IRplus: An Augmented Tool to Detect Inverted Repeats in Plastid Genomes.

High-throughput sequencing methods have increased the accessibility of plastid genomes, which are crucial for clarifying phylogenetic relationships. Current large sequencing efforts require software tools for routine display of their distinctive quadripartite structure, which is denoted by four junction sites. By concentrating on these junctions and their close vicinity, IRscope has emerged as the standard tool for detection of this structure and creating simplified comparative graphical maps of plastid genomes. Here, we provide an augmented version (IRplus) that encompasses a novel set of functions such as integrated error detection, flexible color schemes, and an upgraded method to detect inverted repeats in genomic sequences. Spanning across the plant tree of life, IRplus allows the quick visualization of various sets of plastid genomes and features, next to smooth interoperability with other widely used annotation file formats and platforms. The IRplus can be accessed at https://irscope.shinyapps.io/IRplus/, and source codes are freely available at https://github.com/AmiryousefiLab/IRplus.

Fructooligosaccharides production by immobilized Pichia pastoris cells expressing Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase.

Fructooligosaccharides (FOSs)-fructose-based oligosaccharides-are typical prebiotics with health-promoting effects in humans and animals. The trisaccharide 1-kestotriose is the most attractive inulin-type FOS. We previously reported a recombinant sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4.1.99) from Schedonorus arundinaceus (Sa) that efficiently converts sucrose into 1-kestotriose. In this study, Pichia pastoris PGFT6x-308 constitutively expressing nine copies of the Sa1-SST gene displayed fructosyltransferase activity in undisrupted biomass (49.8 U/ml) and culture supernatant (120.7 U/ml) in fed-batch fermentation (72 hr) with sugarcane molasses. Toluene permeabilization increased 2.3-fold the Sa1-SSTrec activity of whole cells entrapped in calcium-alginate beads. The reaction with refined or raw sugar (600 g/l) yielded 1-kestotriose and 1,1-kestotetraose in a ratio of 8:2 with their sum representing above 55% (wt/wt) of total carbohydrates. The FOSs yield decreased to 45% (wt/wt) when sugarcane syrup and molasses were used as cheaper sucrose sources. The beads retained 80% residual Sa1-SSTrec activity after a 30-day batchwise operation with refined cane sugar at 30°C and pH 5.5. The immobilized biocatalyst is attractive for the continuous production of short-chain FOSs, most particularly 1-kestotriose.

Spanish validation of the Parental Psychological Control Scale and Parental Psychological Control-Disrespect Scale / Validación española de la Escala de Control Psicológico Parental y la Escala Control Psicológico-Falta de Respeto

Abstract The construct "parental psychological control" has been used to refer to those parental behaviors that interfere with the children's thoughts and feelings. Examples of this type of behaviors would be inducing guilt or shame as well as the withdrawal of affection. The objective of this study was to validate and adapt the Spanish version of the Psychological Control Scale-Youth Self-Report (PCS-YSR) and the Psychological Control-Disrespect Scale (PCDS) and to analyze if the parental psychological control is defined as one dimension or two dimensions through manipulative and disrespectful behaviors towards adolescents. Also, the unique contribution of parental psychological control in predicting adolescent's self-concept was examined. In two samples of undergraduate university students (the first, N = 367, mean age = 19.8, women=62.7%, men=37.3%, and the second, N= 312, mean age=19.7, women=73.9%, men=26.1%), exploratory and confirmatory factor analyses confirmed the hypothesized factor structure of the PCS-YSR and the PCDS, for both paternal and maternal ratings. Convergent validity was confirmed by consistent associations between both measures of parental psychological control and other measures of parental autonomy support and parental psychological control. The multi-group analysis confirmed that parental psychological control had a negative influence on self-concept. The results indicated that both scales are useful instruments for assessing parental psychological control.

