LncMachine: a machine learning algorithm for long noncoding RNA annotation in plants.

Following the elucidation of the critical roles they play in numerous important biological processes, long noncoding RNAs (lncRNAs) have gained vast attention in recent years. Manual annotation of lncRNAs is restricted by known gene annotations and is prone to false prediction due to the incompleteness of available data. However, with the advent of high-throughput sequencing technologies, a magnitude of high-quality data has become available for annotation, especially for plant species such as wheat. Here, we compared prediction accuracies of several machine learning algorithms using a 10-fold cross-validation. This study includes a comprehensive feature selection step to refine irrelevant and repeated features. We present a crop-specific, alignment-free coding potential prediction tool, LncMachine, that performs at higher prediction accuracies than the currently available popular tools (CPC2, CPAT, and CNIT) when used with the Random Forest algorithm. Further, LncMachine with Random Forest performed well on human and mouse data, with an average accuracy of 92.67%. LncMachine only requires either a FASTA file or a TAB separated CSV file containing features as input files. LncMachine can deploy several user-provided algorithms in real time and therefore be effortlessly applied to a wide range of studies.

Bloqueo laparoscópico subcostal del plano transverso abdominal (TAP) como analgesia multimodal postquirúrgica en pacientes con patología biliar / Laparoscopic subcostal block of the transverse abdominal plane (TAP) as postoperative multimodal analgesia in patients with biliary pathology

Contexto: el dolor postoperatorio es uno de los principales problemas en la práctica quirúrgica y debe ser tratado de forma adecuada para lograr una evolución favorable en el paciente. Objetivo: evaluar el bloqueo subcostal laparoscópico del plano transverso del abdomen con bupivacaína 0,5%, versus el no bloqueo en pacientes adultos sometidos a cirugía de patología biliar en el hospital San Francisco de Quito. Metodología: estudio epidemiológico de casos y controles de pacientes que ingresaron al hospital San Francisco de Quito. Son 210 pacientes divididos de forma equitativa en dos grupos, en uno de ellos se aplicó bloqueo subcostal, mientras que el otro fue sin intervención. Los dos grupos fueron observados y valorados por 24 horas. Las variables cuantitativas se reportaron como promedios (± DE, desviación estándar) y las cualitativas con sus valores absolutos y relativos (porcentajes). Las comparaciones de variables cuantitativas se realizaron con la prueba t-Student, mientras que para las cualitativas se usaron pruebas de independencia para proporciones x2 y prueba exacta de Fisher. Resultados: comparado con el segundo grupo la tasa de analgesia de rescate fue significativamente menor en los pacientes que recibieron bloqueo laparoscópico con bupivacaína al 0,5% (28,6% vs. 51,4 %, respectivamente). La reducción en el uso de analgésicos fue en promedio del 22,9% (C 95 %:36,7, 9 %; p =0,0012), equivalente a un OR: 0,38 (IC 95 %: 0,21, 0,67; p = 0,004). ORad = 0,38 (IC 95 %: 0,21, 0,67; p <0,01). El uso de opioides fue menor en el grupo con bloqueo, comparado con su control (46,7% vs 59,3 %),mientras que el uso de paracetamol fue mayor (30% vs 18,5 %). Y en el uso de analgésicos no esteroidales (AINES) el resultado fue similar (23,3% vs 22,2 %). Conclusión: el bloqueo subcostal laparoscópico se muestra como una técnica eficaz para control y tratamiento del dolor en pacientes sometidos a cirugía con patología biliar, disminuyendo el uso de analgesia de rescate durante las 6 primeras horas. (AU)

