Descripción: The growth rate of several polyamine-deficient mutants of Escherichia coli was very low in minimal medium and increased markedly upon the addition of putrescine, spermidine, arginine, citrulline, or argininosuccinic acid. The endogenous content of polyamines was not significantly altered by the supplementation of polyamine-starved cultures with arginine or its precursors. In contrast, these compounds as well as putrescine or spermidine caused a 40-fold reduction in intracellular ornithine levels when added to polyamine-depleted bacteria. In vivo experiments with radioactive glutamic acid as a precursor and in vitro assay of the related enzymes showed that the decrease in ornithine levels was due to the inhibition of its biosynthesis rather than to an increase in its conversion to citrulline or Δ1-pyrroline-5-carboxylic acid and proline. High endogenous concentrations of ornithine were toxic for the E. coli strains tested. The described results indicate that the stimulatory effect of putrescine and spermidine on the growth of certain polyamine-starved bacteria may be partially due to the control of ornithine biosynthesis by polyamines.

Descripción: Microbial oncoids have been found in the Oxfordian limestones of the La Manga Formation in the La Vaina section at Potimalal River, Mendoza province. The oncoids ocurr either in packstone or floatstone-rudstone or are scattered in the wackestones. They are mostly elliptical, ameboidal and subordinately spherical in shape. Different types of oncoids were recognized, according to their features of envelopes: (1) micritic laminations, (2) grumose laminations, and (3) organism-bearing laminations. Molluscs and echinoid fragments, peloids, and intraclasts acted as oncoid nuclei. Laminae follow the shape of nuclei, especially in the inner zone of the cortices. However, in the outer zones the laminae contain encrusting organisms dominated by nubeculariids and serpulids. Some oncoids, especially the ameboidal or elliptical forms, are characterized by multiple nuclei, represented by small oncoids. The oncoids are associated with bivalves, echinoderms, forams, and serpulids. The fauna is indicative of calm, shallow conditions and the excellent preservation of echinoderms suggests minimal transport prior to burial. Growth histories are in evidence. The oncoids grew in a shallow, low energy, slightly to moderate agitated subtidal normal sea water environment. The limited rolling growth oncoids was probably accompanished by intermittent currents that reoriented the oncoids parallel to stratification. The discontinuous organisms-bearing laminations reflect periods of non-agitation and litification, which facilitated the growth of encrusting organisms on static oncoid particles during a period of low sedimentation rate. Deposition of the oncoid-bearing limestones took place during shallowing of the carbonate interval and associated with emersion, subaerial exposition and paleokarst as consequence of sea level fluctuations.

Descripción: In flowering plants, the process of pollen germination and tube growth is required for successful fertilization. A pollen receptor kinase from tomato (Solanum lycopersicum), LePRK2, has been implicated in signaling during pollen germination and tube growth as well as in mediating pollen (tube)-pistil communication. Here we show that reduced expression of LePRK2 affects four aspects of pollen germination and tube growth. First, the percentage of pollen that germinates is reduced, and the time window for competence to germinate is also shorter. Second, the pollen tube growth rate is reduced both in vitro and in the pistil. Third, tip-localized superoxide production by pollen tubes cannot be increased by exogenous calcium ions. Fourth, pollen tubes have defects in responses to style extract component (STIL), an extracellular growth-promoting signal from the pistil. Pollen tubes transiently overexpressing LePRK2-fluorescent protein fusions had slightly wider tips, whereas pollen tubes coexpressing LePRK2 and its cytoplasmic partner protein KPP (a Rop-GEF) had much wider tips. Together these results show that LePRK2 positively regulates pollen germination and tube growth and is involved in transducing responses to extracellular growth-promoting signals. © 2008 American Society of Plant Biologists.

Descripción: The population structure, particularly growth, age, mortality and somatic production of the olivid snail Olivancillaria deshayesiana were investigated. Annual formation of internal shell growth marks was confirmed by the record of stable oxygen isotopes in the shell, which reflects seasonal patterns of water temperature. A von Bertalanffy growth model fitted to 81 size-atage data pairs, indicating that O. deshayesiana may attain 31 mm SL in about 10 years. The estimated total mortality Z and natural mortality M were 0.651 y-1 and 0.361 y -1, respectively. Fishing mortality F was 0.290 y -1, and the exploitation rate E was 0.445, indicating that this population was not overexploited at the time of the study. However, this situation may well change in the future, since the important prawn and shrimp fisheries (in intensity and scale) in the Mar del Plata area (38°20'S, 57°37'W) may indirectly affect the exploitation status of the studied population.

