The maize RNA-binding protein, MA16, is a nucleolar protein located in the dense fibrillar component

A developmentally and environmentally regulated gene in maize, MA16, encoding an RNA-binding protein that binds preferentially to uridine and guanosine-rich RNAs has previously been described. To gain some insight into the function of MA16 the distribution of MA16 mRNA and protein during maize development was investigated using in situ hybridization, RNA and protein gel blot analysis and immunocytochemistry. The results show that MA16 is expressed throughout development of the embryo and seedling in different tissues and at different levels. The level of MA16 mRNA is higher in developing and expanding structures such as the root elongation zone and young leaves. After stress treatment MA16 mRNA increases in total and polysomal RNA, but no significant change in the level of the protein was detected. MA16 is a non-ribosomal nucleolar protein. Using immunoelectron microscopy the MA16 protein has been located in the dense fibrillar component and to a lesser extent in the granular component of the nucleolus. It was found that MA16 contains the conserved sequence motifs R(G)_nY(G)_nR and RR(E/D)(G)_nY(G)_n repeated in the C-terminal of the molecule that conforms imperfectly to the GAR motif proposed for nucleolar proteins. In light of these results the stress regulation of MA16 and a likely role for this protein in pre-rRNA processing and/or ribosome assembly is discussed.     

Production of low-molecular weight thiols as a response to cadmium uptake by tumbleweed (Salsola kali).

Tumbleweed (Salsola kali) is a desert plant species that has shown to be a potential Cd hyperaccumulator. In this study, the production of low-molecular weight thiols (LMWT) as a response to cadmium stress was determined in hydroponically grown seedlings exposed to 0, 45, 89, and 178 microM Cd(2+). The treatment of 89 microM Cd(2+) was tested alone and supplemented with an equimolar concentration of ethylenediaminetetraacetic acid (EDTA) to determine the effect of this chelating agent on Cd uptake and thiols production. After 6 days of growth, the Cd concentration in plant tissues was determined by using inductively coupled plasma/optical emission spectroscopy (ICP/OES). Results indicated that Cd uptake by plants was concentration-dependent. Plants treated with 178 microM Cd(2+), had 10+/-0.62, 9.7+/-1.4, and 4.3+/-0.83 mmol Cd kg(-1) dry tissue in roots, stems, and leaves, respectively. The production of thiols was dependent on Cd concentration in tissues. According to the stoichiometry performed, plants treated with Cd concentrations up to 178 muM produced 0.131+/-0.02, and 0.087+/-0.012 mmol SH per mmol Cd present in roots and stems. In leaves, the production of thiols decreased at the highest Cd concentration tested. Thus, up to 89 microM Cd in the media, 0.528+/-0.004 mmol SH per mmol Cd in leaf tissues were produced. EDTA equimolar to Cd reduced both Cd uptake and thiols production. Catalase activity (CAT) (EC 1.11.1.6) was significantly depressed at the lowest Cd concentration. None of the conditions tested affected biomass or plant elongation.     

Dosimetry of the left anterior descending coronary artery in left breast cancer patients treated with postoperative external radiotherapy

Aim

To evaluate the dose distribution to the left anterior descending (LAD) coronary artery in patients treated with postoperative three-dimensional conformal radiotherapy (3DCRT).

Brote de histoplasmosis en la provincia de Neuquén,Patagonia Argentina

BackgroundIn Argentina, there are no reports of autochthonous cases of histoplasmosis in the southern regions of the country.AimTo report a histoplasmosis outbreak in Zapala town, Province of Neuquén, Patagonia Argentina.MethodsWe evaluated the clinical and epidemiological characteristics of 5 patients involved in the outbreak. Environmental studies were conducted to determine the source of infection. The genetic profile of Histoplasma capsulatum strains isolated from the index case (IC) were compared with clinical isolates from Argentinean patients not related to the outbreak, using RAPD-PCR with primers 1281-1283.ResultsThe patients were residents of Zapala, and had not visited other geographical areas before. All patients had an influenza-like syndrome, and X-ray revealed disseminated micronodular images throughout the lung parenchyma. The IC needed specific antifungal therapy; the remaining 4 patients had mild symptoms, and did not require therapy. All of them had a good clinical outcome. Strains of H. capsulatum isolated from blood culture and lung biopsy of the IC showed a genetic profile different from other strains analyzed. The presence of the fungus in the environment was demonstrated by the detection of anti-Histoplasma antibodies in BALB/c mice inoculated with soil obtained in a culvert where workers had dug up earth after a landslide.ConclusionsThis outbreak suggests the histoplasmosis endemic area is under the 38° S parallel. Patients from Neuquén, Patagonia Argentina, with compatible symptoms of histoplasmosis should be tested, regardless of their travel or exposure history.     

