Liver proteome alterations in psychologically distressed rats and a nootropic drug.

BACKGROUND: Chronic psychological distress is considered today a pandemic due to the modern lifestyle and has been associated with various neurodegenerative, autoimmune, or systemic inflammation-related diseases. Stress is closely related to liver disease exacerbation through the high activity of the endocrine and autonomic nervous systems, and the connection between the development of these pathologies and the physiological effects induced by oxidative stress is not yet completely understood. The use of nootropics, as the cognitive enhancer and antioxidant piracetam, is attractive to repair the oxidative damage. A proteomic approach provides the possibility to obtain an in-depth comprehension of the affected cellular processes and the possible consequences for the body. Therefore, we considered to describe the effect of distress and piracetam on the liver proteome. METHODS: We used a murine model of psychological stress by predatory odor as a distress paradigm. Female Sprague-Dawley rats were distributed into four experimental groups (n = 6 - 7/group) and were exposed or not to the stressor for five days and treated or not with piracetam (600 mg/kg) for six days. We evaluated the liver proteome by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1D-SDS-PAGE) followed by liquid chromatography-tandem mass spectrometry (GeLC-MS/MS). Besides, we analyzed the activity of liver antioxidant enzymes, the biochemical parameters in plasma and rat behavior. RESULTS: Our results showed that distress altered a wide range of proteins involved in amino acids metabolism, glucose, and fatty acid mobilization and degradation on the way to produce energy, protein folding, trafficking and degradation, redox metabolism, and its implications in the development of the non-alcoholic fatty liver disease (NAFLD). Piracetam reverted the changes in metabolism caused by distress exposure, and, under physiological conditions, it increased catabolism rate directed towards energy production. These results confirm the possible relationship between chronic psychological stress and the progression of NAFLD, as well as we newly evidenced the controversial beneficial effects of piracetam. Finally, we propose new distress biomarkers in the liver as the protein DJ-1 (PARK7), glutathione peroxidase 1 (GPX), peroxiredoxin-5 (PRDX5), glutaredoxin 5 (GLRX5), and thioredoxin reductase 1 (TXNDR1), and in plasma as biochemical parameters related to kidney function such as urea and blood urea nitrogen (BUN) levels.

Interspecific Variation between the American Quercus virginiana and Mediterranean Quercus Species in Terms of Seed Nutritional Composition, Phytochemical Content, and Antioxidant Activity.

This study aimed to evaluate a complete nutritional composition in the seeds Quercus virginiana to compare this nutritional composition with three Mediterranean Quercus species. We analyzed the seed morphometry, proximate composition, phytochemicals, and antioxidant capacity. The seed of Q. virginiana presented the smaller seed size and weight, while Q. suber presented the highest values. Moreover, Q. virginiana seeds showed the highest amounts of sugar and total lipids, digestibility, energy, palmitic acid, and stearic acid. On the other hand, Q. virginiana seeds showed the lowest values of linoleic acid. Moreover, Q. coccifera seeds presented the highest total phenolics and flavonoids contents and antioxidant activity. The clustering analysis revealed a significant similarity in seed morphometry and nutritional composition between the Mediterranean Q. ilex and Q. suber, grouping with the American Q. virginiana, but to a considerable distance; by contrast, the Mediterranean Q. coccifera was the most distant in the clustering analysis. The content of phenolics and flavonoids and digestibility value were the variables that contributed to the separation to a greater extent in the clustering of the four species. The nutritional and biological activity assessment of plant seed may be considered as an essential mission to find new sustainable sources and novel chemical agents. In this sense, Quercus seeds may be an alternative and a competitive food source for the agri-food industry.

Sensory attributes, physicochemical and antioxidant characteristics, and protein profile of wild prickly pear fruits (O. macrocentra Engelm., O. phaeacantha Engelm., and O. engelmannii Salm-Dyck ex Engelmann.) and commercial prickly pear fruits (O. ficus-indica (L.) Mill.).

