Stimulation of bacterial cytokinesis by bacteriophage predation

Bacteriophage (phage) are ubiquitous in the water column and in the sediments of most natural water. Because of their colloidal nature, they can either aggregate into clumps large enough to settle into the sediment or departing upon the physiochemical conditions or disassociate and reenter the water column. About 80% of the bacterial strains isolated from New River sediment have a virulent phage that can be isolated with them.Liquid cultures of a strain of Pseudomonas aeruginosa isolated from the New River along with its phage were set up. One was infected with the virulent phage and another kept as a control. Daily counts were made of bacterial numbers. After 10 days the control culture was infected and counted for 3 more days.Both cultures divided exponentially at first. The infected culture continued to divide at about half the initial rate. The uninfected culture nearly ceased division, but when phage were added it quickly began to divide.The virulent phage infection clearly stimulated host division. The effect was to establish itself as an endemic infection which did not outpace its host's division rate. Further, the enhanced division rate may act to increase the host's share of available nutrients and benefit its competitive position in the system.     

Quantitative analysis of thymosin alpha1 in human serum by LC-MS/MS

A high performance liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was developed to measure the thymosin alpha 1 (Talpha1) concentration in human serum. Tá1 in human serum was determined by solid phase extraction and reverse phase LC-MS/MS. The high-performance liquid chromatography (HPLC) system interfaced with the MS/MS system with a Turbo Ion spray interface. Positive ion detection and multiple reaction monitoring (MRM) mode were used for this human serum quantitation. Eight different concentration standards were used to establish the detection range. Six quality control (QC) and 2 matrix blanks were checked by calibration curves performed on the same day. The lower quantitation limit was 0.5 ng/mL Talpha1 in human serum. Calibration curves were established between 0.5 to 100 ng/mL by weighted linear regression. The correlation coefficients for different days were 0.9955 or greater. Quantitation of Talpha1 by the LC-MS/MS method is fast, accurate, and precise.     

Penicillium subrubescens, a new species efficiently producing inulinase

Inulin is a reserve carbohydrate in about 15 % of the flowering plants and is accumulated in underground tubers of e.g. chicory, dahlia and Jerusalem artichoke. This carbohydrate consists of linear chains of β-(2,1)-linked fructose attached to a sucrose molecule. Inulinases hydrolyse inulin into fructose and glucose. To find efficient inulin degrading fungi, 126 fungal strains from the Fungal Biotechnology Culture Collection (FBCC) at University of Helsinki and 74 freshly isolated strains from soil around Jerusalem artichoke tubers were screened in liquid cultures with inulin as a sole source of carbon or ground Jerusalem artichoke tubers, which contains up to 19 % (fresh weight) inulin. Inulinase and invertase activities were assayed by the dinitrosalicylic acid (DNS) method and a freshly isolated Penicillium strain originating from agricultural soil (FBCC 1632) was the most efficient inulinase producer. When it was cultivated at pH 6 and 28 °C in 2 litre bioreactors using inulin and Jerusalem artichoke as a carbon source, inulinase and invertase activities were on day 4 7.7 and 3.1 U mL^?1, respectively. The released sugars analysed by TLC and HPLC showed that considerable amounts of fructose were released while the levels of oligofructans were low, indicating an exoinulinase type of activity. Taxonomic study of the inulinase producing strain showed that this isolate represents a new species belonging in Penicillium section Lanata-divaricata. This new species produces a unique combination of extrolites and is phenotypically and phylogenetically closely related to Penicillium pulvillorum. We propose the name Penicillium subrubescens sp. nov. (CBS 132785^T = FBCC 1632^T) for this new species.     

A new species from tropical soils,Eupenicillium tropicum

Forty-three strains of Eupenicillium tropicum sp. nov. were isolated from soils collected in India, Costa Rica and Galapagos, Ecuador. The species is characterized by biverticillate penicilli, slightly rough, subglobose to ovate conidia, brownish cleistothecia that become brown-gray with age, and ascospores with two equatorial flanges and slightly roughened valves. All strains produced a large number of indole alkaloids, and many types of unknown secondary metabolites with characteristic chromophores were produced by a majority of strains. Eupenicillium tropicum is morphologically most similar to E. shearii, but based on ITS-LSU sequences, is most closely related to Penicillium citrinum, P. sartoryi and P. westlingii. Eupenicillium shearii strains consistently produce paxillin, paspalinine and shearinins, while the latter three penicillia all produce citrinin consistently.     

Interaction of the synthetic immunomodulatory dipeptide bestim with murine macrophages and thymocytes

The tritium-labeled dipeptide bestim (γ-D-Glu-L-Trp) with a specific activity of 45 Ci/mmol was obtained by high-temperature solid-state catalytic isotope exchange. It was found that [³H]bestim binds with a high affinity to murine peritoneal macrophages (K _d 2.1 ± 0.1 nM) and thymocytes (K _d 3.1 ± 0.2 nM), as well as with plasma membranes isolated from these cells (K _d 18.6 ± 0.2 and 16.7 ± 0.3 nM, respectively). The specific binding of [³H]bestim to macrophages and thymocytes was inhibited by the unlabeled dipeptide thymogen (L-Glu-L-Trp) (K _i 0.9 ± 0.1 and 1.1 ± 0.1 nM, respectively). After treatment with trypsin, macrophages and thymocytes lost the ability to bind [³H]bestim. Bestim in the concentration range of 10^?10 to 10^?6 M reduced the adenylate cyclase activity in the membranes of murine macrophages and thymocytes.     

Discovery of a series of aminopiperidines as novel iNOS inhibitors

Nitric oxide (NO), a mediator of various physiological and pathophysiological processes, is synthesized by three isozymes of nitric oxide synthase (NOS). Potential candidate clinical drugs should be devoid of inhibitory activity against endothelial NOS (eNOS), since eNOS plays an important role in maintaining normal blood pressure and flow. A new series of aminopiperidines as potent inhibitors of iNOS were identified from a HTS lead. From this study, we identified compound 33 as a potent iNOS inhibitor, with >25-fold selectivity over eNOS and 16-fold selectivity over nNOS.