The carbonic anhydrase of Clostridium autoethanogenum represents a new subclass of β-carbonic anhydrases

Carbonic anhydrase catalyses the interconversion of carbon dioxide and water to bicarbonate and protons. It was unknown if the industrial relevant acetogen Clostridium autoethanogenum possesses these enzymes. We identified two putative carbonic anhydrase genes in its genome, one of the β class and one of the γ class. Carbonic anhydrase activity was found for the purified β class enzyme, but not the γ class candidate. Functional complementation of an Escherichia coli carbonic anhydrase knock-out mutant showed that the β class carbonic anhydrase could complement this activity, but not the γ class candidate gene. Phylogenetic analysis showed that the β class carbonic anhydrase of Clostridium autoethanogenum represents a novel sub-class of β class carbonic anhydrases that form the F-clade. The members of this clade have the shortest primary structure of any known carbonic anhydrase

HST NICMOS Observations of the Polarization of NGC 1068

We have observed the polarized light at 2 micron in the center of NGC 1068 with HST NICMOS Camera 2. The nucleus is dominated by a bright, unresolved source, polarized at a level of 6.0 pm 1.2% with a position angle of 122degr pm 1.5degr. There are two polarized lobes extending up to 8'' northeast and southwest of the nucleus. The polarized flux in both lobes is quite clumpy, with the maximum polarization occurring in the southwest lobe at a level of 17% when smoothed to 0.23'' resolution. The perpendiculars to the polarization vectors in these two lobes point back to the intense unresolved nuclear source to within one 0.076'' Camera 2 pixel, thereby confirming that this is the illuminating source of the scattered light and therefore the probable AGN central engine. Whereas the polarization of the nucleus is probably caused by dichroic absorption, the polarization in the lobes is almost certainly caused by scattering, with very little contribution from dichroic absorption. Features in the polarized lobes include a gap at a distance of about 1'' from the nucleus toward the southwest lobe and a ``knot'' of emission about 5'' northeast of the nucleus. Both features had been discussed by ground-based observers, but they are much better defined with the high spatial resolution of NICMOS. The northeast knot may be the side of a molecular cloud that is facing the nucleus, which cloud may be preventing the expansion of the northeast radio lobe at the head of the radio synchrotron-radiation-emitting jet. We also report the presence of two ghosts in the Camera 2 polarizers. These had not been detected previously (Hines et al. 2000) because they are relatively faint and require observations of a source with a large dynamic range.Comment: 17 pages, 4 figure

Spitzer IRS Observations of the Galactic Center: Shocked Gas in the Radio Arc Bubble

We present Spitzer IRS spectra (R ~600, 10 - 38 micron) of 38 positions in the Galactic Center (GC), all at the same Galactic longitude and spanning plus/minus 0.3 degrees in latitude. Our positions include the Arches Cluster, the Arched Filaments, regions near the Quintuplet Cluster, the ``Bubble'' lying along the same line-of-sight as the molecular cloud G0.11-0.11, and the diffuse interstellar gas along the line-of-sight at higher Galactic latitudes. From measurements of the [O IV], [Ne II], [Ne III], [Si II], [S III], [S IV], [Fe II], [Fe III], and H_2 S(0), S(1), and S(2) lines we determine the gas excitation and ionic abundance ratios. The Ne/H and S/H abundance ratios are ~ 1.6 times that of the Orion Nebula. The main source of excitation is photoionization, with the Arches Cluster ionizing the Arched Filaments and the Quintuplet Cluster ionizing the gas nearby and at lower Galactic latitudes including the far side of the Bubble. In addition, strong shocks ionize gas to O^{+3} and destroy dust grains, releasing iron into the gas phase (Fe/H ~ 1.3 times 10^{-6} in the Arched Filaments and Fe/H ~ 8.8 times 10^{-6} in the Bubble). The shock effects are particularly noticeable in the center of the Bubble, but O$^{+3}$ is present in all positions. We suggest that the shocks are due to the winds from the Quintuplet Cluster Wolf-Rayet stars. On the other hand, the H_2 line ratios can be explained with multi-component models of warm molecular gas in photodissociation regions without the need for H_2 production in shocks.Comment: 51 pages, 17 figures To be published in the Astrophysical Journa

A novel conjugal donor strain for improved DNA transfer into Clostridium spp.

