Central nervous system ependymoma: clinical implications of the new molecular classification, treatment guidelines and controversial issues.

Ependymoma is an uncommon neuroepithelial tumor that may arise anywhere within the neuroaxis, both in children and in adults. It has been classically graded upon histopathological features, yet with limited clinical utility. Recently, DNA methylation profiling has provided a novel classification of ependymoma in nine molecular subgroups. This stratification method harbors prognostic value with supratentorial RELA-fusion and posterior fossa group A tumors showing a significantly shorter survival compared to the rest. Currently, the treatment of choice involves maximal safe resection and, in cases of residual disease, adjuvant conformal radiotherapy. Second-look surgery is also a feasible and recommended option for incompletely resected tumors. The role of chemotherapy is not yet established and can be considered in infants and children with relapsing disease or prior to re-intervention. Although targeted agents do not seem to play a role as adjuvant therapy, they are currently being tested for recurrent disease.

Clinical course and pathogenicity of variant rabbit haemorrhagic disease virus in experimentally infected adult and kit rabbits: Significance towards control and spread.

RHDVb has become the dominant RHDV on the Iberian Peninsula. A better understanding of its pathogenicity is required to aid control measures. Thus, the clinical course, humoral immune response, viraemia and kinetics of RHDV-N11 (a Spanish RHDVb isolate) infection in different tissues at both viral RNA and protein levels were studied in experimentally infected young and adult rabbits. The case fatality rate differed between the two age groups, with 21% of kits succumbing while no deaths were observed in adults. Fever and viremia were strongly associated with death, which occurred 48 h post infection (PI) too fast for an effective humoral immune response to be mounted. A significant effect on the number of viral RNA copies with regard to the variables age, tissue and time PI (p < 0.0001 in all cases) was detected. Histological lesions in infected rabbits were consistently more frequent and severe in liver and spleen and additionally intestine in kits, these tissues containing the highest levels of viral RNA and protein. Although no adults showed lesions or virus antigen in intestine, both kits and adults maintained steady viral RNA levels from days 1 to 7 PI in this organ. Analysis revealed the fecal route as the main dissemination route of RHDV-N11. Subclinically infected rabbits had detectable viral RNA in their faeces for up to seven days and thus may play an important role spreading the virus. This study allows a better understanding of the transmission of this virus and improvement of the control strategies for this disease.

Fast specific field detection of RHDVb.

This work describes a simple and rapid test for field detection of the emerging rabbit pathogen RHDVb. The assay is specific for RHDVb, showing no cross-reactivity with other RHDV types giving a specific result in under 10 min using rabbit liquid exudates or liver homogenate samples taken at necropsy.

ELISA for detection of variant rabbit haemorrhagic disease virus RHDV2 antigen in liver extracts.

The emergence and rapid spread of variant of the rabbit hemorrhagic disease virus (RHDV2) require new diagnostic tools to ensure that efficient control measures are adopted. In the present study, a specific sandwich enzyme-linked immunosorbent assay (ELISA) for detection of RHDV2 antigens in rabbit liver homogenates, based on the use of an RHDV2-specific monoclonal antibody (Mab) 2D9 for antigen capture and an anti-RHDV2 goat polyclonal antibody (Pab), was developed. This ELISA was able to successfully detect RHDV2 and RHDV2 recombinant virions with high sensitivity (100%) and specificity (97.22%). No cross-reactions were detected with RHDV G1 viruses while low cross-reactivity was detected with one of the RHDVa samples analyzed. The ELISA afforded good repeatability and had high analytical sensitivity as it was able to detect a dilution 1:163,640 (6.10ng/mL) of purified RHDV-N11 VLPs, which contained approximately 3.4×108molecules/mL particles. The reliable discrimination between closely related viruses is crucial to understand the epidemiology and the interaction of co-existing pathogens. In the work described here we design and validate an ELISA for laboratory based, specific, sensitive and reliable detection of RHDVb/RHDV2. This ELISA is a valuable, specific virological tool for monitoring virus circulation, which will permit a better control of this disease.

