Beaver effects on macroinvertebrate assemblages in two streams with contrasting morphology.

Beaver populations are increasing throughout Europe and especially in Switzerland. Beaver are major ecological engineers of fluvial systems, dramatically influencing river morphology, ecohydrology and, consequently, aquatic and terrestrial biota. This study compared macroinvertebrate assemblages and trophic structure at two beaver complexes with contrasting topography in Switzerland over an annual cycle. One complex (Marthalen) was in a low gradient open basin, whereas the other complex (Flaach) flowed through a higher gradient ravine-like basin. Both complexes were embedded in an overall agricultural landscape matrix. Water physico-chemistry differed between the two complexes with nitrogen, phosphorus, and DOC being higher at Marthalen than at Flaach. Both complexes showed strong seasonality in physico-chemistry, but retention of nutrients (N, P) was highest in summer and only at Marthalen. Both complexes also showed strong seasonality in macroinvertebrate assemblages, although assemblages differed substantially between complexes. At Marthalen, macroinvertebrate assemblages were predominantly lentic in character at 'pool' sites within the complex. At Flaach, lotic macroinvertebrate assemblages were common at most sites with some lentic taxa also being present. Dietary shifts based on carbon/nitrogen stable isotopes occurred in spring and summer among sites at both complexes (autochthonous resource use increasing over allochthonous resource use downstream), although being most pronounced at Marthalen. In contrast, similar resource use across sites occurred in winter within both complexes. Although beaver significantly influenced fluvial dynamics and macroinvertebrate assemblage structure at both complexes, this influence was most pronounced at Marthalen where beaver caused the system to become more wetland in character, e.g., via higher hydraulic residence time, than at Flaach. We conclude that topography can shape beaver effects on fluvial systems and resident biota.

Immunohistochemical localization of nanog and Oct4 in stem cell compartments of human sacrococcygeal teratomas.

AIMS: To study the range of differentiation and presence of cells positive for stem cell markers in 20 sacrococcygeal teratomas (SCTs) which were consecutively operated on between 1990 and 2000 in the Department of Paediatric Surgery in Tübingen, Germany. METHODS AND RESULTS: Preserved paraffin-embedded material was re-evaluated. In addition to tissues of various organs, caudal organ structures not described before were identified, such as colon with pancreas originating from colonic crypts, Fallopian tube and vaginal epithelia. The derivation of the latter was confirmed by Müllerian duct specific CA125 and CA19-9 antibodies. The expression of stem cell markers was studied with antibodies against nanog, Oct4, SSEA-4, nestin and subtype M3 muscarinic receptors. Cells positive for these markers were encountered in immature end buds and capillary sprouts, and as single cells in neural tissue, gonadal structures, hairs and in the stem cell niches of differentiated epithelia. CONCLUSIONS: Our data indicate that SCTs of the newborn arise from remnants of the epiblast-like tail bud blastema and demonstrate that they contain cells positive for embryonic stem cell markers and may represent a novel source for human embryonic stem cells.

Evaluación del desempeño de una celda de combustible microbiana con electrodo de grafito modificado para el tratamiento de agua residual del procesamiento del café / Evaluation of the performance of a microbial fuel cell with modified graphite electrode for the treatment of wastewater from coffee processing / Avaliação do desempenho de uma célula combustível microbial com eletrodo de grafite modificado para o tratamento de águas residuais do processamento de café

Resumen La actividad cafetalera en Costa Rica procesa aproximadamente 69.000 toneladas de café mediante la técnica de beneficiado húmedo. Esta actividad conlleva un alto impacto ambiental debido a la generación de8Lde agua residual/kg de café oro producido. El presente trabajo tiene como objetivo utilizar el agua residual del procesamiento de café como sustrato en celdas combustibles microbianas (CCM), con el propósito de generar energía eléctrica a través de su uso y, a la vez, disminuir la carga orgánica del residuo. La CCM empleó un cátodo modificado con ftalocianinas de hierro (FePc), generó una eficiencia coulómbica de 0,7% y una densidad de potencia de 89 UW/ cm2 en un ciclo de operación de cinco días. Además, se determinó que la CCM disminuye la demanda química de oxígeno (DQO) del residuo hasta en 27% bajo las condiciones de operación nativas del sustrato, a temperatura ambiente, sin mediadores químicos para la reacción anódica y con el uso de electrodos de platino para el cátodo. El estudio confirma la oportunidad de emplear el sustrato con una flora microbiana nativa apta para la operación de la tecnología de la CCM, y así perfilar el dispositivo como una opción novedosa para el tratamiento de este residuo en Costa Rica.