© 2019 The Authors Importance: The ability to transfer genetic material into a target organism is crucial for the development of a wide range of targeted genetic manipulation techniques. Overcoming the organisms’ native restriction systems which target foreign incoming DNA is one strategy that can increase the efficiency of genetic transfer. The novel E. coli donor strain described here employs this strategy, increasing the frequencies of conjugation into a range of clostridial strains, and therefore opening up the potential to implement novel gene manipulation techniques. Furthermore this novel donor strain has potential applications across a wide range of genetically recalcitrant organisms, and should be beneficial wherever the frequently occurring Type IV restriction systems are possessed by the target in question

Monoamine oxidase A and A/B knockout mice display autistic-like features

This is the published version, also available electronically from http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8876069&fileId=S1461145712000715Converging lines of evidence show that a sizable subset of autism-spectrum disorders (ASDs) is characterized by increased blood levels of serotonin (5-hydroxytryptamine, 5-HT), yet the mechanistic link between these two phenomena remains unclear. The enzymatic degradation of brain 5-HT is mainly mediated by monoamine oxidase (MAO)A and, in the absence of this enzyme, by its cognate isoenzyme MAOB. MAOA and A/B knockout (KO) mice display high 5-HT levels, particularly during early developmental stages. Here we show that both mutant lines exhibit numerous behavioural hallmarks of ASDs, such as social and communication impairments, perseverative and stereotypical responses, behavioural inflexibility, as well as subtle tactile and motor deficits. Furthermore, both MAOA and A/B KO mice displayed neuropathological alterations reminiscent of typical ASD features, including reduced thickness of the corpus callosum, increased dendritic arborization of pyramidal neurons in the prefrontal cortex and disrupted microarchitecture of the cerebellum. The severity of repetitive responses and neuropathological aberrances was generally greater in MAOA/B KO animals. These findings suggest that the neurochemical imbalances induced by MAOA deficiency (either by itself or in conjunction with lack of MAOB) may result in an array of abnormalities similar to those observed in ASDs. Thus, MAOA and A/B KO mice may afford valuable models to help elucidate the neurobiological bases of these disorders and related neurodevelopmental problem

Exponential Random Graph Modeling for Complex Brain Networks

Exponential random graph models (ERGMs), also known as p* models, have been utilized extensively in the social science literature to study complex networks and how their global structure depends on underlying structural components. However, the literature on their use in biological networks (especially brain networks) has remained sparse. Descriptive models based on a specific feature of the graph (clustering coefficient, degree distribution, etc.) have dominated connectivity research in neuroscience. Corresponding generative models have been developed to reproduce one of these features. However, the complexity inherent in whole-brain network data necessitates the development and use of tools that allow the systematic exploration of several features simultaneously and how they interact to form the global network architecture. ERGMs provide a statistically principled approach to the assessment of how a set of interacting local brain network features gives rise to the global structure. We illustrate the utility of ERGMs for modeling, analyzing, and simulating complex whole-brain networks with network data from normal subjects. We also provide a foundation for the selection of important local features through the implementation and assessment of three selection approaches: a traditional p-value based backward selection approach, an information criterion approach (AIC), and a graphical goodness of fit (GOF) approach. The graphical GOF approach serves as the best method given the scientific interest in being able to capture and reproduce the structure of fitted brain networks