FleQ Coordinates Flagellum-Dependent and -Independent Motilities in Pseudomonas syringae pv. tomato DC3000

Motility plays an essential role in bacterial fitness and colonization in the plant environment, since it favors nutrient acquisition and avoidance of toxic substances, successful competition with other microorganisms, the ability to locate the preferred hosts, access to optimal sites within them, and dispersal in the environment during the course of transmission. In this work, we have observed that the mutation of the flagellar master regulatory gene, fleQ, alters bacterial surface motility and biosurfactant production, uncovering a new type of motility for Pseudomonas syringae pv. tomato DC3000 on semisolid surfaces. We present evidence that P. syringae pv. tomato DC3000 moves over semisolid surfaces by using at least two different types of motility, namely, swarming, which depends on the presence of flagella and syringafactin, a biosurfactant produced by this strain, and a flagellum-independent surface spreading or sliding, which also requires syringafactin. We also show that FleQ activates flagellum synthesis and negatively regulates syringafactin production in P. syringae pv. tomato DC3000. Finally, it was surprising to observe that mutants lacking flagella or syringafactin were as virulent as the wild type, and only the simultaneous loss of both flagella and syringafactin impairs the ability of P. syringae pv. tomato DC3000 to colonize tomato host plants and cause disease.     

In vitro reconstitution reveals a key role of human mitochondrial EXOG in RNA primer processing

The removal of RNA primers is essential for mitochondrial DNA (mtDNA) replication. Several nucleases have been implicated in RNA primer removal in human mitochondria, however, no conclusive mechanism has been elucidated. Here, we reconstituted minimal in vitro system capable of processing RNA primers into ligatable DNA ends. We show that human 5′-3′ exonuclease, EXOG, plays a fundamental role in removal of the RNA primer. EXOG cleaves short and long RNA-containing flaps but also in cooperation with RNase H1, processes non-flap RNA-containing intermediates. Our data indicate that the enzymatic activity of both enzymes is necessary to process non-flap RNA-containing intermediates and that regardless of the pathway, EXOG-mediated RNA cleavage is necessary prior to ligation by DNA Ligase III. We also show that upregulation of EXOG levels in mitochondria increases ligation efficiency of RNA-containing substrates and discover physical interactions, both in vitro and in cellulo, between RNase H1 and EXOG, Pol γA, Pol γB and Lig III but not FEN1, which we demonstrate to be absent from mitochondria of human lung epithelial cells. Together, using human mtDNA replication enzymes, we reconstitute for the first time RNA primer removal reaction and propose a novel model for RNA primer processing in human mitochondria.     

Some Pneumococcal Serotypes Are More Frequently Associated with Relapses of Acute Exacerbations in COPD Patients

Objectives

To analyze the role of the capsular type in pneumococci causing relapse and reinfection episodes of acute exacerbation in COPD patients.

Methods

A total of 79 patients with 116 recurrent episodes of acute exacerbations caused by S. pneumoniae were included into this study (1995–2010). A relapse episode was considered when two consecutive episodes were caused by the same strain (identical serotype and genotype); otherwise it was considered reinfection.Antimicrobial susceptibility testing (microdilution), serotyping (PCR, Quellung) and molecular typing (PFGE/MLST) were performed.

Results

Among 116 recurrent episodes, 81 (69.8%) were reinfections, caused by the acquisition of a new pneumococcus, and 35 (30.2%) were relapses, caused by a pre-existing strain. Four serotypes (9V, 19F, 15A and 11A) caused the majority (60.0%) of relapses. When serotypes causing relapses and reinfection were compared, only two serotypes were associated with relapses: 9V (OR 8.0; 95% CI, 1.34–85.59) and 19F (OR 16.1; 95% CI, 1.84–767.20). Pneumococci isolated from relapses were more resistant to antimicrobials than those isolated from the reinfection episodes: penicillin (74.3% vs. 34.6%, p<0.001), ciprofloxacin (25.7% vs. 9.9%, p<0.027), levofloxacin (22.9% vs. 7.4%, p = 0.029), and co-trimoxazole (54.3% vs. 25.9%, p<0.001).

Conclusions

Although the acquisition of a new S. pneumoniae strain was the most frequent cause of recurrences, a third of the recurrent episodes were caused by a pre-existing strain. These relapse episodes were mainly caused by serotypes 9V and 19F, suggesting an important role for capsular type.     

ASIC1a activation by amitriptyline and FMRF-amide is removed by serine proteases

Analgesia induced by certain tricyclic antidepressants has been largely used for decades, yet the mechanisms involved are incompletely understood. Starting from previously reported dual effects of amitriptyline on wild-type ENaC (Pena F, et al. J Pharm Pharmacol 54:1393-8: 2002), we extended our study to ASIC1a by performing a series of whole cell and single-channel recordings of proton-activated currents in HEK293 cells. Acid pulses were applied at 2 or 5 min intervals, and amitriptyline (1-500 microM) was applied at a holding pH of 7.4 or 8.4 between pulses. Dose-response plots were fitted with dual Hill type functions, yielding a half-activatory constant of 0.3 microM and a half-inhibitory constant of 382 microM at pH 7.4. At pH 8.4 both constants were shifted to higher values (0.5 and 444 microM, respectively). In whole-cell experiments, FMRF-amide increased the peak amplitude of ASIC1a transients at 0.1 microM and decreased it at 1 and 100 microM. Single-channel recordings were idealized and fitted using an 8-state linear connectivity model comprising four consecutive activation steps. Both amitriptyline (1 microM) and FMRF-amide (0.1 microM) increased the unitary current amplitude, and modified the opening and closing rates of the first gating mode. They also increased the transition rate from the second to the first gating mode, and the rate of final closure. The activatory effect of both compounds vanished after a mild trypsin pretreatment, suggesting the existence of activatory sites for FMRF-amide and amitriptyline in the outer vestibule of ASIC1a, which can be removed by exo- or endogenous serine-proteases.