Regulation of the Activity of the Dual-Function DnaA Protein in Caulobacter crescentus.

DnaA is a conserved essential bacterial protein that acts as the initiator of chromosomal replication as well as a master transcriptional regulator in Caulobacter crescentus. Thus, the intracellular levels of active DnaA need to be tightly regulated during the cell cycle. Our previous work suggested that DnaA may be regulated at the level of its activity by the replisome-associated protein HdaA. Here, we describe the construction of a mutant DnaA protein [DnaA(R357A)]. The R357 residue in the AAA+ domain of the C. crescentus DnaA protein is equivalent to the R334 residue of the E. coli DnaA protein, which is required for the Regulatory Inactivation of DnaA (RIDA). We found that the expression of the DnaA(R357A) mutant protein in C. crescentus, but not the expression of the wild-type DnaA protein at similar levels, causes a severe phenotype of over-initiation of chromosomal replication and that it blocks cell division. Thus, the mutant DnaA(R357A) protein is hyper-active to promote the initiation of DNA replication, compared to the wild-type DnaA protein. DnaA(R357A) could not replace DnaA in vivo, indicating that the switch in DnaA activity once chromosomal replication has started may be an essential process in C. crescentus. We propose that the inactivation of DnaA is the main mechanism ensuring that chromosomal replication starts only once per cell cycle. We further observed that the R357A substitution in DnaA does not promote the activity of DnaA as a direct transcriptional activator of four important genes, encoding HdaA, the GcrA master cell cycle regulator, the FtsZ cell division protein and the MipZ spatial regulator of cell division. Thus, the AAA+ domain of DnaA may play a role in temporally regulating the bifunctionality of DnaA by reallocating DnaA molecules from initiating DNA replication to transcribing genes within the unique DnaA regulon of C. crescentus.

Ethyl glucuronide in plant extracts :O16

Introduction: The specificity of ethyl glucuronide (EtG) in hair as marker of alcohol consumption exceeds by far those of fatty acid ethyl esters. False positive cases are therefore very rare but not excluded as recent publications have shown. Especially, the use of plant extracts containing high percentages of ethanol can lead to EtG hair concentrations typically found in cases of chronic alcohol consumption. As proposed by Baumgartner et al., a nucleohilic substitution could most likely explain this phenomenon. Fresh and dried plants as well as commercial hair lotions based on plants extracts have been analysed for EtG presence or EtG formation. Methods: Urtica dioica, Plantago lanceolata, Cortex Quercus, Sempervivum, Armoracia rusticana, Juniperus communis, Brassica alba, Thymian vulgaris, Salvia officinalis, Majorana hortensis, Aloe vera, birch gingko and green tea leafs, ginger, lemon grass were extracted in water, water/ethanol (50/50) and ethanol (100%). The extracts as well as diluted hair lotions were measured by immunological test (Microgenics DRI® EtG assay) and by LC-MS/MS on Shimadzu Nexera UHPLC coupled with an AB Sciex 4500 QTrap. Results: EtG could not be detected in water extracts of all tested plants. However, DRI® EtG assay indicated the presence of EtG in 66% of the tested ethanolic plant extracts. That could only be confirmed by mass spectrometry in the cases of fresh thyme as well as in dried birch, oak and plantain extracts where EtG concentrations between of 0.25 and 2,09 mg/l were measured. In one hair lotion, the EtG concentration was 0,76 mg/l. Conclusion: Ethanolic plant extracts represents a non-negligible risk for false positive EtG hair tests, especially when applied as lotion without following washing out. The use of hair care products must therefore be evaluated at every hair sampling. In case of doubt, the product should be analysed by mass spectrometric methods since the presence of EtG can't be proven by use of the DRI® EtG assay, only. Our results support Baumgartner's assumption of a nucleophilic substitution in presence of ethanol because EtG was only measured in the ethanolic extracts.

Fideïcomís de residu versus substitució preventiva de residu: la voluntat expressa

El plet que origina la STSJC té per objecte la interpretació de la clàusula testamentària que instituïa hereva a la vídua del causant, subjectant-la, segons la interpretació dels demandants, a un fideïcomís de residu, i segons la demandada, a una substitució preventiva de residu. El TSJC va considerar que el testador havia ordenat un fideïcomís de residu. La qüestió havia estat abordada en diverses ocasions pel Tribunal Suprem, però tan sols existia una sentència del TSJC. Aquesta sentència aplica el mateix criteri que la STSJC 28.10.1991, instaurant així una línia jurisprudencial pròpia.