Abstract In Costa Rica coffee production is the most traditional agroindustrial activity, each year approximately 69,000 tons of coffee are processed through the technique of wet processing. The process has a high environmental impact since it generates eight liters of wastewater/kg of produced coffee. Consequently, the main goal of this research was to evaluate the electric generation of a Microbial Fuel Cell (MFC) with two chambers, using coffee wastewater as a substrate, which would generate a sustainable solution with an added economic value to this waste in Costa Rica. The MFC with a cathode modified with iron phthalocyanines (FePc) generated a coulombic efficiency of 0.7% and a power density of 89 -uW/cm2 in a 5-day operation cycle. In addition, it was determined that the MFC decreases the COD of the waste by up to 27% under native substrate conditions, without the use of high temperatures, or chemical mediators for the anodic reaction and platinum electrodes for the cathode chamber. The efficiency of the device can be improved with changes at design level that reduce the ohmic internal resistance and improve electrical generation, the study confirms the potential of the substrate with a native microorganism suitable for the use of MFC technology, shaping the device as a novelty option for the treatment of the waste in Costa Rica.

Resumo A indústria do café na Costa Rica processa cerca de 69 000 toneladas de café por meio da técnica de moagem úmida, o que acarreta um alto impacto ambiental devido à geração de 8 L de água residual / kg de café dourado. O objetivo deste trabalho era usar águas residuais do processamento do café como substrato em Células de Combustível Microbianas (CCM) a fim de gerar energia elétrica por meio do seu aproveitamento e ao mesmo tempo reduzir a carga orgânica do resíduo. CCM usando cátodo modificado com ftalocianinas de ferro (FePc) gerou uma eficiência coulômbica de 0,7% e uma densidade de potência de 89 uW/cm2 em um ciclo operacional de cinco dias. Além disso, foi determinado que o CCM reduz a Demanda Química de Oxigénio (DQO) do resíduo em até 27% nas condições nativas de operação do substrato, à temperatura ambiente, sem mediadores químicos para a reação anódica e com a utilização de eletrodos de platina para o cátodo. O estudo confirma a oportunidade de utilizar o substrato com flora microbiana nativa adequada para o funcionamento da tecnologia CCM e, assim, delinear o dispositivo como uma nova opção para o tratamento desses resíduos na Costa Rica.

Lack of MSH2 and MSH6 characterizes endometrial but not colon carcinomas in hereditary nonpolyposis colorectal cancer.

Hereditary nonpolyposis colorectal cancer syndrome is associated with an inherited predisposition to primarily colorectal cancer (CRC) and endometrial cancer (EC); however, the biological basis of the organ involvement remains unknown. As an attempt to explore whether the expression levels of MLH1, MSH2, and MSH6 may play a role, we used immunohistochemistry to study 42 ECs and 35 CRCs from patients carrying the same predisposing mutations. Among MSH2 mutation carriers, MLH1 was expressed in both tumor types, whereas MSH2 and, in many cases, also MSH6, were absent. Remarkably, among MLH1 mutation carriers, 54% of ECs (21 of 39), but none of the CRCs (0 of 32), lacked the MSH2 and/or MSH6 protein in addition to lacking MLH1 protein expression. These results demonstrate a marked difference between hereditary nonpolyposis colorectal cancer-related CRCs and ECs and suggest that the development of the latter tumors is selectively associated with the MSH2/MSH6 protein complex deficiency.

A pattern recognition tool for quantitative analysis of in planta hyphal growth of powdery mildew fungi.

