EvoLaps 2: Advanced phylogeographic visualization.

EvoLaps is a user-friendly web application designed to visualize the spatial and temporal spread of pathogens. It takes an annotated tree as entry, such as a maximum clade credibility tree obtained through continuous phylogeographic inference. By following a 'Top-Down' reading of a tree recursively, transitions (latitude/longitude changes from a node to its children) are represented on a cartographic background using graphical paths. The complete set of paths forms the phylogeographic scenario. EvoLaps offers several features to analyze complex scenarios: (1) enhanced path display using multiple graphical variables with time-dependent gradients, (2) cross-highlighting and selection capabilities between the phylogeographic scenario and the phylogenetic tree, (3) production of specific spatio-temporal scales and synthetic views through dynamic and iterative clustering of localities into spatial clusters, (4) animation of the phylogeographic scenario using tree brushing, which can be done manually or automatically, gradually over time or at specific time intervals, and for the entire tree or a specific clade, and (5) an evolving library of additional tools. EvoLaps is freely available for use at evolaps.org.

Epidemiology, medical outcomes and costs of catheter-related bloodstream infections in intensive care units of four European countries: literature- and registry-based estimates.

Despite high incidence rates, little information is available on the burden of illness of catheter-related bloodstream infections (CRBSIs) in Europe. A review of the available data was performed to estimate the clinical outcomes and costs associated with CRBSIs during intensive care unit (ICU) stays in four European countries (France, Germany, Italy and the UK). Based on these data we have estimated the CRBSI-related mortality and the annual costs associated with CRBSIs in the aforementioned countries. Results show large variation between countries: 1.12-4.2 CRBSI per 1000 catheter days, 8400-14,400 CRBSIs episodes per year, 1000-1584 deaths per year, 15,960-201,600 ICU days caused by CRBSIs and euro35.9 to euro163.9 million associated costs. Discrepancies are mainly explained by the heterogeneous quality of epidemiological studies, as well as the variety of national clinical practices.

CEACAM1, a SOX9 direct transcriptional target identified in the colon epithelium.

A deletion of the transcription factor SOX9 gene in the mice intestine affects the morphology of the colon epithelium and leads to hyperplasia. Nevertheless, direct transcriptional targets of SOX9 in this tissue are still unknown. A microarray analysis identified the tumor suppressor carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) as a possible SOX9 target gene and we demonstrate here that SOX9 upregulates CEACAM1 in human colonic cells. Moreover, CEACAM1 expression is reduced in colon of SOX9-deficient mouse, suggesting an important function for SOX9 in the transcriptional activation of the CEACAM1 gene. We further identified SOX9-binding sequences in the human and rat CEACAM1 promoters, and an electrophoretic mobility shift together with a chromatin immunoprecipitation provided an additional evidence of the SOX9 binding to the human promoter. In addition, we established that histone acyl-transferase p300 behaves as an SOX9 co-activator of the rat and human CEACAM1promoters. These results highlight CEACAM1 as the first direct target of SOX9 identified in the colon epithelium.

Mitochondrial Haplotype Influences Mycelial Growth of Agaricus bisporus Heterokaryons.

We evaluated the influence of mitochondrial haplotype on growth of the common button mushroom Agaricus bisporus. Ten pairs of heterokaryon strains, each pair having the same nuclear genome but different mitochondrial genomes, were produced by controlled crosses among a group of homokaryons of both wild and commercial origins. Seven genetically distinct mitochondrial DNA (mtDNA) haplotypes were evaluated in different nuclear backgrounds. The growth of heterokaryon pairs differing only in their mtDNA haplotypes was compared by measuring mycelial radial growth rate on solid complete yeast medium (CYM) and compost extract medium and by measuring mycelial dry weight accumulation in liquid CYM. All A. bisporus strains were incubated at temperatures similar to those utilized in commercial production facilities (18, 22, and 26(deg)C). Statistically significant differences were detected in 8 of the 10 heterokaryon pairs evaluated for one or two of the three growth parameters measured. Some heterokaryon pairs showed differences in a single growth parameter at all three temperatures of incubation, suggesting a temperature-independent difference. Others showed differences at only a single temperature, suggesting a temperature-dependent difference. The influence of some mtDNA haplotypes on growth was dependent on the nuclear genetic background. Our results show that mtDNA haplotype can influence growth of A. bisporus heterokaryons in some nuclear backgrounds. These observations demonstrate the importance of including a number of mitochondrial genotypes and evaluating different nuclear-mitochondrial combinations of A. bisporus in strain improvement programs.

Mechanisms for the development of genetically variable mycorrhizal mycelia in the ectomycorrhizal fungus Laccaria bicolor.

