An inhibitor of FtsZ with potent and selective anti-staphylococcal activity

FtsZ is an essential bacterial guanosine triphosphatase and homolog of mammalian beta-tubulin that polymerizes and assembles into a ring to initiate cell division. We have created a class of small synthetic antibacterials, exemplified by PC190723, which inhibits FtsZ and prevents cell division. PC190723 has potent and selective in vitro bactericidal activity against staphylococci, including methicillin- and multi-drug-resistant Staphylococcus aureus. The putative inhibitor-binding site of PC190723 was mapped to a region of FtsZ that is analogous to the Taxol-binding site of tubulin. PC190723 was efficacious in an in vivo model of infection, curing mice infected with a lethal dose of S. aureus. The data validate FtsZ as a target for antibacterial intervention and identify PC190723 as suitable for optimization into a new anti-staphylococcal therapy.     

Information science. Going, going, gone: lost Internet references

The use of Internet references in academic literature is common, and Internet references are frequently inaccessible. The extent of Internet referencing and Internet reference activity in medical or scientific publications was systematically examined in more than 1000 articles published between 2000 and 2003 in the New England Journal of Medicine, The Journal of the American Medical Association, and Science. Internet references accounted for 2.6% of all references (672/25548) and in articles 27 months old, 13% of Internet references were inactive. Publishers, librarians, and readers need to reassess policies, archiving systems, and other resources for addressing Internet reference attrition to prevent further information loss.     

Distinguishing inchworm and hand-over-hand processive kinesin movement by neck rotation measurements

The motor enzyme kinesin makes hundreds of unidirectional 8-nanometer steps without detaching from or freely sliding along the microtubule on which it moves. We investigated the kinesin stepping mechanism by immobilizing a Drosophila kinesin derivative through the carboxyl-terminal end of the neck coiled-coil domain and measuring orientations of microtubules moved by single enzyme molecules at submicromolar adenosine triphosphate concentrations. The kinesin-mediated microtubule-surface linkage was sufficiently torsionally stiff (>/=2.0 +/- 0.9 x 10(-20) Newton meters per radian2) that stepping by the hypothesized symmetric hand-over-hand mechanism would produce 180 degree rotations of the microtubule relative to the immobilized kinesin neck. In fact, there were no rotations, a finding that is inconsistent with symmetric hand-over-hand movement. An alternative "inchworm" mechanism is consistent with our experimental results.     

Structure and mechanism of the lactose permease of Escherichia coli

Membrane transport proteins that transduce free energy stored in electrochemical ion gradients into a concentration gradient are a major class of membrane proteins. We report the crystal structure at 3.5 angstroms of the Escherichia coli lactose permease, an intensively studied member of the major facilitator superfamily of transporters. The molecule is composed of N- and C-terminal domains, each with six transmembrane helices, symmetrically positioned within the permease. A large internal hydrophilic cavity open to the cytoplasmic side represents the inward-facing conformation of the transporter. The structure with a bound lactose homolog, beta-D-galactopyranosyl-1-thio-beta-D-galactopyranoside, reveals the sugar-binding site in the cavity, and residues that play major roles in substrate recognition and proton translocation are identified. We propose a possible mechanism for lactose/proton symport (co-transport) consistent with both the structure and a large body of experimental data.     

Development of A dormancy release technology: A review

Tuber dormancy can be released immediately in many commercially important potato cultivars by brief treatment (1-2 days) with bromoethane (BE) vapor at room temperature. The development of a large scale technology for BE application and safe removal through a capturing technique is necessary for successful application of this dormancy release method. Ideally, BE treatment of seed tubers would occur in a closed environment that would capture BE vapor in an unaltered form and allow controlled release for treatment of subsequent tuber lots. Results of screening studies for adsorbents indicate that the medium capacity activated carbon adsorbent, YAO has: i) a high capacity for BE; ii) a low capacity for water; and, iii) adsorbs and de-adsorbs BE quickly and easily. A plausible design of a large scale, dormancy release facility is presented. The proposed facility should meet present goals of the seed potato industry in an environmentally responsible manner.     

13-Acetoxysarcocrassolide Induces Apoptosis on Human Gastric Carcinoma Cells Through Mitochondria-Related Apoptotic Pathways: p38/JNK Activation and PI3K/AKT Suppression

13-acetoxysarcocrassolide (13-AC), an active compound isolated from cultured Formosa soft coral Sarcophyton crassocaule, was found to possess anti-proliferative and apoptosis-inducing activities against AGS (human gastric adenocarcinoma cells) gastric carcinoma cells. The anti-tumor effects of 13-AC were determined by MTT assay, colony formation assessment, cell wound-healing assay, TUNEL/4,6-Diamidino-2-phenylindole (DAPI) staining, Annexin V-fluorescein isothiocyanate/propidium iodide (PI) staining and flow cytometry. 13-AC inhibited the growth and migration of gastric carcinoma cells in a dose-dependent manner and induced both early and late apoptosis as assessed by flow cytometer analysis. 13-AC-induced apoptosis was confirmed through observation of a change in ΔΨm, up-regulated expression levels of Bax and Bad proteins, down-regulated expression levels of Bcl-2, Bcl-xl and Mcl-1 proteins, and the activation of caspase-3, caspase-9, p38 and JNK. Furthermore, inhibition of p38 and JNK activity by pretreatment with SB03580 (a p38-specific inhibitor) and SP600125 (a JNK-specific inhibitor) led to rescue of the cell cytotoxicity of 13-AC-treated AGS cells, indicating that the p38 and the JNK pathways are also involved in the 13-AC-induced cell apoptosis. Together, these results suggest that 13-AC induces cell apoptosis against gastric cancer cells through triggering of the mitochondrial-dependent apoptotic pathway as well as activation of the p38 and JNK pathways.     