Mexico presents the highest richness of Opuntia Mill. species. These species are an important economic factor for the country, and source of nutrients, bioactive compounds, pigments, and nutraceuticals which can be of interest for the food and pharmaceutical industry. However, there are some wild Opuntia species in the Chihuahua desert, that have not been analyzed to establish their properties and potential use. The aim of study was to evaluate the sensory, physicochemical and protein profile in wild prickly pear fruits (O. macrocentra Engelm. (OM), O. phaeacantha Engelm. (OP), and O. engelmannii Salm-Dyck ex Engelmann. (OE)) from Samalayuca, Chihuahua and compare them with two commercial prickly pear fruits (O. ficus-indica (L.) Mill. (green-OFG, red-OFR). The sensory profile of wild species was characterized by highest color, odor, and sour taste compared to the commercial fruits. Pulp, peel, and seeds from wild prickly pear fruits showed lower pH, and higher titratable total acidity, total phenolic compounds, total flavonoids, antioxidant capacity, protein, lipids, ash, carbohydrates (only peel), and crude fiber content than commercial Opuntia species. Furthermore, O. engelmannii showed a tendency to present the highest betacyanins, betaxanthins, and betalains contents. A total of 181, 122, 113, 183 and 140 different proteins were identified in OM, OP, OE, OFG, OFR species, respectively. All species showed the highest enrichment in three main pathways such as amino acids biosynthesis, glycolysis (dark)/gluconeogenesis (light), and the citric acid cycle. The wild prickly pear fruits of this study showed important nutritional, protein, and antioxidant properties with biological interest, and can be a potential source of functional ingredients and nutraceuticals.

Effect of Elicitation on Polyphenol and Carotenoid Metabolism in Butterhead Lettuce (Lactuca sativa var. capitata).

The effect of elicitation in butterhead lettuce on carotenoid and polyphenol metabolism was evaluated. Different concentrations of arachidonic acid (AA), salicylic acid (SA), methyl jasmonate (MJ) (15, 45, and 90 µM) and Harpin protein (HP) (30, 60, and 120 mg/L) were applied on red and green butterhead lettuces. Total phenolic and flavonoid content were incremented by MJ (90 µM) in green and red lettuce. Carotenoids were increased in red lettuce (AA; 45 µM). Green lettuce modifies their phenolic acid profile after elicitation with AA and MJ; meanwhile, red lettuce incremented mainly in hydroxycinnamic acids and flavonols, MJ being the elicitor with the highest effect. There was an impact on secondary metabolite enzyme gene transcript concentration. Phenylalanine ammonia-lyase (PAL) and lycopene beta cyclase (LBC) increased in both varieties after elicitation. A relationship between phytochemical increase and the activation of the metabolic pathways after elicitation in butterhead lettuce was observed.

Seed Characteristics and Nutritional Composition of Pine Nut from Five Populations of P. cembroides from the States of Hidalgo and Chihuahua, Mexico.

The aim of this study was to analyze the seed characteristics and nutritional composition of five pine nut P. cembroides samples from two Mexican states. Morphometry, proximal composition, phenolic compounds, and antioxidant capacity were determined. Samples differed in several morphometric trails, but important differences were documented between SMCH and JCZH samples from Hidalgo State. JCZH and FMH had the highest contents of water, lipids, protein, flavonoids, and antioxidant activity, while CMCC population from Chihuahua State had presented the highest content of ash and carbohydrates. Morphometry and chemical composition data were subjected to clustering analysis. This analysis showed that SMCH and LFCH from Hidalgo State were well separated from the JCZH and FMH populations from Hidalgo State, which showed a strong similarity between them, while the CMCC from Chihuahua State was the most distant population. Principal components analysis showed that the variables that strongly contributed to PC1 were the antioxidant activity determined by FRAP assay, flavonoids, and water content. These data have provided biochemical markers that could help to establish phylogenetic associations between populations, and also to reveal potentially account as an alternative source for dietary nutrition.

Unraveling the in vitro secretome of the phytopathogen Botrytis cinerea to understand the interaction with its hosts.

