Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp

The transition from marine/brackish waters to freshwater habitats constitutes a severe osmotic and ionic challenge, and successful invasion has demanded the selection of morphological, physiological, biochemical and behavioral adaptations. We evaluated short-term (1 to 12 h exposure) and long-term (5 d acclimation), anisosmotic extracellular (osmolality, [Na(+), Cl(-)]) and long-term isosmotic intracellular osmoregulatory capability in Palaemon northropi, a neotropical intertidal shrimp. F northropi survives well and osmo- and ionoregulates strongly during short- and long-term exposure to 5-45 parts per thousand salinity, consistent with its rocky tide pool habitat subject to cyclic salinity fluctuations, Muscle total free amino acid (FAA) concentrations decreased by 63% in shrimp acclimated to 5%. salinity, revealing a role in hypoosmotic cell volume regulation; this decrease is mainly a consequence of diminished glycine, arginine and proline. Total FAA contributed 31% to muscle intracellular osmolality at 20 parts per thousand, an isosmotic salinity, and decreased to 13% after acclimation to 5 parts per thousand. Gill and nerve tissue FAA concentrations remained unaltered. These tissue-specific responses reflect efficient anisosmotic and anisoionic extracellular regulatory mechanisms, and reveal the dependence of muscle tissue on intracellular osmotic effectors. FAA concentration is higher in P. northropi than in diadromous and hololimnetic palaemonids, confirming muscle FAA concentration as a good parameter to evaluate the degree of adaptation to dilute media. The osmoregulatory capability of P. northropi may reflect the potential physiological capacity of ancestral marine palaemonids to penetrate into dilute media, and reveals the importance of evaluating osmoregulatory processes in endeavors to comprehend the invasion of dilute media by ancestral marine crustaceans.

Metabolic labeling and protein linearization technology allow the study of proteins secreted by cultured cells in serum-containing media.

Supernatants from cell cultures (also called conditioned media, CMs) are commonly analyzed to study the pool of secreted proteins (secretome). To reduce the exogenous protein background, serum-free media are often used to obtain CMs. Serum deprivation, however, can severely affect cell viability and phenotype, including protein secretion. We present a strategy to analyze the proteins secreted by cells in fetal bovine serum-containing CMs, which combines the advantage of metabolic labeling and protein concentration linearization techniques. Incubation of CMs with a hexapeptide ligand library was used to reduce the dynamic range of the samples and led to the identification of 3 times more proteins than in untreated CM samples. Labeling with a deuterated amino acid was used to distinguish between cellular proteins and homologous bovine proteins contained in the medium. Application of the strategy to two breast cancer cell lines led to the identification of proteins secreted in different amounts and which could correlate with their varying degree of aggressiveness. Selected reaction monitoring (SRM)-based quantitation of three proteins of interest in the crude samples yielded data in good agreement with the results from concentration-equalized samples.

Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Gene Content Analysis of Sugarcane Public ESTs Reveals Thousands of Missing Coding-Genes and an Unexpected Pool of Grasses Conserved ncRNAs

Sugarcane is the most important crop for sugar industry and raw material for bioethanol. Here we present a quantitative analysis of the gene content from publicly available sugarcane ESTs. The current sugarcane EST collection sampled orthologs for ~58 % of the closely-related sorghum proteome, suggesting that more than 10,000 sugarcane coding-genes remain undiscovered. Moreover the existence of more than 2,000 ncRNAs conserved between sugarcane and sorghum was revealed, among which over 500 are also detected in rice, supporting the existence of hundreds of conserved ncRNAs in grasses. New efforts towards sugarcane transcriptome sequencing were needed to sample the missing coding-genes as well as to expand the catalog of ncRNAs. © 2012 Springer Science+Business Media, LLC.

Production and action of an Aspergillus phoenicis enzymatic pool using different carbon sources

Aspergillus phoenicis is an interesting heat tolerant fungus that can synthesize enzymes with several applications in the food industry due to its great hydrolytic potential. In this work, the fungus produced high enzymatic levels when cultivated on inexpensive culture media consisting of flakes from different origins such as cassava flour, wheat fibre, crushed soybean, agro-industrial wastes, starch, glucose or maltose. Several enzymatic systems were produced from these carbon sources, but amylase was the most evident, followed by pectinase and xylanase. Traces of CMCases, avicelase, lipase, β-xylosidase, β-glucosidase and α-glucosidase activities were also detected. Amylases were produced on rye flakes, starch, oat flakes, corn flakes, cassava flour and wheat fibre. Significant amylolytic levels were produced in the culture medium with glucose or when this sugar was exhausted, suggesting an enzyme in the constitutive form. Cassava flour, rye, oats, barley and corn flakes were also used as substrates in the hydrolytic reactions, aiming to verify the liberation potential of reducing sugars. Corn flakes induced greater liberation of reducing sugars as compared to the others. Thin layer chromatography of the reaction end products showed that the hydrolysis of cassava flour liberated maltooligosaccharides, but cassava flour and corn, rye, oats and barley flakes were hydrolyzed to glucose. These results suggested the presence of glucoamylase and α-amylase as part of the enzymatic pool of A. phoencis.

