Studies on the Bacteriophage 2 Receptors of Pseudomonas aeruginosa

The lysogenization of Pseudomonas aeruginosa strain BI with phage 2 resulted in the loss of the capacity to adsorb the same phage. The absence of phage 2 receptors on the surface of the lysogenized strain BI(2)(8) was confirmed by the failure of purified slime polysaccharide (SPB) or lipopolysaccharide (LPS) to inactivate phage 2. SPB and LPS from a phage 2-resistant strain also failed to inactivate phage 2 in contrast to the phage inactivation exhibited by the SPB and LPS obtained from the wild-type strain BI. Chemically, quantitative differences were apparent when the SPB and LPS of strains BI(2)(8) and BI/2S(2) were compared with those of the wild-type strain BI. The most striking difference noted was the absence of amino sugars in the SPB of strain BI/2S(2). The SPB of strain BI(2)(8) also contained a lower percentage of amino sugars compared with the SPB of the wild-type strain BI.     

Nitrate reductase from Spinacea oleracea. Effects of sulfhydryl-group reagents on the activities of the complex and the inactivation by NADH.

Sulfhydryl-group reagents inactive the nitrate reductase complex from Spinacea oleracea. Most of the reagents used inactivate selectively the NADH-diaphorase moiety. However, at higher concentrations of reagent the FNH₂-nitrate reductase is also affected. Enzyme preparations inactivated by p-hydroxy-mercuribenzoate can be reactivated by dithioerythritol. Nitrate reductase lacking NADH-diaphorase activity, after treatment with p-hydroxymercuribenzoate, is inactivated in its FNH₂-nitrate reductase moiety by NADH in the same way as the untreated preparation. This apparent independence of the NADH-inactivation process from NADH-diaphorase activity supports the postulated existence of a binding site for pyridine nucleotides implicated in NADH-inactivation and different from the diaphorase catalytic site.     

X-ray microanalysis of toluidine blue stained chromosomes: a quantitative study of the metachromatic reaction of chromatin

The stoichiometry of metachromatic staining of chromatin by toluidine blue was investigated in isolated metaphase chromosomes from L929 cells using X-ray microanalysis. Microspectrophotometric measurements revealed that a hypsochromic shift (from 595 to 570 nm) occurs in toluidine blue stained chromosomes in relation to the staining solution. Under the electron microscope, stained chromosomes. After toluidine blue staining, X-ray microanalysis of chromosomes revealed a large increase for sulphur counts and a considerable increase for Fe and Cu counts, while the signal of Mg, Ca, Cl, K and Zn was reduced. After subtraction of the intrinsic sulphur signal, S/P ratios of 0.82--for euchromatic arms--and 0.85--for centromeric heterochromatin--were obtained. They are considered representative of dye/DNA phosphate ratios. These results indicate the occurrence of a nearly stoichiometric binding of toluidine blue to chromatin DNA and suggest that an external dye stacking is responsible for the metachromatic staining of metaphase chromosomes.     

Regional Integration and Household Resilience: Infrastructure Connectivity and Livelihood Diversity in the Southwestern Amazon

The Inter-Oceanic Highway is among the first wave of large infrastructure projects under the auspices of the Initiative for the Integration of Regional Infrastructure in South America, which proposes regional integration as a means of economic development. Such projects have reignited debates over infrastructure impacts, which in many ways center on the ramifications for natural resource management. We pursue an analysis of the implications of highway paving for local livelihoods by focusing on the effects of market connectivity on livelihood diversity. Given that infrastructure brings shocks to affected regions, we argue that livelihood diversity is usefully interpreted in terms of household resilience to such shocks. We draw on rural household surveys from the tri-national frontier where Bolivia, Brazil and Peru meet in the southwestern Amazon, where the Inter-Oceanic Highway has recently been paved. The findings show that households more connected to markets in terms of travel time and road paving have less diverse livelihoods. This confirms concerns about regional integration and rural household vulnerability.     

IL2/IL21 region polymorphism influences response to rituximab in systemic lupus erythematosus patients

To determine whether the IL2/IL21 region, a general autoimmunity locus, contributes to the observed variation in response to rituximab in patients with systemic lupus erythematosus as well as to analyze its influence in a cohort including other autoimmune diseases. rs6822844 G/T polymorphism at the IL2–IL21 region was analyzed by TaqMan assay in 84 systemic lupus erythematosus (SLE) and 60 different systemic autoimmune diseases Spanish patients receiving rituximab. Six months after the first infusion patients were classified, according to the EULAR criteria, as good responders, partial responders and non-responders. A statistically significant difference was observed in GG genotype frequency between responder (total and partial response) (83.56 %) and non-responder (45.45 %) SLE patients (p = 0.010, odds ratio (OR) = 6.10 [1.28–29.06]). No association with the response was evident in the group of patients with autoimmune diseases other than lupus. Furthermore, when both groups of patients were pooled in a meta-analysis, a reduced statistical significance of the association was observed (p = 0.024, OR = 3.53 [1.06–11.64]). Our results show for a first time that IL2–IL21 region seems to play a role in the response to rituximab in SLE patients but not in other autoimmune diseases.     