Resumen El constructo "control psicológico parental" se ha utilizado para referirse a los comportamientos de los progenitores que interfieren con los pensamientos y sentimientos de los niños y niñas. Ejemplos de este tipo de comportamientos son la inducción de culpa o vergüenza, así como la retirada de afecto. Nuestro estudio tuvo como objetivo validar y adaptar la versión española de la Escala de Control Psicológico-Autoinforme Adolescente (PCS-YSR) y la Escala Control Psicológico-Falta de Respeto (PCDS), y analizar si el control psicológico parental se define como una dimensión o dos dimensiones a través de conductas manipuladoras e irrespetuosas hacia los adolescentes. Además, examinamos la contribución única del control psicológico parental en la predicción del autoconcepto del adolescente. En dos muestras de estudiantes universitarios de pregrado (la primera n = 367, edad media = 19.8, mujeres = 62.7%, hombres = 37.3%, y la segunda, n = 312, edad media = 19.7, mujeres = 73.9%, hombres = 26.1%), los análisis factoriales exploratorios y confirmatorios confirmaron la estructura factorial hipotética del PCS-YSR y del PCDS, tanto para las puntuaciones paterna como materna. La validez convergente fue confirmada por asociaciones consistentes entre ambas medidas de control psicológico parental y otras medidas de apoyo parental a la autonomía y control psicológico parental. El análisis multigrupo confirmó que el control psicológico de los progenitores influyó negativamente en el autoconcepto. Los resultados indicaron que ambas escalas son instrumentos útiles para evaluar el control psicológico parental.

Removal of bacterial dextran in sugarcane juice by Talaromyces minioluteus dextranase expressed constitutively in Pichia pastoris.

A gene construct encoding the mature region of Talaromyces minioluteus dextranase (EC 3.2.1.11) fused to the Saccharomyces cerevisiae SUC2 signal sequence was expressed in Pichia pastoris under the constitutive glyceraldehyde 3-phosphate dehydrogenase promoter (pGAP). The increase of the transgene dosage from one to two and four copies enhanced proportionally the extracellular yield of the recombinant enzyme (r-TmDEX) without inhibiting cell growth. The volumetric productivity of the four-copy clone in fed batch fermentation (51 h) using molasses as carbon source was 1706 U/L/h. The secreted N-glycosylated r-TmDEX was optimally active at pH 4.5-5.5 and temperature 50-60 °C. The addition of sucrose (600 g/L) as a stabilizer retained intact the r-TmDEX activity after 1-h incubation at 50-60 °C and pH 5.5. Bacterial dextran in deteriorated sugarcane juice was completely removed by applying a crude preparation of secreted r-TmDEX. The high yield of r-TmDEX in methanol-free cultures and the low cost of the fed batch fermentation make the P. pastoris pGAP-based expression system appropriate for the large scale production of dextranase and its use for dextran removal at sugar mills.

Validation of the Dependency-Oriented and Achievement-Oriented Psychological Control Scale (DAPCS) in a Spanish-speaking late adolescent sample / Validación de la Escala de Control Psicológico Orientado a la Dependencia y al Logro (DAPCS) en una Muestra de adolescentes tardíos que hablan español

Parental psychological control refers to those parental behaviors that interfere with the child's thoughts and feelings and are characterized by excessive use of manipulative techniques, such as inducing guilt or shame and love-withdrawal. This study was aimed at validating the Spanish version of the Dependency-oriented and Achievement-oriented Psychological Control Scale (DAPCS), a new measure designed to assess an adolescent's perception of psychological control. Another objective was to determine the influence of perceived parental psychological control on adolescents' self-concept. We evaluated the factor structure, reliability, and convergent validity in a sample of Spanish late adolescents (N = 310, mean age = 19.75). Exploratory and confirmatory factor analyses showed that the DAPCS factor structure is best described by a 2-factor solution, for both paternal and maternal ratings, indicating the distinction between dependency-oriented and achievement-oriented psychological control. High indices of internal consistency indicated that DAPCS produced reliable scores. Moreover, convergent validity was confirmed by consistent associations between the DAPCS and other measures of psychological control and other parenting dimensions. Hierarchical regression also showed that DAPCS predicted late adolescents' self-concept. The results of this study indicate that the Spanish version of the DAPCS is a useful instrument for studying both types of psychological control