Context: the postoperative pain is one of the main problems in surgical practices; it must be adequately treated to achieve a favorable evolution in the patient. Objective: To evaluate the laparoscopic subcostal block of the transverse plane of the abdomen with 0.5% of bupivacaine, versus the non-block in adult patients that had surgery of biliary pathology at San Francisco de Quito Hospital. Methodology: epidemiological study of cases and controls of patients that were admitted to the San Francisco de Quito Hospital. There are 210 patients equally divided in two groups, in the first group the subcostal block was applied, while in the other group was without intervention. The two groups were observed and assessed for 24 hours. The quantitative variables were reported as averages (± SD, Standard Deviation) and qualitative variables with their absolute and relative values (percentages) were also reported as well. The comparisons of quantitative variables were performed with a t-Student test, while the comparison of qualitative variables was performed with independent tests for proportions x2 and Fisher's exact test. Results: compared with the second group, the rate of analgesia for rescue was significantly lower in patients who received laparoscopic block with 0.5% bupivacaine (28.6% vs. 51.4%, respectively). The reduction in the use of analgesics was of 22.9% on average (95% CI: 36.7, 9%, p = 0.0012), equivalent to an OR: 0.38 (95% CI: 0.21, 0.67, p = 0.004). ORad = 0.38 (95% CI: 0.21, 0.67, p <0.01). The use of opioids was lower in the group with block compared to its control (46.7% vs 59.3%), the use of paracetamol was higher (30% vs 18.5%) and in the use of non-steroidal analgesics (NSAIDs), the result was similar (23.3% vs 22.2%). Conclusion: laparoscopic subcostal block is shown as an effective technique for pain control and treatment in patients that had surgery with biliary pathology, reducing the use of analgesia on average during the first 6 hours. (AU)

Approach, attitudes, and knowledge of general practitioners in relation to Helicobacter pylori is inadequate. There is much room for improvement! / El abordaje, las actitudes y el conocimiento acerca de Helicobacter pylori en médicos generales es deficiente. ¡Hay mucho que mejorar!

BACKGROUND: Helicobacter pylori (Hp) infection is one of the most common infections in humans. Due to its worldwide prevalence, a series of guidelines and recommendations has been developed for the appropriate screening, diagnosis, and management of patients with Hp infection. AIM: To evaluate the approach, attitudes, and knowledge of a group of Mexican general practitioners in relation to Hp. MATERIALS AND METHODS: A cross-sectional study was conducted that evaluated the knowledge of Hp diagnosis and treatment through the application of a questionnaire. RESULTS: From the total of 430 questionnaires delivered, information was obtained from 411 (95% response rate). The most frequent indications for eradication treatment were peptic ulcer (48.4%), gastroesophageal reflux disease (41.8%), and dyspepsia (39.2%). Thirty-one percent of the physicians surveyed used more than one test to look for Hp, and serology was the most commonly employed method. The most widely used antibiotic regimen was clarithromycin plus amoxicillin (63.8%), followed by metronidazole plus tetracycline (16%). Ninety-two percent of the physicians confirmed eradication through endoscopy for the histologic analysis and only 23% utilized the breath test. CONCLUSIONS: Our results show the lack of knowledge about the established diagnostic and treatment recommendations for Hp infection. The implementation of workshops, courses, and conferences, or the formulation of practical guidelines should be directed at primary care physicians to strengthen the practice of medicine based on scientific evidence.

Estudio para determinar los niveles de calcio y paratohormona en pacientes con y sin vaciamiento central profiláctico, con diagnóstico de cáncer papilar de tiroides / Study to determine calcium and parathyroid hormone levels in patients with and without prophylactic central emptying, with diagnosis of papillary thyroid cancer

Contexto: El cáncer papilar de tiroides representa una patología que va en aumento, y el manejo quirúrgico es complejo. Una disección profiláctica cervical central para los pacientes con cáncer papilar de tiroides es controvertida. Dado que los riesgos de complicaciones transitorias probablemente son altas, y los beneficios parecen ser pequeños, de tal forma, que para realizarlo se utiliza un enfoque selectivo, en función de factores de riesgo del paciente y el nivel de experiencia del cirujano. Objetivo: determinar en el postquirúrgico los niveles de calcio y paratohormona en pacientes adultos, de ambos sexos con sospecha alta de cáncer papilar de tiroides, a los cuales se les realizó o no, vaciamiento central en los hospital Eugenio Espejo de la ciudad de Quito, durante un año. Sujetos y métodos: observacional, analítico, epidemiológico transversal, de cohortes. Se estudiaron todos los pacientes con sospecha citológica de cáncer papilar de tiroides, sometidos a tiroidectomía total, con o sin vaciamiento central profiláctico; se midió a las 24 horas los niveles de calcio y paratohormona. Las variables cuantitativas se reportaron como promedios y las cualitativas con sus valores absolutos y relativos. Las comparaciones de variables cuantitativas se realizaron con la prueba t-Student, mientras que para las cualitativas se usaron pruebas de independencia para proporciones x2 y prueba exacta de Fisher. Se hizo un análisis multivariados con regresión logística reportada como Odds Ratios ajustados (ORadj). Resultados: con respecto al calcio el grupo con vaciamiento central presentó valores significativamente menores que el grupo sin vaciamiento, esta diferencia fue de -0.07 mmol/L (IC 95 %: -0.036, -0.097 mmol/L;p < 0.0001). Los pacientes sometidos a vaciamiento central tuvieron tasas significativamente menores de paratohormona (para valores inferiores a los 10pg/ml) hasta el 53.9%, mientras que en el grupo sin vaciamiento solo el 11.1%(IC 95%: 29.5, 56.2%; p < 0.0001). Conclusión: se observó que en el grupo de tiroidectomía con vaciamiento central profiláctico, hubo una tasa importante de hipocalcemia e hipoparatiroidismo, todos estos pacientes presentaron signos de hipocalcemia durante su periodo postquirúrgico, lo que prolongo su estadía hospitalaria, (AU)