Botrytis cinerea is a necrotrophic fungus with high adaptability to different environments and hosts. It secretes a large number of extracellular proteins, which favor plant tissue penetration and colonization, thus contributing to virulence. Secretomics is a proteomics sub-discipline which study the secreted proteins and their secretion mechanisms, so-called secretome. By using proteomics as experimental approach, many secreted proteins by B. cinerea have been identified from in vitro experiments, and belonging to different functional categories: (i) cell wall-degrading enzymes such as pectinesterases and endo-polygalacturonases; (ii) proteases involved in host protein degradation such as an aspartic protease; (iii) proteins related to the oxidative burst such as glyoxal oxidase; (iv) proteins which may induce the plant hypersensitive response such as a cerato-platanin domain-containing protein; and (v) proteins related to production and secretion of toxins such as malate dehydrogenase. In this mini-review, we made an overview of the proteomics contribution to the study and knowledge of the B. cinerea extracellular secreted proteins based on our current work carried out from in vitro experiments, and recent published papers both in vitro and in planta studies on this fungi. We hypothesize on the putative functions of these secreted proteins, and their connection to the biology of the B. cinerea interaction with its hosts.

Evolución clínica de los enfermos renales crónicos en tratamiento sustitutivo con infección por VIH / Clinical evolution of chronic renal patients with HIV infection in replacement therapy

Los pacientes con infección por el virus de la inmunodeficiencia humana (VIH) y enfermedad renal que terminan en tratamiento sustitutivo renal constituyen un grupo especial con interés creciente para la nefrología. Con el objetivo de conocer datos epidemiológicos de los pacientes VHI+ en España, recogimos información individualizada durante los años 2004 a 2011 (periodo de uso de tratamiento antiviral de alta eficacia) en las comunidades autónomas (CCAA) de Andalucía, Aragón, Asturias, Cataluña, Comunidad Valenciana, Castilla-La Mancha, Castilla y León, Galicia, Madrid, La Rioja y País Vasco, que comprendían un 85% de la población española. Se analizó a un total de 271 pacientes incidentes y 209 prevalentes. Se compararon con el resto de pacientes en tratamiento sustitutivo durante el mismo periodo de tiempo. La incidencia anual fue de 0,8 pacientes por millón de habitantes, con un aumento significativo a lo largo del periodo de seguimiento. La proporción de pacientes prevalentes VIH+ fue de 5,1/1.000 pacientes en tratamiento sustitutivo, intervalo de confianza (IC) del 95%: 4,4-5,8. Las causas glomerulares constituyeron la mayoría (42%), aunque hubo un 14% de nefropatía diabética. En el total de España, esos porcentajes son 13 y 25%, respectivamente. Comparando frente al total de pacientes en tratamiento, el riesgo de muerte fue significativamente mayor en el grupo VIH+: hazard ratio (HR) ajustado por edad, sexo y presencia de diabetes: 2,26 (IC 95%: 1,74-2,91). La coinfección por hepatitis C aumentó el riesgo de muerte dentro del grupo VIH+: HR 1,77 (IC 95%: 1,10-2,85). La probabilidad de recibir trasplante renal en los VIH+ solo alcanzó el 17% a los 7 años, comparando con el total de pacientes en diálisis HR: 0,15 (IC 95%: 0,10-0,24). A pesar del uso de las nuevas combinaciones de antivirales, la incidencia de pacientes VIH+ en diálisis se ha incrementado, su mortalidad supera todavía al resto de pacientes, y tienen una tasa de trasplante muy baja. Se hace necesario profundizar en el conocimiento de esta enfermedad para mejorar los resultados (AU)