El control psicológico parental se refiere a los comportamientos de los padres que interfieren con los Pensamientos y sentimientos del niño y se caracterizan por el uso excesivo de técnicas de manipulación, Como la inducción de culpa o vergüenza y la retirada del afecto. Este estudio tuvo como objetivo validar. La versión en español de la Dependency-oriented and Achievement - oriented Psychological Control Scale (DAPCS) (Escala de control psicológico orientado a la dependencia y control psicológico orientado Al logro), una nueva medida diseñada para evaluar la percepción del control psicológico de un adolescente. Otro objetivo fue determinar la influencia del control psicológico parental percibido sobre el Autoconcepto de los adolescentes. Evaluamos la estructura factorial, la fiabilidad y la validez convergente en una muestra de adolescentes tardíos españoles (N=310, edad media = 19.75). Los análisis de factores Exploratorios y confirmatorios mostraron que la estructura factorial de la escala DAPCS se describe mejor con una solución de 2 factores, tanto para padres como para madres, lo que indica la distinción entre control psicológico orientado a la dependencia y el orientado al logro. Los altos índices de consistencia interna indicaron que la escala DAPCS produjo óptimas puntuaciones. Además, la validez convergente fue confirmada por la correlación entre el DAPCS y otras medidas de control psicológico, así como con otras medidas del análisis de la crianza. La regresión jerárquica también mostró que DAPCS predijo. El autoconcepto de los adolescentes tardíos. Los resultados de este estudio indican que la versión en Español del DAPCS es un instrumento útil para estudiar ambos tipos de control psicológico

Epigenetic Deregulation of Protocadherin PCDHGC3 in Pheochromocytomas/Paragangliomas Associated With SDHB Mutations.

CONTEXT: SDHB mutations are found in an increasing number of neoplasms, most notably in paragangliomas and pheochromocytomas (PPGLs). SDHB-PPGLs are slow-growing tumors, but ∼50% of them may develop metastasis. The molecular basis of metastasis in these tumors is a long-standing and unresolved problem. Thus, a better understanding of the biology of metastasis is needed. OBJECTIVE: This study aimed to identify gene methylation changes relevant for metastatic SDHB-PPGLs. DESIGN: We performed genome-wide profiling of DNA methylation in diverse clinical and genetic PPGL subtypes, and validated protocadherin γ-C3 (PCDHGC3) gene promoter methylation in metastatic SDHB-PPGLs. RESULTS: We define an epigenetic landscape specific for metastatic SDHB-PPGLs. DNA methylation levels were found significantly higher in metastatic SDHB-PPGLs than in SDHB-PPGLs without metastases. One such change included long-range de novo methylation of the PCDHA, PCDHB, and PCDHG gene clusters. High levels of PCDHGC3 promoter methylation were validated in primary metastatic SDHB-PPGLs, it was found amplified in the corresponding metastases, and it was significantly correlated with PCDHGC3 reduced expression. Interestingly, this epigenetic alteration could be detected in primary tumors that developed metastasis several years later. We also show that PCDHGC3 down regulation engages metastasis-initiating capabilities by promoting cell proliferation, migration, and invasion. CONCLUSIONS: Our data provide a map of the DNA methylome episignature specific to an SDHB-mutated cancer and establish PCDHGC3 as a putative suppressor gene and a potential biomarker to identify patients with SDHB-mutated cancer at high risk of metastasis who might benefit from future targeted therapies.

Engineered thermostable ß-fructosidase from Thermotoga maritima with enhanced fructooligosaccharides synthesis.

The thermostable ß-fructosidase (BfrA) from the bacterium Thermotoga maritima converts sucrose into glucose, fructose, and low levels of short-chain fructooligosaccharides (FOS) at high substrate concentration (1.75 M) and elevated temperatures (60-70 °C). In this research, FOS produced by BfrA were characterized by HPAE-PAD analysis as a mixture of 1-kestotriose, 6G-kestotriose (neokestose), and to a major extent 6-kestotriose. In order to increase the FOS yield, three BfrA mutants (W14Y, W14Y-N16S and W14Y-W256Y), designed from sequence divergence between hydrolases and transferases, were constructed and constitutively expressed in the non-saccharolytic yeast Pichia pastoris. The secreted recombinant glycoproteins were purified and characterized. The three mutants synthesized 6-kestotriose as the major component of a FOS mixture that includes minor amounts of tetra- and pentasaccharides. In all cases, sucrose hydrolysis was the predominant reaction. All mutants reached a similar overall FOS yield, with the average value 37.6% (w/w) being 3-fold higher than that of the wild-type enzyme (12.6%, w/w). None of the mutations altered the enzyme thermophilicity and thermostability. The single mutant W14Y, with specific activity of 841 U mg-1, represents an attractive candidate for the continuous production of FOS-containing invert syrup at pasteurization temperatures.