An in vitro study investigated mechanisms for the development of genetically variable mycorrhizal mycelia for Laccaria bicolor. Seedlings of jack pine (Pinus banksiana) grown nonaseptically in an autoclaved soil substrate were given different L. bicolor inoculum treatments. These included (i) a dikaryotic mycelium genotype (D); (ii) D and basidiospores collected from one group of five sporophores (T1); (iii) D and basidiospores collected from 10 sporophores, two from each of five different groups (T5); (iv) T1 alone; (v) T5 alone; and (vi) a noninoculated control. Dikaryotic mycelial inoculum was provided at the time of sowing, while basidiospore inoculum was added at 10 weeks after seed germination. Sporophore formation was induced after 20 weeks of growth, and dikaryotic cultures were isolated from their tissue. Seedlings were harvested, and growth and mycorrhization were assessed. Levels of both were generally lower for T1-treated seedlings, compared with seedlings receiving D, while levels for T5-treated seedlings were intermediate. Sporophore genotype variability was assessed for inoculum treatments by using the isoenzymatic marker leucine aminopeptidase. The greatest genetic variability was seen with the basidiospore treatments T1 and T5, with up to four leucine aminopeptidase patterns per seedling. The mixed treatments D plus T1 and D plus T5 produced most frequently, but not exclusively, the inoculated dikaryon genotype. After isoenzyme results were assessed, variable sporophore isolates of mixed treatments were analyzed with randomly amplified polymorphic DNA and PCR mitochondrial DNA markers to determine if they were formed by dikaryon-monokaryon crosses between the inoculated dikaryon and monosporous mycelia.(ABSTRACT TRUNCATED AT 250 WORDS)

Behavioural profile of two potential antidepressant pyridazine derivatives including arylpiperazinyl moieties in their structure, in mice.

The potential antidepressant effects of two pyridazine derivatives, 5-benzyl 6-methyl 2-[4-(3-trifluoro-methyl phenyl) piperazin-1-yl] methylpyridazin-3-one (PC4) and 5-benzyl 6-methyl 2-[4-(3-chlorophenyl) piperazin-1-yl] methylpyridazin-3-one (PC13), were evaluated using classical psychopharmacological tests in mice. The intraperitoneal LD50 values of PC4 and PC13 were respectively 1125.8 and 429.6 mg kg-1. Only at intraperitoneal doses of 100 mg kg-1 did PC4 or PC13 significantly decrease locomotor activity. Both compounds (5-20 mg kg-1, i.p.) reduced the duration of immobility of mice in the forces swimming test, antagonized reserpine (2.5 mg kg-1, i.p.)-induced ptosis, and potentiated reserpine (2.5 mg kg-1, i.p.)-induced hypothermia. PC4 and PC13 (20 mg kg-1, i.p.) partly reversed hypothermia induced by low dose apomorphine (5 mg kg-1, s.c.) but were less effective for higher doses of apomorphine (16 mg kg-1, s.c.). At 200 mg kg-1, intraperitoneal PC13 enhanced the toxic effects of yohimbine (30 mg kg-1, s.c.), while PC4 was inactive. Head twitches produced either by L-5-hydroxytryptophan (4 mg kg-1, i.p.) in mice pretreated with pargyline (100 mg kg-1, i.p.) or by 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (3 mg kg-1, i.p.) were antagonized by both pyridazine derivatives (20 mg kg-1, i.p.). PC4 and PC13 showed analgesic properties in the phenylbenzoquinone-induced abdominal constriction test (5.0 < ED50 < 5.5 mg kg-1, i.p.) and in the hot-plate test (10 to 37% of analgesia at 10 mg kg-1, i.p.).(ABSTRACT TRUNCATED AT 250 WORDS)

Procyanidins from tormentil: fractionation and study of the anti-radical activity towards superoxide anion.

A standardised water-soluble extract was prepared from rhizomes of Potentilla tormentilla. The procyanidins in the extract were fractionated according to their degree of polymerisation by chromatography on Sephadex LH20. The anti-radical activities of the different fractions towards superoxide anion were compared when pentamers and hexamers were found to be the most active.

[The interaction of Cupressus sempervirens L. proanthocyanidolic oligomers with elastase and elastins]. / Etude de l'interaction des oligomères proanthocyanidoliques de Cupressus sempervirens L. sur l'élastase et les élastines.

