Lidocaine and pinacidil added to blood versus crystalloid cardioplegic solutions: study in isolated hearts

Abstract Objective: The present study aimed the functional recovery evaluation after long term of cardiac arrest induced by Custodiol (crystalloid-based) versus del Nido (blood-based) solutions, both added lidocaine and pinacidil as cardioplegic agents. Experiments were performed in isolated rat heart perfusion models. Methods: Male rat heart perfusions, according to Langendorff technique, were induced to cause 3 hours of cardiac arrest with a single dose. The hearts were assigned to one of the following three groups: (I) control; (II) Custodiol-LP; and (III) del Nido-LP. They were evaluated after ischemia throughout 90 minutes of reperfusion. Left ventricular contractility function was reported as percentage of recovery, expressed by developed pressure, maximum dP/dt, minimum dP/dt, and rate pressure product variables. In addition, coronary resistance and myocardial injury marker by alpha-fodrin degradation were also evaluated. Results: At 90 minutes of reperfusion, both solutions had superior left ventricular contractile recovery function than the control group. Del Nido-LP was superior to Custodiol-LP in maximum dP/dt (46%±8 vs. 67%±7, P<0.05) and minimum dP/dt (31%±4 vs. 51%±9, P<0.05) variables. Coronary resistance was lower in del Nido-LP group than in Custodiol-LP (395%±50 vs. 307%±13, P<0.05), as well as alpha-fodrin degradation, with lower levels in del Nido-LP group (P<0.05). Conclusion: Del Nido-LP cardioplegia showed higher functional recovery after 3 hours of ischemia. The analysis of alpha-fodrin degradation showed del Nido-LP solution provided greater protection against myocardial ischemia and reperfusion (IR) in this experimental model.

Effects of different extracts of the mushroom Agaricus blazei Murill on the hematologic profile of mice with Ehrlich tumor / Efeitos de diferentes extratos do cogumelo Agaricus blazei Murill sobre o perfil hematológico de camundongos com tumor de Ehrlich

The aim of this study was to investigate the effect of the mushroom Agaricus blazeii Murril (ABM) extracts on the hematological profile of Swiss mice bearing an Ehrlich solid tumor. Three fractions (total extract, polysaccharides, and supernatant) of ABM extracts obtained by four methods (ultrasonic or water bath, at pH 4 or pH 7) were administered to mice over 21 days. Polysaccharide solutions were analyzed by gas and liquid chromatography that showed both mannose and glucose concentrations. The method of extraction influenced the degree of glucose polymerization and the mannose/glucose relationship. The treatment with ABM supernatant at pH 7 and water bath was associated with reduced concentrations of leukocytes and lymphocytes and altered the percentage of CD4+ and CD8+ lymphocytes in Ehrlich tumor-bearing mice. The treatment with the ABM extract in water bath and ultrasound at pH 4 resulted in lower lymphocyte counts, regardless of tumor presence, and greater granulocyte values in mice with Ehrlich tumor than in controls. We concluded that different fractions and methods of extraction of A. blazei produced differing blood profiles in mice inoculated with the Ehrlich tumor.

O objetivo deste estudo foi investigar o efeito de diferentes extratos do cogumelo Agaricus blazeii Murril (ABM) sobre o perfil hematológico de camundongos Swiss portadores de tumor de Ehrlich sólido. Três frações (extrato total, polissacarídeos e sobrenadante) dos extratos de ABM foram obtidas por quatro métodos (sonificador, banho-maria, em pH 4 ou pH 7) e administradas para camundongos durante 21 dias. Soluções de polissacarídeos foram analisadas por cromatografia gasosa e líquida, que mostraram concentrações de glucose e manose. O método de extração influenciou o grau de polimerização da glicose e a relação manose/glucose. O tratamento com o sobrenadante de ABM (em pH 7 e banho-maria) estava associado com reduzidas concentrações de leucócitos e linfócitos, além de alterar a porcentagem de linfócitos CD4+ e CD8+ em camundongos portadores de tumor sólido de Ehrlich. O tratamento com extratos de ABM, obtidos tanto em banho-maria como no sonificador em pH 4, resultou nas mais baixas contagens de linfócitos, independentemente da presença do tumor, e nos maiores valores de granulócitos em camundongos com tumor de Ehrlich. Conclui-se que os diferentes métodos de extração com as respectivas frações de A. blazei são capazes de intereferir no perfil hematológico de camundongos com tumor sólido de Ehrlich.