The development of fungal pathogens can be quantified easily at the level of spore germination or penetration. However, the exact quantification of hyphal growth rates after initial, successful host invasion is much more difficult. Here, we report on the development of a new pattern recognition software (HyphArea) for automated quantitative analysis of hyphal growth rates of powdery mildew fungi on plant surfaces that usually represent highly irregular and noisy image backgrounds. By using HyphArea, we measured growth rates of colonies of the barley powdery mildew, Blumeria graminis f. sp. hordei, on susceptible and induced-resistant host plants. Hyphal growth was not influenced by the resistance state of the plants up to 48 h postinoculation. At later time points, growth rate increased on susceptible plants, whereas it remained restricted on induced-resistant plants. This difference in hyphal growth rate was accompanied by lack of secondary haustoria formation on induced-resistant plants, suggesting that induced resistance in barley against Blumeria graminis is caused mainly by reduced penetration rates of primary as well as secondary appressoria leading, finally, to fewer and less-developed fungal colonies. No evidence was found for reduced nutrient-uptake efficiency of the primary haustoria in induced-resistant leaves, which would be expected to have resulted in reduced hyphal growth rates during the first 48 h of the interaction.

A high-throughput gene-silencing system for the functional assessment of defense-related genes in barley epidermal cells.

Large-scale gene silencing by RNA interference (RNAi) offers the possibility to address gene function in eukaryotic organisms at a depth unprecedented until recently. Although genome-wide RNAi approaches are being carried out in organisms like Caenorhabditis elegans, Drosophila spp. or human after the corresponding tools had been developed, knock-down of only single or a few genes by RNAi has been reported in plants thus far. Here, we present a method for high-throughput, transient-induced gene silencing (TIGS) by RNAi in barley epidermal cells that is based on biolistic transgene delivery. This method will be useful to address gene function of shoot epidermis resulting in cell-autonomous phenotypes such as resistance or susceptibility to the powdery-mildew fungus Blumeria graminis f. sp. hordei. Gene function in epidermal cell elongation, stomata regulation, or UV resistance might be addressed as well. Libraries of RNAi constructs can be built up by a new, cost-efficient method that combines highly efficient ligation and recombination by the Gateway cloning system. This method allows cloning of any blunt-ended DNA fragment without the need of adaptor sequences. The final RNAi destination vector was found to direct highly efficient RNAi, as reflected by complete knock-down of a cotransformed green fluorescent protein reporter gene as well as by complete phenolcopy of the recessive loss-of-function mlo resistance gene. By using this method, a role of the t-SNARE protein HvSNAP34 in three types of durable, race-nonspecific resistance was observed.

Effect of Jasmonic Acid on the Interaction of Barley (Hordeum vulgare L.) with the Powdery Mildew Erysiphe graminis f.sp. hordei.

Jasmonic acid or its methyl ester induce de novo synthesis of a number of proteins of mostly unknown function in barley (Hordeum vulgare L.). In a topical spray application, 30 [mu]g of jasmonic acid per plant effectively protected barley against subsequent infection by Erysiphe graminis f.sp. hordei. Examination of jasmonic acid-induced barley proteins revealed the presence of several acid-soluble (pH 2.8) proteins. Two prominent groups of 25 kD and 10 to 12 kD apparent molecular mass were present in the intercellular washing fluid. The set of extracellular, induced proteins showed no similarity to barley pathogenesis-related proteins. An in vivo test against E. graminis revealed no antifungal activity of the extracellular jasmonic acid-induced proteins. Experiments with the transcription inhibitor cordycepin showed no correlation between accumulation of jasmonic acid-induced proteins and protection. The application of jasmonic acid and E. graminis simultaneously resulted in independent extracellular accumulation of both jasmonic acid-induced proteins and of pathogenesis-related proteins. The data suggest that jasmonic acid directly inhibits appressoria differentiation of the fungus, and that it is not involved in the signal transduction mechanism leading to induction of pathogenesis-related proteins.

Gene-Expression Patterns and Levels of Jasmonic Acid in Rice Treated with the Resistance Inducer 2,6-Dichloroisonicotinic Acid.