Levansucrase from Halomonas smyrnensis AAD6T: first halophilic GH-J clan enzyme recombinantly expressed, purified, and characterized.

Fructans, homopolymers of fructose produced by fructosyltransferases (FTs), are emerging as intriguing components in halophiles since they are thought to be associated with osmotic stress tolerance and overall fitness of microorganisms and plants under high-salinity conditions. Here, we report on the full characterization of the first halophilic FT, a levansucrase from Halomonas smyrnensis AAD6T (HsLsc; EC 2.4.1.10). The encoding gene (lsc) was cloned into a vector with a 6xHis Tag at its C-terminus, then expressed in Escherichia coli. The purified recombinant enzyme (47.3 kDa) produces levan and a wide variety of fructooligosaccharides from sucrose, but only in the presence of high salt concentrations (> 1.5 M NaCl). HsLsc showed Hill kinetics and pH and temperature optima of 5.9 and 37 °C, respectively. Interestingly, HsLsc was still very active at salt concentrations close to saturation (4.5 M NaCl) and was selectively inhibited by divalent cations. The enzyme showed high potential in producing novel saccharides derived from raffinose as both fructosyl donor and acceptor and cellobiose, lactose, galactose, and ʟ-arabinose as fructosyl acceptors. With its unique biochemical characteristics, HsLsc is an important enzyme for future research and potential industrial applications in a world faced with drought and diminishing freshwater supplies.

Fructooligosaccharides production by Schedonorus arundinaceus sucrose:sucrose 1-fructosyltransferase constitutively expressed to high levels in Pichia pastoris.

The non-saccharolytic yeast Pichia pastoris was engineered to express constitutively the mature region of sucrose:sucrose 1-fructosyltransferase (1-SST, EC 2.4.1.99) from Tall fescue (Schedonorus arundinaceus). The increase of the transgene dosage from one to nine copies enhanced 7.9-fold the recombinant enzyme (Sa1-SSTrec) yield without causing cell toxicity. Secretion driven by the Saccharomyces cerevisiae α-factor signal peptide resulted in periplasmic retention (38%) and extracellular release (62%) of Sa1-SSTrec to an overall activity of 102.1 U/ml when biomass reached (106 g/l, dry weight) in fed-batch fermentation using cane sugar for cell growth. The volumetric productivity of the nine-copy clone PGFT6x-308 at the end of fermentation (72 h) was 1422.2 U/l/h. Sa1-SSTrec purified from the culture supernatant was a monomeric glycoprotein optimally active at pH 5.0-6.0 and 45-50 °C. The removal of N-linked oligosaccharides by Endo Hf treatment decreased the enzyme stability but had no effect on the substrate and product specificities. Sa1-SSTrec converted sucrose (600 g/l) into 1-kestose (GF2) and nystose (GF3) in a ratio 9:1 with their sum representing 55-60% (w/w) of the total carbohydrates in the reaction mixture. Variations in the sucrose (100-800 g/l) or enzyme (1.5-15 units per gram of substrate) concentrations kept unaltered the product profile. Sa1-SSTrec is an attractive candidate enzyme for the industrial production of short-chain fructooligosaccharides, most particularly 1-kestose.

Parental control in interpersonal acceptance-rejection theory: a study with a Spanish sample using Parents' Version of Parental Acceptation-Rejection/Control Questionnaire / Control parental en la teoría de la aceptación-rechazo interpersonal: un estudio con una muestra española usando la Versión de los Padres del Cuestionario de Aceptación / Control Parental