Introduction: Papillary thyroid cancer is on the rise and surgical treatment is complex. Prophylactic central neck dissection for papillary thyroid cancer patients is controversial because of the possibility of higher risk of transient complications and the benefits of the surgery are low. As a result, the surgery is performed based of the risk factors of the patient and the experience of the surgeon. Objective: to determine the levels of calcium and parathyroid hormone post-surgery in adult male and female patients with suspected risk of papillary thyroid cancer, to who prophylactic central drainage, was and was not performed. Subjects and methods: this is an epidemiological, observational, analytic, cross-sectional, of cohorts. The research will be done on all patients suspected of cytological papillary thyroid cancer in Hospital Eugenio Espejo, and who underwent total thyroidectomy with and without prophylactic central drainage. Calcium and parathyroid hormone levels will be checked 24 hours post-surgery. The quantitative variables will be reported as averages and the qualitative variables as absolute and relative data. t-Student test was used to compare qualitative data. Qualitative data was compared using independent test with scales of x2 and the Fisher exact test. A logistic regression analysis was done and reported as Adjusted Odds ratios (AOR). Results: calcium level on the group with central drainage had significant lower levels compare to the group without central drainage, the difference was of -0.07 mmol/L (IC 95 %: -0.036, -0.097 mmol/L; p<0.0001). The patients with central drainage had significantly lower levels of parathyroid hormone (levels lower than 10pg/ml) up to 53.9%; while the group without central drainage only had 11.1% %(IC 95%: 29.5, 56.2%; p < 0.0001). Conclusion: though the study the thyroidectomy with prophylactic central drainage group had a significant rate of hypocalcemia and hypoparathyroidism. These patients had symptoms of hypocalcemia post-surgery, which prolong their hospital stay. (AU)

[Clinical utility and psychometric properties of Prefrontal Symptoms Inventory (PSI) in acquired brain injury and degenerative dementias]. / Utilidad clínica y propiedades psicométricas del inventario de síntomas prefrontales (ISP) en el daño cerebral adquirido y las demencias degenerativas.

INTRODUCTION: The cognitive, emotional and behavioural alterations secondary to acquired brain injury and degenerative dementias can be quantitatively and quantitatively appraised by administering self-reports that ask both patients and reliable informants about the difficulties patients have in their everyday life. SUBJECTS AND METHODS: The Prefrontal Symptoms Inventory (PSI) and the Modified Memory Failures in Everyday Life Questionnaire (MFE-30) were administered to 174 paired participants: 87 patients with brain damage or degenerative dementias and their 87 reliable informants. In addition to the psychometric goodness of the tests, the study also explored the clinical usefulness of applying these questionnaires to patients and their informants in order to obtain a rate of discrepancy in the scores as a measure of anosognosia. RESULTS: The results show how applying the PSI-20 (20 items) or the PSI (46 items), whether administered together with the MFE-30 (30 items) or not, is a very useful procedure for assessing the symptoms in individuals with acquired brain injury or degenerative dementias, since it yields a great deal of information about patients' difficulties in their daily life. CONCLUSIONS: We recommend that, in addition to the compulsory neuropsychological assessment, questionnaires or inventories of symptoms like those proposed here should be carried out, due to the fact that they offer a number of advantages from the clinical point of view, as well as being efficacious and effective in economic terms.