Evolutionary trajectories of primate genes involved in HIV pathogenesis.

The current availability of five complete genomes of different primate species allows the analysis of genetic divergence over the last 40 million years of evolution. We hypothesized that the interspecies differences observed in susceptibility to HIV-1 would be influenced by the long-range selective pressures on host genes associated with HIV-1 pathogenesis. We established a list of human genes (n = 140) proposed to be involved in HIV-1 biology and pathogenesis and a control set of 100 random genes. We retrieved the orthologous genes from the genome of humans and of four nonhuman primates (Pan troglodytes, Pongo pygmaeus abeli, Macaca mulatta, and Callithrix jacchus) and analyzed the nucleotide substitution patterns of this data set using codon-based maximum likelihood procedures. In addition, we evaluated whether the candidate genes have been targets of recent positive selection in humans by analyzing HapMap Phase 2 single-nucleotide polymorphisms genotyped in a region centered on each candidate gene. A total of 1,064 sequences were used for the analyses. Similar median K(A)/K(S) values were estimated for the set of genes involved in HIV-1 pathogenesis and for control genes, 0.19 and 0.15, respectively. However, genes of the innate immunity had median values of 0.37 (P value = 0.0001, compared with control genes), and genes of intrinsic cellular defense had K(A)/K(S) values around or greater than 1.0 (P value = 0.0002). Detailed assessment allowed the identification of residues under positive selection in 13 proteins: AKT1, APOBEC3G, APOBEC3H, CD4, DEFB1, GML, IL4, IL8RA, L-SIGN/CLEC4M, PTPRC/CD45, Tetherin/BST2, TLR7, and TRIM5alpha. A number of those residues are relevant for HIV-1 biology. The set of 140 genes involved in HIV-1 pathogenesis did not show a significant enrichment in signals of recent positive selection in humans (intraspecies selection). However, we identified within or near these genes 24 polymorphisms showing strong signatures of recent positive selection. Interestingly, the DEFB1 gene presented signatures of both interspecies positive selection in primates and intraspecies recent positive selection in humans. The systematic assessment of long-acting selective pressures on primate genomes is a useful tool to extend our understanding of genetic variation influencing contemporary susceptibility to HIV-1.

Industrial agglomerations and wage gradients: the Spanish economy in the interwar period

This paper gives new evidence on the relationship between integration and industrial agglomeration in the presence of scale economies, by testing directly one of the predictions that can be derived from Krugman (1991), that is, the existence of regional nominal wage gradients and its transformation following changes in trade regimes. Our case study analyzes the effects of the substitution of an open economy by a closed economy regime, exactly the opposite process studied by Hanson (1996, 1997). In Spain, during the interwar period, protectionist policies would have favored the loss of centrality of the coastal location (Barcelona) and the relative rise of central locations (such as Madrid). Our results indicate the existence of a wage gradient centered in Barcelona during the interwar period (1914-1930) and its weakening after 1925.

Nickel incorporation in Fe(II, III) hydroxysulfate Green Rust: effect on crystal lattice spacing and oxidation products

Ni(II)-Fe(II)-Fe(III) layered double hydroxides (LDH) or Ni-containing sulfate green rust (GR2) samples were prepared from Ni(II), Fe(II) and Fe(III) sulfate salts and analyzed with X ray diffraction. Nickel is readily incorporated in the GR2 structure and forms a solid solution between GR2 and a Ni(II)-Fe(III) LDH. There is a correlation between the unit cell a-value and the fraction of Ni(II) incorporated into the Ni(II)-GR2 structure. Since there is strong evidence that the divalent/trivalent cation ratio in GR2 is fixed at 2, it is possible in principle to determine the extent of divalent cation substitution for Fe(II) in GR2 from the unit cell a-value. Oxidation forms a mixture of minerals but the LDH structure is retained if at least 20 % of the divalent cations in the initial solution are Ni(II). It appears that Ni(II) is incorporated in a stable LDH structure. This may be important for two reasons, first for understanding the formation of LDHs, which are anion exchangers, in the natural environment. Secondly, this is important for understanding the fate of transition metals in the environment, particularly in the presence of reduced Fe compounds.