Patients on renal replacement therapy (RRT) infected with the human immunodeficiency virus (HIV) are a special group with growing interest. In order to study the epidemiological data of HIV+ patients on RRT in Spain, we collected individual information from 2004-2011 (period of use of highly active antiretroviral therapy [HAART] in the Autonomous Communities of Andalusia, Aragon, Asturias, Catalonia, Valencia, Castilla la Mancha, Castilla León, Galicia, Madrid, La Rioja and the Basque Country, comprising 85% of the Spanish population. A total of 271 incident and 209 prevalent patients were analysed. They were compared with the remaining patients on RRT during the same period. The annual incidence was 0.8 patients per one million inhabitants, with a significant increase during the follow-up period. The proportion of prevalent HIV+ patientswas 5.1 per 1,000 patients on RRT (95% confidence interval [CI] 4.4-5.8. Although glomerular diseases constituted the majority of cases (42%), diabetic nephropathy was the cause in 14% of patients. The nation-wide totals for these percentages were 13 and 25%, respectively. Compared to the total of patients in treatment, the risk of death was significantly higher in the HIV+ group: hazard ratio (HR) adjusted for age, sex and diabetes was 2.26 (95% CI 1.74 - 2.91). Hepatitis C coinfection increased the risk of death in the HIV+ group (HR 1.77; 95% CI 1.10 - 2.85). The probability of kidney transplantation in HIV+ was only 17% after 7 years, comparing with total RTT patients (HR 0.15; 95% CI: 0.10-0.24). Despite the use of HAART, the incidence of HIV+ patients on dialysis has increased; their mortality still exceeds non-HIV patients, and they have a very low rate of transplantation. It is necessary to further our knowledge of this disease in order to improve results (AU)

Clinical evolution of chronic renal patients with HIV infection in replacement therapy.

Patients on renal replacement therapy (RRT) infected with the human immunodeficiency virus (HIV) are a special group with growing interest. In order to study the epidemiological data of HIV+ patients on RRT in Spain, we collected individual information from 2004-2011 (period of use of highly active antiretroviral therapy [HAART] in the Autonomous Communities of Andalusia, Aragon, Asturias, Catalonia, Valencia, Castilla la Mancha, Castilla León, Galicia, Madrid, La Rioja and the Basque Country, comprising 85% of the Spanish population. A total of 271 incident and 209 prevalent patients were analysed. They were compared with the remaining patients on RRT during the same period. The annual incidence was 0.8 patients per one million inhabitants, with a significant increase during the follow-up period. The proportion of prevalent HIV+ patients was 5.1 per 1,000 patients on RRT (95% confidence interval [CI] 4.4-5.8. Although glomerular diseases constituted the majority of cases (42%), diabetic nephropathy was the cause in 14% of patients. The nation-wide totals for these percentages were 13 and 25%, respectively. Compared to the total of patients in treatment, the risk of death was significantly higher in the HIV+ group: hazard ratio (HR) adjusted for age, sex and diabetes was 2.26 (95% CI 1.74 - 2.91). Hepatitis C coinfection increased the risk of death in the HIV+ group (HR 1.77; 95% CI 1.10 - 2.85). The probability of kidney transplantation in HIV+ was only 17% after 7 years, comparing with total RTT patients (HR 0.15; 95% CI: 0.10-0.24). Despite the use of HAART, the incidence of HIV+ patients on dialysis has increased; their mortality still exceeds non-HIV patients, and they have a very low rate of transplantation. It is necessary to further our knowledge of this disease in order to improve results.

Renal replacement therapy in Europe: a summary of the 2012 ERA-EDTA Registry Annual Report.

BACKGROUND: This article summarizes the 2012 European Renal Association-European Dialysis and Transplant Association Registry Annual Report (available at www.era-edta-reg.org) with a specific focus on older patients (defined as ≥65 years). METHODS: Data provided by 45 national or regional renal registries in 30 countries in Europe and bordering the Mediterranean Sea were used. Individual patient level data were received from 31 renal registries, whereas 14 renal registries contributed data in an aggregated form. The incidence, prevalence and survival probabilities of patients with end-stage renal disease (ESRD) receiving renal replacement therapy (RRT) and renal transplantation rates for 2012 are presented. RESULTS: In 2012, the overall unadjusted incidence rate of patients with ESRD receiving RRT was 109.6 per million population (pmp) (n = 69 035), ranging from 219.9 pmp in Portugal to 24.2 pmp in Montenegro. The proportion of incident patients ≥75 years varied from 15 to 44% between countries. The overall unadjusted prevalence on 31 December 2012 was 716.7 pmp (n = 451 270), ranging from 1670.2 pmp in Portugal to 146.7 pmp in the Ukraine. The proportion of prevalent patients ≥75 years varied from 11 to 32% between countries. The overall renal transplantation rate in 2012 was 28.3 pmp (n = 15 673), with the highest rate seen in the Spanish region of Catalonia. The proportion of patients ≥65 years receiving a transplant ranged from 0 to 35%. Five-year adjusted survival for all RRT patients was 59.7% (95% confidence interval, CI: 59.3-60.0) which fell to 39.3% (95% CI: 38.7-39.9) in patients 65-74 years and 21.3% (95% CI: 20.8-21.9) in patients ≥75 years.