Interpersonal Acceptance-Rejection theory (IPARTheory) provides a good theoretical framework to explain the parenting dimensions that influence children's social, cognitive and emotional adjustment. This theory develops one main dimension, warmth-rejection, where warmth would be one pole of the dimension and rejection the opposite one. Besides, the theory has also defined behavioral control dimension with two poles: permissiveness-strictness. In the context of this theory, our study was conducted with a sample of Spanish parents to (a) examine whether parental perceived acceptance-rejection was related to parental behavioral control; (2) analyze how behavioral control dimension worked in acceptance-rejection theory; and (3) examine whether there were differences in the relations obtained between fathers and mothers. We have to conclude that control behavior correlates positively with warmth behaviors. Fathers' and mothers' correlations among these dimensions show that parents can develop certain control behaviors but that they are compatible with affection conducts (AU)

Control parental en la teoría de la aceptación-rechazo interpersonal: un estudio con una muestra española usando la Versión de los Padres del Cuestionario de Aceptación / Control Parental. La teoría de la aceptación - rechazo interpersonal (IPART) proporciona un adecuado marco teórico para explicar las diversas dimensiones parentales que influencian el ajuste social, cognitivo y emocional de los niños. En efecto, esta teoría desarrolla una dimensión principal, cariño - rechazo, en la que el cariño se encontraría situado en un extremo del continuo y el rechazo en el opuesto. Además, esta teoría también define el control parental tomando como referencia dos polos: el control permisivo y el restrictivo. En el contexto de esta teoría, nuestro estudio ha sido llevado a cabo con una muestra de padres para (a) examinar en qué medida los padres y las madres perciben la aceptación - rechazo como elementos integrantes de su comportamiento de control parental (b) analizar cómo funciona la dimensión control dentro de la teoría IPART (c) examinar si se observan diferencias en las relaciones observadas en los padres y en las madres. Se concluye que el control de conductas correlaciona positivamente con las conductas de cariño. Las correlaciones de padres y madres en estas dimensiones muestran que los padres pueden desarrollar ciertos comportamientos de control pero que éstos son compatibles con las conductas afectivas (AU)

Cervix tuberculosis simulating cancer / Tuberculosis de cérvix: simulando una neoplasia

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Scaling-up batch conditions for efficient sucrose hydrolysis catalyzed by an immobilized recombinant Pichia pastoris cells in a stirrer tank reactor

Background: Invert sugar is used greatly in food and pharmaceutical industries. This paper describes scaling-up batch conditions for sucrose inversion catalyzed by the recombinant Pichia pastoris BfrA4X whole cells expressing Thermotoga maritima invertase entrapped in calcium alginate beads. For the first time, we describe the application of a kinetic model to predict the fractional conversion expected during sucrose hydrolysis reaction in both, a model and a prototype bioreactor with 0.5- and 5-L working volume, respectively. Results: Different scaled-up criteria used to operate the 0.5-L bioreactor were analyzed to explore the invert sugar large scale production. After model inversion studies, a 5-L scaled-up reaction system was performed in a 7-L stirred reactor. Both scaled-up criteria, immobilized biocatalyst dosage and stirring speed, were analyzed in each type of bioreactors and the collected data were used to ensure an efficient scale-up of this biocatalyst. Conclusions: To date, there is not enough information to describe the large-scale production of invert sugar using different scaled-up criteria such as dose of immobilized biocatalyst and stirring speed effect on mass transfer. The present study results constitute a valuable tool to successfully carry out this type of high-scale operation for industrial purposes.

T1 high-grade bladder carcinoma outcome: the role of p16, topoisomerase-IIα, survivin, and E-cadherin.

High-grade papillary urothelial carcinoma with subepithelial connective tissue invasion (T1HG) is an aggressive disease at high risk of progression after transurethral resection/Bacillus Calmette-Guerin standardized therapy. The European Organization for Research and Treatment of Cancer has identified T1HG bladder carcinoma that is single and ≤3 cm in the largest dimension at first diagnosis as a category in which the prognosis cannot be further stratified based on conventional criteria. This category may benefit from biomarker analysis as a valuable tool to determine the patient's outcome. To further the issue of biomarkers in predicting aggressiveness in single T1HG bladder carcinoma ≤3 cm in greatest dimension at first diagnosis, we have conducted a validation study of the biomarker risk score set previously reported by our group. The study set included immunohistochemical detection of galectin-3, CD44, E-cadherin (E-CAD), CD138, p16, survivin, HYAL-1, and topoisomerase-IIα in 92 randomly selected specimens at participating institutions. Topoisomerase-IIα expression was identified as a predictor of disease-free survival. p16, survivin, and E-CAD expression predicted progression-free survival, but p16 and E-CAD also predicted overall survival. The current study validates a panel of immunohistochemical markers with the potential of being implemented in practice and supports the use of biomarkers in predicting aggressiveness in patients with first diagnosis of single T1HG bladder carcinoma ≤3 cm in greatest dimension and therefore in identifying patients who need closer surveillance or earlier aggressive treatment.