Carbon content in Amazonian Oxisols after forest conversion to pasture

Soil plays an important role in the C cycle, and substitution of tropical forest by cultivated land affects C dynamic and stock. This study was developed in an area of expansion of human settlement in the Eastern Amazon, in Itupiranga, State of Pará, to evaluate the effects of native forest conversion to Brachiaria brizantha pasture on C contents of a dystrophic Oxisol. Soil samples were collected in areas of native forest (NF), of 8 to 10 year old secondary forest (SF), 1 to 2 year old SF (P1-2), 5 to 7 year old SF (P5-7), and of 10 to 12 year old SF (P10-12), and from under pastures, in the layers 0-2, 2-5 and 5-10 cm, to evaluate C levels and stocks and carry out separation of OM based on particle size. After deforestation, soil density increased to a depth of 5 cm, with greater increase in older pastures. Variation in C levels was greatest in the top soil layer; C contents increased with increasing pasture age. In the layers 2-5 and 5-10 cm, C content proved to be stable for the types of plant cover evaluated. Highest C concentrations were found in the silt fraction; however, C contents were highest in the clay fraction, independent of the plant cover. An increase in C associated with the sand fraction in the form of little decomposed organic residues was observed in pastures, confirming greater sensitivity of this fraction to change in soil use.

A common ancestor DNA motif for invertebrate and vertebrate hormone response elements.

The ecdysone-responsive DNA sequence of the Drosophila hsp27 gene promoter contains four direct and inverted repeats reminiscent of those that compose the vertebrate palindromic estrogen response element (ERE) and the thyroid hormone/retinoic acid response element (TRE/RRE). Interestingly, a 3 bp substitution in the wild-type Hsp27 ecdysone response element (EcdRE) increases both its similarity with the vertebrate ERE and TRE/RRE and its capacity to confer ecdysone responsiveness to a heterologous promoter. Remarkably, increasing the spacing between the inverted repeats of this strong EcdRE by two nucleotides converts it into an ERE. Inversely, decreasing the spacing between the two inverted repeats of the vertebrate consensus palindromic ERE, from three to one nucleotide, converts it into a functional EcdRE. Thus, the only difference between an invertebrate EcdRE and a vertebrate palindromic ERE or TRE/RRE is in the spacing between the conserved inverted repeated motifs forming these palindromic HREs. The finding that the sequence motif 5'-GGTCA-3' present in the vertebrate ERE and TRE/RRE is also a functionally important characteristic of an invertebrate HRE, suggests that a common ancestor regulatory DNA sequence gave rise to all HREs known so far. We discuss the possibility that this progenitor motif is the GGTCA sequence.