Chemical salivary composition and its relationship with periodontal disease and dental calculus

Aim: To determine the relationship between the chemical composition of saliva, periodontal disease and dental calculus. Methods: An observational analytical cross-sectional study was conducted with patients over 55 years of age. Ethical principles of autonomy and risk protection were applied according to the international standards. Sociodemographic and diagnosis variables (presence of dental calculus and periodontal status) were considered to measure salivary concentrations of glucose (by the glucose oxidase/peroxidase method, amylase (by the colorimetric test), urea (by the amount of indophenol), total protein (by the Bradford method) and albumin (by the nephelometric method). Patients chewed a sterile rubber band and 3 mL of stimulated saliva were collected. The samples were stored at -5 °C, centrifuged at 2,800 rpm for 10 min, and the supernatant was removed and stored at -20 °C. Data were presented as frequencies and proportions for qualitative variables and measures of central tendency and dispersion for quantitative variables. Data were analyzed by either analysis of variance or Kruskal Wallis test . A p value <0.05 was considered statistically significant. Results: Significant relationships were observed between the concentration of salivary urea and periodontal status (p = 0.03) and the presence of dental calculus and urea (p = 0.04) was demonstrated. Conclusions: A relationship between the salivary urea concentration and the presence of periodontal disease and dental calculus is suggested.

Development of cardioplegic solution without potassium: experimental study in rat / Desenvolvimento de solução cardioplégica sem potássio: estudo experimental em ratos

INTRODUCTION: Myocardial preservation during open heart surgeries and harvesting for transplant are of great importance. The heart at the end of procedure has to resume its functions as soon as possible. All cardioplegic solutions are based on potassium for induction of cardioplegic arrest. OBJECTIVE: To assess a cardioplegic solution with no potassium addition to the formula with two other commercially available cardioplegic solutions. The comparative assessment was based on cytotoxicity, adenosine triphosphate myocardial preservation, and caspase 3 activity. The tested solution (LIRM) uses low doses of sodium channel blocker (lidocaine), potassium channel opener (cromakalin), and actin/myosin cross bridge inhibitor (2,3-butanedione monoxime). METHODS: Wistar rats underwent thoracotomy under mechanical ventilation and three different solutions were used for "in situ" perfusion for cardioplegic arrest induction: Custodiol (HTK), Braile (G/A), and LIRM solutions. After cardiac arrest, the hearts were excised and kept in cold storage for 4 hours. After this period, the hearts were assessed with optical light microscopy, myocardial ATP content and caspase 3 activity. All three solutions were evaluated for direct cytotoxicity with L929 and WEHI-164 cells. RESULTS: The ATP content was higher in the Custodiol group compared to two other solutions (P<0.05). The caspase activity was lower in the HTK group compared to LIRM and G/A solutions (P<0.01). The LIRM solution showed lower caspase activity compared to Braile solution (P<0.01). All solutions showed no cytotoxicity effect after 24 hours of cells exposure to cardioplegic solutions. CONCLUSION: Cardioplegia solutions without potassium are promised and aminoacid addition might be an interesting strategy. More evaluation is necessary for an optimal cardioplegic solution development.

INTRODUÇÃO: Preservação do miocárdio durante cirurgias cardíacas abertas e de colheita para transplante são de grande importância. O coração ao final do processo tem de retomar as suas funções, logo que possível. Todas as soluções cardioplégicas são baseadas em potássio, para indução de parada cardioplégica. OBJETIVO: Comparar a uma solução cardioplégica sem adição de potássio à sua fórmula com duas outras soluções cardioplégicas disponíveis comercialmente. A avaliação comparativa foi baseada na citotoxicidade, preservação miocárdica (adenosina trifosfato, ATP) e atividade da caspase 3. A solução testada (LIRM) utiliza baixas doses de bloqueador de canal de sódio (lidocaína), abridor do canal de potássio (cromacalina) e inibidor da ponte actina/miosina (2,3-butanodiona monoxima). MÉTODOS: Ratos Wistar foram submetidos à toracotomia sob ventilação mecânica e três soluções diferentes foram utilizadas para perfusão in situ para a indução de parada cardioplégica: soluções Custodiol (HTK) Braile (G/A) e LIRM. Após parada cardíaca, os corações foram retirados e mantidos em câmara fria por 4 horas. Após esse período, o coração foi avaliado com microscopia de luz ótica, o conteúdo de ATP miocárdico e atividade da caspase 3. Todas as três soluções foram avaliadas quanto à citotoxicidade direta com células L929 e WEHI-164. RESULTADOS: A quantidade de ATP foi maior no grupo Custodiol em comparação às com outras duas soluções (P<0,05). A atividade de caspase foi menor no grupo HTK quando comparado às soluções LIRM e G/A (P<0,01). A solução LIRM demonstrou menor atividade da caspase em comparação à solução Braile (P<0,01). Todas as soluções não mostraram qualquer efeito de citotoxicidade após 24 horas de exposição das células às soluções cardioplégicas. CONCLUSÃO: Soluções cardioplégicas sem potássio são uma perspectiva e a adição de aminoácido pode ser uma estratégia interessante. Mais avaliações são necessárias para o desenvolvimento ideal da solução cardioplégica.