Conventional and real time RT-PCR assays for the detection and differentiation of variant rabbit hemorrhagic disease virus (RHDVb) and its recombinants.

Since its emergence, variant RHDV (RHDVb/RHDV2) has spread throughout the Iberian Peninsula aided by the apparent lack of cross protection provided by classic (genogroup 1; G1) strain derived vaccines. In addition to RHDVb, full-length genome sequencing of RHDV strains has recently revealed the circulation of recombinant viruses on the Iberian Peninsula. These recombinant viruses contain the RHDVb structural protein encoding sequences and the non-structural coding regions of either pathogenic RHDV-G1 strains or non-pathogenic (np) rabbit caliciviruses. The aim of the work was twofold: firstly to validate a diagnostic real time RT-PCR developed in 2012 for the detection of RHDVb strains and secondly, to design a conventional RT-PCR for the differentiation of RHDVb strains from RHDVb recombinants by subsequent sequencing of the amplicon.

Fluorescence spectroscopy and parallel factor analysis as a dissolved organic monitoring tool to assess treatment performance in drinking water trains.

Fluorescence excitation emission matrix (FEEM) spectroscopy was used to evaluate its applicability as a tool to track dissolved organic matter (DOM) in a drinking water treatment plant (DWTP) that incorporates a conventional line (consisting in ozonation and GAC filtration) and a membrane-based line (consisting in ultrafiltration, reverse osmosis and mineralization) working in parallel. Seven sampling points within the different process stages were characterized monthly during 2014. A global Parallel Factor Analysis (PARAFAC) was used to pull out underlying organic fractions from the fluorescence spectra. Accordingly a five components model was selected to describe the system and the pros and cons of the model were discussed by analysis of the residuals. Among the five fluorescent components, those associated to humic-like matter (C1, C3 and C4) showed a similar season variability in the river water feeding the DWTP (which resembled that of UV254 and TOC), whereas the two components associated to protein-like matter (C2 and C5) exhibited a different behavior. The maximum fluorescence intensity values (Fmax) were used to quantify DOM removals across the plant. Compared to the conventional line, water from the UF/RO membrane-based line showed between 6 and 14 times lower fluorescence intensity signal for the humic-like components and between 1 and 3 for the protein-like components as compared to the conventional line. The differences in DOM composition due to seasonal variations and along the treatment trains point out the suitability of using fluorescence measurements over other parameters such as UV254 as a monitoring tool to help optimize operation conditions of each treatment stage and improve produced water quality in a DWTP.

Illegal discharges in Spanish waters. Analysis of the profile of the Alleged Offending Vessel.

There is at present a growing concern, on an international level, over environmental offences caused by oil discharges into the sea from vessels. The objective of the Spanish Maritime Administration is to prevent the illegal discharges of polluting substances in Spanish maritime waters by vessels in transit. To combat such discharges, since 2007 Spain has reinforced its means of response with the use of aircrafts that provide services of maritime surveillance, identifying the Alleged Offending Vessels and acting as a deterrent. The objective of the present study is both to introduce the concept and to analyze certain aspects of the so-called "Alleged Offending Vessel" (AOV) that have been detected within Spanish Search and Rescue (SAR) jurisdiction waters in the period 2008-2012, in order to build a profile of such a vessel. For this purpose, an analysis methodology is formalized based on the GINI index and Lorenz curves, associated with certain aspects of vessels: type, flag and sailing area.

[Critical care nurses and continuous renal replacement therapy]. / Enfermería de cuidados intensivos y terapias continuas de reemplazo renal.

Continuous renal replacement therapy (CRRT) has evolved within the space of a few years and is currently as widely used in intensive care units as, for example, mechanical ventilation. However, for this treatment to be useful to the critically ill patient, the nurse must have the knowledge required to solve the complications that may arise when this technique is used in order to achieve the desired objectives and to avoid complications arising from its misuse. In some units and countries, this has led to conflict about the nurses best qualified to apply these therapies. However, advances in technology have simplified the machines used. The purpose of this article was firstly, to provide a basic approximation to knowledge of the continuous extrarenal purification system and, secondly, to describe the nursing care, which can be divided into three parts: catheter care, care of the circuit, and patient care for the appropriate use of CRRT. Nurses initially perceive CRRT as being difficult to apply but with proper training these treatments become one more therapy within the framework of advanced life support treatment of critically the ill.

