Avaliação de atividades endoglucanase, exoglucanase, lacase e lignina peroxidase em dez fungos da podridão branca

Este trabajo presenta una vía de rastreo de producción de enzimas lignocelulolíticas en diez especies de hongos de pudrición blanca: Lentinula edodes, Schizophyllum commune, Trametes trogii, Coriolus versicolor, Pycnoporus sanguineus, Ganoderma applanatum, Ganoderma lucidum, Grifola frondosa, Pleurotus ostreatus y Auricularia delicata. Estas especies primero fueron rastreadas sobre medios de cultivo sólido que contienen carboximetil celulosa, celulosa cristalina, ABTS (2,2´-azino-bis(3-etilbenzotiazolina-6-sulfonato)) y azure B, los cuales evidenciaron la producción de las enzimas endoglucanasa, exoglucanasa, lacasa y lignina peroxidasa (LiP). Las actividades celulolíticas fueron detectadas a los cinco días de incubación con el indicador rojo congo, formándose un halo claro-blanco en las zonas donde se degrada la celulosa. Para las ligninasas, este rastreo consistió en el seguimiento a la formación de halos verdes por oxidación del ABTS para lacasa y halos de decoloración sobre el azure B para la LiP durante 14 días de incubación. De este rastreo cualitativo, se seleccionaron cuatro cepas (G. lucidum, L. edodes, C. versicolor y T. Trogii), como las mejores productoras de enzimas celulolíticas y ligninolíticas. Estas cuatro especies fueron inoculadas sobre un sustrato de aserrín de roble, obteniéndose 51,8% de lignina degradada por L. edodes y 22% de celulosa degrada por C. versicolor.This paper presents a way of tracking the production of lignocellulolytic enzymes in ten species of white rot fungi: Lentinula edodes, Schizophyllum commune, Trametes trogii, Coriolus versicolor, Pycnoporus sanguineus, Ganoderma applanatum, Ganoderma lucidum, Grifola frondosa, Pleurotus ostreatus and Auricularia delicata. These species were first screened on solid culture media containing carboxymethyl cellulose, crystalline cellulose, ABTS (2,2´-azino-bis(3-ethylbenzothiazoline-6-sulphonate)) and azure B, which showed the production of endoglucanase, exoglucanase, laccase and lignin peroxidase (LiP) enzymes. Cellulolytic activities were detected after five days of incubation with congo red indicator, forming a clear-white halo in areas where cellulose was degraded. For ligninases, the tracking consisted of the monitoring in the formation of green halos due to ABTS oxidation for laccase, and decolorization halos on azure B for LiP during 14 days of incubation. From this qualitative screening, four strains were selected (G. lucidum, L. edodes, C. versicolor and T. trogii) as the best producers of cellulolytic and ligninolytic enzymes. These four species were inoculated on a substrate of sawdust oak, yielding 51,8% of lignin degraded by L. edodes and 22% of cellulose degraded by C. versicolor.Este trabalho apresenta uma maneira de seguir produção da enzimas lignocelulolíticas em 10 espécies de fungos da podridão branca: Lentinula edodes, Schizophyllum commune, Trametes trogii, Coriolus versicolor, Pycnoporus sanguineus, Ganoderma applanatum, Ganoderma lucidum, Grifola frondosa, Pleurotus ostreatus e Auricularia delicata. Estas espécies foram primeiramente rastreados em meio de cultura sólido contendo os reagentes de carboximetil celulose, celulose cristalina, ABTS (2,2 ‘-azinobis (3-etilbenztiazoline-6-sulfonato)) e azure B que mostrou a produção de enzimas de endoglucanase, exoglucanase lacase, peroxidase de lignina e (LiP). Actividades celulolíticas foram detectados após cinco dias de incubação com o indicador vermelho congo, formando um claro halo-branco em áreas degrada a celulose. Para ligninases, esta consistia de rastreamento faixa formação de halo verde por oxidação de ABTS a lacase e descoloração halos no lábio azure B, durante 14 dias de incubação. Esta triagem qualitativa, foram selecionados quatro cepas: G. lucidum, L. edodes, C. versicolor e T. trogii, cepas como os melhores produtores de enzimas celulolíticas e ligninolíticas. Estas quatro espécies foram inoculados sobre um substrato de madeira de carvalho serradura, obtendo-se 51,8% de lignina por L. edodes degradad e 22% de celulose degradada por C. versicolor.Fil: Montoya, Sandra. Universidad de Caldas; ColombiaFil: Sánchez, Oscar Julián. Universidad de Caldas; ColombiaFil: Levin, Laura Noemi. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Micología y Botánica. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Micología y Botánica; Argentin

Development and evaluation of a new survey instrument to measure the quality of colorectal cancer screening decisions

