo deiodination. These findings suggest that the binding of [(125)I]BP-I to σ(1) receptors in vivo is specific.     

Deletion of putative apoptosis factors leads to lifespan extension in the fungal ageing model Podospora anserina

Podospora anserina is a filamentous fungus with a limited lifespan. After a strain-specific period of growth, cultures turn to senescence and ultimately die. Here we provide evidence that the last step in the ageing of P. anserina is not accidental but programmed. In this study, PaAMID1, a homologue of a mammalian 'AIF-homologous mitochondrion-associated inducer of death', was analysed as a putative member of a caspase-independent signalling pathway. In addition, two metacaspases, PaMCA1 and PaMCA2, were investigated. While deletion of PaAmid1 as well as of PaMca2 was found to result in a moderate lifespan extension (59% and 78%, respectively), a 148% increase in lifespan was observed after deletion of PaMca1. Measurement of arginine-specific protease activity demonstrates a metacaspase-dependent activity in senescent but not in juvenile cultures, pointing to an activation of these proteases in the senescent stage of the life cycle. Moreover, treatment of juvenile wild-type cultures with hydrogen peroxide leads to a PaMCA1-dependent activity. The presented data strongly suggest that death of senescent wild-type cultures is triggered by an apoptotic programme induced by an age-dependent increase of reactive oxygen species during ageing of cultures and is executed after metacaspase activation.     

A benefit-finding intervention for family caregivers of persons with Alzheimer disease: study protocol of a randomized controlled trial

Background

Patients with ST-elevation myocardial infarction (STEMI) not treated with primary or rescue percutaneous coronary intervention (PCI) are at risk for recurrent ischemia, especially when viability in the infarct-area is present. Therefore, an invasive strategy with PCI of the infarct-related coronary artery in patients with viability would reduce the occurrence of a composite end point of death, reinfarction, or unstable angina (UA).

Methods

Patients admitted with an (sub)acute myocardial infarction, who were not treated by primary or rescue PCI, and who were stable during the first 48 hours after the acute event, were screened for the study. Eventually, we randomly assigned 216 patients with viability (demonstrated with low-dose dobutamine echocardiography) to an invasive or a conservative strategy. In the invasive strategy stenting of the infarct-related coronary artery was intended with abciximab as adjunct treatment. Seventy-five (75) patients without viability served as registry group. The primary endpoint was the composite of death from any cause, recurrent myocardial infarction (MI) and unstable angina at one year. As secondary endpoint the need for (repeat) revascularization procedures and anginal status were recorded.

Results

The primary combined endpoint of death, recurrent MI and unstable angina was 7.5% (8/106) in the invasive group and 17.3% (19/110) in the conservative group (Hazard ratio 0.42; 95% confidence interval [CI] 0.18-0.96; p = 0.032). During follow up revascularization-procedures were performed in 6.6% (7/106) in the invasive group and 31.8% (35/110) in the conservative group (Hazard ratio 0.18; 95% CI 0.13-0.43; p < 0.0001). A low rate of recurrent ischemia was found in the non-viable group (5.4%) in comparison to the viable-conservative group (14.5%). (Hazard-ratio 0.35; 95% CI 0.17-1.00; p = 0.051).

Conclusion

We demonstrated that after acute MI (treated with thrombolysis or without reperfusion therapy) patients with viability in the infarct-area benefit from a strategy of early in-hospital stenting of the infarct-related coronary artery. This treatment results in a long-term uneventful clinical course. The study confirmed the low risk of recurrent ischemia in patients without viability.

Trial registration

ClinicalTrials.gov: NCT00149591.     

3-(4-(6-Fluoroalkoxy-3,4-dihydroisoquinoline-2(1H)-yl)cyclohexyl)-1H-indole-5-carbonitriles for SERT imaging: chemical synthesis, evaluation in vitro and radiofluorination

Aminocyclohexyl indoles bind with high affinity and specificity toward the serotonin transporter (SERT). Based on this structural lead, we designed fluoroalkoxydihydroisoquinoline-cyclohexyl indole carbonitriles for future application as (18)F-labeled tracers for SERT imaging by PET. Six compounds, three pairs of cis- and trans-isomer derivatives, respectively, were synthesized and evaluated in vitro. The chemistry of the new compounds, their affinity and specificity data, the general route to the phenolic precursor for labeling, and the successful (18)F-fluoroalkylation of one pair of compounds are described herein.