Effects of fasting and refeeding on the metabolic functions of the turtle Kinosternon scorpioides (Linnaeus, 1766) raised in captivity / Efeitos do jejum e da realimentação sobre as funções metabólicas da tartaruga Kinosternon scorpioides (Linnaeus, 1766) criada em cativeiro

The metabolic responses of adult and young freshwater Kinosternon scorpioides turtles raised in captivity were evaluated. Two experiments were performed: a) blood metabolite changes caused by food deprivation, and b) liver and muscle glycogen and total lipid differences after fasting and refeeding. Blood glucose concentration of young animals was susceptible to food deprivation. In both groups this metabolite decreased after 30 days of fasting. Feeding for 15 days did not recover blood glucose. Total seric proteins were not affected by food deprivation. Fasting decreased blood urea nitrogen and the highest difference was found around 30 days. Uric acid increased in young animals after 60 days of fasting. Triacylglicerol decreased after 15 days of fasting and refeeding for 15 days recovered the pre-fasting levels. Free fatty acid plasma tended to increase around 15 days of fasting. Liver glycogen decreased at day 15 of fasting, being stable thereafter while muscle glycogen decreased at a slower rate. Total liver lipid stabilized after 30 days and then decreased 70% after 60 days of fasting. Muscle lipids remained stable throughout fasting. It could be concluded that fasting of Kinosternon scorpioides led to metabolic adaptations similar to the one reported from reptiles and fish.

Neste trabalho foi avaliada as respostas metabólicas da tartaruga Kinosternon scorpioides criada em cativeiro, nas fases, adultos e jovens nos estados: alimentado, jejuado e realimentado. O estudo compreendeu dois experimentos: (a) mudanças metabólicas no sangue causadas por privação alimentar e realimentação e (b) diferenças nas concentrações de glicogênio e lipídeos totais no fígado e no músculo após jejum e realimentação. No experimento dos animais jovens a concentração de glicose no sangue apresentou mudanças significativas. Entretanto, nos dois experimentos esse metabólito reduziu significativamente aos 30 dias de jejum. Realimentados por um período de 15 dias foi observado que a concentração de glicose não recuperou os níveis de pré-jejum. Concentração de glicose no sangue de animais jovens foi mais suscetível à privação de alimentos. Em ambos os grupos os metabólitos analisados decresceu após 30 dias de jejum. Retomando a alimentação por 15 dias foi observado que a concentração de glicose não recuperou. As concentrações de proteínas séricas totais não foram afetadas pela privação alimentar. O jejum decresceu a concentração de uréia no sangue e a maior diferença ocorreu aos 30 dias. O ácido úrico decresceu nos animais jovens após 60 dias de jejum. O triacilglicerol diminuiu após 15 dias de jejum e a realimentação por 15 dias recuperou os níveis de pré-jejum. O glicogênio hepático diminuiu aos 15 dias de jejum, e estabilizou a partir daí, enquanto o glicogênio muscular diminuiu a um ritmo mais lento. O lipídio total hepático total se manteve estável até os 30 dias de jejum, diminuindo até 70% aos 60 dias de jejum. E, em seguida, diminuiu 70% após 60 dias de jejum. Os lipídeos musculares permaneceram estáveis durante o jejum. Conclui-se que o jejum na espécie Kinosternon scorpioides apresentou adaptações metabólicas semelhantes aos relatados para outros répteis e peixes.