Background: Guidelines for colorectal cancer screening recommend that patients be informed about options and be able to select preferred method of screening; however, there are no existing measures available to assess whether this happens. Methods: Colorectal Cancer Screening Decision Quality Instrument (CRC-DQI) includes knowledge items and patients' goals and concerns. Items were generated through literature review and qualitative work with patients and providers. Hypotheses relating to the acceptability, feasibility, discriminant validity and retest reliability of the survey were examined using data from three studies: (1) 2X2 randomized study of participants recruited online, (2) cross-sectional sample of patients recruited in community health clinics, and (3) cross-sectional sample of providers recruited from American Medical Association Master file. Results: 338 participants were recruited online, 94 participants were recruited from community health centers, and 115 physicians were recruited. The CRC-DQI was feasible and acceptable with low missing data and high response rates for both online and paper-based administrations. The knowledge score was able to discriminate between those who had seen a decision aid or not (84% vs. 64%, p < 0.001) and between providers, online patients and clinic patients (89% vs. 74% vs. 41%, p < 0.001 for all comparisons). The knowledge score and most of the goals had adequate retest reliability. About half of the participants received a test that matched their goals (47% and 51% in online and clinic samples respectively). Many respondents who had never been screened had goals that indicated a preference for colonoscopy. A minority of respondents in the online (21%) and in clinic (2%) samples were both well informed and received a test that matched their goals. Conclusions: The CRC-DQI demonstrated good psychometric properties in diverse samples, and across different modes of administration. Few respondents made high quality decisions about colon cancer screening

Molecular diagnosis and typing of Trypanosoma cruzi populations and lineages in cerebral Chagas disease in a patient with AIDS

Trypanosoma cruzi DNA was amplified from an intracranial biopsy and peripheral blood of an HIV patient with encephalitis; this episode was indicative of AIDS and congenital Chagas disease. The analysis of a microsatellite locus revealed a multiclonal parasite population at the brain lesion with a more complex minicircle signature than that profiled in blood using restriction fragment length polymorphism (RFLP)-PCR and low stringency single primer (LSSP) PCR. Interestingly, different sublineages of T. cruzi II were detected in blood and brain by means of spliced-leader and 24s ribosomal-DNA amplifications. Quantitative-competitive PCR monitored the decrease of parasitic load during treatment and secondary prophylaxis with benznidazole. The synergy between parasiticidal plus antiretroviral treatments probably allowed the patient a longer survival than usually achieved in similar episodes. This is the first case report demonstrating a differential distribution of natural parasite populations and sublineages in Chagas disease reactivation, showing the proliferation of cerebral variants not detectable in peripheral blood.Fil: Burgos, Juan Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Bergher, Sandra B.. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Freitas, Jorge M.. Universidade Federal de Minas Gerais; BrasilFil: Bisio, Margarita María Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Altcheh, Jaime Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; ArgentinaFil: Teijeiro, Ricardo. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Begher, Sandra B.. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Freilij, Hector León. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; ArgentinaFil: Deccarlini, Florencia. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Levalle, Jorge. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Lopez Alcoba, Horacio. Gobierno de la Ciudad de Buenos Aires. Hospital "Ignacio Pirovano"; ArgentinaFil: Burgos, Juan Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Levin, Mariano Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Duffy, Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Macedo, Andrea M.. Universidade Federal de Minas Gerais; BrasilFil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentin

The thrombophilic network of autoantibodies in celiac disease

BACKGROUND: Celiac disease is a life-long autoimmune condition, affecting genetically susceptible individuals that may present with thromboembolic phenomena. This thrombophilia represents a puzzle with multiple constituents: hyperhomocysteinemia, B12 and\or folate deficiency, methylenetetrahydrofolate reductase mutations, and protein C and S deficiency due to vitamin K deficiency. However, the well known thrombogenic factors, antiphosphatidylserine/prothrombin and antiprothrombin have never been explored in celiac disease. METHODS: The serum autoantibody levels were determined in 248 individuals, classified into three groups. Group 1 comprised 70 children with definitive celiac disease (age: 7.04 ±4.3 years, male to female ratio 1.06) and group 2 comprised 88 normal children (age: 6.7 ±4.17 years, male to female ratio 0.87), representing controls. The pediatric populations were compared to group 3, which included 90 adults who were family members (parents) of group 1 (age: 34.6 ±11.35 years, male to female ratio 1.2). Antibodies were checked by enzyme-linked immunosorbent assay. RESULTS: Mean optical density levels of serum antiphosphatidylserine/prothrombin immunoglobulin G antibodies were 32.4 ±19.4, 3.6 ±2.5 and 16.1 ±15.8 absorbance units in groups 1, 2 and 3 respectively (P <0.0001), with 45.7%, 0% and 7.8% of groups 1, 2 and 3 respectively positive for the antibody (P <0.01). Mean optical density levels of serum antiphosphatidylserine/prothrombin immunoglobulin M antibodies were 14.2 ±8.7, 6.7 ±6.4 and 12.4 ±15.5 absorbance units in groups 1, 2 and 3 respectively (P <0.0001), with 7.1%, 3.4% and 9.9% of groups 1, 2 and 3 positive for the antibody. Mean optical density levels of serum antiprothrombin and antiphospholipid immunoglobulin G antibodies were higher in groups 1 and 3 compared with 2 (P <0.005) and in groups 1 and 2 compared with 3 (P <0.01), respectively. Groups 1, 2 and 3 were positive for antiphospholipid immunoglobulin G antibodies (groups 1 and 2 compared with 3) . Celiac disease sera harbor a higher antiprothrombin immunoglobulin G level compared with controls. CONCLUSIONS: It is suggested that the intestinal injury, endothelial dysfunction, platelet abnormality and enhanced apoptosis recently described in celiac disease are at the origin of the increased exposure of phospholipids or new epitopes representing autoantigens. Those autoantibodies might play a pathogenic role in the thrombophilia associated with celiac disease and represent markers for potential anticoagulant preventive therapy