A comparative molecular dynamics study of thermophilic and mesophilic ß-fructosidase enzymes.

Arabidopsis thaliana cell wall invertase 1 (AtcwINV1) and Thermotoga maritima ß-fructosidase (BfrA) are among the best structurally studied members of the glycoside hydrolase family 32. Both enzymes hydrolyze sucrose as the main substrate but differ strongly in their thermal stability. Mesophilic AtcwINV1 and thermophilic BfrA have divergent sequence similarities in the N-terminal five bladed ß-propeller catalytic domain (31 %) and the C-terminal ß-sandwich domain (15 %) of unknown function. The two enzymes were subjected to 200 ns molecular dynamics simulations at 300 K (27 °C) and 353 K (80 °C). Regular secondary structure regions, but not loops, in AtcwINV1 and BfrA showed no significant fluctuation differences at both temperatures. BfrA was more rigid than AtcwINV1 at 300 K. The simulation at 353 K did not alter the structural stability of BfrA, but did increase the overall flexibility of AtcwINV1 exhibiting the most fluctuating regions in the ß-propeller domain. The simulated heat treatment also increased the gyration radius and hydrophobic solvent accessible surface area of the plant enzyme, consistent with the initial steps of an unfolding process. The preservation of the conformational rigidity of BfrA at 353 K is linked to the shorter size of the protein loops. Shortening of BfrA loops appears to be a key mechanism for thermostability.

Complicaciones asociadas al catéter venoso central en pacientes hematológicos / Complications associated to central venous catheters in hematology patients

OBJETIVO: Conocer la incidencia de complicaciones asociadas a los catéteres venosos centrales (tunelizados, reservorios y PICC), en pacientes con patología oncohematológica, ingresados en Unidades de Hematología o Trasplantes de Progenitores Hematopoyéticos, en dos hospitales terciarios. METODOLOGÍA: Se desarrolló un estudio descriptivo transversal donde se recogieron datos sociodemográficos, clínicos, complicaciones y seguimiento del protocolo de cuidados. A cada catéter se le asignó un número de identificación correlativo. Se recogió información de 366 catéteres: 185 en el Hospital Universitario Ramón y Cajal (HURYC): 80 tunelizados, 40 reservorios y 65 PICC; 181 en el Hospital Universitario Gregorio Marañón (HUGM): 101 tunelizados y 80 reservorios. RESULTADOS: Las principales complicaciones en los tunelizados fueron las infecciones (13,7% en el HURYC vs.6,8% en el HUGM; p < 0,001) y las oclusiones (al menos una vez 3,8% vs.21,8%). En los reservorios se confirmaron un 5% de infecciones en el HURYC frente a 1,2% en el HUGM; se ocluyeron al menos una vez un 10% en el HUGM. No se detectaron otras complicaciones significativas. Respecto a los PICC solo se recogió información en el HURYC, donde las complicaciones fueron: flebitis 10,8%; trombosis 7,7%; infección o sospecha 4,6%; oclusión al menos una vez 7,7%. CONCLUSIONES: La diferencia entre hospitales respecto a la infección y oclusión, puede asociarse a las distintas pautas de cuidados. Destaca la alta incidencia de flebitis y trombosis en PICC