Produção de glicose e frutose por invertase de Saccharomyces cerevisiae imobilizada em suporte MANAE-Agarose / Glucose and Fructose Production by Saccharomyces cerevisiae Invertase Immobilized on MANAE-Agarose Support

Invertase from Saccharomyces cerevisiae was immobilized on agarose beads, activated with various groups (glyoxyl, MANAE or glutaraldehyde), and on some commercial epoxy supports (Eupergit and Sepabeads). Very active and stable invertase derivatives were produced by the adsorption of the enzyme on MANAE-agarose, MANAE-agarose treated with glutaraldhyde and glutaraldehyde-agarose supports. At pH 5.0, these derivatives retained full activity after 24h at 40 ºC and 50 ºC. When assayed at 40 °C and 50 °C, with the pH adjusted to 7.0, the invertase-MANAE-agarose derivative treated with glutaraldehyde retained 80% of the initial activity. Recovered activities of the derivatives produced with MANAE, MANAE treated with glutaraldehyde and glutaraldehyde alone were 73.5%, 44.4% and 36.8%, respectively. These three preparations were successfully employed to produce glucose and fructose in 3 cycles of sucrose hydrolysis.

Invertase de Saccharomyces cerevisiae foi imobilizada em agarose ativada com diferentes grupos (glioxil, MANAE ou glutaraldeído) e suportes epóxidos comerciais (Eupergit e Sepabeads). Derivados de invertase ativos e estabilizados foram produzidos pela adsorção da enzima em suportes MANAE-agarose, MANAE-agarose tratado com glutaraldeído e glutaraldeído-agarose. Em pH 5,0 estes derivados retiveram total atividade até 24h a 40 ºC e 50 ºC. Quando os ensaios foram a 40 °C e 50 °C com o pH alterado para 7,0, o derivado invertase-MANAE-agarose tratado com glutaraldeído apresentou 80% da atividade inicial. As atividades recuperadas dos derivados foram 73,5%, 44,4% e 36,8%, respectivamente para MANAE, MANAE tratado com glutaraldeído e glutaraldeído. Essas três preparações foram empregadas com sucesso em 3 ciclos de hidrólise da sacarose para produzir glicose e frutose.

Beneficial effects of hyperosmotic perfusion in the myocardium after ischemia/reperfusion injury in isolated rat hearts / Efeitos benéficos da perfusão hiperosmótica no miocárdio após lesão isquemia/reperfusão em corações isolados de ratos

OBJECTIVE: A simple method to reduce the ischemia/reperfusion injury that can accompany cardiac surgery would have great clinical value. This study was to investigate the effect of hyperosmotic perfusion on ischemia/reperfusion injury in isolated perfused rat hearts. METHOD: Forty male Sprague-Dawley rats were randomly divided either to have their isolated hearts perfused with normal osmotic buffer or buffer made hyperosmotic by addition of glucose. Hearts were then subjected to 30 min ischemia followed by 30 min reperfusion. Coronary flow, time to ischemic arrest, reperfusion arrhythmia, and ventricular function were recorded. Creatine phosphokinase leakage into the coronary artery, and myocardial content and activity of superoxide dismutase and catalase were also examined. RESULTS: Rat hearts with hyperosmotic perfusion showed higher coronary flow, a prolonged time to ischemic arrest (10.60 vs. 5.63 min, P<0.005), a lower reperfusion arrthythmia score (3.2 vs. 5.3, P<0.001), better ventricular function, and less creatine phosphokinase leakage (340.1 vs. 861.9, P<0.001) than normal osmotic controls. Myocardial catalase content and activity were increased significantly (1435 vs. 917 U/g wet weight, P<0.001) in hearts perfused with hyperosmotic solution in comparison to the normal osmotic controls. CONCLUSION: Pretreatment with hyperosmotic perfusion in normal rat hearts, which is attributed partly to the increased antioxidative activity, could provide beneficial effects from ischemia and reperfusion-induced injury by increasing coronary flow, and decreasing reperfusion arrhythmia.