Musculoskeletal pain in female asylum seekers and hypovitaminosis D3

Résumé de l'article L'hypovitaminose D3 est bien connue et courante chez la population âgée en Occident. Toutefois, elle est probablement sous-diagnostiquée chez les jeunes femmes immigrantes, bien qu'elle soit bien documentée, principalement en Angleterre dans la population Indo-Pakistanaise. Lorsque ce déficit est diagnostiqué, le traitement substitutif est simple et bon marché. Nous avons suspecté une haute prévalence chez de jeunes femmes requérantes d'asile, surtout chez celles provenant de cultures différant quant à l'exposition solaire et la diète. Nous publions donc une série de 11 cas de patientes avec une hypovitaminose D symptomatique issues de la consultation générale de la Policlinique Médicale Universitaire. Toutes les patientes présentaient une anamnèse d'une faible exposition solaire et de douleurs osseuses diffuses, d'une fatigue, d'une faiblesse musculaire ou des modifications de la marche. Toutefois, les premiers diagnostics évoqués par les médecins étaient une possible somatisation (3 patientes), des douleurs dorso-lombaires chroniques (4 patientes) et des symptômes somatiques multiples et inexpliqués (3 patientes). Le diagnostic a été posé d'emblée chez une patiente seulement. La durée moyenne des plaintes avant la pose du diagnostic était de 38 mois et 5 jours. Avec le traitement de cholecalciférol et de calcium, les symptômes disparaissaient entre 1 et 3 mois, chez une patiente ils ont duré sept mois. Le taux moyen de 250H vitamine D3 était de 10.9 nmol/l (IR 21-131). Le taux moyen de calcium était de 2.19 mmol/l (2.15-2.55) et quatre patientes présentaient une hypocalcémie. Nous pensons donc que les femmes requérantes d'asile sont à risque d'une durée prolongée de symptômes, de part la possible haute prévalence de cette condition et la difficulté chez les médecins à la reconnaître. Le diagnostic d'hypovitaminose devrait être recherché chez les femmes requérantes d'asile souffrant de douleurs musculo-squelettiques de longue durée. Le premier diagnostic souvent évoqué, dans un contexte psychosocial souvent difficile, était de l'ordre d'un trouble somatoforme douloureux ou de somatisations. Toutefois, les douleurs liées à l'hypovitaminose D3 sont relativement bien définies ; elles sont symétriques, osseuses, débutent souvent dans la région lombaire pour ensuite s'étendre au bassin, aux membres inférieures proximalement et à la cage thoracique. Le traitement substitutif est simple, peu onéreux et il serait judicieux d'initier d'autres études pour évaluer le besoin d'une substitution de routine dans la population présentée. Abstract: Deficiency of vitamin D, which can lead to osteomalacia, is common in elderly patients in Western countries. However, it is still widely underdiagnosed in young immigrant women, even though the condition has been extensively reported in the immigrant Indo- Asian population in the United Kingdom since the 1960s. A recent study reports an average 59 months before diagnosis was established, and another study found a prevalence of 78% of hypovitaminosis D3 (compared with 58% in controls) in an Indo-Asian population attending a UK rheumatology clinic. When recognised, hypovitaminosis D3 is easily treatable. A study on osteomalacic myopathy in veiled Arabic women in Denmark found that muscle strength returned to normal (except in maximal voluntary con-traction) after six months' treatment. We expected to see this disease in female asylum seekers, especially in those from societies with different customs regarding exposure to sunlight and diet We report 11 cases of symptomatic hypovitaminosis D3 in female asylum seekers (table 1). We focus on the pathology encountered by the primary care doctors caring for these 11 patients, the length of time between the appearance of symptoms, and the establishment of the diagnosis of hypovitaminosis D3 as well as the women's response to treatment by the improvement of a wide range of clinical symptoms-bone pain, muscular weakness, and fatigue.

Method for retinal gene repair in neonatal mouse.

Gene correction at the site of the mutation in the chromosome is the absolute way to really cure a genetic disease. The oligonucleotide (ODN)-mediated gene repair technology uses an ODN perfectly complementary to the genomic sequence except for a mismatch at the base that is mutated. The endogenous repair machinery of the targeted cell then mediates substitution of the desired base in the gene, resulting in a completely normal sequence. Theoretically, it avoids potential gene silencing or random integration associated with common viral gene augmentation approaches and allows an intact regulation of expression of the therapeutic protein. The eye is a particularly attractive target for gene repair because of its unique features (small organ, easily accessible, low diffusion into systemic circulation). Moreover therapeutic effects on visual impairment could be obtained with modest levels of repair. This chapter describes in details the optimized method to target active ODNs to the nuclei of photoreceptors in neonatal mouse using (1) an electric current application at the eye surface (saline transpalpebral iontophoresis), (2) combined with an intravitreous injection of ODNs, as well as the experimental methods for (3) the dissection of adult neural retinas, (4) their immuno-labelling, and (5) flat-mounting for direct observation of photoreceptor survival, a relevant criteria of treatment outcomes for retinal degeneration.

Industrial agglomerations and wage gradients: the Spanish economy in the interwar period

This paper gives new evidence on the relationship between integration and industrial agglomeration in the presence of scale economies, by testing directly one of the predictions that can be derived from Krugman (1991), that is, the existence of regional nominal wage gradients and its transformation following changes in trade regimes. Our case study analyzes the effects of the substitution of an open economy by a closed economy regime, exactly the opposite process studied by Hanson (1996, 1997). In Spain, during the interwar period, protectionist policies would have favored the loss of centrality of the coastal location (Barcelona) and the relative rise of central locations (such as Madrid). Our results indicate the existence of a wage gradient centered in Barcelona during the interwar period (1914-1930) and its weakening after 1925.