OBJECTIVE: To discover the incidence of central venous catheters (tunnelled, subcutaneous and PICC) in patients with onco-hematological conditions, hospitalized in the Hematology or Transplantations of Hematopoietic Stem Cells Units, in two tertiary care hospitals. METHODOLOGY: A cross-sectional, descriptive study form was developed in order to gather sociodemographic, clinical data as well as complications and follow-up of the care protocol. Each catheter was assigned a correlative identification number. Information was collected on 366 catheters: 185 in the University Hospital Ramón y Cajal (HURYC), 80 tunnelled, 40 subcutaneous venous access and 65 PICC, and 181 in the University Hospital Gregorio Marañón (HUGM), 101 tunnelled and 80 subcutaneous venous access. Findings: Major complications in the tunnellized were infections (13.7% in HURYC vs.6.8% in HUGM - p < 0 .001) and occlusions (at least once in 3.8% vs.21.8%). In subcutaneous venous access, infections were confirmed in 5% in HURYC vs.1.2% in HUGM. There were occlusions at least once in 10% in HUGM and no other significant complications were detected. Regarding PICC, information was only collected in HURYC, where complications were phlebitis 10.8%, thrombosis 7.7%, confirmed or suspected infection 4.6%, occlusion at least once 7.7%. CONCLUSIONS: Differences between hospitals with regard to major complications, infection and occlusion may be related to different care protocol. We need to stress the high incidence of phlebitis and thrombosis in PICC catheters, compared with data of lower incidence of other papers

[Complications associated to central venous catheters in hematology patients]. / Complicaciones asociadas al catéter venoso central en pacientes hematológicos.

OBJECTIVE: To discover the incidence of central venous catheters (tunnelled, subcutaneous and PICC) in patients with onco-hematological conditions, hospitalized in the Hematology or Transplantations of Hematopoietic Stem Cells Units, in two tertiary care hospitals. METHODOLOGY: A cross-sectional, descriptive study form was developed in order to gather sociodemographic, clinical data as well as complications and follow-up of the care protocol. Each catheter was assigned a correlative identification number. Information was collected on 366 catheters: 185 in the University Hospital Ramón y Cajal (HURYC), 80 tunnelled, 40 subcutaneous venous access and 65 PICC, and 181 in the University Hospital Gregorio Marañón (HUGM), 101 tunnelled and 80 subcutaneous venous access. FINDINGS: Major complications in the tunnellized were infections (13.7% in HURYC vs. 6.8% in HUGM - p<0.001) and occlusions (at least once in 3.8% vs. 21.8%). In subcutaneous venous access, infections were confirmed in 5% in HURYC vs. 1.2% in HUGM. There were occlusions at least once in 10% in HUGM and no other significant complications were detected. Regarding PICC, information was only collected in HURYC, where complications were phlebitis 10.8%, thrombosis 7.7%, confirmed or suspected infection 4.6%, occlusion at least once 7.7%. CONCLUSIONS: Differences between hospitals with regard to major complications, infection and occlusion may be related to different care protocol. We need to stress the high incidence of phlebitis and thrombosis in PICC catheters, compared with data of lower incidence of other papers.

The flavonoid quercetin reverses pulmonary hypertension in rats.

Quercetin is a dietary flavonoid which exerts vasodilator, antiplatelet and antiproliferative effects and reduces blood pressure, oxidative status and end-organ damage in humans and animal models of systemic hypertension. We hypothesized that oral quercetin treatment might be protective in a rat model of pulmonary arterial hypertension. Three weeks after injection of monocrotaline, quercetin (10 mg/kg/d per os) or vehicle was administered for 10 days to adult Wistar rats. Quercetin significantly reduced mortality. In surviving animals, quercetin decreased pulmonary arterial pressure, right ventricular hypertrophy and muscularization of small pulmonary arteries. Classic biomarkers of pulmonary arterial hypertension such as the downregulated expression of lung BMPR2, Kv1.5, Kv2.1, upregulated survivin, endothelial dysfunction and hyperresponsiveness to 5-HT were unaffected by quercetin. Quercetin significantly restored the decrease in Kv currents, the upregulation of 5-HT2A receptors and reduced the Akt and S6 phosphorylation. In vitro, quercetin induced pulmonary artery vasodilator effects, inhibited pulmonary artery smooth muscle cell proliferation and induced apoptosis. In conclusion, quercetin is partially protective in this rat model of PAH. It delayed mortality by lowering PAP, RVH and vascular remodeling. Quercetin exerted effective vasodilator effects in isolated PA, inhibited cell proliferation and induced apoptosis in PASMCs. These effects were associated with decreased 5-HT2A receptor expression and Akt and S6 phosphorylation and partially restored Kv currents. Therefore, quercetin could be useful in the treatment of PAH.