OBJETIVO: Um método simples para reduzir a lesão de isquemia/reperfusão que pode acompanhar a cirurgia cardíaca teria grande valor clínico. O objetivo deste estudo foi investigar o efeito da perfusão hiperosmótica na isquemia/reperfusão em corações isolados de ratos perfundidos. MÉTODOS: Quarenta ratos machos Sprague-Dawley foram divididos aleatoriamente e tiveram os seus corações isolados perfundidos com tampão osmótico normal ou tampão hiperosmótico com a adição de glucose. Os corações foram então submetidos a 30 minutos de isquemia, seguida de 30 min de reperfusão. O fluxo coronariano, tempo de parada isquêmica, arritmia de reperfusão e da função ventricular foram registrados. Vazamento creatinofosfoquinase na artéria coronária, o miocárdio e atividade de superóxido dismutase e catalase foram também examinados. RESULTADOS: Crações de ratos com perfusão hiperosmótica apresentaram maior fluxo coronariano, tempo prolongado de parada isquêmica (10,60 vs. 5,63 min, P<0,005), menor pontuação de reperfusão arritmica (3,2 vs. 5,3, P<0,001), melhor função ventricular e menos vazamento de creatina fosfoquinase (340,1 vs. 861,9, P<0,001) do que controles normais osmóticos. Teor de catalase e atividade do miocárdio também tiveram aumento significativo (1435 vs. 917 peso U/g de peso fresco, P<0,001) em corações perfundidos com solução hiperosmótica em comparação com os controles normais osmóticos. CONCLUSÃO: O pré-tratamento com perfusão hiperosmótica em corações de ratos normais, o que é atribuído, em parte, ao aumento da atividade antioxidante, pode oferecer efeitos benéficos de isquemia e reperfusão induzida por lesão, aumentando o fluxo coronário e diminuindo a arritmia de reperfusão.

Influencia de la dieta sobre las actividades de fosforiladoras de glucosa en hígado e intestino de rata / Influence of diet on glucose phosphorylating activity of rat liver and small intestine

La fosforilación de la glucosa en los mamíferos, es catalizada por una familia de isoenzimas (hexoquinasas I-IV; HQ) de diferente Km para el azúcar. En los hepatocitos y células b-pancreáticas se encuentra la glucoquinasa (GQ; HQ IV) de Km elevado (12-20 mM). Hemos observado que GQ está presente en el intestino delgado y podría contribuir a la producción de lactato durante la absorción del azúcar. En este trabajo se determinó el efecto de la dieta (ratarina R; 60% de glucosa G; sacarosa S; almidón A; caseína C), suministrada ad libitum, sobre las actividades de HQ y GQ en homogenatos de hígado y mucosa intestinal de rata. El suministro de glucosa (5%) en el agua de beber (SG) también fue evaluado en las dietas con R y G. Las actividades de HQ (Glucosa 1 mM) y la capacidad fosforilativa total (CFT: Glucosa = 100 mM) se determinaron enzimáticamente. GQ se estimó por diferencia. En el grupo control (R) y en S, A y C, la GQ hepática fue un 85% de la CFT, mientras que en G, GSG y RSG un 66%. La HQ intestinal alcanzó en los grupos R, GSG, A y C un 87% y en RSG un 30% de la CFT. La GQ en G, S, aumentó, pero una menor magnitud. La presencia de GQ en el intestino delgado y su expresión diferencial de acuerdo a la dieta, abren la posibilidad de que dicho órgano contribuya al metabolismo inicial de la glucosa procedente de la dieta y provea al hígado de un precursor (lactato) muy eficaz para sus procesos anabólicos.

Glucose phosphorylation in mammals, is catalyzed by a family of isoenzymes (hexokinases I- IV; HQ) of different Km for the sugar. In hepatocytes and pancreatic b- cells are glucokinase (GQ ; HQ IV) of high Km (12-20 mM). We observed that GQ is present in the small intestine and may contribute to the production of lactate during the absorption of sugar. In this work, the effect of diet (ratarina R, G 60% glucose, sucrose S; starch A; casein C) provided ad libitum , on the activities of HQ and GQ in liver homogenates of rat intestinal mucosa . The supply of glucose (5%) in the drinking water ( SG ) was also evaluated in the diets with R and G. HQ activities (Glucose 1 mM) and phosphorylating full capacity ( CFT : Glucose = 100 mM ) were determined enzymatically . GQ was estimated by difference. In the control group (R) and S, A and C, the GQ liver was about 85% of CFT, whereas G, GSG and RSG 66%. The intestinal HQ reached in the R groups, GSG, A and C by 87% and 30% RSG the CFT. The GQ in G, S , increased , but a lower magnitude. the presence of glucokinase in the small intestine and its differential expression according to diet, open the possibility that this structure contributes to initial metabolism of glucose and provide to the liver a precursor (lactate) very effective for their anabolic processes.