Descripción: In the Beagle Channel, southern South America (ca. 55°S 67°W), about 20% of false king crabs (Paralomis granulosa) >80 mm carapace length are fouled with the barnacle Notobalanus flosculus. To evaluate differences in growth rates of barnacles attached to artificial and live substrates, clay tiles were anchored as collectors to the bottom at two different sites in the Beagle Channel in September 1996: in Ushuaia harbour (low currents and high levels of suspended matter) and around the Bridges Islands (strong currents and low level of suspended matter). Another set of collectors was deployed at the same sites in October 1998 to collect barnacles for histological studies. Tiles were removed from each place, approximately, on a monthly basis. Carapaces of P. granulosa with the epizoic N. flosculus were sampled between November 1996 and 1997, and between March 1998 and September 1999, to study sexual maturation of barnacles. Growth of barnacles was compared between the collectors and P. granulosa carapaces following a qualitative approach. A sexual maturity scale was defined, based on the stage of development of the female reproductive apparatus of N. flosculus. Growth rate of barnacles was highest in the harbour, intermediate on P. granulosa, and lowest around the Bridges Islands. Presence of oocytes was registered only in epizoic barnacles, suggesting that at least a proportion of these individuals is able to spawn on the carapaces. The potential advantages of settling on a living substrate, namely increased availability of food particles and decreased predation risks are discussed. © Springer-Verlag and AWI 2005.

Descripción: We assessed the effects of different arcA mutations on poly(3- hydroxybutyrate) (PHB) synthesis in recombinant Escherichia coli strains carrying the pha synthesis genes from Azotobacter sp. strain FA8. The arcA mutations used were an internal deletion and the arcA2 allele, a leaky mutation for some of the characteristics of the Arc phenotype which confers high respiratory capacity. PHB synthesis was not detected in the wild-type strain in shaken flask cultures under low-oxygen conditions, while ArcA mutants gave rise to polymer accumulation of up to 24% of their cell dry weight. When grown under microaerobic conditions in a bioreactor, the arcA deletion mutant reached a PHB content of 27% ± 2%. Under the same conditions, higher biomass and PHB concentrations were observed for the strain bearing the arcA2 allele, resulting in a PHB content of 35% ± 3%. This strain grew in a simple medium at a specific growth rate of 0.69 ± 0.07 h-1, whereas the deletion mutant needed several nutritional additives and snowed a specific growth rate of 0.56 ± 0.06 h-1. The results presented here suggest that arcA mutations could play a role in heterologous PHB synthesis in microaerobiosis. Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Descripción: We have previously described a murein hydrolase activity for the surface layer (S-layer) of Lactobacillus acidophilus ATCC 4356. Here we show that, in combination with nisin, this S-layer acts synergistically to inhibit the growth of pathogenic Gram-negative Salmonella enterica and potential pathogenic Gram-positive bacteria, Staphylococcus aureus and Bacillus cereus. In addition, bacteriolytic effects were observed for the Gram-positive species tested. We postulate that the S-layer enhances the access of nisin into the cell membrane by enabling it to cross the cell wall, while nisin provides the sudden ion-nonspecific dissipation of the proton motive force required to enhance the S-layer murein hydrolase activity. Copyright © 2010, American Society for Microbiology. All Rights Reserved.

Descripción: Heme oxygenase-1 (HO-1), the inducible enzyme responsible for the rate-limiting step in the heme catabolism, is expressed in AIDS-Kaposi sarcoma (KS) lesions. Its expression is up-regulated by the Kaposi sarcoma-associated herpesvirus (KSHV) in endothelial cells, but the mechanisms underlying KSHV-induced HO-1 expression are still unknown. In this study we investigated whether the oncogenic G protein-coupled receptor (KSHV-GPCR or vGPCR), one of the key KSHV genes involved in KS development, activated HO-1 expression. Here we show that vGPCR induces HO-1 mRNA and protein levels in fibroblasts and endothelial cells. Moreover, targeted knock-down gene expression of HO-1 by small hairpin RNA and chemical inhibition of HO-1 enzymatic activity by tin protoporphyrin IX (SnPP), impaired vGPCR-induced survival, proliferation, transformation, and vascular endothelial growth factor (VEGF)-A expression. vGPCR-expressing cells implanted in the dorsal flank of nude mice developed tumors with elevated HO-1 expression and activity. Chronic administration of SnPP to the implanted mice, under conditions that effectively blocked HO-1 activity and VEGF-A expression in the transplanted cells, strikingly reduced tumor growth, without apparent side effects. On the contrary, administration of the HO-1 inducer cobalt protoporphyrin (CoPP) further enhanced vGPCR-induced tumor growth. These data postulate HO-1 as an important mediator of vGPCR-induced tumor growth and suggest that inhibition of intratumoral HO-1 activity by SnPP may be a potential therapeutic strategy. © 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