Transitional effects of a pension system change in Spain

This paper studies the output effects, transition costs and the change in pension benefits derived from the substitution of the current unfunded pension system by a fully funded pension system financed through mandatory savings.These effects are estimated by using reduced versions of the neoclassical and endogenous growth frameworks. Because of the greater capital accumulation during the transition phase, final output increases by 23,6% (neoclassicalframework); and a 24,5-31,5% (endogenous growth framework). The initial revenue loss for the government would represent a 4,8% of the GDP, raising very slowly during the transition period. Given the new growth rates, rates of return ofphysical capital, and financial intermediation costs, we have that the capitalization pension benefits obtained by all 30-contribution-year worker would be more than twice than those that guarantee the financial sustainability of thepublic pension system

Trauma et mémoire - quand la douleur infiltre le corps et l'âme: un regard sur les toxicodépendances graves

This article describes the long-lasting psychological after-effects of a traumatic experience. There is growing lmowledge of the biomedical underpinnings of these phenomena: the underlying mechanisms belong to an implicit learning process whereby the victim remains under the influence of painful past experiences. One of these mechanisms concerns the development of a traumatic bonding which Iioticeably impedes the establishment of interpersona! relationships. The other mechanism, called "contextualisation deficit", is the difficulty of adjusting a person's emotional and behavioural reactivity to the context of present day !ife. This capacity of a traumatic experience to become incrusted long-termina human being's mind, and to haunt the victim with various forms of psychological and physical suffering, can be compared with the presence of a tumour or an abscess in somatic medicine. Th us, severe drug addiction can be conceptualised as a disorder in which the patient tries - in most cases ineffectively - to soothe the pain of today's world in cmmection with the trauma of the past. In conclusion, this article urges the development of psychiatrie care programmes which operate at the centre of the suffering encountered by the se patients, as a complement to the already well-established offers such as harm reduction, substitution therapy and social support.

Choice of Initial Combination Antiretroviral Therapy in Individuals With HIV Infection: Determinants and Outcomes.

BACKGROUND Current guidelines give recommendations for preferred combination antiretroviral therapy (cART). We investigated factors influencing the choice of initial cART in clinical practice and its outcome. METHODS We analyzed treatment-naive adults with human immunodeficiency virus (HIV) infection participating in the Swiss HIV Cohort Study and starting cART from January 1, 2005, through December 31, 2009. The primary end point was the choice of the initial antiretroviral regimen. Secondary end points were virologic suppression, the increase in CD4 cell counts from baseline, and treatment modification within 12 months after starting treatment. RESULTS A total of 1957 patients were analyzed. Tenofovir-emtricitabine (TDF-FTC)-efavirenz was the most frequently prescribed cART (29.9%), followed by TDF-FTC-lopinavir/r (16.9%), TDF-FTC-atazanavir/r (12.9%), zidovudine-lamivudine (ZDV-3TC)-lopinavir/r (12.8%), and abacavir/lamivudine (ABC-3TC)-efavirenz (5.7%). Differences in prescription were noted among different Swiss HIV Cohort Study sites (P < .001). In multivariate analysis, compared with TDF-FTC-efavirenz, starting TDF-FTC-lopinavir/r was associated with prior AIDS (relative risk ratio, 2.78; 95% CI, 1.78-4.35), HIV-RNA greater than 100 000 copies/mL (1.53; 1.07-2.18), and CD4 greater than 350 cells/μL (1.67; 1.04-2.70); TDF-FTC-atazanavir/r with a depressive disorder (1.77; 1.04-3.01), HIV-RNA greater than 100 000 copies/mL (1.54; 1.05-2.25), and an opiate substitution program (2.76; 1.09-7.00); and ZDV-3TC-lopinavir/r with female sex (3.89; 2.39-6.31) and CD4 cell counts greater than 350 cells/μL (4.50; 2.58-7.86). At 12 months, 1715 patients (87.6%) achieved viral load less than 50 copies/mL and CD4 cell counts increased by a median (interquartile range) of 173 (89-269) cells/μL. Virologic suppression was more likely with TDF-FTC-efavirenz, and CD4 increase was higher with ZDV-3TC-lopinavir/r. No differences in outcome were observed among Swiss HIV Cohort Study sites. CONCLUSIONS Large differences in prescription but not in outcome were observed among study sites. A trend toward individualized cART was noted suggesting that initial cART is significantly influenced by physician's preference and patient characteristics. Our study highlights the need for evidence-based data for determining the best initial regimen for different HIV-infected persons.