Kidney Transplantation from a Donor Following Cardiac Death Supported with Extracorporeal Membrane Oxygenation

To expand the donor pool, organ donation after cardiac death (DCD) has emerged. However, kidneys from DCD donors have a period of long warm ischemia between cardiac arrest and the harvesting of the organs. Recently, we used extracorporeal membrane oxygenation (ECMO) to minimize ischemic injury during 'no touch' periods in a Maastricht category II DCD donor and performed two successful kidney transplantations. The kidneys were procured from a 49-yr-old male donor. The warm ischemia time was 31 min, and the time of maintained circulation using ECMO was 7 hr 55 min. The cold ischemia time was 9 hr 15 min. The kidneys were transplanted into two recipients and functioned immediately after reperfusion. The grafts showed excellent function at one and three months post-transplantation; serum creatinine (SCr) levels were 1.0 mg/dL and 0.8 mg/dL and the estimated glomerular filtration rates (eGFR) were 63 mL/min/1.73 m2 and 78 mL/min/1.73 m2 in the first recipient, and SCr levels were 1.1 mg/dL and 1.0 mg/dL and eGFR were 56 mL/min/1.73 m2 and 64 mL/min/1.73 m2 in the second recipient. In conclusion, it is suggested that kidney transplantation from a category II DCD donor assisted by ECMO is a reasonable modality for expanding donor pool.

Marcación de un derivado de glucosa con 99mTc mediante la formación de un complejo Tc(V)-nitruro: estudios preliminares / Labelling of a glucose derivative with 99mTc through the formation of a Tc(V)-nitride symmetrical complex: preliminary studies

La preparación de derivados de glucosa marcados con 99mTc reviste gran interés para la evaluación del consumo de glucosa in vivo en oncología y cardiología nuclear. Este trabajo presenta la marcación de un análogo de glucosa (GLU-DTC) mediante la formación de un complejo Tc(V)-nitruro simétrico. Para ello se incorporó a la biomolécula un grupo ditiocarbamato capaz de coordinar al metal. La marcación fue realizada mediante sustitución de ligandos, obteniéndose una única especie con pureza radioquímica superior al 90 por cientp, la que se mantuvo durante al menos 4 hs. La caracterización fisicoquímica y biológica muestra que el complejo 99mTc(V)-nitruro(GLU-DTC)2 es un compuesto estable y altamente hidrofílico, aunque su unión a proteínas plasmáticas es mayor a la esperada, hecho que justificaría la alta actividad retenida en sangre y en hígado durante la evaluación biológica en ratones CD1 normales. Estos resultados indican que la marcación con 99mTc de este derivado de glucosa produce una alteración significativa de su comportamiento biológico.

The preparation of 99mTc-labeled glucose derivatives is of great interest to evaluate the in vivo glucose uptake in nuclear oncology and cardiology. This paper presents the labelling of a glucose analogue (GLU-DTC) through the formation of a Tc(V)-nitride symmetrical complex. For this purpose, a dithiocarbamate group was incorporated to the biomolecule, in order to coordinate the metal. The labelling reaction was carried out by substitution yielding a single complex with radiochemical purity above 90 percent. This complex was stable for at least 4 hours. The physicochemical and biological characterization shows that the 99mTc(V)-nitride(GLU-DTC)2 complex is a stable and highly hydrophilic compound, although its plasma protein binding is greater than expected, a fact which justifies the high activity retained in blood and liver during the biological evaluation in normal CD1 mice. These results indicate that 99mTc labelling of this glucose derivate alters significantly its biological behaviour.

Efecto del oxígeno y la glucosa en la metanogénesis y mineralización de resinas poliméricas / The effect of oxygen and glucose on polymeric resin methanogenesis and mineralisation

The effect of dissolved oxygen concentration and glucose on polymeric resin compounds’ methanogenesisand mineralisation was examined in batch cultures. They were inoculated with sludge from an upflow anaerobicsludge blanket reactor fed with polymeric resin compounds and 1.0 mg L-1 steady-state dissolved oxygen.All tests were carried out with 1,500 mg L-1 chemical oxygen demand (COD), at 30±2ºC, with 23.8 g L-1 volatile suspended solids (VSS) as inoculum and 1:1 COD/VSS ratio. The effect of different dissolved oxygen concentrations showed that COD efficiently removed glucose whilst methanogenic activity remained constant at low concentrations (0.6 and 1.0 mg L-1), but polymeric resin compounds’ COD removal efficiency increased 58.1±1% whilst methane yield decreased, due to the higher aerobic mineralisation of carbon to carbon dioxide. The result of different glucose/polymeric resin compound ratios in the presence of 0.6 mg L-1 dissolved oxygen showed that glucose did not improve polymeric resin compound removal. However, methanogenic activity decreased by 75% with polymeric resin compounds as substrate compared to methanogenic activity with glucose as sole carbon source, suggesting that the presence of glucose promotes conditions for highertolerance to oxygen. The presence of low dissolved oxygen concentrations therefore promotes polymeric resin compounds’ methanogenesis and mineralisation.

