Azotobacter genomes: the genome of Azotobacter chroococcum NCIMB 8003 (ATCC 4412)

The genome of the soil-dwelling heterotrophic N2-fixing Gram-negative bacterium Azotobacter chroococcum NCIMB 8003 (ATCC 4412) (Ac-8003) has been determined. It consists of 7 circular replicons totalling 5,192,291 bp comprising a circular chromosome of 4,591,803 bp and six plasmids pAcX50a, b, c, d, e, f of 10,435 bp, 13,852, 62,783, 69,713, 132,724, and 311,724 bp respectively. The chromosome has a G+C content of 66.27% and the six plasmids have G+C contents of 58.1, 55.3, 56.7, 59.2, 61.9, and 62.6% respectively. The methylome has also been determined and 5 methylation motifs have been identified. The genome also contains a very high number of transposase/inactivated transposase genes from at least 12 of the 17 recognised insertion sequence families. The Ac-8003 genome has been compared with that of Azotobacter vinelandii ATCC BAA-1303 (Av-DJ), a derivative of strain O, the only other member of the Azotobacteraceae determined so far which has a single chromosome of 5,365,318 bp and no plasmids. The chromosomes show significant stretches of synteny throughout but also reveal a history of many deletion/insertion events. The Ac-8003 genome encodes 4628 predicted protein-encoding genes of which 568 (12.2%) are plasmid borne. 3048 (65%) of these show > 85% identity to the 5050 protein-encoding genes identified in Av-DJ, and of these 99 are plasmid-borne. The core biosynthetic and metabolic pathways and macromolecular architectures and machineries of these organisms appear largely conserved including genes for CO-dehydrogenase, formate dehydrogenase and a soluble NiFe-hydrogenase. The genetic bases for many of the detailed phenotypic differences reported for these organisms have also been identified. Also many other potential phenotypic differences have been uncovered. Properties endowed by the plasmids are described including the presence of an entire aerobic corrin synthesis pathway in pAcX50f and the presence of genes for retro-conjugation in pAcX50c. All these findings are related to the potentially different environmental niches from which these organisms were isolated and to emerging theories about how microbes contribute to their communities.

Active Transport of Glutamate in Leishmania (Leishmania) amazonensis

Leishmania spp. are the causative agents of leishmaniasis, a complex of diseases with a broad spectrum of clinical manifestations. Leishmania (Leishmania) amazonensis is a main etiological agent of diffuse cutaneous leishmaniasis. Leishmania spp., as other trypanosomatids, possess a metabolism based significantly on the consumption of amino acids. However, the transport of amino acids in these organisms remains poorly understood with few exceptions. Glutamate transport is an important biological process in many organisms. In the present work, the transport of glutamate is characterized. This process is performed by a single kinetic system (K-m=0.59 +/- 0.04 mM, V-max=0.123 +/- 0.003 nmol/min per 20 x 10(6) cells) showing an energy of activation of 52.38 +/- 4.7 kJ/mol and was shown to be partially inhibited by analogues, such as glutamine, aspartate, alpha-ketoglutarate and oxaloacetate, methionine, and alanine. The transport activity was sensitive to the extracellular concentration of H+ but not to Na+ or K+. However, unlike other amino acid transporters presently characterized, the treatment with specific ionophores confirmed the participation of a K+, and not H+ membrane gradient in the transport process.

Salmonella enterica Serovar Typhimurium Vaccine Strains Expressing a Nontoxic Shiga-Like Toxin 2 Derivative Induce Partial Protective Immunity to the Toxin Expressed by Enterohemorrhagic Escherichia coli

