Causes and biophysical consequences of cellulose production by Pseudomonas fluorescens SBW25 at the air-liquid interface

Cellulose over-producing wrinkly spreader mutants of Pseudomonas fluorescens SBW25 have been the focus of much investigation, but conditions promoting the production of cellulose in ancestral SBW25, its effects and consequences have escaped in-depth investigation through lack of in vitro phenotype. Here, using a custom built device, we reveal that in static broth microcosms ancestral SBW25 encounters environmental signals at the air-liquid interface that activate, via three diguanylate cyclase-encoding pathways (Wsp, Aws and Mws), production of cellulose. Secretion of the polymer at the meniscus leads to modification of the environment and growth of numerous micro-colonies that extend from the surface. Accumulation of cellulose and associated microbial growth leads to Rayleigh-Taylor instability resulting in bioconvection and rapid transport of water-soluble products over tens of millimetres. Drawing upon data we build a mathematical model that recapitulates experimental results and captures the interactions between biological, chemical and physical processes.IMPORTANCE This work reveals a hitherto unrecognized behaviour that manifests at the air-liquid interface, which depends on production of cellulose, and hints to undiscovered dimensions to bacterial life at surfaces. Additionally, the study links activation of known diguanylate cyclase-encoding pathways to cellulose expression and to signals encountered at the meniscus. Further significance stems from recognition of the consequences of fluid instabilities arising from surface production of cellulose for transport of water-soluble products over large distances

Understanding the Wolbachia-mediated inhibition of arboviruses in mosquitoes: progress and challenges

Arthropod-borne viruses (arboviruses) pose a considerable threat to human and animal health, yet effective control measures have proven difficult to implement, and novel means of controlling their replication in arthropod vectors, such as mosquitoes, are urgently required. One of the most exciting approaches to emerge from research on arthropods is the use of the endosymbiotic intracellular bacterium Wolbachia to control arbovirus transmission from mosquito to vertebrate. These α-proteobacteria propagate through insects, in part through modulation of host reproduction, thus ensuring spread through species and maintenance in nature. Since it was discovered that Wolbachia endosymbiosis inhibits insect virus replication in Drosophila species, these bacteria have also been shown to inhibit arbovirus replication and spread in mosquitoes. Importantly, it is not clear how these antiviral effects are mediated. This review will summarize recent work and discuss determinants of antiviral effectiveness that may differ between individual Wolbachia/vector/arbovirus interactions. We will also discuss the application of this approach to field settings and the associated risks

A submillimetre wavelength spectral line search of the Orion molecular cloud core

A submillimetre wavelength molecular line search of the Orion molecular cloud has been made covering a total of about 5 percent of the frequency range 342.8 - 358.6 GHz. This search, coupled with the authors' previous observations of submillimetre transitions in this cloud, has led to the detection of 22 transitions of 14 molecular species, of which 16 are reported here for the first time. No unidentified lines have been detected in the present search. Mapping observations have been obtained for several of the lines and, in the case of H2CO the authors have been able to compare the present data with that obtained from other telescopes, to estimate the density and abundance in the emitting region

The cell adhesion molecule Fasciclin2 regulates brush border length and organization in Drosophila renal tubules

Multicellular organisms rely on cell adhesion molecules to coordinate cell–cell interactions, and to provide navigational cues during tissue formation. In Drosophila, Fasciclin 2 (Fas2) has been intensively studied due to its role in nervous system development and maintenance; yet, Fas2 is most abundantly expressed in the adult renal (Malpighian) tubule rather than in neuronal tissues. The role Fas2 serves in this epithelium is unknown. Here we show that Fas2 is essential to brush border maintenance in renal tubules of Drosophila. Fas2 is dynamically expressed during tubule morphogenesis, localizing to the brush border whenever the tissue is transport competent. Genetic manipulations of Fas2 expression levels impact on both microvilli length and organization, which in turn dramatically affect stimulated rates of fluid secretion by the tissue. Consequently, we demonstrate a radically different role for this well-known cell adhesion molecule, and propose that Fas2-mediated intermicrovillar homophilic adhesion complexes help stabilize the brush border

Perturbed cholesterol and vesicular trafficking associated with dengue blocking in Wolbachia-infected Aedes aegypti cells

Wolbachia are intracellular maternally inherited bacteria that can spread through insect populations and block virus transmission by mosquitoes, providing an important approach to dengue control. To better understand the mechanisms of virus inhibition, we here perform proteomic quantification of the effects of Wolbachia in Aedes aegypti mosquito cells and midgut. Perturbations are observed in vesicular trafficking, lipid metabolism and in the endoplasmic reticulum that could impact viral entry and replication. Wolbachia-infected cells display a differential cholesterol profile, including elevated levels of esterified cholesterol, that is consistent with perturbed intracellular cholesterol trafficking. Cyclodextrins have been shown to reverse lipid accumulation defects in cells with disrupted cholesterol homeostasis. Treatment of Wolbachia-infected Ae. aegypti cells with 2-hydroxypropyl-β-cyclodextrin restores dengue replication in Wolbachia-carrying cells, suggesting dengue is inhibited in Wolbachia-infected cells by localised cholesterol accumulation. These results demonstrate parallels between the cellular Wolbachia viral inhibition phenotype and lipid storage genetic disorders

