The Nitric Oxide Production in the Moss Physcomitrella patens Is Mediated by Nitrate Reductase

During the last 20 years multiple roles of the nitric oxide gas (•NO) have been uncovered in plant growth, development and many physiological processes. In seed plants the enzymatic synthesis of •NO is mediated by a nitric oxide synthase (NOS)-like activity performed by a still unknown enzyme(s) and nitrate reductase (NR). In green algae the •NO production has been linked only to NR activity, although a NOS gene was reported for Ostreococcus tauri and O. lucimarinus, no other Viridiplantae species has such gene. As there is no information about •NO synthesis neither for non-vascular plants nor for non-seed vascular plants, the interesting question regarding the evolution of the enzymatic •NO production systems during land plant natural history remains open. To address this issue the endogenous •NO production by protonema was demonstrated using Electron Paramagnetic Resonance (EPR). The •NO signal was almost eliminated in plants treated with sodium tungstate, which also reduced the NR activity, demonstrating that in P. patens NR activity is the main source for •NO production. The analysis with confocal laser scanning microscopy (CLSM) confirmed endogenous NO production and showed that •NO signal is accumulated in the cytoplasm of protonema cells. The results presented here show for the first time the •NO production in a non-vascular plant and demonstrate that the NR-dependent enzymatic synthesis of •NO is common for embryophytes and green algae.     

Magnolia nuevoleonensis sp. nov. (Magnoliaceae) from northeastern Mexico and a key to species of section Macrophylla

We describe and illustrate Magnolia nuevoleonensis A. Vázquez & Domínguez‐Yescas, a new deciduous tree species that belongs to M. sect. Macrophylla Figlar & Nooteboom. It is morphologically similar to M. ashei Weath., but differs from the latter in being a larger tree, with smaller flowers, larger and broadly ovoid (vs cylindrical to ellipsoid) fruits, prominently beaked (vs shortly beaked) carpels, and firm and short (vs flexible and elongated) styles.     

Ozone exposure alters 5-hydroxy-indole-acetic acid contents in dialysates from dorsal raphe and medial preoptic area in freely moving rats. Relationships with simultaneous sleep disturbances

Ozone (O3) has been reported to affect sleep patterns and also striatal and mesencephalic contents of 5-hydroxy-indole-acetic acid (5-HIAA) in rats. The aim of this work was to elucidate the effects of O3 exposure in rats upon extracellular 5-HIAA levels in the dorsal raphe (DR) and the hypothalamic medial preoptic area (MPO), two structures involved in sleep-wake homeostasis. Exposure to O3 followed a bell-shaped diurnal pattern, similar to that observed in cities with high air pollution levels. The highest O3 concentration employed was 0.5 ppm. Simultaneous polygraphic records were performed to evaluate the concomitant effects of this exposure model on sleep patterns. Results showed that extracellular 5-HIAA levels increased by 28% in the DR (P=0.0213) while paradoxical sleep (PS) decreased by 56% (P=0.0000) during the light O3 exposure phase. A decrease of 32% in 5-HIAA levels in the MPO (P=0.0450), and of 22% in slow wave sleep (SWS) (P=0.0002) and an increase of 21% in wakefulness (P=0.0430) during the dark post-exposure (Dpost) phase were also observed. We propose that the decrease in PS is the behavioral expression of disruptions of serotonergic DR modulation and, that post-exposure effects observed in the MPO can be explained on the basis of the hypothalamic role in the sleep-wake cycle.     

Production of the Escherichia coli Common Pilus by Uropathogenic E. coli Is Associated with Adherence to HeLa and HTB-4 Cells and Invasion of Mouse Bladder Urothelium