Shiga-like toxin 2 (Stx2)-producing enterohemorrhagic Escherichia coli (referred to as EHEC or STEC) strains are the primary etiologic agents of hemolytic-uremic syndrome (HUS), which leads to renal failure and high mortality rates. Expression of Stx2 is the most relevant virulence-associated factor of EHEC strains, and toxin neutralization by antigen-specific serum antibodies represents the main target for both preventive and therapeutic anti-HUS approaches. In the present report, we describe two Salmonella enterica serovar Typhimurium aroA vaccine strains expressing a nontoxic plasmid-encoded derivative of Stx2 (Stx2 Delta AB) containing the complete nontoxic A2 subunit and the receptor binding B subunit. The two S. Typhimurium strains differ in the expression of flagellin, the structural subunit of the flagellar shaft, which exerts strong adjuvant effects. The vaccine strains expressed Stx2 Delta AB, either cell bound or secreted into the extracellular environment, and showed enhanced mouse gut colonization and high plasmid stability under both in vitro and in vivo conditions. Oral immunization of mice with three doses of the S. Typhimurium vaccine strains elicited serum anti-Stx2B (IgG) antibodies that neutralized the toxic effects of the native toxin under in vitro conditions (Vero cells) and conferred partial protection under in vivo conditions. No significant differences with respect to gut colonization or the induction of antigen-specific antibody responses were detected in mice vaccinated with flagellated versus nonflagellated bacterial strains. The present results indicate that expression of Stx2 Delta AB by attenuated S. Typhimurium strains is an alternative vaccine approach for HUS control, but additional improvements in the immunogenicity of Stx2 toxoids are still required.

Current situation of practical work in secondary school biology (VCE biology) in Victoria state, Australia

In secondary school biology in Victoria State, Australia, practical work including laboratory exercises, fieldwork and other research activities is carried out more frequently than in Japanese senior high school biology. The authors examined the contents of the practical work and how often such practical work is carried out in some urban and rural secondary schools in Victoria. The topics of biology practical work were based on the VCE Biology Study Design which was published by the Victorian Board of Studies. Some of the activities continued for some weeks. Sometimes students went out from their school for fieldwork for a few days. The average number of practical work per credit was about 4. This number is consider ably larger than the value (2.3 per credit) which was reported on senior high schools in Osaka Prefecture. Why so often can the practical work be carried out? The main reason is that as well as the scores of ordinary paper tests, the evaluation of each practical work is taken into consideration at the entrance examination of universities and other tertiary education institutes in Victoria State.

Crown ethers : applications in inorganic synthesis

The ability of macroheterocyclic compounds to complex with ionic species has led to the synthesis and investigation of many multidentate macroheterocyclic species. The most stable complexes are formed between macrocyclic polyetheral ligands (crown ethers) with alkali or alkaline earth metal iona. There is an excellent correlation of the stability of these complexes with the size of the cation and the site of the cavity in the macrocyclic ligand. Additional factors, such as the basicity of the ligand and the solvating ability of the solvent, also play important roles in the stabilization of the complex. The stability of such complexes has been advantageously used to increase anionic reactivity and has been successfully applied to several organic fluorinations, oxidations, and similar reactions. The use of macrocyclic ligands in inorganic syntheses of otherwise difficult to obtain fluoro compounds has not been reported. O-carborane and m-carborane, C2BlOHl2, are icosahedral cage systems derived from Bl2H122- by replacement of BH with the isoelectronic CH group. These stable molecules exhibit electron-deficient bonding which can best be explained by delocalization of electrons. This delocalization gives rise to stability similar to that found in aromatic hydrocarbons. Crown ether activated potassium fluoride has been successfully employed in the conversion of alkyl, acyl and aryl halides to their respective fluorides. Analogously halide substituted carboranes were prepared, but their fluoro-derivatives were not obtained. The application of crown ethers in the synthesis of transition metal complexes is relatively unexplored. The usual synthesis of fluoro-derivative transition metal complexes involves highly reactive and toxic fluorinating agents such as antimony trifluoride, antimony penta fluoride. bromine trifluoride and hydrogen fluoride, An attempted preparation of the hexafluoroosmate (IV) ion via a crown activated, or naked fluoride~was unsuccessful. Potassium hexafluoroosmate (IV), K208F6. was eventually prepared using bromine trifluoride as a fluorinating and oxidizing agent .

Factors affecting the site of investment, and the reliance on savings for arctic breeders : the capital–income dichotomy revisited

The extent to which migratory birds that breed in the Arctic and winter in southern biomes rely on residual body stores for reproduction is unresolved. The short arctic summer and the limited availability of food early in the season constrain the time available for successful reproduction. Birds that are able to bring sufficient endogenous reserves to the breeding ground to meet, at least partially, the demands of egg-laying can initiate clutch production soon after arrival, thereby shortening the length of the breeding season and improving the chances of reproductive success. The amount of reserves available will be influenced by body size, the increased energetic and predation costs associated with carrying large stores, distances between staging sites and the location of the breeding grounds within the Arctic. Birds need not fly directly to the breeding grounds from the established temperate staging sites. Extensive feeding by migrants may occur in the Arctic, even within a few kilometres of the breeding sites as the birds track the retreating snowline. Irrespective of their size, birds are thus able to store some resources necessary for egg laying at local or regional scales. It is thus important to make a distinction between local capital and distant capital breeding. The extent to which a bird is characterized as a distant capital, local capital, or an income breeder not only varies between species, but also between individuals and seasons.