Anderson Localization, Non-linearity and Stable Genetic Diversity

In many models of genotypic evolution, the vector of genotype populations satisfies a system of linear ordinary differential equations. This system of equations models a competition between differential replication rates (fitness) and mutation. Mutation operates as a generalized diffusion process on genotype space. In the large time asymptotics, the replication term tends to produce a single dominant quasispecies, unless the mutation rate is too high, in which case the populations of different genotypes becomes de-localized. We introduce a more macroscopic picture of genotypic evolution wherein a random replication term in the linear model displays features analogous to Anderson localization. When coupled with non-linearities that limit the population of any given genotype, we obtain a model whose large time asymptotics display stable genotypic diversityComment: 25 pages, 8 Figure

MM and subMM molecular line observations of the southwest lobe of L1551: Evidence of a shell structure

Observations have been made of the southwest outflow lobe of L1551 in several millimeter and submillimeter molecular lines. Maps have been made in the J=3-2 and J=2-1 transitions of CO over areas of 7.5 by 2.5 arc minutes and 5 by 5 arc minutes respectively at UKIRT. More detailed maps have also been made in the J=2-1 CO transition over an area of about 6 by 3.5 arc minutes at the NRAO 12m telescope. Additional observations of the J=4-3 transitions of HCN, HCO(+) abd H(13)CO(+) were made at selected positions. The HC(+) J=4-3 transition was detected at several positions along the outflow axis and at the position of IRS 5. Similarly the HCN J=4-3 transition was detected at the position of IRS 5 and also at a position close to HH29. However, the J=4-3 transition of H(13)CO(+) was bit detected at the position of IRS 5 even through it was observed at the position close to HH29 with a peak corrected antenna temperature of 0.23K at a V(LSR) of 1 km s(-1). The detection of the J=4-3 transitions of both HCO(+) and H(13)CO(+) close to the position of HH29 suggest the presence of very dense gas in this region. LVG analysis of the various molecular lines observed give a kinetic temperature between 10 and 15K and a density from 10(5) to 10(6) cm(-3) at the position of IRS 5 at the ambient cloud velocity. At the position close to HH29 LVG analysis of the CO observations gives a density between 10(3) and 10(4) cm(-3) at a kinetic temperature of 25k for a V(LSR) of 0 km s(-1). To the southwest of HH29 there is a large decrease in both the linewidth and intensity of CO emission. This may be due to the interaction between the outflow and a dense clump of gas which gives rise to HH29. The maps of the CO J=3-2 and CO J=2-1 emission integrated in 3.25 km s intervals show the shell structure postulated by Snell and Schloerb (1985)

Characterization of the monocyte-specific esterase (MSE) gene

Carboxylic esterases are widely distributed in hematopoietic cells. Monocytes express the esterase isoenzyme (termed 'monocyte-specific esterase', MSE) that can be inhibited by NaF in the alpha-naphthyl acetate cytochemical staining. We examined the expression of MSE in normal cells and primary and cultured leukemia-lymphoma cells. The MSE protein was demonstrated by isoelectric focusing (IEF); MSE mRNA expression was investigated by Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). The following samples were positive for MSE protein and Northern mRNA expression: 20/24 monocytic, 4/32 myeloid, and 1/20 erythroid-megakaryocytic leukemia cell lines, but none of the 112 lymphoid leukemia or lymphoma cell lines; of the normal purified cell populations only the monocytes were positive whereas, T, B cells, and granulocytes were negative; of primary acute (myelo) monocytic leukemia cells (CD14-positive, FAB M4/M5 morphology) 14/20 were Northern mRNA and 11/14 IEF protein positive. RT-PCR revealed MSE expression in 29/49 Northern-negative lymphoid leukemia-lymphoma cell lines. The RT-PCR signals in monocytic cell lines were on average 50-fold stronger than the mostly weak trace expression in lymphoid specimens. On treatment with various biomodulators, only all-trans retinoic acid significantly upregulated MSE message and protein levels but could not induce new MSE expression in several leukemia cell lines; lipopolysaccharide and interferon-gamma increased MSE expression in normal monocytes. Analysis of DNA methylation with sensitive restriction enzymes showed no apparent regulation of gene expression by differential methylation; the MSE gene is evolutionarily conserved among mammalian species; the half-life of the human MSE transcripts was about 5-6 h. The extent of MSE expression varied greatly among different monocytic leukemia samples. However, the MSE overexpression in a significant number of specimens was not associated with gene amplification, gross structural rearrangements or point mutations within the cDNA region. Taken together, the results suggest that MSE expression is not absolutely specific for, but strongly associated with cells of the monocytic lineage; MSE is either not expressed at all or expressed at much lower levels in cells from other lineages. The biological significance, if any, of rare MSE messages in lymphoid cells detectable only by the hypersensitive RT-PCR remains unclear. Further studies on the regulation of this gene and on the physiological function of the enzyme will no doubt be informative with respect to its striking overexpression in some malignant cells and to a possible role in the pathobiology of monocytic leukemias

Anthrax Toxin Receptor 2 Determinants that Dictate the pH Threshold of Toxin Pore Formation

The anthrax toxin receptors, ANTXR1 and ANTXR2, act as molecular clamps to prevent the protective antigen (PA) toxin subunit from forming pores until exposure to low pH. PA forms pores at pH ∼6.0 or below when it is bound to ANTXR1, but only at pH ∼5.0 or below when it is bound to ANTXR2. Here, structure-based mutagenesis was used to identify non-conserved ANTXR2 residues responsible for this striking 1.0 pH unit difference in pH threshold. Residues conserved between ANTXR2 and ANTXR1 that influence the ANTXR2-associated pH threshold of pore formation were also identified. All of these residues contact either PA domain 2 or the neighboring edge of PA domain 4. These results provide genetic evidence for receptor release of these regions of PA as being necessary for the protein rearrangements that accompany anthrax toxin pore formation