Organic anion-to-acid ratio influences pH change of soils differing in initial pH

Purpose

This study aimed to investigate the effect of initial soil pH and organic anion-to-acid ratio on changes in soil pH.

Materials and methods

Two soils (Podosol and Tenosol) along with two carboxylic acids (malic and citric acid) and their anions (sodium malate and citrate), commonly found in plant residues, were used in this study. Stock solutions of either malic acid and disodium malate or citric acid and trisodium citrate were added to pre-incubated soils at anion-to-acid ratios of 0:100, 10:90, 25:75, 50:50, 75:25, 90:10, 100:0 and at 0.25 g C kg^?1 soil. Soils were adjusted to 80 % field capacity and mixed thoroughly, and three replicates of 50 g of each soil were transferred into individual plastic cores and incubated at 25 °C in the dark for 30 days. Soil pH, respiration, NH₄ ⁺, and NO₃ ^? were determined.

Results and discussion

Soil pH increased linearly with increasing organic anion-to-acid ratio. The addition of organic anions to soil resulted in net alkalinisation. However, the addition of organic acids immediately decreased soil pH. During subsequent incubation, soil pH increased when the organic anions were decomposed. Alkalinity generation was lower in the Podosol (initial pH 4.5) than in the Tenosol (initial pH 6.2), and was proportional to anion-to-acid ratio across all the treatments. Cumulative CO₂-C release was approximately three times lower in the Podosol than the Tenosol at day 2 due to lower microbial activity in the low-pH Podosol.

Conclusions

Increasing anion-to-acid ratio of organic compounds increased soil pH. Increases in soil pH were mainly attributed to direct chemical reactions and decomposition of organic anions. Low pH decreased the amount of alkalinity generated by addition of organic compounds due to incomplete decomposition of the added compounds. This study implies that organic anion-to-acid ratio in plant residues plays an important role in soil pH change.     

Real-time polymerase chain reaction monitoring of recombinant DNA entry into soil from decomposing roundup ready leaf biomass

Glyphosate-tolerant, Roundup Ready (RR) soybeans account for about 57% of all genetically modified (GM) crops grown worldwide. The entry of recombinant DNA into soil from GM crops has been identified as an environmental concern due to the possibility of their horizontal transfer to soil microorganisms. RR soybeans contain recombinant gene sequences that can be differentiated from wild-type plant and microbial genes in soil by using a sequence-specific molecular beacon and real-time polymerase chain reaction (PCR). A molecular beacon-based real-time PCR system to quantify a wild-type soybean lectin ( le1) gene was designed to compare amounts of endogenous soybean genes to recombinant DNA in soil. Microcosm studies were carried out to develop methodologies for the detection of recombinant DNA from RR soybeans in soil. RR soybean leaf litterbags were imbedded in the soil under controlled environmental conditions (60% water holding capacity, 10/15 degrees C, and 8/16 h day/night) for 30 days. The soybean biomass decomposition was described using a single-phase exponential equation, and the DNA concentration in planta and in soil was quantified using real-time PCR using sequence-specific molecular beacons for the recombinant cp4 epsps and endogenous soybean lectin ( le1) genes. The biomass of RR soybean leaves was 8.6% less than nontransgenic (NT) soybean leaves after 30 days. The pooled half-disappearance time for cp4 epsps and le1 in RR and of le1 in NT soybean leaves was 1.4 days. All genes from leaves were detected in soil after 30 days. This study provides a methodology for monitoring the entry of RR and NT soybean DNA into soil from decomposing plant residues.     

Mercury Partitioning in Surface Sediments of the Upper St. Lawrence River (Canada): Evidence of the Importance of the Sulphur Chemistry

An intensive survey of mercury speciation was performed at a site on the Upper St. Lawrence River near Cornwall, Ontario, Canada with a history mercury contamination in sediments. Surface sediments were collected every 1.50 h. Total mercury (Hg_total), methylmercury (MeHg), organic carbon, inorganic and organic sulphur were determined in the solid fraction. Dissolved Hg_total, MeHg and dissolved organic carbon (DOC) were measured in pore waters. Concentrations of Hg_total in the upper layers (first 5 cm) were high, ranging from 1.42 to 25.8 nmol g^?1 in solids and from 125 to 449 pM in pore waters. MeHg levels were also high, ranging from 4.34 to 34.1 pmol g^?1 in solids and from 40 to 96 pM in pore waters. This amounts to up to 1.4% of Hg_total present as MeHg in solids and 64% in pore waters. A daily pattern for Hg_total was observed in the solid fraction. The MeHg distribution in solids and pore waters was not correlated with Hg_total or DOC_, suggesting that the concentrations of MeHg are probably more influenced by the relative rates of methylation/demethylation reactions in the sediment–water interface. Acid volatile sulphide levels and DOC were inversely correlated with organic sulphur (S_org) levels suggesting that both parameters are involved in the rapid production of S_org. A positive correlation was also observed between Hg_total and S_org in solids (R?=?0.87, p?<?0.01) illustrating the importance of organic sulphur in the retention and distribution of Hg in the solid fraction of the sediments. The results suggest that variations of Hg_total concentrations in Upper St. Lawrence River surface sediments were strongly influenced by the formation/deposition/retention of organic sulphur compounds in the sediment–water interface.