Descripción: An atomistic model for Cu electrodeposition under nonequilibrium conditions is presented. Cu electrodeposition takes place with a height-dependent deposition rate that accounts for fluctuations in the local [formula presented] ions concentration at the interface, followed by surface diffusion. This model leads to an unstable interface with the development of protrusions and grooves. Subsequently the model is extended to account for the presence of organic additives, which compete with [formula presented] for adsorption at protrusions, leading to a stable interface with scaling exponents consistent with those of the Edwards-Wilkinson equation. The model reproduces the interface evolution experimentally observed for Cu electrodeposition in the absence and in the presence of organic additives. © 2002 The American Physical Society.

Descripción: Ornithine decarboxylase (ODC) of Crithidia fasciculata extracts shows maximal activity during exponential growth of the parasite and decreases markedly in the stationary phase. The inhibition of protein synthesis by cycloheximide evoked a rapid loss of enzyme activity with a half-life of about 30 min. Upon removal of DFMO from Crithidia cultures treated with the drug for 24 h, the ODC activity increased at the same rate as total protein synthesis. The addition of putrescine at high concentrations to parasites cultivated in a synthetic medium showed that Crithidia CDC levels were not reduced by polyamines. © 1992.

Descripción: The present work was aimed at testing the hypothesis that mycorrhizal Prosopis alba, an economically important tree species worldwide, presents increased salt-tolerance compared with non-mycorrhizal ones and at gaining insight into the possible mechanisms underlying that improvement. For this purpose, a randomized complete block experiment with two factors: mycorrhizal treatments with or without the arbuscular fungus Glomus intraradices and two salinity levels, 0 and 200 mM NaCl was performed. Plant growth in P. alba plants colonized by G. intraradices was less affected by salinity than that in non-arbuscular mycorrhizal (AM) plants, indicating that mycorrhizal colonization turned P. alba more tolerant to salinity. Photosynthesis was reduced by salinity in non-AM plants but not in AM ones. Salinity caused a significant decrease in mean stomatal conductance and transpiration rate, in mycorrhizal plants, but not in uninoculated ones. In this work, we detected two main mechanisms intervening in the salt tolerance enhancement of P. alba by the inoculation with G. intraradices: a- maintaining the net photosynthesis level and b- control of the transpiration rate. Taken together, the results suggest that inoculation with G. intraradices improves P. alba survival rates during the implantation period and seems to be a promising strategy to improve P. alba cultivation in saline lands.

Descripción: Ciliates from sub-surface waters of the Argentine shelf and the Drake Passage under austral summer and autumn conditions were examined and compared for the first time. In both environments, the taxonomic structure of ciliates was related to temperature and salinity, and aloricate oligotrichs dominated in density (80%) over loricate oligotrichs, litostomatids and prostomatids, while the microplanktonic fraction prevailed in terms of biomass (90%) over the nanociliates. Myrionecta rubra was found all along the Argentine shelf only in autumn, but showed isolated peaks of abundance (103 ind. L -1) during summer. Mean values of density and biomass of total ciliates decreased ca. 2-fold from the shelf-slope to oceanic waters, while potential maximum production of aloricate oligotrichs decreased 9-fold, in relation with the drop in chlorophyll a concentration and the latitudinal decline of temperature, also reflected in maximum growth rates. Fifty percent of total ciliate abundance was represented by local increases (maximum: 20 000 ind. L-1 and 25 μg C L-1), which were spatially superimposed with ranges of seawater temperature and chlorophyll a concentrations of 10-15°C and 0.6-6 μg L-1, respectively, and were found in the nearby of fronts located on the shelf and the slope.