Enfermería de cuidados intensivos y terapias continuas de reemplazo renal / Critical care nurses and continuous renal replacement therapy

La terapia continua de reemplazo renal (TCRR) es un tratamiento que ha ido evolucionando en pocos años, de manera que hoy día constituye un tratamiento tan habitual en las unidades de cuidados intensivos como puede ser la ventilación mecánica. Pero para que sea de utilidad para el enfermo crítico, enfermería debe tener los conocimientos necesarios para resolver las complicaciones que se presenten durante la aplicación de la técnica, con el fin de conseguir los objetivos deseados y evitar complicaciones derivadas de su uso. Esto ha generado en ciertas unidades y países algunos conflictos en cuanto a cuál es la enfermería mejor cualificada en aplicar estas terapias, pero lo cierto es que los avances en la tecnología ha llevado a que las máquinas utilizadas tengan un funcionamiento más sencillo El objetivo de este artículo es, en primer lugar, realizar un acercamiento básico al conocimiento del sistema de depuración extrarrenal continuo y, en segundo, describir los cuidados de enfermería, que se pueden dividir en 3 apartados: cuidados del catéter, cuidados del circuito y cuidados del paciente para la utilización adecuada de las TCRR. Las TCRR son unas terapias que, en principio, parecen de un complicado manejo por parte del personal de enfermería, pero con un adecuado entrenamiento pasan a ser una terapia más dentro del entramado de tratamiento de soporte vital avanzado del enfermo crítico(AU)

Continuous renal replacement therapy (CRRT) has evolved within the space of a few years and is currently as widely used in intensive care units as, for example, mechanical ventilation. However, for this treatment to be useful to the critically ill patient, the nurse must have the knowledge required to solve the complications that may arise when this technique is used in order to achieve the desired objectives and to avoid complications arising from its misuse. In some units and countries, this has led to conflict about the nurses best qualified to apply these therapies. However, advances in technology have simplified the machines used. The purpose of this article was firstly, to provide a basic approximation to knowledge of the continuous extrarenal purification system and, secondly, to describe the nursing care, which can be divided into three parts: catheter care, care of the circuit, and patient care for the appropriate use of CRRT. Nurses initially perceive CRRT as being difficult to apply but with proper training these treatments become one more therapy within the framework of advanced life support treatment of critically the ill(AU)

Managing an odour episode in Barcelona's water supply: strategies adopted, the causative agent (diacetyl) and determination of its organoleptic properties.

The Llobregat River is a basic drinking water resource for the city of Barcelona, meeting 40% of the demand. The river runs through a densely populated industrial area, so that it has in its history experienced various episodes due to industrial spills. The present work deals with a recent episode involving diacetyl. To the best of our knowledge this episode is the first time that this compound has been identified as an odour-causing compound in water. In a previous work the analytical method used for the identification of the compound causing the episode was described. The present work focuses its attention on how the episode was handled: action taken at the Sant Joan Despi water treatment plant and the sampling strategy on the river that led to the identification of the company responsible for the spill, a paper mill. It also deals with the role played by the FPA panel in the resolution of the episode: organoleptic description of samples of the problem in the initial phase of the episode (from the treatment plant, distribution network and customer complaints) and, once the compound responsible was identified, the determination of its organoleptic characteristics (odour threshold, odour descriptors, Weber-Fechner curve).

A review of taste and odour events in Barcelona's drinking water area (1990-2004).

The main, relevant, solved problems associated with taste and odour incidents in Barcelona's drinking water area in the last 14 years are reviewed. Events produced by creosote, dioxanes and dioxolanes, dicylopentadiene, and diacetyl, among the anthropogenic compounds; geosmin, MIB and iodinated trihalomethanes and chlorobromoanisoles as examples of compounds of natural origin are exemplified. The determination of the odour threshold concentrations of selected odorous compounds is also shown as a tool to gain a better knowledge of future taste and odour events.