Acquired disease resistance can be induced in rice (Oryza sativa) by a number of synthetic or natural compounds, but the molecular mechanisms behind the phenomenon are poorly understood. One of the synthetic inducers of resistance, 2,6-dichloroisonicotinic acid (INA), efficiently protected rice leaves from infection by the rice blast fungus Magnaporthe grisea (Hebert) Barr. A comparison of gene-expression patterns in plants treated with INA versus plants inoculated with the compatible pathogen M. grisea or the incompatible pathogen Pseudomonas syringae pv syringae revealed only a marginal overlap: 6 gene products, including pathogenesis-related proteins (PR1-PR9), accumulated in both INA-treated and pathogen-attacked leaves, whereas 26 other gene products accumulated only in INA-treated or only in pathogen-attacked leaves. Lipoxygenase enzyme activity and levels of nonconjugated jasmonic acid (JA) were enhanced in leaves of plants treated with a high dose of INA (100 ppm). Exogenously applied JA enhanced the gene induction and plant protection caused by lower doses of INA (0.1 to 10 ppm) that by themselves did not give rise to enhanced levels of endogenous (-)-JA. These data suggest that INA, aside from activating a pathogen-induced signaling pathway, also induces events that are not related to pathogenesis. JA acts as an enhancer of both types of INA-induced reactions in rice.

Jasmonate-Inducible Genes Are Activated in Rice by Pathogen Attack without a Concomitant Increase in Endogenous Jasmonic Acid Levels.

The possible role of the octadecanoid signaling pathway with jasmonic acid (JA) as the central component in defense-gene regulation of pathogen-attacked rice was studied. Rice (Oryza sativa L.) seedlings were treated with JA or inoculated with the rice blast fungus Magnaporthe grisea (Hebert) Barr., and gene-expression patterns were compared between the two treatments. JA application induced the accumulation of a number of pathogenesis-related (PR) gene products at the mRNA and protein levels, but pathogen attack did not enhance the levels of (-)-JA during the time required for PR gene expression. Pathogen-induced accumulation of PR1-like proteins was reduced in plants treated with tetcyclacis, a novel inhibitor of jasmonate biosynthesis. There was an additive and negative interaction between JA and an elicitor from M. grisea with respect to induction of PR1-like proteins and of an abundant JA-and wound-induced protein of 26 kD, respectively. Finally, activation of the octadecanoid signaling pathway and induction of a number of PR genes by exogenous application of JA did not confer local acquired resistance to rice. The data suggest that accumulation of nonconjugated (-)-JA is not necessary for induction of PR genes and that JA does not orchestrate localized defense responses in pathogen-attacked rice. Instead, JA appears to be embedded in a signaling network with another pathogen-induced pathway(s) and may be required at a certain minimal level for induction of some PR genes.

Salicylic Acid in Rice (Biosynthesis, Conjugation, and Possible Role).

Salicylic acid (SA) is a natural inducer of disease resistance in some dicotyledonous plants. Rice seedlings (Oryza sativa L.) had the highest levels of SA among all plants tested for SA content (between 0.01 and 37.19 [mu]g/g fresh weight). The second leaf of rice seedlings had slightly lower SA levels than any younger leaves. To investigate the role of SA in rice disease resistance, we examined the levels of SA in rice (cv M-201) after inoculation with bacterial and fungal pathogens. SA levels did not increase after inoculation with either the avirulent pathogen Pseudomonas syringae D20 or with the rice pathogens Magnaporthe grisea, the causal agent of rice blast, and Rhizoctonia solani, the causal agent of sheath blight. However, leaf SA levels in 28 rice varieties showed a correlation with generalized blast resistance, indicating that SA may play a role as a constitutive defense compound. Biosynthesis and metabolism of SA in rice was studied and compared to that of tobacco. Rice shoots converted [14C]cinnamic acid to SA and the lignin precursors p-coumaric and ferulic acids, whereas [14C]benzoic acid was readily converted to SA. The data suggest that in rice, as in tobacco, SA is synthesized from cinnamic acid via benzoic acid. In rice shoots, SA is largely present as a free acid; however, exogenously supplied SA was converted to [beta]-O-D-glucosylSA by an SA-inducible glucosyltransferase (SA-GTase). A 7-fold induction of SA-GTase activity was observed after 6 h of feeding 1 mM SA. Both rice roots and shoots showed similar patterns of SA-GTase induction by SA, with maximal induction after feeding with 1 mM SA.