Linajes de Trypanosoma cruzi en pacientes con enfermedad de Chagas y coinfección por VIH

Introducción. Las poblaciones naturales de T. cruzi han sido clasificadas en seis linajes filogenéticos o unidades de tipificación discreta: T. cruzi I, IIa, IIb, IIc, IId y IIe, que pueden jugar un rol en el tropismo tisular y patogénesis de la enfermedad de Chagas. El impacto de la infección por VIH en la diversidad genética de T. cruzi en pacientes coinfectados es un campo poco explorado en parasitología. Objetivo. Caracterizar linajes de poblaciones parasitarias naturales en muestras clínicas de pacientes coinfectados por T. cruzi y VIH. Materiales y Métodos. Se analizaron muestras de sangre y/o lesiones de 25 pacientes residentes en Argentina: 8 pediátricos nacidos de 7 madres coinfectadas, 3 adultos con Chagas indeterminado y VIH y 7 con encefalitis chagásica por SIDA. El diagnóstico molecular y seguimiento de tratamiento etiológico se realizó por PCR hacia secuencias del minicírculo y/o satélite. Los linajes de T. cruzi fueron identificados por PCR para fragmentos de genes para miniexón y ARN ribosomal 24s. La diversidad infra-linaje fue caracterizada por polimorfismo de fragmentos de restricción de las regiones variables del minicírculo. Resultados. De las 7 madres coinfectadas, 2 transmitieron tanto VIH como T. cruzi a sus hijos y 4 sólo transmitieron T. cruzi. El otro caso fue una mujer embarazada que al entrar en coma por presentar un cuadro de Chagas cerebral fue tratada con Benznidazol y no transmitió ni Chagas ni VIH a su hija. En los casos tratados se observó la negativización de la PCR. La mayoría de las poblaciones parasitarias sanguíneas fueron T. cruzi IId, con perfiles de minicírculos particulares de cada paciente, excepto en pares madre-niño infectados, en que resultaron idénticas. Se hallaron poblaciones mixtas con T. cruzi I-IId. En pacientes con reactivación chagásica se encontró tropismo diferencial de T. cruzi IIb y T. cruzi I en lesiones. En estos pacientes los perfiles de minicírculos mostraron patrones complejos sugiriendo poblaciones policlonales. Conclusiones. La elevada proporción de muestras PCR positivas es indicativa de cargas parasitarias más elevadas que en población chagásica sin VIH. Esta exacerbación estaría también implicada en la alta tasa de transmisión vertical. La prevalencia de linaje IId en sangre periférica concuerda con lo hallado en población chagásica en la región. La asociación de linajes infrecuentes en lesiones asociadas a encefalitis chagásica sugiere tropismo diferencial. El análisis directo de linajes parasitarios en muestras clínicas permitió detectar una mayor prevalencia de infecciones mixtas que la detectada a partir de aislamientos en cultivo.Background. Natural populations of T. cruzi have been classified into six phylogenetic lineages or discrete typing units, T. cruzi I, IIa, IIb, IIc, IId, and IIe, believed to play a role in tissue tropism and disease pathogenesis. The impact of HIV infection in the T. cruzi genetic diversity in coinfected patients is a scarcely explored field of parasitology. Objective. To characterize parasitic lineages in clinical samples from patients co-infected with T. cruzi and HIV Materials and Methods. We analyzed blood and lesions samples from 25 patients residing in Argentina, namely 8 infants born to 7 HIV - T. cruzi co-infected mothers, 3 indeterminate adult chagasic patients with HIV co-infection and 7 presenting cerebral Chagas due to AIDS. Molecular diagnosis and monitoring of etiological treatment was carried out by PCR targeted to kinetoplastid (kDNA) and/or satellite sequences. T. cruzi lineages were identified by means of PCR targeted to the intergenic spacer of miniexon gene and 24s ribosomal ARN genes. To characterize the infra-lineage diversity, restriction fragment length polymorphism (RFLP) of KDNA amplicons was carried out. Results. Out of the 7 co-infected mothers, two transmitted both HIV and T. cruzi to their siblings, four transmitted only T. cruzi. The remaining case was a pregnant woman with cerebral Chagas disease who entered into a coma being treated with benznidazole; she did not transmit congenital Chagas disease nor HIV to her newborn. Most bloodstream populations belonged to T. cruzi IId, with unique minicircle signatures for each patient´s strain, but identical signatures between strains from mothers and their congenitally infected infants. Mixtures of lineages T. cruzi I and T. cruzi IId were also detected. Differential tissue tropism of T. cruzi IIb and T. cruzi I was found in patients with cerebral chagas. Minicircle signatures showed complex patterns suggestive of polyclonal populations. Conclusions. The higher proportion of PCR positive samples suggests higher parasite loads that in chagasic population without HIV. The higher prevalence of T. cruzi IId in bloodstream is in agreement with previous findings in this region. The association of rare lineages at sites of encephalytis suggests differential tropism. The direct characterization of parasite lineages in clinical samples allowed identification of a higher prevalence of mixed infections, than previously assumed, from studies based on culture isolates.Fil: Bisio, Margarita María Catalina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Cura, Carolina Inés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Duffy, Tomás. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Altcheh, Jaime Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; ArgentinaFil: Giganti, Salvador Óscar. Servicios de Neurocirugía y Clínica Médica; ArgentinaFil: Begher, Sandra. Gobierno de la Ciudad de Buenos Aires. Hospital de Agudos "Ignacio Pirovano"; ArgentinaFil: Scapellato, Pablo Gustavo. Gobierno de la Ciudad de Buenos Aires. Hospital de Agudos "D. F. Santojanni"; ArgentinaFil: Burgos, Juan Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Levin, Mariano Jorge. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; ArgentinaFil: Schreck, Ricardo. Servicios de Neurocirugía y Clínica Médica; ArgentinaFil: Freilij, Hector León. Gobierno de la Ciudad de Buenos Aires. Hospital General de Niños "Ricardo Gutiérrez"; ArgentinaFil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentin

Treatment of Uremic Diabetic Patients: Hemodialysis or Transplantation?

Eighty-one patients with chronic renal failure associated with or secondary to diabetic nephropathy were treated with dialysis and/or transplant. Twenty-five had juvenile diabetes and 56 had adult onset diabetes. Juvenile diabetics did poorly on hemodialysis with 13 patients having a 19% four-year survival rate, whereas those who had cadaveric transplantation did well with a four-year patient and graft survival rate of 56% in nine patients. The three juvenile diabetics who received related kidney grafts are presently alive with good function. Patients with adult onset diabetes did well on hemodialysis with a four-year survival rate of 63% in 45 patients. In this last group 11 patients received cadaveric transplants, but none survived 18 months

Catches, bycatches and stock indicators of fisheries targeting cyprinids along the Swedish Baltic Sea coast

Decreasing abundance of many traditionally exploited fish stocks in the Baltic Sea force small-scale fisheries to find new ways to make a living. In line with Swedish national strategies on food supply there is an interest to develop commercial cyprinid fisheries. In the Bothnian Bay in the northern part of the Baltic Sea, annual catches have increased from zero-catches 2018-30 tonnes 2021. To aid a sustainable development of these cyprinid fisheries that target mainly bream (Abramis brama) and ide (Leuciscus idus), we study catch efficiency of target species and bycatch in different gears and seasons using logbook data from the Bothnian Bay. Using cameras, we also assessed bycatch rates. To assist the sustainability of the fishery we develop potential stock indicators. Our results suggests that larger gear (pound-nets) are more effective in catching bream, and that the proportion of bycatch decreased with gear size, being < 10% in the largest gear, which is similar or lower than many other Baltic Sea fisheries. By-catches of salmon is of concern in the Bothnian Bay, but the camera study indicates that salmon bycatches are sporadic. Catch per unit effort (CPUE) of bream was highest in spring and fall, and we conclude that site specific median CPUE is the most suitable stock abundance indicator. The size indicator L90, the 90th percentile of the length distribution, was similar among areas and we propose it as a suitable indicator of the demographic structure of the targeted bream stocks. Our results provide reference points for relatively unfished conditions, but as the study was based on mainly fishery dependent data, it is important to also include fishery independent data to assess ecosystem effects of a future and intensified cyprinid fishery