A physiological, biochemical and proteomic characterization of Saccharomyces cerevisiae trk1,2 transport mutants grown under limiting potassium conditions.

Saccharomyces cerevisiae mutants lacking both isoforms of the main plasma membrane potassium transporter display impaired potassium transport and defective growth at limiting concentrations of the cation. Moreover, they are hyperpolarized and have a lower intracellular pH than wild-type. In order to unravel global physiological processes altered in trk1,2 mutants, we have established conditions at which both wild-type and mutants can grow at different rates. Using a combination of physiological, biochemical and proteomic approaches, we show that during growth at suboptimal potassium concentrations, double trk1,2 mutants accumulate less potassium and reach lower yields. In contrast, the mutants maintain increased viability in the stationary phase and retain more potassium. Moreover, the mutants show increased expression of stress-related proteins such as catalase T, thioredoxin peroxidase or hexokinase 2, suggesting that they are better adapted to the additional stress factors associated with entry into stationary growth phase.

Metabolite and proteome changes during the ripening of Syrah and Cabernet Sauvignon grape varieties cultured in a nontraditional wine region in Brazil.

Grapevines are an important fruit crop from economic and cultural point of views in many countries, including Brazil, where the practice of vitiviniculture is being developed in different regions. We compared the anthocyanin concentration, the main organic acids and sugars, and the proteome profiles during berry ripening of Syrah and Cabernet Sauvignon grapes from two distinct geographical sources in São Paulo State. The proteome was mapped by two-dimensional gel electrophoresis and differentially abundant proteins during the ripening process were subjected to MALDI-TOF/TOF-MS analysis. An increase in sugar concentration and in anthocyanin content was observed, as well a decrease in the tartaric and malic acid concentration. A total of 128 spots varied with geographical origin, grape variety, and ripening stage, with 108 being identified. The identified proteins resulted in 80 gene products. A multivariate analysis of protein abundance clustered the samples according to grape variety, geographical origin, and stage of ripening, and showed the possibility of using proteomics to characterize three variables: variety, area where grown, and the ripening process. The changes observed during the ripening process corresponded to enzymes involved in sugar and organic acid metabolism. These results are in accordance with the metabolic profile reported for the process. BIOLOGICAL SIGNIFICANCE: Given the importance of discriminating grapes, thus making the adulteration of wines more difficult, in this paper we showed the possibility of differentiating varieties of grapes, geographical area of cultivation and stage of ripening by combining the results of differentially abundant protein determinations and multivariate analysis.

Proteomic analysis of mycelium and secretome of different Botrytis cinerea wild-type strains.

The necrotrophic fungus Botrytis cinerea is a very damaging phytopathogen of wide host range and environmental persistence. It is difficult to control because of its genetic versatility, expressed in the many phenotypical differences among isolates. The genomes of the B. cinerea B05.10 and T4 strains have been recently sequenced, becoming a model system for necrotrophic pathogens, and thus opening new alternatives for functional genomics analysis. In this work, the mycelium and secreted proteome of six wild-type strains with different host range, and grown in liquid minimal medium, have been analyzed by using complementary gel-based (1-DE and 2-DE) and gel-free/label-free (nUPLC-MS(E)) approaches. We found differences in the protein profiles among strains belonging to both the mycelium and the secretome. A total of 47 and 51 variable proteins were identified in the mycelium and the secretome, respectively. Some of them, such as malate dehydrogenase or peptidyl-prolyl cis-trans isomerase from the mycelium, and endopolygalacturonase, aspartic protease or cerato-platanin protein from the secretome have been reported as virulence factors, which are involved in host-tissue invasion, pathogenicity or fungal development. BIOLOGICAL SIGNIFICANCE: The necrotrophic fungus Botrytis cinerea is an important phytopathogen of wide host range and environmental persistence, causing substantial economic losses worldwide. In this work, the mycelium and secreted proteome of six B. cinerea wild-type strains with different host range have been analyzed by using complementary gel-based and gel-free/label-free approaches. Fungal genetic versatility was confirmed at the proteome level for both mycelium proteome and secreted proteins. A high number of hypothetical proteins with conserved domains related to toxin compounds or to unknown functions were identified, having qualitative differences among strains. The identification of hypothetical proteins suggests that the B. cinerea strains differ mostly in processes involved in adaptation to a particular environment or a growth condition, rather than in essential metabolic reactions. Proteomics can help in the identification of variable proteins related to the infection and colonization of host plant tissues, as well as of virulence and aggressiveness factors among different B. cinerea wild-type strains. This article is part of a Special Issue entitled: Trends in Microbial Proteomics.