TITLE: Utilidad clinica y propiedades psicometricas del inventario de sintomas prefrontales (ISP) en el daño cerebral adquirido y las demencias degenerativas.Introduccion. Las alteraciones cognitivas, emocionales y comportamentales secundarias al daño cerebral adquirido y las demencias degenerativas pueden valorarse cuantitativa y cualitativamente mediante la administracion de autoinformes que interroguen ­a pacientes e informadores fiables­ sobre las dificultades de los pacientes en la vida cotidiana. Sujetos y metodos. Se administro el inventario de sintomas prefrontales (ISP) y el cuestionario de fallos de memoria en la vida cotidiana modificado (MFE-30) a 174 participantes emparejados: 87 pacientes con daño cerebral o demencias degenerativas y sus 87 informadores fiables. Se exploro, junto con la bondad psicometrica de las pruebas, la utilidad clinica de la aplicacion de estos cuestionarios a pacientes e informadores para obtener un indice de discrepancia de las puntuaciones como medida de la anosognosia. Resultados. Los resultados muestran como aplicar el ISP-20 (20 items) o el ISP (46 items), sean o no administrados conjuntamente con el MFE-30 (30 items), resulta un procedimiento muy util para la valoracion de la sintomatologia en los individuos con daño cerebral adquirido o demencias degenerativas, al proporcionar una gran cantidad de informacion sobre las dificultades de los pacientes en la vida cotidiana. Conclusiones. Se recomienda, junto con la obligada evaluacion neuropsicologica, la cumplimentacion de cuestionarios o inventarios de sintomas como los propuestos, dado que presentan ventajas desde el punto de vista clinico, ademas de resultar eficaces, efectivos y eficientes en terminos economicos.

Tumores estromales de potencial maligno incierto (STUMP): descripción epidemiológica y manejo durante el período 2004-2013 / Stromal tumors of an uncertain malignant potential (STUMP): epidemiological description and handling during the 2004-2013 period

Los tumores estromales de potencial maligno incierto (STUMP) se enmarcan dentro de las lesiones de células fusadas de la próstata, entidades de relativa infrecuencia en la práctica urológica habitual, contabilizándose entre 0,1-0,2 por ciento 1 de todas los tumores malignos prostáticos. Corresponden a una proliferación no epitelial mesenquimática de células fusadas estromales, que pueden adquirir la capacidad de infiltrar el epitelio glandular prostático. Representan un desafío tanto para el patólogo, por la dificultad diagnóstica, como para el clínico, pues no existe consenso respecto a su manejo. Materiales y métodos: estudio retrospectivo descriptivo. Se filtró la base de datos del servicio de Anatomía Patología del Hospital San Borja Arriarán con los términos clave “STUMP”, “Tumor estromal próstata”, tanto de biopsias transrrectales como de piezas operatorias. De los pacientes con diagnóstico histológico e inmunohistoquímico compatible, se registraron características epidemiológicas, estudio, manejo y sobrevida global. Resultados: se encontraron 3(tres) pacientes con diagnóstico histológico de STUMP. Edad promedio al diagnóstico 66años. Dos de los casos (66,6 ´po0r ciento) correspondieron a hallazgo en biopsia diferida por cirugía benigna prostática, (una por Adenomectomía transvesical y otra por RTU-P). El tercer caso correspondió a un hallazgo en biopsia por punción transrrectal ecodirigida, en contexto de PSA elevado (100ng/mL) y 4 biopsias previas: 3 normales, y una con informe de posible sarcoma prostático. La terapia de elección en 2 casos fue seguimiento y en uno se indicó cirugía radical. En los casos descritos no se registra mortalidad, con seguimiento promedio de 32 meses. Conclusión: Los tumores estromales de potencial maligno incierto son un diagnóstico histológico infrecuente, descrito como hallazgo tanto en la biopsia transrrectal como de pieza quirúrgica. No existen consensos internacionales ni guías de manejo. En nuestra experiencia...