Imaging of pulmonary embolism and t-PA therapy effects using MDCT and liposomal iohexol blood pool agent – preliminary results in a rabbit model

Hypothesis and Objectives PEGylated liposomal blood pool contrast agents maintain contrast enhancement over several hours. This study aimed to evaluate (long-term) imaging of pulmonary arteries, comparing conventional iodinated contrast with a liposomal blood pool contrast agent. Secondly, visualization of the (real-time) therapeutic effects of tissue-Plasminogen Activator (t-PA) on pulmonary embolism (PE) was attempted. Materials and Methods Six rabbits (approximate 4 kg weight) had autologous blood clots injected through the superior vena cava. Imaging was performed using conventional contrast (iohexol, 350 mg I/ml, GE HealthCare, Princeton, NJ) at a dose of 1400 mgI per animal and after wash-out, animals were imaged using an iodinated liposomal blood pool agent (88 mg I/mL, dose 900 mgI/animal). Subsequently, five animals were injected with 2mg t-PA and imaging continued for up to 4 ½ hours. Results Both contrast agents identified PE in the pulmonary trunk and main pulmonary arteries in all rabbits. Liposomal blood pool agent yielded uniform enhancement, which remained relatively constant throughout the experiments. Conventional agents exhibited non uniform opacification and rapid clearance post injection. Three out of six rabbits had mistimed bolus injections, requiring repeat injections. Following t-PA, Pulmonary embolus volume (central to segmental) decreased in four of five treated rabbits (range 10–57%, mean 42%). One animal showed no response to t-PA. Conclusions Liposomal blood pool agents effectively identified acute PE without need for re-injection. PE resolution following t-PA was quantifiable over several hours. Blood pool agents offer the potential for repeated imaging procedures without need for repeated (nephrotoxic) contrast injections

High Resolution Vascular Imaging of the Rat Spine using Liposomal Blood Pool MR Agent

High resolution, vascular magnetic resonance imaging of the spine region in small animals poses several challenges. The small anatomical features, extravascular diffusion, and the low signal-to-noise ratio limit the use of conventional contrast agents. We hypothesize that a long circulating, intravascular liposomal-encapsulated MR contrast agent (liposomal-Gd) would facilitate visualization of small anatomical features of the perispinal vasculature not visible with conventional contrast agent (Gd-DTPA).

High-resolution vascular imaging of the rat spine using liposomal blood pool MR agent.

BACKGROUND AND PURPOSE: High-resolution, vascular MR imaging of the spine region in small animals poses several challenges. The small anatomic features, extravascular diffusion, and low signal-to-noise ratio limit the use of conventional contrast agents. We hypothesize that a long-circulating, intravascular liposomal-encapsulated MR contrast agent (liposomal-Gd) would facilitate visualization of small anatomic features of the perispinal vasculature not visible with conventional contrast agent (gadolinium-diethylene-triaminepentaacetic acid [Gd-DTPA]). METHODS: In this study, high-resolution MR angiography of the spine region was performed in a rat model using a liposomal-Gd, which is known to remain within the blood pool for an extended period. The imaging characteristics of this agent were compared with those of a conventional contrast agent, Gd-DTPA. RESULTS: The liposomal-Gd enabled acquisition of high quality angiograms with high signal-to-noise ratio. Several important vascular features, such as radicular arteries, posterior spinal vein, and epidural venous plexus were visualized in the angiograms obtained with the liposomal agent. The MR angiograms obtained with conventional Gd-DTPA did not demonstrate these vessels clearly because of marked extravascular soft-tissue enhancement that obscured the vasculature. CONCLUSIONS: This study demonstrates the potential benefit of long-circulating liposomal-Gd as a MR contrast agent for high-resolution vascular imaging applications.

Imaging of pulmonary embolism and t-PA therapy effects using MDCT and liposomal iohexol blood pool agent: preliminary results in a rabbit model.