Web-building spiders attract prey by storing decaying matter

The orb-weaving spider Nephila edulis incorporates into its web a band of decaying animal and plant matter. While earlier studies demonstrate that larger spiders utilise these debris bands as caches of food, the presence of plant matter suggests additional functions. When organic and plastic items were placed in the webs of N. edulis, some of the former but none of the latter were incorporated into the debris band. Using an Y-maze olfactometer, we show that sheep blowflies Lucilia cuprina are attracted to recently collected debris bands, but that this attraction does not persist over time. These data reveal an entirely novel foraging strategy, in which a sit-and-wait predator attracts insect prey by utilising the odours of decaying organic material. The spider's habit of replenishing the debris band may be necessary to maintain its efficacy for attracting prey.

Does the rate of foraging attempts predict ingestion rate for young-of-the-year brook trout (Salvelinus fontinalis) in the field?

To assess the costs and benefits of young fish adopting different behavioural tactics, field studies of juvenile salmonines have assumed that (but did not test whether) the rate of foraging attempts predicts ingestion rate. We tested this assumption by quantifying capture, ingestion, and rejection rates of potential prey items for individual young-of-the-year brook trout (Salvelinus fontinalis) in a lake. Overall, capture rate (a conservative estimate of the rate of foraging attempts) was only a fair predictor of overall ingestion rate (Kendall's 1 = 0.54) and only 46% of captured items (number/minute) were ingested. Surface capture rate was a poor predictor of surface ingestion rate (T = 0.27) and only 1% of captured items were ingested. In contrast, subsurface capture rate was an excellent predictor of subsurface ingestion rate (T = 0.75) and 93% of captured items were ingested. No benthic prey captures were observed. Fish that ingested a low proportion of captured items spent a greater proportion of time moving, moved faster, and pursued prey further than fish that ingested a higher proportion of captured items. Rejection of captured items can represent a significant and little appreciated component of the foraging cycle for young salmonid fishes.

The international society for bipolar disorders (ISBD) task force report on antidepressant use in bipolar diosrders

Objective:
The risk-benefit profile of antidepressant medications in bipolar disorder is controversial. When conclusive evidence is lacking, expert consensus can guide treatment decisions. The International Society for Bipolar Disorders (ISBD) convened a task force to seek consensus recommendations on the use of antidepressants in bipolar disorders.

Method:  
An expert task force iteratively developed consensus through serial consensus-based revisions using the Delphi method. Initial survey items were based on systematic review of the literature. Subsequent surveys included new or reworded items and items that needed to be rerated. This process resulted in the final ISBD Task Force clinical recommendations on antidepressant use in bipolar disorder.

Results:  
There is striking incongruity between the wide use of and the weak evidence base for the efficacy and safety of antidepressant drugs in bipolar disorder. Few well-designed, long-term trials of prophylactic benefits have been conducted, and there is insufficient evidence for treatment benefits with antidepressants combined with mood stabilizers. A major concern is the risk for mood switch to hypomania, mania, and mixed states. Integrating the evidence and the experience of the task force members, a consensus was reached on 12 statements on the use of antidepressants in bipolar disorder.

Conclusions:
Because of limited data, the task force could not make broad statements endorsing antidepressant use but acknowledged that individual bipolar patients may benefit from antidepressants. Regarding safety, serotonin reuptake inhibitors and bupropion may have lower rates of manic switch than tricyclic and tetracyclic antidepressants and norepinephrine-serotonin reuptake inhibitors. The frequency and severity of antidepressant-associated mood elevations appear to be greater in bipolar I than bipolar II disorder. Hence, in bipolar I patients antidepressants should be prescribed only as an adjunct to mood-stabilizing medications.

The central repeat domain 1 of Kaposi's sarcoma-associated herpesvirus (KSHV) latency associated-nuclear antigen 1 (LANA1) prevents cis MHC class I peptide presentation

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