Stromal tumors of uncertain malignant potential (STUMP) are included in the spindle cell lesions of the adult prostate, a very rare diagnostic entity in the clinical practice, accounting no more than 0,1-0,2 percent of all prostate malignancies. STUMP is defined as a non epithelial proliferation of spindle cells, which can acquire the ability of infiltrate the prostate glands. This lesions represent a challenge for both clinical and pathology physician. Material and methods: retrospective cohort study. A research was performed at the San Borja Hospital Pathology Service’s database, using keywords “STUMP”, “Stromal prostate tumor”. From eligible patients we took epidemiology and clinical data. Results: a total of three patients were included. Average age was 66 years. STUMP was founded in two patients in the pathology report after benign prostatic surgery (Simple prostatectomy and TUR-P). A third patient was diagnosed by a needle biopsy (TRUS Biopsy). This patient had an elevated PSA (>100 ng/mL), 4 previous biopsies, three normal and a fourth suspicious of prostate sarcoma. Surveillance was the therapy chosen for those patients diagnosed with STUMP after benign surgery. Radical prostatectomy was offered to the third patient. Average follow up was 32 month Conclusion: STUMP is a rare diagnostic, and is a challenge for both clinical and pathology physician. There are no guidelines for the management. In our series we confirmed the low frequency of this condition, and the management in our service is similar as other series described in literature...

Uso de agonista de GnRH para inducir ovulación en ciclos de hiperestimulación ovárica controlada: experiencia de Clínica Monteblanco, Santiago, Chile / Use of GnRH agonist to induce ovulation cycles controlled ovarian hyperstimulation: experience of Monteblanco Clinic, Santiago, Chile

Objetivo: Presentar la experiencia de la Unidad de Medicina Reproductiva de Clínica Monteblanco con el uso de análogos GnRh para la inducción final de la maduración ovocitaria. Método: Se registraron los casos de IVF/ICSI durante el año 2012 en los que se indujo la maduración final ovocitaria con análogos GnRh (Lupron®). Todos los ciclos fueron estimulados con FSHr (Puregon®) y gonadotrofina urinaria altamente purificada (Menopur®), para la prevención del alza prematura de LH, el día 5° de estimulación se agregó diariamente antagonista de GnRh. La maduración ovocitaria final se realizó con 1,25 mg de acetato de leuprolide (Lupron®), posteriormente se realizó aspiración folicular bajo guía ecográfica. Todos los embriones obtenidos fueron vitrificados y transferidos en ciclos posteriores. Resultados: Entre enero y diciembre del año 2012 se registraron 110 pacientes cuya inducción de maduración final ovocitaria se realizó con acetato de leuprolide. El promedio de ovocitos recuperados fue de 21, la proporción de ovocitos maduros fue de 72 por ciento y la frecuencia de fecundación fue de 64 por ciento. No hubo ningún caso de síndrome de hiperestimulación ovárico severo. Conclusiones: En los casos presentados de inducción de la maduración ovocitaria final con acetato de leuprolida, los resultados obtenidos son óptimos en términos de número de ovocitos en metafase II recuperados y en frecuencia de fecundación, mostrando ser una alternativa eficiente en la prevención del síndrome de hiperestimulación ovárico severo, sin alterar el pronóstico de las pacientes.

Objective: To present the experience of the Reproductive Medicine Unit of Clinica Monteblanco inducing oocyte final maturation by GnRh analogue administration. Methods: We analysed all IVF/ICSI cases performed in 2012, in which final oocyte maturation was induced by administration of GnRH analogue (Lupron®). Controlled ovarian hyperstimulation was achieved bydaily rFSH (Puregon®) and highly purified urinary gonadotropin (Menopur®) administration. In order to prevent premature LH rise, on the 5th day of stimulation daily GnRH antagonist (Orgalutran®) was added. Final oocyte maturation was induced by the administration of 1.25 mg leuprolide acetate (Lupron®). Follicular aspiration was subsequently performed under ultrasound guidance. All embryos were vitrified and transferred in a subsequent cycle. Results: We registered 110 patients. The mean number of recovered oocytes was 21; the proportion of mature oocytes was 72 percent, and the fecundation rate reached was 64 percent. No case of severe ovarian hyperstimulation syndrome (OHSS) was recorded. Conclusions: In this cohort, the use of leuprolide acetate for induce final oocyte maturation demonstrated to be an efficient alternative to induce oocyte final maturation, while preventing OHSS.