Cutin monomers and surface wax constituents elicit H2O2 in conditioned cucumber hypocotyl segments and enhance the activity of other H2O2 elicitors

Hypocotyls from etiolated cucumber (Cucumis sativus L.) seedlings were gently abraded at their epidermal surface and cut segments were conditioned to develop competence for H2O2 elicitation. Alkaline hydrolysates of cutin from cucumber, tomato, and apple elicited H2O2 in such conditioned segments. The most active constituent of cucumber cutin was identified as dodecan-1-ol, a novel cutin monomer capable of forming hydrophobic terminal chains. Additionally, the cutin hydrolysates enhanced the activity of a fungal H2O2 elicitor, similar to cucumber surface wax, which contained newly identified alkan-1,3-diols. The specificity of elicitor and enhancement activity was further elaborated using some pure model compounds. Certain saturated hydroxy fatty acids were potent H2O2 elicitors as well as enhancers. Some unsaturated epoxy and hydroxy fatty acids were also excellent H2O2 elicitors but inhibited the fungal elicitor activity. Short-chain alkanols exhibited good elicitor and enhancer activity, whereas longer-chain alkan-1-ols were barely active. The enhancement effect was also observed for H2O2 elicitation by ergosterol and chitosan. The physiological significance of these observations might be that once the cuticle is degraded by fungal cutinase, the cutin monomers may act as H2O2 elicitors. Corrosion of cutin may also bring surface wax constituents in contact with protoplasts and enhance elicitation.

Tracheal surgery in children.

BACKGROUND: The aim of this study is to show that five distinct types of tracheal anomalies should be differentiated with respect to therapy and prognosis. METHODS: The records of 12 infants and children seen over a period of 20 years for different tracheal anomalies such as laryngotracheal stenosis (n = 3), long or short-segment stenosis of the upper (n = 2), middle (n = 6), and lower (n = 1) trachea were reviewed. In addition to these 12 patients with congenital stenosis, 3 other patients needed tracheal resections because of oncologic or traumatic disease: in 2, the trachea was infiltrated by a papillary carcinoma of the thyroid gland and in one, the upper part of the trachea was injured by an oral explosion accident. 25 patients presenting during the same period for other tracheal pathologies including esophagotracheal cleft syndrome (n = 7), tracheomalacia (n = 4), total tracheal agenesis (n = 3), or for placement of a tracheostomy (n = 11) due to other diseases were excluded from this study. RESULTS: There was 1 early death after repair of a laryngotracheal stenosis by cricoid-split and cricoid-splint due to both cerebral hemorrhage and cardiac insufficiency secondary to Fallot's tetralogy. Another child died four weeks after slide tracheoplasty as a result of hypoxic cerebral lesions induced by severe central catheter-related septicemia. One child with therapy-resistant obstructing granulation tissue which developed after a slide tracheoplasty required a tracheostomy. The patient with the tracheal injury died after another accident one year after discharge. All other patients (n = 11) are doing well. CONCLUSIONS: With respect to therapy of congenital and post-intubation tracheal stenosis, four types should be distinguished. Each of these types requires an adequate surgical procedure. The most important postoperative problem in tracheal surgery is the development of granulation tissue. However, the pathogenesis of granulation is still unknown.

Pathogenesis of extrahepatic bile duct atresia (EHBA): comprehension from a surgical point of view.

INTRODUCTION: The aim of the study is to establish a complete comprehension of the pathogenesis of Biliary Atresia, and to explain both the variable and redundant pathomorphological, as well as, histological findings. MATERIALS AND METHODS: The pathomorphological and histological findings in 223 patients with histologically evident EHBA were recorded retrospectively (72 patients) or prospectively (151 patients), according to a projected ascending study. These findings were compared with histological findings in human and rat embryos. RESULTS: 1) The pathomorphological findings recorded in patients with EHBA were also found in stages of normal embryogenesis of the bile duct system in human and rat embryos. 2) Each histological finding in Biliary Atresia corresponds to a finding in an interrupted stage of the normal development in human and rat embryos. 3) The findings in patients and embryos can be explained completely by a disturbed intrinsic epithelium/mesoderm interaction. 4) Some findings in Biliary Atresia cannot be explained easily by the assumption of an extrinsic factor. CONCLUSION: There is no finding in Biliary Atresia which cannot be completely explained as the result of an intrinsic developmental error, probably due to disturbances or interruption of epithelium/mesoderm interaction during embryogenesis.

It's all in your head: sinus node dysfunction secondary to a sphenoid wing meningioma.