Dietary supplements for lactating goats by polyunsaturated fatty acid-rich protected fat. Effects after supplement withdrawal.

The objective of this study was to investigate the persistence of the effects of supplements after they were withdrawn. Two groups of 12 goats were maintained under semiextensive breeding conditions; they were fed indoors with a concentrate with alfalfa hay and olivetree leaves. Goats were in their sixth month of lactation at initiation of the study. During the first month of the trials, the concentrate supplied to the goats was either nonsupplemented (group 1) or supplemented with 9% polyunsaturated fatty acids (PUFA)-rich protected fat (group 2). After this period, the 2 groups were given the nonsupplemented until the average daily production of milk per animal within each group had decreased to 300 g or less. The fat supplementation supplied increased milk production and also improved fat and protein yield. These effects persisted after the supplement was withdrawn. The supplement, moreover, produced noticeable changes in the fatty acids profile of the milk fat, namely a reduction in the concentration of saturated fatty acids and an increase in that of PUFA. In contrast to the effects on milk production and on the yield of its main constituents, the effects on fat composition disappeared when the supplement was withdrawn.

Heterologous expression and functional characterization of thioredoxin from Fasciola hepatica.

The full thioredoxin coding sequence from Fasciola hepatica has been cloned into the pGEX-2T expression vector and produced in Escherichia coli as a fusion protein. The recombinant protein proved to be biologically active, using an insulin reduction assay, and was also able to activate thioredoxin peroxidase from F. hepatica. These observations suggest that this protein could participate in a redox cascade involved in the maintenance of cell homeostasis as well as in parasite protection against reactive oxygen species produced by the host.

Identification of the amino acid residue involved in rabbit hemorrhagic disease virus VPg uridylylation.

The virus genome-linked protein (VPg) coding region from rabbit hemorrhagic disease virus (RHDV) (isolate AST/89) was expressed in Escherichia coli by using a glutathione S-transferase-based vector. The recombinant polypeptide could be purified in good yields and was uridylylated in vitro from [alpha-32P]UTP in a reaction catalyzed by the recombinant RNA-dependent RNA polymerase from RHDV in the absence of added template RNA. The use of deletion and point mutants allowed the identification of Tyr-21 as the residue involved in uridylylation and consequently in the linkage between VPg and the viral genome. These data constitute the first report on the identity of the amino acid residue involved in VPg uridylylation in a member of the Caliciviridae family.

Characterisation of the RNA-dependent RNA polymerase from Rabbit hemorrhagic disease virus produced in Escherichia coli.

All positive-strand RNA viruses encode a RNA-dependent RNA polymerase which in most cases has been only identified on the basis of its sequence conservation. Catalytic activity has been experimentally demonstrated in only a handful of these viral proteins, including that from Rabbit hemorrhagic disease virus. Studies from our laboratory have reported that RHDV RNA polymerase produced in Escherichia coli was enzymatically active showing poly(A)-dependent poly(U) polymerase as well as RNA polymerase activity on heteropolymeric substrates. In this work, we have investigated the in vitro activity of the recombinant 3Dpol from RHDV, including ion requirements, resistance to inhibitors, substrate specificity as well as data on the initiation mechanism of the template-linked products derived from heteropolymeric RNA substrates. Our study demonstrates that in an in vitro reaction recombinant RHDV RNA polymerase generated the minus strand of the heteropolymeric RNA substrates by a "copy-back" mechanism that initiated at the template 3'-terminal OH.

Fasciola hepatica: heterologous expression and functional characterization of a thioredoxin peroxidase.