Biodesulfurization of gas oil using inorganic supports biomodified with metabolically active cells immobilized by adsorption.

The immobilization of Pseudomonas stutzeri using adsorption on different inorganic supports was studied in relation to the number of adsorbed cells, metabolic activity and biodesulfurization (BDS). The electrophoretic migration (EM) measurements and Tetrazolioum (TTC) method were used to evaluate adsorption and metabolic activity. Results indicate that maximal immobilization was obtained with an initial load of 14 x 10(8) cells mL(-1) for Al and Sep, whereas Ti requires 20 x 10(8) cells mL(-1). The highest interaction was observed in the P. stutzeri/Si and P. stutzeri/Sep biocatalysts. The IEP values and metabolic activities indicate that P. stutzeri change the surface of supports and maintains metabolic activity. A direct relation between BDS activity and the adsorption capacity of the bacterial cells was observed at the adsorption/desorption equilibrium level. The biomodification of inorganic supports by the adsorption process increases the bioavailability of sulphur substrates for bacterial cells, improving BDS activity.

Oxygen regulation of a nodule-located carbonic anhydrase in alfalfa.

Control of the permeability to oxygen is critical for the function of symbiotic nitrogen fixation in legume nodules. The inner cortex (IC) seems to be a primary site for this regulation. In alfalfa (Medicago sativa) nodules, expression of the Msca1 gene encoding a carbonic anhydrase (CA) was previously found to be restricted to the IC. We have now raised antibodies against recombinant Msca1 protein and used them, together with antibodies raised against potato leaf CA, to demonstrate the presence of two forms of CA in mature nodules. Each antibody recognizes a different CA isoform in nodule tissues. Immunolocalization revealed that leaf-related CAs were localized primarily in the nitrogen-fixing zone, whereas the Msca1 protein was restricted exclusively to the IC region, in indeterminate and determinate nodules. In alfalfa nodules grown at various O(2) concentrations, an inverse correlation was observed between the external oxygen pressure and Msca1 protein content in the IC, the site of the putative diffusion barrier. Thus Msca1 is a molecular target of physiological processes occurring in the IC cells involved in gas exchange in the nodule.

Molecular Mechanisms in Root Nodule Development.

Under nitrogen-limiting conditions, bacteria from the family Rhizobiaceae establish a symbiosis with leguminous plants to form nitrogen-fixing root nodules. These organs require a coordinated control of the spatiotemporal expression of plant and bacterial genes during morphogenesis. Both plant and bacterial signals are involved in this regulation in the plant host. Plant genes induced during nodule development, the so-called nodulin genes, have been extensively characterized. Products of several of these genes show homologies to known regulators of signal transduction pathways in other plant or animal systems. Initial functional analysis of the molecular mechanisms implicated in nodulation have been undertaken using model legumes. Insertion mutagenesis and transgenic technologies to modify nodulin gene expression, as well as pharmacologic approaches, have been used to analyze molecular mechanisms involved in morphologic responses induced by the bacterial symbiont in the plant. G protein-mediated transduction mechanisms have been implicated, and the nin transcription factor appears to be required for early steps in nodule development. ENOD40, a gene coding for an RNA that contains only short ORFs, seems to be closely tied to nodule primordium formation. In addition, a vascular-associated Krüppel-like transcription factor and small Rab type G-proteins affect bacteroid differentiation and the function of the nitrogen-fixing zone. These initial results presage a wealth of information that will be obtained from the application of genomic approaches to legumes.

The effects of sevoflurane on serum creatinine and blood urea nitrogen concentrations: a retrospective, twenty-two-center, comparative evaluation of renal function in adult surgical patients.