Descripción: Two possible patterns of bias in primary sex ratio have been proposed for size-dimorphic brood parasites that do not evict host chicks: (1) larger males should be laid at greater frequency in hosts larger than the parasite because they compete better (increasing their survival) than females with large host nest-mates, and (2) more costly males (i.e. the larger sex) should be laid at greater frequency in hosts smaller than the parasite because, in these hosts, parasite nestlings are provisioned at a higher rate and grow faster than in larger hosts. We tested these hypotheses in two hosts of the sexually size-dimorphic shiny cowbird, Molothrus bonariensis, one smaller (house wren, Troglodytes aedon) and one larger (chalk-browed mockingbird, Mimus saturninus) than the parasite. We measured: (1) sex ratio at laying; (2) development of sexual differences in body mass during the nestling stage; and (3) chick survival and sex ratio of chicks before fledging. In both hosts, we found sexual differences in body mass of nestlings from 7 days of age onwards, although we did not find a bias in the sex ratio of eggs laid and chicks fledged. The results of the present study do not support the hypothesis that shiny cowbird females benefit from biasing the primary sex ratio depending on the size of the hosts they parasitize. © 2013 The Linnean Society of London.

Descripción: Addition of affinity tags to bacteriophage particles facilitates a variety of applications, including vaccine construction and diagnosis of bacterial infections. Addition of tags to phage capsids is desirable, as modification of the tails can lead to poor adsorption and loss of infectivity. Although tags can readily be included as fusions to head decoration proteins, many phages do not have decoration proteins as virion components. The addition of a small (10-amino-acid) Strep-tag II (STAG II) to the mycobacteriophage TM4 capsid subunit, gp9, was not tolerated as a genetically homogenous recombinant phage but could be incorporated into the head by growth of wild-type phage on a host expressing the capsid-STAG fusion. Particles with capsids composed of wild-type and STAG-tagged subunit mixtures could be grown to high titers, showed good infectivities, and could be used to isolate phage-bacterium complexes. Preparation of a STAG-labeled fluoromycobacteriophage enabled capture of bacterial complexes and identification of infected bacteria by fluorescence. © 2013, American Society for Microbiology.

Descripción: Aims: To establish a reliable and rapid protocol to simultaneously obtain high quality DNA from an infected host plant and the infecting pathogen. To develop an accurate and sensitive low-cost assay for the quantification and in planta monitoring of Phytophthora infestans growth.Methods and Results: In this study, we describe a SYBR Green-based quantitative real-time PCR (qPCR) method for the quantification of P. infestans. The method is based on a simultaneous plant-pathogen DNA purification followed by a qPCR in which the relative quantification of pathogen and plant DNA is performed. Besides assuring an accurate quantification, the use of a plant gene provides a reliable indicator of sample quality, allowing the exclusion of inappropriate samples. By applying this methodology, we were able to detect P. infestans in potato leaf and tuber tissue before the first symptoms of the disease were observed and to monitor the in planta growth of the pathogen for 6 days.Conclusions: This is a reliable low-cost assay that provides rapid, accurate and sensitive quantification of the late blight pathogen, allowing the in planta monitoring of P. infestans growth.Significance and Impact of the Study: The quantitative nature of the assay described in this study may be useful in plant breeding programmes and basic research. The method is appropriate for the comparison of cultivars with different, and even subtle, degrees of pathogen resistance and in the screening of new anti-oomycete compounds. The method can be easily adapted to tomato and the model plant Nicotiana benthamiana. © No claim to Argentinean Government works. Letters in Applied Microbiology 51, 603-610 © 2010 The Society for Applied Microbiology.

Descripción: Aims: To evaluate the antagonistic activity of Fusarium oxysporum nonpathogenic fungal strain S6 against the phytopathogenic fungus Sclerotinia sclerotiorum and to identify the antifungal compounds involved. Methods and Results: The antagonistic activity of Fusarium oxysporum strain S6 was determined in vitro by dual cultures. The metabolite responsible for the activity was isolated by chromatographic techniques, purified and identified by spectroscopic methods as cyclosporine A. The antifungal activity against the pathogen was correlated with the presence of this metabolite by a dilution assay and then quantified. Cyclosporine A caused both growth inhibition and suppression of sclerotia formation. In a greenhouse assay, a significant increase in the number of surviving soybean (Glycine max) plants was observed when S. sclerotiorum and F. oxysporum (S6) were inoculated together when compared with plants inoculated with S. sclerotiorum alone. Conclusion: Fusarium oxysporum (S6) may be a good fungal biological control agent for S. sclerotiorum and cyclosporine A is the responsible metabolite involved in its antagonistic activity in vitro. Significance and Impact of the Study: Cyclosporine A has not been previously described as an inhibitor of S. sclerotiorum. Its minimum inhibitory concentration (MIC) of 0·1 μg disc-1 makes it suitable to use as a biofungicide. In vivo experiments showed that F. oxysporum (S6) is a good candidate for the biocontrol of S. sclerotiorum in soybean. © 2006 The Society for Applied Microbiology.