A Fasciola hepatica cDNA clone of 779 bp was isolated from an adult worm cDNA expression library by immunological screening using a rabbit serum against the excretory-secretory antigens. The nucleotide sequence of the cDNA revealed the presence of an open reading frame of 582 bp which encoded a 194-amino-acid-residue polypeptide (M(r) 21,723 Da) showing a high degree of homology to thioredoxin peroxidases. This putative antioxidant protein gene was expressed in Escherichia coli as a GST fusion protein. The recombinant fusion protein showed in vitro antioxidant properties and protected rabbit muscle enolase and E. coli glutamine synthetase from inactivation by nonenzymatic Fe(3+)/O(2)/DTT or Fe(3+)/O(2)/ascorbate metal-catalyzed oxidation systems.

Production of healthier goat milk. Use of a concentrate supplemented with a "protected" fat rich in PUFA.

In order to obtain healthier goat milk as far as its fat composition is concerned, milk production and composition trials were carried out with Granadina goats kept in semi-extensive breeding conditions. The animals were feeding indoors with a concentrate supplemented or not with 7% of fat conveniently protected against the action of rumen, which had 35% of polyunsaturated fatty acids (PUFA). The amount of milk produced was higher (p < 0.05) with consumption of the fat supplemented concentrate. At the same time, and although there was no difference in the milk concentration of fat and protein nitrogen (p > 0.05), the corresponding yields were higher for consumption of the fat supplemented concentrate. As for the composition of milk fat, the use of the supplemented concentrate resulted in fat with a higher (p < 0.05) concentration of PUFA and a lower (p < 0.05) concentration of stearic acid. At the same time, the concentration of medium chain triglycerides, which is the specific nutritional value particular to goats' milk, was the same whichever concentrate was consumed.

Cloning and expression in Escherichia coli of a Fasciola hepatica gene encoding a calcium-binding protein.

A Fasciola hepatica cDNA clone of 994 bp was isolated from an adult worm cDNA expression library using a rabbit serum against the excretory-secretory antigens. The nucleotide sequence of the cDNA clone revealed the presence of an open reading frame of 572 bp which encoded a 22 kDa polypeptide (Fh22) showing putative EF-hand domains. This gene was expressed in Escherichia coli and the recombinant protein used for the production of specific antibodies. Immunoblotting studies using the anti-Fh22 serum showed the presence of a polypeptide of similar molecular mass in the excretory-secretory extract of the adult parasite. The recombinant Fh22 polypeptide showed calcium-dependent electrophoretic mobility (decreased with Ca2(+)-ions and increased with EGTA). The observed behaviour of recombinant Fh22 in gel filtration experiments also suggested calcium-induced conformational changes.

Macrophage tropism of rabbit hemorrhagic disease virus is associated with vascular pathology.

To delineate the interactions between rabbit hemorrhagic disease virus (RHDV) and host cells, organ and cellular targets of infection were identified in vivo. Viral specific antigens were detected by immunohistochemistry in liver, lung, spleen and lymph nodes cells. Also, intravascular infected cells were detected in most organs including kidneys, myocardium, thymus and central nervous system. To further characterize infected target cells, viral proteins and cell-specific surface antigens were identified simultaneously in double labeling experiments. Numerous lymphoid organ macrophages, from the splenic red pulp, circulating monocytes, alveolar macrophages and Kupffer cells were double labeled, demonstrating that cells of the mononuclear phagocyte lineage are major hosts for RHDV. Double labeling for other specific cell markers were negative. The distribution of viral antigens in these tissues coincided with those areas where cells presented morphology of apoptosis. Association of intravascular monocyte infection and apoptosis, could represent a possible mechanism to develop disseminated intravascular coagulation.

Immunization with potato plants expressing VP60 protein protects against rabbit hemorrhagic disease virus.

The major structural protein VP60 of rabbit hemorrhagic disease virus (RHDV) has been produced in transgenic potato plants under the control of a cauliflower mosaic virus 35S promoter or a modified 35S promoter that included two copies of a strong transcriptional enhancer. Both types of promoters allowed the production of specific mRNAs and detectable levels of recombinant VP60, which were higher for the constructs carrying the modified 35S promoter. Rabbits immunized with leaf extracts from plants carrying this modified 35S promoter showed high anti-VP60 antibody titers and were fully protected against the hemorrhagic disease.