UNLABELLED: Despite mounting clinical evidence that supports its safety, the question of the potential adverse effects of sevoflurane on renal function continues to generate some controversy. This study retrospectively evaluated pooled renal laboratory data from 22 different clinical trials that compared sevoflurane with three widely used anesthetics. The trials examined postoperative changes in serum creatinine and blood urea nitrogen levels from a total of 3, 436 ASA physical status I-IV adult surgical patients administered either sevoflurane (n = 1941) or a control drug (isoflurane, enflurane, or propofol; n = 1495) as the maintenance anesthetic. The incidences of increased serum creatinine and blood urea nitrogen concentrations were similar among patients administered sevoflurane and those administered control drugs. Additionally, no trends specific to sevoflurane were observed with respect to postoperative serum creatinine concentration and fresh gas flow rate, concurrent treatment with nephrotoxic antibiotics, or type of carbon dioxide absorbent. IMPLICATIONS: Our data for changes in serum creatinine and blood urea nitrogen indicate that, for exposures of less than 4 minimum alveolar anesthetic concentration/h, sevoflurane is not associated with an increased risk of renal toxicity compared with other commonly used anesthetics. For clinical purposes, the pre- to postoperative changes in serum creatinine and blood urea nitrogen are appropriate measures of renal function in surgical patients.

Are isocitrate dehydrogenases and 2-oxoglutarate involved in the regulation of glutamate synthesis?

In plants, nitrogen assimilation into amino acids relies on the availability of the reduced form of nitrogen, ammonium. The glutamine synthetase-glutamate synthase pathway, which requires carbon skeletons in the form of 2-oxoglutarate, achieves this. To date, the exact enzymatic origin of 2-oxoglutarate for plant ammonium assimilation is unknown. Isocitrate dehydrogenases synthesize 2-oxoglutarate. Recent efforts have concentrated on evaluating the involvement of different isocitrate dehydrogenases, distinguished by co-factor specificity and sub-cellular localization. Furthermore, several observations indicate that 2-oxoglutarate is likely to be a metabolic signal that regulates the coordination of carbon:nitrogen metabolism. This is discussed in the context of recent advances in bacterial signalling processes.

[Immune humoral response in pigs infected with eggs and posoncospheres of Taenia solium]. / Estudio de la respuesta inmune humoral en cerdos infectados con huevos y posoncosferas de Taenia solium.

Due to the importance of cysticercosis in Mexico and Latin America and to the fact that in the last years another mechanism of infection for this disease has been proposed, i.e. through postoncospheres and immunosuppression of the host, we have considered relevant to perform the present work, which consisted in assessing the immune response induced by dexamethasone as well as that produced by parasites in pigs infected with T. solium eggs, or postoncosphere-infected, and in postoncosphere-infected and dexamethasone-treated animals. We used 10 recently weaned pigs, three were used as controls, two of them without the drug and one with it; two were infected with T. solium eggs; five with postoncospheres receiving also dexamethasone three of them. We evaluated the humoral response against parasite antigen using indirect haemagglutination (IH) and ELISA methods. Results of the immune humoral response revealed titres of up to 1:128 in T. solium eggs infected animals, of 1:16 in postoncosphere infected animals, and of 1:32 towards the end of the experiment in postoncosphere plus dexamethasone animals. Absorbance titres with ELISA confirmed these findings. Data obtained by IH show that the antibody titres of the pigs challenged with postoncospheres and postoncospheres plus dexamethasone are positive as compared to the titres obtained in the pigs infected with T. solium eggs. Results from the ELISA confirmed this finding, since, from weeks 14 to 17, the pigs became positive, behaving as those pigs that developed cysticercosis. This is relevant as it indicates that the antiposcosphere antibodies recognized antigens of T. solium larvae.

Eucalypt NADP-dependent isocitrate dehydrogenase. cDNA cloning and expression in ectomycorrhizae.

NADP-dependent isocitrate dehydrogenase (NADP-ICDH) activity is increased in roots of Eucalyptus globulus subsp. bicostata ex Maiden Kirkp. during colonization by the ectomycorrhizal fungus Pisolithus tinctorius Coker and Couch. To investigate the regulation of the enzyme expression, a cDNA (EgIcdh) encoding the NADP-ICDH was isolated from a cDNA library of E. globulus-P. tinctorius ectomycorrhizae. The putative polypeptide sequence of EgIcdh showed a high amino acid similarity with plant NADP-ICDHs. Because the deduced EgICDH protein lacks an amino-terminal targeting sequence and shows highest similarity to plant cytosolic ICDHs, it probably represents a cytoplasmic isoform. RNA analysis showed that the steady-state level of EgIcdh transcripts was enhanced nearly 2-fold in ectomycorrhizal roots compared with nonmycorrhizal roots. Increased accumulation of NADP-ICDH transcripts occurred as early as 2 d after contact and likely led to the observed increased enzyme activity. Indirect immunofluorescence microscopy indicated that NADP-ICDH was preferentially accumulated in the epidermis and stele parenchyma of nonmycorrhizal and ectomycorrhizal lateral roots. The putative role of cytosolic NADP-ICDH in ectomycorrhizae is discussed.