Hubble Space Telescope NICMOS Polarization Measurements of OMC-1

We present 2micron polarization measurements of positions in the BN region of the Orion Molecular Cloud (OMC-1) made with NICMOS Camera 2 (0.2'' resolution) on HST. Our results are as follows: BN is sim 29% polarized by dichroic absorption and appears to be the illuminating source for most of the nebulosity to its north and up to sim 5'' to its south. Although the stars are probably all polarized by dichroic absorption, there are a number of compact, but non-point-source, objects that could be polarized by a combination of both dichroic absorption and local scattering of star light. We identify several candidate YSOs, including an approximately edge-on bipolar YSO 8.7'' east of BN, and a deeply-embedded variable star. Additional strongly polarized sources are IRc2-B, IRc2-D, and IRc7, all of which are obviously self-luminous at mid-infrared wavelengths and may be YSOs. None of these is a reflection nebula illuminated by a star located near radio source I, as was previously suggested. Other IRc sources are clearly reflection nebulae: IRc3 appears to be illuminated by IRc2-B or a combination of the IRc2 sources, and IRc4 and IRc5 appear to be illuminated by an unseen star in the vicinity of radio source I, or by Star n or IRc2-A. Trends in the magnetic field direction are inferred from the polarization of the 26 stars that are bright enough to be seen as NICMOS point sources. The most polarized star has a polarization position angle different from its neighbors by sim 40^o, but in agreement with the grain alignment inferred from millimeter polarization measurements of the cold dust cloud in the southern part of OMC-1.Comment: 41 pages, 8 figures, 4 tables, to appear in The Astrophysical Journa

The One with the Feminist Critique: Revisiting Millennial Postfeminism with Friends

In the aftermath of its initial broadcast run, iconic millennial sitcom Friends (NBC, 1994–2004) generated some quality scholarship interrogating its politics of gender. But as a site of analysis, it remains a curious, almost structuring absence from the central canon of the first wave of feminist criticism of postfeminist culture. This absence is curious not only considering the place of Friends at the forefront of millennial popular culture but also in light of its long-term syndication in countries across the world since that time. And it is structuring in the sense that Friends was the stage on which many of the familiar tropes of postfeminism interrogated across the body of work on it appear in retrospect to have been tried and tested. This article aims to contribute toward redressing this absence through interrogation and contextualization of the series’ negotiation of a range of structuring tropes of postfeminist media discourse, and it argues for Friends as an unacknowledged ur-text of millennial postfeminism

Engineering of vitamin prototrophy in Clostridium ljungdahlii and Clostridium autoethanogenum

Clostridium autoethanogenum and Clostridium ljungdahlii are physiologically and genetically very similar strict anaerobic acetogens capable of growth on carbon monoxide as sole carbon source. While exact nutritional requirements have not been reported, we observed that for growth, the addition of vitamins to media already containing yeast extract was required, an indication that these are fastidious microorganisms. Elimination of complex components and individual vitamins from the medium revealed that the only organic compounds required for growth were pantothenate, biotin and thiamine. Analysis of the genome sequences revealed that three genes were missing from pantothenate and thiamine biosynthetic pathways, and five genes were absent from the pathway for biotin biosynthesis. Prototrophy in C. autoethanogenum and C. ljungdahlii for pantothenate was obtained by the introduction of plasmids carrying the heterologous gene clusters panBCD from Clostridium acetobutylicum, and for thiamine by the introduction of the thiC-purF operon from Clostridium ragsdalei. Integration of panBCD into the chromosome through allele-coupled exchange also conveyed prototrophy. C. autoethanogenum was converted to biotin prototrophy with gene sets bioBDF and bioHCA from Desulfotomaculum nigrificans strain CO-1-SRB, on plasmid and integrated in the chromosome. The genes could be used as auxotrophic selection markers in recombinant DNA technology. Additionally, transformation with a subset of the genes for pantothenate biosynthesis extended selection options with the pantothenate precursors pantolactone and/or beta-alanine. Similarly, growth was obtained with the biotin precursor pimelate combined with genes bioYDA from C. acetobutylicum. The work raises questions whether alternative steps exist in biotin and thiamine biosynthesis pathways in these acetogens