Renal replacement therapy in Europe: a summary of the 2011 ERA-EDTA Registry Annual Report.

BACKGROUND: This article provides a summary of the 2011 ERA-EDTA Registry Annual Report (available at www.era-edta-reg.org). METHODS: Data on renal replacement therapy (RRT) for end-stage renal disease (ESRD) from national and regional renal registries in 30 countries in Europe and bordering the Mediterranean Sea were used. From 27 registries, individual patient data were received, whereas 17 registries contributed data in aggregated form. We present the incidence and prevalence of RRT, and renal transplant rates in 2011. In addition, survival probabilities and expected remaining lifetimes were calculated for those registries providing individual patient data. RESULTS: The overall unadjusted incidence rate of RRT in 2011 among all registries reporting to the ERA-EDTA Registry was 117 per million population (pmp) (n = 71.631). Incidence rates varied from 24 pmp in Ukraine to 238 pmp in Turkey. The overall unadjusted prevalence of RRT for ESRD on 31 December 2011 was 692 pmp (n = 425 824). The highest prevalence was reported by Portugal (1662 pmp) and the lowest by Ukraine (131 pmp). Among all registries, a total of 22 814 renal transplantations were performed (37 pmp). The highest overall transplant rate was reported from Spain, Cantabria (81 pmp), whereas the highest rate of living donor transplants was reported from Turkey (39 pmp). For patients who started RRT between 2002 and 2006, the unadjusted 5-year patient survival on RRT was 46.8% [95% confidence interval (CI) 46.6-47.0], and on dialysis 39.3% (95% CI 39.2-39.4). The unadjusted 5-year patient survival after the first renal transplantation performed between 2002 and 2006 was 86.7% (95% CI 86.2-87.2) for kidneys from deceased donors and 94.3% (95% CI 93.6-95.0) for kidneys from living donors.

Physiological and proteomic analyses of drought stress response in Holm oak provenances.

Responses to drought stress by water withholding have been studied in 1 year old Holm oak (Quercus ilex subsp. ballota [Desf.] Samp.) seedlings from seven provenances from Andalusia (southern Spain). Several physiological parameters, including predawn xylem water potentials and relative water content in soil, roots, and leaves as well as maximum quantum efficiency and yield of PSII were evaluated for 28 days in both irrigated and nonirrigated seedlings. The leaf proteome map of the two provenances that show the extreme responses (Seville, GSE, is the most susceptible, while Almería, SSA, is the least susceptible) was obtained. Statistically significant variable spots among provenances and treatments were subjected to MALDI-TOF/TOF-MS/MS analysis for protein identification. In response to drought stress, ~12.4% of the reproducible spots varied significantly depending on the treatment and the population. These variable proteins were mainly chloroplastic and belonged to the metabolism and defense/stress functional categories. The 2-DE protein profile of nonirrigated seedlings was similar in both provenances. Physiological and proteomics data were generally in good agreement. The general trend was a decrease in protein abundance upon water withholding in both provenances, mainly in those involved in ATP synthesis and photosynthesis. This decrease, moreover, was most marked in the most susceptible population compared with the less susceptible one.

Application of label-free shotgun nUPLC-MS(E) and 2-DE approaches in the study of Botrytis cinerea mycelium.