Descripción: Trypanosoma cruzi, etiological agent of Chagas' disease, has a complex life cycle which involves the invasion of mammalian host cells, differentiation and intracellular replication. Here we report the first insights into the biological role of a poly(ADP-ribose) glycohydrolase in a trypanosomatid (TcPARG). In silico analysis of the TcPARG gene pointed out the conservation of key residues involved in the catalytic process and, by Western blot, we demonstrated that it is expressed in a life stage-dependant manner. Indirect immunofluorescence assays and electron microscopy using an anti-TcPARG antibody showed that this enzyme is localized in the nucleus independently of the presence of DNA damage or cell cycle stage. The addition of poly(ADP-ribose) glycohydrolase inhibitors ADP-HPD (adenosine diphosphate (hydroxymethyl) pyrrolidinediol) or DEA (6,9-diamino-2-ethoxyacridine lactate monohydrate) to the culture media, both at a 1 μM concentration, reduced in vitro epimastigote growth by 35% and 37% respectively, when compared to control cultures. We also showed that ADP-HPD 1 μM can lead to an alteration in the progression of the cell cycle in hydroxyurea synchronized cultures of T. cruzi epimastigotes. Outstandingly, here we demonstrate that the lack of poly(ADP-ribose) glycohydrolase activity in Vero and A549 host cells, achieved by chemical inhibition or iRNA, produces the reduction of the percentage of infected cells as well as the number of amastigotes per cell and trypomastigotes released, leading to a nearly complete abrogation of the infection process. We conclude that both, T. cruzi and the host, poly(ADP-ribose) glycohydrolase activities are important players in the life cycle of Trypanosoma cruzi, emerging as a promising therapeutic target for the treatment of Chagas' disease. © 2013 Vilchez Larrea et al.

Descripción: A new biosensor was designed for the assessment of aquatic environment quality. Three microalgae were used as toxicity bioindicators: Chlorella vulgaris, Pseudokirchneriella subcapitata and Chlamydomonas reinhardtii. These microalgae were immobilized in alginate and silica hydrogels in a two step procedure. After studying the growth rate of entrapped cells, chlorophyll fluorescence was measured after exposure to (3-(3,4-dichlorophenyl)-1,1-dimethylurea) (DCMU) and various concentrations of the common herbicide atrazine. Microalgae are very sensitive to herbicides and detection of fluorescence enhancement with very good efficiency was realized. The best detection limit was 0.1 μM, obtained with the strain C. reinhardtii after 40 minutes of exposure. © 2012 by the authors; licensee MDPI, Basel, Switzerland.

Descripción: The effect of chronic exposure to Escherichia coli on morphometrical parameters, different antioxidant defenses, lipid peroxidation and protein oxidation has been studied in digestive gland of the freshwater mussel Diplodon chilensis in the laboratory during 6 weeks. In, a parallel field study, we evaluated morphometrical and oxidative stress parameters in digestive glands of mussels collected from a sewage polluted area (SMA) and from a pristine area (control). Both sites were characterized by analyzing bacteriological and physic-chemical parameters. In the laboratory D. chilensis was able to filter bacteria at high concentrations with a clearing rate of 0.510 ± 0.036. L/h per gram of dry soft tissue mass. The digestive gland mass to shell length ratio (DGM/SL), reduced glutathione (GSH), lipid peroxidation, as concentration of thiobartituric acid reactive substances (TBARS), and glutathione-S-transferase (GST) activity of mussels fed with bacteria were significantly higher than those of control mussels after the fourth week. Fecal bacteria in lake water samples were undetectable in the control, and higher than 24,000. MPN (most probable number)/100. mL in SMA. DGM/SL was higher in SMA. No differences between sites were observed in total lipid and protein content, neither in superoxide dismutase activity. GSH content was higher in SMA, with no difference in the oxidized form. GST activity and MDA were significantly higher in SMA but protein oxidation was not affected. © 2011 Elsevier GmbH.