Mitochondrial localization of a NADP-dependent [corrected] isocitrate dehydrogenase isoenzyme by using the green fluorescent protein as a marker.

In this work, we describe the isolation of a new cDNA encoding an NADP-dependent isocitrate dehydrogenase (ICDH). The nucleotide sequence in its 5' region gives a deduced amino acid sequence indicative of a targeting peptide. However, even if this cDNA clearly encodes a noncytosolic ICDH, it is not possible to say from the targeting peptide sequence to which subcellular compartment the protein is addressed. To respond to this question, we have transformed tobacco plants with a construct containing the entire targeting signal-encoding sequence in front of a modified green fluorescent protein (GFP) gene. This construct was placed under the control of the cauliflower mosaic virus 35S promoter, and transgenic tobacco plants were regenerated. At the same time, and as a control, we also have transformed tobacco plants with the same construct but lacking the nucleotide sequence corresponding to the ICDH-targeting peptide, in which the GFP is retained in the cytoplasm. By optical and confocal microscopy of leaf epiderm and Western blot analyses, we show that the putative-targeting sequence encoded by the cDNA addresses the GFP exclusively into the mitochondria of plant cells. Therefore, we conclude that this cDNA encodes a mitochondrial ICDH.

Identification of a tobacco cDNA encoding a cytosolic NADP-isocitrate dehydrogenase.

A cDNA which encodes a specific member of the NADP-dependent isocitrate dehydrogenase (ICDH) multi-isoenzyme family has been isolated from a tobacco cell suspension library, and the expression pattern of ICDH transcripts examined in various plant tissues. To assign this cDNA to a specific ICDH isoenzyme, the major, cytosolic ICDH isoenzyme of tobacco leaves (ICDH1) was purified to homogeneity and its N-terminus as well as several tryptic peptides, representing 30% of the protein, were sequenced. The comparison of these amino acid sequences with the deduced protein sequence of the cDNA confirmed that this clone encodes for ICDH1. The total ICDH specific activity and protein content were higher in vascular-enriched tobacco leaf tissue than in deveined (depleted in midrib and first-order veins) leaves. Taking advantage of antibodies raised against either ICDH1 or the chloroplastic ICDH2 isoenzyme from tobacco cell suspensions, an immuno-cytochemical approach indicated that the ICDH1 isoenzyme, located in the cytosolic compartment of tobacco leaf cells, is responsible for this expression pattern. This observation was confirmed by northern blot analyses, using a specific probe obtained from the 3' non-coding region of the ICDH1 cDNA. A comparison of ICDH protein sequences shows a large degree of similarity between eukaryotes (> 60%) but a poor homology is observed when compared to Escherichia coli ICDH (< 20%). However, it was found that the amino acids implicated in substrate binding, deduced from the 3-dimensional structure of the E. coli NADP-ICDH, appear to be conserved in all the deduced eukaryotic ICDH proteins reported until now.

Purification and characterization of a fully active recombinant tobacco cytosolic NADP-dependent isocitrate dehydrogenase in Escherichia coli: evidence for a role for the N-terminal region in enzyme activity.

The recently isolated full-length NADP-dependent isocitrate dehydrogenase (ICDH) cDNA encoding the tobacco cytosolic isoenzyme has been cloned into the expression vector pET8c and used to transform Escherichia coli strain BL21 (DE3). The recombinant protein was purified to electrophoretic homogeneity and used to raise polyclonal antibodies. Its kinetic properties were found to be identical to those of the cytosolic ICDH isoenzyme purified from tobacco cell cultures. The recombinant and the endogenous bacterial ICDH could be easily distinguished by their different behaviors during anion-exchange column chromatography and immunological response. An incomplete ICDH-encoding cDNA clone, encoding a protein lacking the first 36 amino acids at the N-terminus, was cloned into the expression vector pKK233-2 and used to transform ICDH-lacking E. coli cells (strain 2004). The truncated, recombinant ICDH produced by the bacteria was found to be inactive.