The Measurement of the Effect on Citation Inequality of Differences in Citation Practices across Scientific Fields

This paper has two aims: (i) to introduce a novel method for measuring which part of overall citation inequality can be attributed to differences in citation practices across scientific fields, and (ii) to implement an empirical strategy for making meaningful comparisons between the number of citations received by articles in 22 broad fields. The number of citations received by any article is seen as a function of the article’s scientific influence, and the field to which it belongs. A key assumption is that articles in the same quantile of any field citation distribution have the same degree of citation impact in their respective field. Using a dataset of 4.4 million articles published in 1998–2003 with a five-year citation window, we estimate that differences in citation practices between the 22 fields account for 14% of overall citation inequality. Our empirical strategy is based on the strong similarities found in the behavior of citation distributions. We obtain three main results. Firstly, we estimate a set of average-based indicators, called exchange rates, to express the citations received by any article in a large interval in terms of the citations received in a reference situation. Secondly, using our exchange rates as normalization factors of the raw citation data reduces the effect of differences in citation practices to, approximately, 2% of overall citation inequality in the normalized citation distributions. Thirdly, we provide an empirical explanation of why the usual normalization procedure based on the fields’ mean citation rates is found to be equally successful.     

Global Distribution of Bartonella Infections in Domestic Bovine and Characterization of Bartonella bovis Strains Using Multi-Locus Sequence Typing

Bartonella bovis is commonly detected in cattle. One B. bovis strain was recently isolated from a cow with endocarditis in the USA, suggesting its role as an animal pathogen. In the present study, we investigated bartonella infections in 893 cattle from five countries (Kenya, Thailand, Japan, Georgia, and Guatemala) and 103 water buffaloes from Thailand to compare the prevalence of the infection among different regions and different bovid hosts. We developed a multi-locus sequence typing (MLST) scheme based on nine loci (16S rRNA, gltA, ftsZ, groEL, nuoG, ribC, rpoB, ssrA, and ITS) to compare genetic divergence of B. bovis strains, including 26 representatives from the present study and two previously described reference strains (one from French cows and another from a cow with endocarditis in the USA). Bartonella bacteria were cultured in 6.8% (7/103) of water buffaloes from Thailand; all were B. bovis. The prevalence of bartonella infections in cattle varied tremendously across the investigated regions. In Japan, Kenya, and the Mestia district of Georgia, cattle were free from the infection; in Thailand, Guatemala, and the Dusheti and Marneuli districts of Georgia, cattle were infected with prevalences of 10–90%. The Bartonella isolates from cattle belonged to three species: B. bovis (n=165), B. chomelii (n=9), and B. schoenbuchensis (n=1), with the latter two species found in Georgia only. MLST analysis suggested genetic variations among the 28 analyzed B. bovis strains, which fall into 3 lineages (I, II, and III). Lineages I and II were found in cattle while lineage III was restricted to water buffaloes. The majority of strains (17/28), together with the strain causing endocarditis in a cow in the USA, belonged to lineage I. Further investigations are needed to determine whether B. bovis causes disease in bovids.     

Organelle Transport in Cultured Drosophila Cells: S2 Cell Line and Primary Neurons.

Drosophila S2 cells plated on a coverslip in the presence of any actin-depolymerizing drug form long unbranched processes filled with uniformly polarized microtubules. Organelles move along these processes by microtubule motors. Easy maintenance, high sensitivity to RNAi-mediated protein knock-down and efficient procedure for creating stable cell lines make Drosophila S2 cells an ideal model system to study cargo transport by live imaging. The results obtained with S2 cells can be further applied to a more physiologically relevant system: axonal transport in primary neurons cultured from dissociated Drosophila embryos. Cultured neurons grow long neurites filled with bundled microtubules, very similar to S2 processes. Like in S2 cells, organelles in cultured neurons can be visualized by either organelle-specific fluorescent dyes or by using fluorescent organelle markers encoded by DNA injected into early embryos or expressed in transgenic flies. Therefore, organelle transport can be easily recorded in neurons cultured on glass coverslips using living imaging. Here we describe procedures for culturing and visualizing cargo transport in Drosophila S2 cells and primary neurons. We believe that these protocols make both systems accessible for labs studying cargo transport.