The phytopathogenic fungus Botrytis cinerea infects more than different 200 plant species and causes substantial losses in numerous crops. The B05.10 and T4 wild-type strain genomes have been recently sequenced, becoming a model system for necrotrophic pathogens, as well as opening up new alternatives in functional genomics, such as proteomics. We analyzed B. cinerea mycelium from these two wild-type strains, introducing label-free shotgun nUPLC-MS(E) methodology to complement the 2-DE-MS-based approach. We assessed the label-free nUPLC-MS(E) methodology for protein identification and quantification using five mycelium protein dilutions. A total of 225 and 170 protein species were identified by nUPLC-MS(E) in the B05.10 and T4 strains, respectively. Moreover, 129 protein species were quantified in both strains. Significant differences in protein abundance were found in 15 more abundant and 16 less abundant protein species in the B05.10 strain compared to the T4 strain. Twenty-nine qualitative and 15 significant quantitative differences were found using 2-DE. The label-free nUPLC-MS(E) was a reliable, reproducible and sensitive method for protein identification and quantification to study the B. cinerea mycelial proteome. Results obtained by gel-based and gel-free complementary approaches allow a deeper characterization of this fungus, as well as the identification of potential virulence factors.

Renal replacement therapy in Europe-a summary of the 2010 ERA-EDTA Registry Annual Report.

BACKGROUND: This study provides a summary of the 2010 European Renal Association-European Dialysis and Transplant Association (ERA-EDTA) Registry Annual Report (available at www.era-edta-reg.org). METHODS: This report includes data on renal replacement therapy (RRT) using data from the national and regional renal registries in 29 countries in Europe and bordering the Mediterranean Sea. Individual patient data were received from 27 registries, whereas 18 registries contributed data in aggregated form. We present incidence and prevalence of RRT, transplant rates, survival probabilities and expected remaining lifetimes. The latter two are solely based on individual patient records. RESULTS: In 2010, the overall incidence rate of RRT for end-stage renal disease (ESRD) among all registries reporting to the ERA-EDTA Registry was 123 per million population (pmp) (n = 91 798). The highest incidence rate was reported by Turkey (252pmp) and the lowest reported by Montenegro (21 pmp). The overall prevalence of RRT for ESRD at 31 December 2010 among all registries reporting to the ERA-EDTA Registry was 741 pmp (n = 551 005). The prevalence varied from 124 pmp in Ukraine to 1580 pmp in Portugal. The overall number of renal transplantations performed in 2010 among all registries was 29.2 pmp (n = 21 740). The highest overall transplant rate was reported from Spain, Cantabria (73 pmp), whereas the highest transplant rate for living donor kidneys was reported from the Netherlands (28 pmp). For patients who started RRT between 2001 and 2005, the unadjusted 5-year patient survival on RRT was 46.2% [95% confidence interval (CI) 46.0-46.3], and on dialysis 38.6% (95% CI 38.5-38.8). The unadjusted 5-year patient survival after the first renal transplantation performed between 2001 and 2005 was 86.6% (95% CI 86.1-87.1) for deceased donor kidneys and 94.1% (95% CI 93.4-94.8) for living donor kidneys.

Proteomic analysis of Holm oak (Quercus ilex subsp. ballota [Desf.] Samp.) pollen.

This paper presents an analysis of Holm oak pollen proteome, together with an evaluation of the potentiality that a proteomic approach may have in the provenance variability assessment. Proteins were extracted from pollen of four Holm oak provenances, and they were analyzed by gel-based (1- and 2-DE in combination with MALDI-TOF/TOF) and gel-free (nLC-LTQ Orbitrap MS) approaches. A comparison of 1- and 2-DE protein profiles of the four provenances revealed significant differences, both qualitative and quantitative, in abundance (18 bands and 16 spots, respectively). Multivariate statistical analysis carried out on bands and spots clearly showed distinct associations between provenances, which highlight their geographical origins. A total of 100 spots selected from the 402 spots observed on 2-DE gels were identified by MALDI-TOF/TOF. Moreover, a complementary gel-free shotgun approach was performed by nLC-LTQ Orbitrap MS. The identified proteins were classified according to biological processes, and most proteins in both approaches were related to metabolism and defense/stress processes. The nLC-LTQ Orbitrap MS analysis allowed us the identification of proteins belonging to the cell wall and division, transport and translation categories. Besides providing the first reference map of Holm oak pollen, our results confirm previous studies based on morphological observations and acorn proteomic analysis. Moreover, our data support the valuable use of proteomic techniques as phylogenetic tool in plant studies.