Cupressus sempervirens L. proanthocyanidolic (O.P.C.) oligomers inhibited the esterolytic activity of pancreatic elastase with a Cl50 of 0.0075 mg/ml when a sap substrate suc(Al)3NA was used in a Tris-HCl 0.05 M buffer with a pH of 7.5. Inhibition was slightly lower when the ionic strength of the buffer was increased. Elastolytic activity was inhibited using an elastinorcein substrate with a Cl50 of 0.05 mg/ml, whatever the pH or the ionic strength of the buffer. The oligomers bound with the elastase to form a precipitant complex where a 2 mg/ml concentration of oligomers precipitated the elastase at 1 mg/ml. Insoluble elastin fixed few 150 micrograms oligomers for 1 mg of elastin but the latter was partly protected by the subsequent action of the elastase. Soluble elastin fixed a greater number of oligomers but it was the peptids of elastin enzyme hydrolysis which fixed the largest amount: around 1500 micrograms per mg. The oligomers-elastin complex seems to be more stable than that of oligomers-elastase which regains part of its esterase activity. The elastic fibers seem to be protected by the O.P.C.

Synthesis and anticonvulsant properties of new benzylpyridazine derivatives.

Several 3-substituted pyridazines and a series of imidazo- and triazolopyridazines were synthesized and tested for anticonvulsant activity against maximal electroshock-induced seizures in mice. The most active derivatives, 3-ureidopyridazine 7 and triazolopyridazines 16, 18, 21, and 25 with oral ED50's that ranged from 6.2 to 22.0 mg/kg, were more extensively investigated by evaluating their ability to prevent chemically induced seizures and were compared with phenytoin, phenobarbital, sodium valproate, carbamazepine, and diazepam. 3-amino-7-(2,6-dichlorobenzyl)-6-methyltriazolo-[4,3-b]pyridazine (25) was also protective in the pentylenetetrazole-induced seizures test (ED50 = 76 mg/kg per os) and blocked strychnine-induced tonic extensor seizures (ED50 = 34.5 mg/kg per os). Furthermore, derivative 25 showed anticonvulsant effects on bicuculline- and yohimbine-induced seizures tests in mice. All these results suggest that the pharmacological activity of 25 is partly due to modifications of glycinergic and GABAergic transmission. Moreover, molecular modeling studies based on the antiepileptic drug lamotrigine and the most stable conformer of 25 show structural similarities between these two molecules. This conformer also agrees with the electronic tolerances and volume of benzodiazepine pharmacophore models.

[Propolis extract. I. Acute toxicity and determination of acute primary cutaneous irritation index]. / Extrait de propolis: I. Etude de la toxicité aiguë et détermination de l'indice d'irritation primaire cutanée.

In first part, the aim of this work is to study the orally subacute toxicity of a propolis extract on conscious mice. LD50 is more than 7.34 g/kg propolis extract: this product is not toxic. In second part, cutaneous primary irritation of several products: excipients and propolis extract alone or in ointments, is evaluated in the rabbit. The cutaneous reactions after reiterated applications for 14 days are observed. The index of cutaneous primary irritation is calculated by evaluation of the erythematous and oedematous lesions. The propolis extract is non irritant.

[Propolis extract. II. Wound healing the the rat and rabbit]. / Extrait de propolis: II. Etude de la cicatrisation de plaies chez le lapin et chez le rat.

This work is related to wounds healing properties of a propolis extract. In first study on the Albinos Rabbit, the activity of a propolis extract is compared with these of a Peru balsam. Optimal concentrations of them in ointments are evaluated by applications on deep cutaneous scarifications. In order to go further into details, we have chosen in second part, another assay on the Rat, allowing the obtention of deeper wounds; By this way, more complete quantification of retained parameters and a better appraising of the wounds healing process evolution are possible.

Synthesis of new 5-substituted pyrazolo[1,5-d][1,2,4]-triazine derivatives and their analgesic, anti-inflammatory and antipyretic properties.

A series of 2-aryl-4-oxo-pyrazolo[1,5-d][1,2,4]triazines substituted in the 5-position by aminoalkyl or benzoyl moieties was synthesized and evaluated for analgesic activity. The structures of new triazine derivatives were confirmed by IR, 1H-NMR spectra and by elementary analysis. In the phenylbenzoquinone induced writhing test, only 3,3a-dihydropyrazolo triazines substituted by an arylpiperazinylmethyl group exhibited potent analgesic effect. In addition, these compounds possessed significant anti-inflammatory and antipyretic properties. A desaturation in 3,3a positions or other groups than arylpiperazinylmethyl moieties notably decreased analgesic effects.

Synthesis and aldose reductase inhibitory activity of triazine derivatives possessing acetic acid group.

N-Acetic acid derivatives of 6-aryl-pyrazolo-triazin-4-ones were synthesized for evaluation as new aldose reductase inhibitors. The intrinsic activity of each compound was assessed by measuring the inhibition of enzymatic activity in an isolated pig lens enzyme preparation. All the prepared compounds exhibited a significant in vitro aldose reductase inhibitory effect (10(-6) M less than or equal to IC50 less than or equal to 10(-4) M). Furthermore, biological activity (log 1/IC50) for most of the data sets could be correlated directly to electronic and steric parameters. Finally, spatial configuration of the most active derivative 6c (IC50 = 2 x 10(-6) M) was compared with that of tolrestat and with pharmacophor requirements of the aldose reductase inhibitor site using a molecular modeling system.