RATIONALE AND OBJECTIVES: Polyethylene glycol-coated liposomal blood pool contrast agents maintain contrast enhancement over several hours. This study aimed to evaluate (long-term) imaging of pulmonary arteries, comparing conventional iodinated contrast with a liposomal blood pool contrast agent. Also, visualization of the (real-time) therapeutic effects of tissue plasminogen activator (t-PA) on pulmonary embolism (PE) was attempted. MATERIALS AND METHODS: Six rabbits (weight approximately 4 kg) had autologous blood clots injected through the superior vena cava. Imaging was performed using conventional contrast (iohexol, 350 mg I/ml; GE HealthCare, Princeton, NJ) at a dose of 1400 mg I per animal, and after wash-out, animals were imaged using an iodinated liposomal blood pool agent (88 mg I/mL, dose 900 mg I/animal). Subsequently, five animals were injected with 2 mg of t-PA and imaging continued for up to 4(1/2) hours. RESULTS: Both contrast agents identified PE in the pulmonary trunk and main pulmonary arteries in all rabbits. Liposomal blood pool agent yielded uniform enhancement, which remained relatively constant throughout the experiments. Conventional agents exhibited nonuniform opacification and rapid clearance postinjection. Three of six rabbits had mistimed bolus injections, requiring repeat injections. Following t-PA, pulmonary embolus volume (central to segmental) decreased in four of five treated rabbits (range 10-57%, mean 42%). One animal showed no response to t-PA. CONCLUSIONS: Liposomal blood pool agents effectively identified acute PE without need for reinjection. PE resolution following t-PA was quantifiable over several hours. Blood pool agents offer the potential for repeated imaging procedures without need for repeated (nephrotoxic) contrast injections.

Detection of a variable intracellular acid-labile carbon pool in Thalassiosira weissflogii (Heterokontophyta) and Emiliania huxleyi (Haptophyta) in response to changes in the seawater carbon system

Accumulation of an intracellular pool of carbon (C(i) pool) is one strategy by which marine algae overcome the low abundance of dissolved CO2 (CO2 (aq) ) in modern seawater. To identify the environmental conditions under which algae accumulate an acid-labile C(i) pool, we applied a (14) C pulse-chase method, used originally in dinoflagellates, to two new classes of algae, coccolithophorids and diatoms. This method measures the carbon accumulation inside the cells without altering the medium carbon chemistry or culture cell density. We found that the diatom Thalassiosira weissflogii [(Grunow) G. Fryxell & Hasle] and a calcifying strain of the coccolithophorid Emiliania huxleyi [(Lohmann) W. W. Hay & H. P. Mohler] develop significant acid-labile C(i) pools. C(i) pools are measureable in cells cultured in media with 2-30 µmol/l CO2 (aq), corresponding to a medium pH of 8.6-7.9. The absolute C(i) pool was greater for the larger celled diatoms. For both algal classes, the C(i) pool became a negligible contributor to photosynthesis once CO2 (aq) exceeded 30 µmol/l. Combining the (14) C pulse-chase method and (14) C disequilibrium method enabled us to assess whether E. huxleyi and T. weissflogii exhibited thresholds for foregoing accumulation of DIC or reduced the reliance on bicarbonate uptake with increasing CO2 (aq) . We showed that the C(i) pool decreases with higher CO2 :HCO3 (-) uptake rates.

Fractal Behaviour and Irregularity of Wetting Fronts in Heterogeneous Porous Media

The wetting front is the zone where water invades and advances into an initially dry porous material and it plays a crucial role in solute transport through the unsaturated zone. Water is an essential part of the physiological process of all plants. Through water, necessary minerals are moved from the roots to the parts of the plants that require them. Water moves chemicals from one part of the plant to another. It is also required for photosynthesis, for metabolism and for transpiration. The leaching of chemicals by wetting fronts is influenced by two major factors, namely: the irregularity of the fronts and heterogeneity in the distribution of chemicals, both of which have been described by using fractal techniques. Soil structure can significantly modify infiltration rates and flow pathways in soils. Relations between features of soil structure and features of infiltration could be elucidated from the velocities and the structure of wetting fronts. When rainwater falls onto soil, it doesn?t just pool on surfaces. Water ?or another fluid- acts differently on porous surfaces. If the surface is permeable (porous) it seeps down through layers of soil, filling that layer to capacity. Once that layer is filled, it moves down into the next layer. In sandy soil, water moves quickly, while it moves much slower through clay soil. The movement of water through soil layers is called the the wetting front. Our research concerns the motion of a liquid into an initially dry porous medium. Our work presents a theoretical framework for studying the physical interplay between a stationary wetting front of fractal dimension D with different porous materials. The aim was to model the mass geometry interplay by using the fractal dimension D of a stationary wetting front. The plane corresponding to the image is divided in several squares (the minimum correspond to the pixel size) of size length ". We acknowledge the help of Prof. M. García Velarde and the facilities offered by the Pluri-Disciplinary Institute of the Complutense University of Madrid. We also acknowledge the help of European Community under project Multi-scale complex fluid flows and interfacial phenomena (PITN-GA-2008-214919). Thanks are also due to ERCOFTAC (PELNoT, SIG 14)