Uropathogenic Escherichia coli (UPEC) strains cause urinary tract infections and employ type 1 and P pili in colonization of the bladder and kidney, respectively. Most intestinal and extra-intestinal E. coli strains produce a pilus called E. coli common pilus (ECP) involved in cell adherence and biofilm formation. However, the contribution of ECP to the interaction of UPEC with uroepithelial cells remains to be elucidated. Here, we report that prototypic UPEC strains CFT073 and F11 mutated in the major pilin structural gene ecpA are significantly deficient in adherence to cultured HeLa (cervix) and HTB-4 (bladder) epithelial cells in vitro as compared to their parental strains. Complementation of the ecpA mutant restored adherence to wild-type levels. UPEC strains produce ECP upon growth in Luria-Bertani broth or DMEM tissue culture medium preferentially at 26°C, during incubation with cultured epithelial cells in vitro at 37°C, and upon colonization of mouse bladder urothelium ex vivo. ECP was demonstrated on and inside exfoliated bladder epithelial cells present in the urine of urinary tract infection patients. The ability of the CFT073 ecpA mutant to invade the mouse tissue was significantly reduced. The presence of ECP correlated with the architecture of the biofilms produced by UPEC strains on inert surfaces. These data suggest that ECP can potentially be produced in the bladder environment and contribute to the adhesive and invasive capabilities of UPEC during its interaction with the host bladder. We propose that along with other known adhesins, ECP plays a synergistic role in the multi-step infection of the urinary tract.     

De novo identification of binding sequences for antibody replacement molecules

A new in silico method has been developed that automatically identifies peptide sequences that can bind to targets of known three‐dimensional structure. The method is potentially faster and more economical than traditional methods of raising antibodies by means of hybridomas or biopanning technology. The current algorithm creates an initial peptide library that is either completely random or that is constrained by the user. This library represents only a small fraction of possible sequence space and the peptides are created with a specified torsional geometry. The library is used as input to any number of available molecular docking programs and the library is docked and scored. The final rank ordering is then used to create a new library by constraining that library to the sequence conservation pattern deduced from the top N‐scoring peptides in the first round. Successive rounds of screening, scoring, and new library creation ultimately results in the system converging to a final solution set of peptides. To test the method, a family of novel peptides that can bind to, and inhibit the enzyme Deoxyribonuclease I has been discovered. The peptides inhibit the enzyme either alone or when placed into a protein backbone structure as has been confirmed experimentally. Proteins 2009. © 2009 Wiley‐Liss, Inc.     

Primary Cutaneous Mucormycosis Caused by <Emphasis Type="Italic">Rhizopus oryzae</Emphasis>: A Case Report and Review of Literature

Mucormycosis is an invasive infection caused by opportunistic fungi. Rhizopus, Lichtheimia, Mucor and Rhizomucor are the most common isolated genera. Primary cutaneous mucormycosis is usually related to traumatic injuries, but immunocompromised cases are associated with underlying conditions such as diabetes mellitus and malignancies. The treatment of choice is surgical debridement and liposomal amphotericin B. We present a 40-year-old male with fever and a painful necrotic lesion on the middle back and history of poorly controlled diabetes mellitus. Rhizopus oryzae was isolated and identified using an internal transcribed spacer regions ITS1 and ITS2. An initial good response to treatment was observed; however, 7 days later a diabetic ketoacidosis due to poor adherence to treatment caused a lethal outcome.     

An enhanced genome-scale metabolic reconstruction of Streptomyces clavuligerus identifies novel strain improvement strategies

Bioprocess and Biosystems Engineering - In this work, we expanded and updated a genome-scale metabolic model of Streptomyces clavuligerus. The model includes 1021 genes and 1494 biochemical...     

Prolactin-like hormone in the nematode Trichinella spiralis larvae

Expression of prolactin (PRL) or prolactin-like hormone has been reported in invertebrates. We investigated the larval phase of Trichinella spiralis: (a) to express 23 kDa PRL, (b) to define its localization and (c) to test its possible biological activity. Immunostaining in isolated larvae demonstrated positive material to 23 kDa PRL by all along the stichosome, specifically in the stichocytes. Homogenized immunoblot larvae showed a 23 kDa protein band. To assess PRL release and its biological activity, larvae were incubated in culture medium and the excretory/secretory products were analyzed by the Nb2 cells bioassay. A cellular growth equivalent until 10 nM PRL and using antibody against 23 kDa PRL, the growth was blocked. In conclusion our result provides evidence that PRL-like hormone is expressed and secreted by the larvae of T. spiralis.