Synthesis and aldose reductase inhibitory activity of pyridazine derivatives possessing acetic acid group.

N-acetic acid and S-acetic acid derivatives of 5-arylidene pyridazines were synthesized for evaluation as new aldose reductase inhibitors. Intrinsic activity for each compound was assessed by measuring inhibition of enzymatic activity in an isolated pig lens enzyme preparation. All prepared compounds exhibited a significant in vitro aldose reductase inhibitory effect (10(-5) M less than or equal IC50 less than or equal to 10(-4) M). It was found that lipophilicity was important in increasing activity. Furthermore, this activity (log 1/IC50) could be correlated directly to a lipophilic parameter (log kw) for the whole data set.

Compared plasma and brain pharmacokinetics of clomipramine and its metabolite demethylclomipramine in two strains of mice (NMRI and CD1).

The fate of clomipramine (CMI) and its main demethylated metabolite demethylclomipramine (DCMI) was studied in two strains of Swiss mice (NMRI and CD1) after intraperitoneal injection. A study of its distribution among various tissues showed that fixation was most marked in lungs, perirenal fat and kidneys, and only moderate in the brain. The pharmacokinetic parameters of both molecules were determined in brain tissue and plasma. Absorption was rapid (tmax CMI = 14 min), metabolism prompt (tmax DCMI = 17 or 18 min according to the breed) and elimination rapid from both plasma and brain tissue. The first two stages were similar in the two strains, but elimination of CMI from both plasma and brain was faster in the NMRI mice (plasma t1/2 = 53 min against 165 min in the CD1 mice). Both values were well below that reported for man (mean plasma t1/2 = 24 h). The data presented can serve as a basis for designing true chronic administration protocol in animals.

Solid-phase extraction of midazolam and two of its metabolites from plasma for high-performance liquid chromatographic analysis.

A rapid, sensitive and selective assay of midazolam and two of its metabolites in plasma, based on high-performance liquid chromatography, has been developed. The compounds are subjected to solid-phase extraction, using C18 cartridges (Bond-Elut). Recoveries are in excess of 90% for midazolam and its metabolites. The limit of quantitation of the assay is 50 ng/ml of plasma for each compound.

[Synthesis and evaluation of the aldose reductase inhibitory activity of new diaryl pyridazine-3-ones]. / Synthèse et évaluation de l'activité inhibitrice d'aldose réductase de nouvelles diaryl pyridazin-3-ones.

It has been possible to prepare from 4,6-diaryl pyridazinones a series of derivatives substituted in the 2-position by chains of various lengths bearing a carboxylic acid function. Pig lens aldose reductase inhibitory activity was evaluated for all compounds. N-acetic acid derivative 3c with a chlorine atom on the phenyl nucleus at the 6-position on the pyridazin ring was the most active pyridazinone with an IC50 value of 1.2 x 10(-5) M. Furthermore, it has been shown that lipophilicity and spatial configuration of the synthesized compounds took a prominent part on enzymatic activity.

[Synthesis and evaluation of central nervous system activity of new triaza-spirodecanediones]. / Synthèse et évaluation de l'activité sur le système nerveux central de nouvelles triaza-spirodécanediones.

It has been possible to prepare from 8-(2-phenylethyl)-1,3,8-triaza [4, 5] spirodecane-2,4-dione a new series of derivatives substituted on the nitrogen atom in the 3-position of the hydantoïn ring. Several compounds exhibited sedative, anticonvulsant and anxiolytic activities. The aryl or heteroaryl-piperazinomethyl substituted compounds were the most active derivatives. "Buspirone-like" anxiolytic effects were found in some compounds from 5 mg/kg dose per os.

Anticonvulsant activity of 3-oxo-5-substituted benzylidene-6-methyl-(4H)-2-pyridazinylacetamides and 2-pyridazinylacetylhydrazides.

A series of 3-oxo-5-substituted-benzylidene-6-methyl-(4H)-2- pyridazinylacetamides and 2-pyridazinylacetylhydrazides were synthesized and evaluated for anticonvulsant activity against electrically and chemically induced seizures. In the maximal electroshock-induced seizures test, most of the derivatives showed an anticonvulsant effect better than that of sodium valproate, a commonly used anticonvulsant drug. At 100 mg/kg orally, compounds 5a and 5b respectively protected 50 and 60% of the mice against pentylentetrazole-induced seizures. In addition, these two derivatives showed significant anticonvulsant properties at doses that did not produce ataxia or sedation. The title compounds were also tested for their ability to antagonize convulsions induced by bicuculline and strychnine. Their effect on tremors induced by oxotremorine in mice was also evaluated.