Portrayal of depression and other mental illnesses in Australian nonfiction media

This study describes Australian media portrayal of mental illnesses, focusing on depression. A random sample of 1,123 items was selected for analysis from a pool of 13,389 nonfictional media items about mental illness collected between March 2000 and February 2001. Depression was portrayed more frequently than other mental illnesses. Items about depression, eating disorders, and substance use disorders most commonly described policies or programs, whereas items about schizophrenia most frequently portrayed individuals or symptoms and treatment. A minority of items about depression presented information about symptoms, causes, treatment, or prognosis. Although such information was generally accurate, a proportion of items conveyed misleading messages. There is therefore scope for increasing the level of accurate information provided about depression in the Australian media. (c) 2005 Wiley Periodicals, Inc.

From icon to naturalised icon:a linguistic analysis of media representations of the BP Deepwater Horizon crisis

This research explores how news media reports construct representations of a business crisis through language. In an innovative approach to dealing with the vast pool of potentially relevant texts, media texts concerning the BP Deepwater Horizon oil spill are gathered from three different time points: immediately after the explosion in 2010, one year later in 2011 and again in 2012. The three sets of 'BP texts' are investigated using discourse analysis and semi-quantitative methods within a semiotic framework that gives an account of language at the semiotic levels of sign, code, mythical meaning and ideology. The research finds in the texts three discourses of representation concerning the crisis that show a movement from the ostensibly representational to the symbolic and conventional: a discourse of 'objective factuality', a discourse of 'positioning' and a discourse of 'redeployment'. This progression can be shown to have useful parallels with Peirce's sign classes of Icon, Index and Symbol, with their implied movement from a clear motivation by the Object (in this case the disaster events), to an arbitrary, socially-agreed connection. However, the naturalisation of signs, whereby ideologies are encoded in ways of speaking and writing that present them as 'taken for granted' is at its most complete when it is least discernible. The findings suggest that media coverage is likely to move on from symbolic representation to a new kind of iconicity, through a fourth discourse of 'naturalisation'. Here the representation turns back towards ostensible factuality or iconicity, to become the 'naturalised icon'. This work adds to the study of media representation a heuristic for understanding how the meaning-making of a news story progresses. It offers a detailed account of what the stages of this progression 'look like' linguistically, and suggests scope for future research into both language characteristics of phases and different news-reported phenomena.

Evolutionary transition to freshwater by ancestral marine palaemonids: evidence from osmoregulation in a tide pool shrimp

The transition from marine/brackish waters to freshwater habitats constitutes a severe osmotic and ionic challenge, and successful invasion has demanded the selection of morphological, physiological, biochemical and behavioral adaptations. We evaluated short-term (1 to 12 h exposure) and long-term (5 d acclimation), anisosmotic extracellular (osmolality, [Na(+), Cl(-)]) and long-term isosmotic intracellular osmoregulatory capability in Palaemon northropi, a neotropical intertidal shrimp. F northropi survives well and osmo- and ionoregulates strongly during short- and long-term exposure to 5-45 parts per thousand salinity, consistent with its rocky tide pool habitat subject to cyclic salinity fluctuations, Muscle total free amino acid (FAA) concentrations decreased by 63% in shrimp acclimated to 5%. salinity, revealing a role in hypoosmotic cell volume regulation; this decrease is mainly a consequence of diminished glycine, arginine and proline. Total FAA contributed 31% to muscle intracellular osmolality at 20 parts per thousand, an isosmotic salinity, and decreased to 13% after acclimation to 5 parts per thousand. Gill and nerve tissue FAA concentrations remained unaltered. These tissue-specific responses reflect efficient anisosmotic and anisoionic extracellular regulatory mechanisms, and reveal the dependence of muscle tissue on intracellular osmotic effectors. FAA concentration is higher in P. northropi than in diadromous and hololimnetic palaemonids, confirming muscle FAA concentration as a good parameter to evaluate the degree of adaptation to dilute media. The osmoregulatory capability of P. northropi may reflect the potential physiological capacity of ancestral marine palaemonids to penetrate into dilute media, and reveals the importance of evaluating osmoregulatory processes in endeavors to comprehend the invasion of dilute media by ancestral marine crustaceans.