Pyonephrosis as a sign of sarcomatoid carcinoma of the renal pelvis.

We report the case of an urgent nephrectomy because of a pyonephrosis and sepsis due to an unsuspected sarcomatoid transitional cell carcinoma, an infrequent subtype with a bad oncological prognosis. We present a 58-year-old man assessed by internal medicine for a general syndrome and weakness many months previously. A pyonephrotic kidney was observed at abdominal computed tomography in the context of septic shock, without suspecting the underlying cause. The pathology report described a sarcomatoid transitional cell carcinoma. Sarcomatoid transitional cell carcinoma is an invasive and infrequent subtype of urothelial tumors. The symptoms are often the same as other renal masses; however, in this case, sepsis and pyonephrosis were the rare initial symptoms.

Complete sucrose hydrolysis by heat-killed recombinant Pichia pastoris cells entrapped in calcium alginate.

BACKGROUND: An ideal immobilized biocatalyst for the industrial-scale production of invert sugar should stably operate at elevated temperatures (60-70°C) and high sucrose concentrations (above 60%, w/v). Commercial invertase from the yeast Saccharomyces cerevisiae is thermolabile and suffers from substrate inhibition. Thermotoga maritima ß-fructosidase (BfrA) is the most thermoactive and thermostable sucrose-hydrolysing enzyme so far identified and allows complete inversion of the substrate in highly concentrated solutions. RESULTS: In this study, heat-killed Pichia pastoris cells bearing N-glycosylated BfrA in the periplasmic space were entrapped in calcium alginate beads. The immobilized recombinant yeast showed maximal sucrose hydrolysis at pH 5-7 and 90°C. BfrA was 65% active at 60°C and had no activity loss after incubation without the substrate at this temperature for 15 h. Complete inversion of cane sugar (2.04 M) at 60°C was achieved in batchwise and continuous operation with respective productivities of 4.37 and 0.88 gram of substrate hydrolysed per gram of dry beads per hour. The half-life values of the biocatalyst were 14 and 20 days when operated at 60°C in the stirred tank and the fixed-bed column, respectively. The reaction with non-viable cells prevented the occurrence of sucrose fermentation and the formation of by-products. Six-month storage of the biocatalyst in 1.46 M sucrose (pH 5.5) at 4°C caused no reduction of the invertase activity. CONCLUSIONS: The features of the novel thermostable biocatalyst developed in this study are more attractive than those of immobilized S. cerevisiae cells for application in the enzymatic manufacture of inverted sugar syrup in batch and fixed-bed reactors.

Clinicopathological characteristics and outcome of nested carcinoma of the urinary bladder.

We present the clinicopathological features of 56 cases of the nested variant of urothelial bladder carcinoma. This is an uncommon variant of bladder cancer, recognized by the current WHO classification of urologic tumors. The nested component represented 100 % of the tumor in 24 cases. The architectural pattern of the tumor varied from solid expansile to infiltrative nests characterized by deceptively bland histologic features resembling von Brunn nests. Typical features of high-grade conventional urothelial carcinoma were present in 32 cases. Most neoplastic cells had nuclei of low to intermediate nuclear grade with occasional nuclear enlargement, most frequently seen in deep areas of tumor. The nested component expressed cytokeratins 7, 20, CAM5.2, and high molecular weight (34ßE12), p63, Ki67, p53, p27, and GATA3. Tumor extension was T1 (n = 9), minimally T2 (n = 10), T2a (n = 1), T2b (n = 4), T3a (n = 8), T3b (n = 13), and T4a (n = 11). On follow-up, 36 of patients died of or were alive with disease from 2 to 80 months (mean 21 months). Four patients died of other causes. Eleven other patients remained disease free. Univariate survival analysis showed no differences for nested carcinoma compared with conventional urothelial carcinoma. As in conventional urothelial carcinoma, in nested carcinoma of the bladder pT category defined different survival groups. In summary, nested variant of urothelial bladder carcinoma is typically associated with advanced stage. In samples of limited volume, it may be misdiagnosed as proliferation of von Brunn nests or other nested-like bladder lesions, delaying definitive therapy.