Contribution of proteomics to the study of plant pathogenic fungi.

Phytopathogenic fungi are one of the most damaging plant parasitic organisms, and can cause serious diseases and important yield losses in crops. The study of the biology of these microorganisms and the interaction with their hosts has experienced great advances in recent years due to the development of moderm, holistic and high-throughput -omic techniques, together with the increasing number of genome sequencing projects and the development of mutants and reverse genetics tools. We highlight among these -omic techniques the importance of proteomics, which has become a relevant tool in plant-fungus pathosystem research. Proteomics intends to identify gene products with a key role in pathogenicity and virulence. These studies would help in the search of key protein targets and in the development of agrochemicals, which may open new ways for crop disease diagnosis and protection. In this review, we made an overview on the contribution of proteomics to the knowledge of life cycle, infection mechanisms, and virulence of the plant pathogenic fungi. Data from current, innovative literature, according to both methodological and experimental systems, were summarized and discussed. Specific sections were devoted to the most studied fungal phytopathogens: Botrytis cinerea, Sclerotinia sclerotiorum, and Fusarium graminearum.

Proteomics of plant pathogenic fungi.

Plant pathogenic fungi cause important yield losses in crops. In order to develop efficient and environmental friendly crop protection strategies, molecular studies of the fungal biological cycle, virulence factors, and interaction with its host are necessary. For that reason, several approaches have been performed using both classical genetic, cell biology, and biochemistry and the modern, holistic, and high-throughput, omic techniques. This work briefly overviews the tools available for studying Plant Pathogenic Fungi and is amply focused on MS-based Proteomics analysis, based on original papers published up to December 2009. At a methodological level, different steps in a proteomic workflow experiment are discussed. Separate sections are devoted to fungal descriptive (intracellular, subcellular, extracellular) and differential expression proteomics and interactomics. From the work published we can conclude that Proteomics, in combination with other techniques, constitutes a powerful tool for providing important information about pathogenicity and virulence factors, thus opening up new possibilities for crop disease diagnosis and crop protection.

Immunohistochemical study of matrix metalloproteinases and their inhibitors in pure and mixed invasive and in situ ductal carcinomas of the breast.

We assessed differences in the patterns of expression of matrix metalloproteases and their inhibitors (tissue inhibitors of metalloproteases) in ductal carcinoma in situ alone and admixed with invasive ductal carcinomas (n = 40), as well as in pure invasive ductal carcinomas (n = 40), immunohistochemically and using tissue arrays. The invasive ductal carcinoma components showed higher expression of matrix metalloprotease-9 and -13 than did the admixed ductal carcinoma in situ, whereas stromal fibroblasts of the invasive components showed higher expression of matrix metalloprotease-2, -7, -9, -13, and -14 and tissue inhibitor of metalloprotease-1 and -3 than did fibroblasts around the neoplastic ducts of the admixed ductal carcinoma in situ. Expression of matrix metalloprotease-14 and tissue inhibitor of metalloprotease-3 was significantly higher in the mononuclear inflammatory cells of the invasive components. By contrast, matrix metalloprotease-1 expression was significantly higher in stromal cells of the ductal carcinoma in situ admixed with invasive ductal carcinoma. The pure invasive ductal carcinomas had significantly higher expression of matrix metalloprotease-1, -9, -11, and -14 and tissue inhibitor of metalloprotease-1 and -3 than the invasive ductal carcinomas admixed with ductal carcinoma in situ. Our findings indicate a significant association of matrix metalloprotease expression by the periductal stromal cells of the ductal carcinoma in situ component of mixed tumors and the occurrence of distant metastasis. Our data suggest that the molecular matrix metalloprotease/tissue inhibitor of metalloprotease profile can contribute to better characterization of early breast carcinomas.