BACKGROUND: A 57-year-old man presented with recurrent episodes of dizziness, weakness of legs, and presyncope. The electrocardiogram showed a sinus bradycardia and recurrent sinus pauses. RESULTS: Cardiac evaluation revealed a normal left ventricular ejection fraction without ischemic, structural, or valvular heart disease. Pronounced limb weakness prompted neurological consultation. Cranial magnetic resonance imaging showed a large right-sided intracranial tumor adjacent to the medial sphenoid wing. Surgical removal of the tumor was accomplished successfully after application of a transient cardiac pacemaker, while decision upon permanent pacemaker implantation was postponed. Histopathology provided evidence of a meningothelial meningioma. Postoperative assessment displayed the absence of sinus node dysfunction after tumor removal. CONCLUSION: Careful differential diagnostic assessment of patients with symptomatic bradycardias needs to rule out reversible causes before implantation of permanent devices.

Effects of heart rate changes on left ventricular volume and ejection fraction: a 2-dimensional echocardiographic study.

The influence of heart rate on left ventricular (LV) volumes and ejection fraction (EF) using 2-dimensional (2-D) echocardiography during atrial pacing was analyzed. The study was performed in 13 normal control subjects, 23 patients with coronary heart disease and 8 patients with dilated cardiomyopathy. An electronic sector scanner (2.25 MHz, 84 degrees) was used. Under constant scanning of the left ventricle, heart rate was increased, in steps of 20 beats/min, from 80 to 140 beats/min. The 2-D echocardiograms were stored on videotape and analyzed off-line. The end-diastolic and end-systolic volumes (EDV and ESV) were determined using a disc method. Stroke volume (SV) and EF were calculated. Constant LV scanning was possible during atrial stimulation, as shown by the analysis of simultaneously recorded 2-D echocardiograms and cineventriculograms at different heart rates, revealing a constant position of the echocardiographic transducer. Simultaneous recordings of cineventriculography and 2-D echocardiography at 80 and 120 beats/min showed that despite differences in absolute values, percent changes of LV volumes and EF determined with both methods were similar. Thus, changes of LV function can be analyzed by 2-D echocardiography. In normal control subjects, an increase in heart rate of 10 beats/min reduced EDV by 4 ml, ESV by 2 ml, SV by 2 ml and EF by 1%, corresponding to percent reductions of 4, 2, 5 and -2%, respectively. In contrast, the absolute decreases in the patients were 6 ml, 1 ml, 5 ml and 2% and the percent changes 2%, 1%, 8% and 5%.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth inhibition of neuroblastoma cells by lovastatin and L-ascorbic acid is based on different mechanisms.

Hydroxymethyl-glutaryl-CoA-reductase (HMG-CoA-reductase), the key enzyme for cholesterol synthesis and essential for the synthesis of the precursor for p21ras farnesylation, was inhibited in neuroblastoma cells by lovastatin or L-ascorbic acid. Both compounds inhibited clonogenic colony formation of neuroblastoma cells in soft agar. However, while the addition of mevalonate, the product of HMG-CoA-reductase, circumvented the inhibition by lovastatin it had no reversing effect on the inhibition by L-ascorbic acid. The role of reactive oxygen compounds generated by the degradation of catecholamines, and the pro-oxidative effects of L-ascorbic acid are discussed as mechanisms of action of L-ascorbic acid.

Toxic reactions of oxidized LDL on cells of acute myeloid leukemia.

In AML patients LDL concentration of the serum is reduced due to the high LDL receptor activity of the AML cells. This phenomenon enables the use of LDL particles as vehicles for drug targeting. Toxic lipid peroxides and aldehydes were introduced into LDL particles by the simple but effective oxidation with 10 microM CuSO4. Up to 250 nmol peroxides and 6 nmol malondialdehyde were formed per mg LDL protein within 30 h of oxidation. This oxidized LDL is effectively taken up by AML cells of the FAB type M3 and M5 indicating the presence of scavenger receptors on these cells. Within 96 h 61-84% of the AML cells are killed by the oxidized LDL. Our results open a possibility to achieve specificity for targeting lipophilic antineoplastic drugs towards AML cells using oxidized LDL as vehicles. The use of oxidized LDL as drug carrier is recommended for purging of AML bone marrow because hematopoietic stem cells that don't possess scavenger receptors are protected from toxic action.