En el presente trabajo se evaluó el efecto de la concentración de oxígeno disuelto (OD) y la presencia de laglucosa en la metanogénesis y mineralización de los compuestos de resinas poliméricas, mediante cultivos en lote. El inóculo utilizado para los cultivos fueron lodos procedentes de un reactor de lecho de lodos con flujoascendente UASB (por sus siglas en inglés), alimentado con glucosa y aguas residuales de resinas poliméricas,y 1,0 mg L-1 d-1 de oxígeno disuelto en estado estacionario. Todas las pruebas fueron realizadas con 1500 mgL-1 de demanda química de oxígeno, a 30±2° C, con 23,8 g L-1 de sólidos suspendidos volátiles como inóculoy una relación demanda química de oxígeno / sólidos suspendidos volátiles de 1. El efecto de diferentesconcentraciones de oxígeno disuelto mostró que, para la glucosa, con bajas concentraciones (0,6 y 1,0 mgL-1) la eficiencia de remoción de la demanda química de oxígeno y la actividad metanogénicas permanecieronconstantes, pero la eficiencia de remoción de la demanda química de oxígeno para los compuestos de resinaspoliméricas aumentó 58,1±1% y el rendimiento de metano disminuyó, debido a que una mayor fracción delcarbono alimentado fue mineralizada a dióxido de carbono, posiblemente por una ruta aerobia. Los resultadoscon diferentes proporciones de glucosa / compuestos de resinas poliméricas (CRP) en presencia de 0,6 mg L-1 de oxígeno disuelto mostraron que la presencia de glucosa no mejoró la eliminación de los compuestosde resinas poliméricas. Sin embargo, la actividad metanogénica disminuyó en un 75% cuando solamente se usó a los compuestos de resinas poliméricas como sustrato, al compararla con la actividad metanogénica con glucosa como única fuente de carbono, sugiriendo que la glucosa no favorece la eliminación de los compuestosde resinas poliméricas, pero proporciona una mejor tolerancia al oxígeno..

Sequential use of ammonium and leucine as nitrogen sources during growth of Geotrichum candidum on a glucose based medium / Uso secuencial del amonio y leucina como fuentes de nitrógeno durante el crecimiento de Geotrichum candidum en un medio basado en glucosa

Geotrichum candidum growth on ammonium and leucine as nitrogen sources and glucose as a carbon source was examined. A clear preference of G. candidum for ammonium over leucine as a nitrogen source was shown. Indeed, ammonium was completely exhausted at the end of exponential growth after less than 35 hrs of culture; in contrast only 5 percent of leucine was concomitantly assimilated. Growth continued at slower rates on glucose and leucine as carbon and nitrogen sources respectively, and at the end of culture (185 hrs), leucine was completely exhausted.

Influenza de la concentración de oxígeno en la producción de etanol a partir de la fermentación de mezclas D-glucosa/D-xilosa e hidrolizados de aserrín de Curupaú por Pichia stipitis CBS 5773 / Influence of oxygen concentration in ethanol production by Pichia stipitis CBS 5773 through fermentation of D-glucose/D-sylose mixtures and Curupaú hydrolyzed

El aserrin de Curupaú, un residuo de la industria maderera en Bolivia, es un substrato potencial para la producción de etanol de segunda generación.

Decolorization of pulp paper mill effluent by Pleurotus sajorcaju.

Pleurotus sajorcaju MTCC-141 procured from Microbial Type Culture Collection Centre and Gene Bank, Institute of Microbial Technology, Chandigarh has been used for color removal from paper mill effluent. The paper mill effluent amended with basal medium supports the growth of Pleurrotus sajorcaju and removes the colour. The optimum concentrations of carbon source (glucose) and nitrogen source (NH4NO3) for the maximum decolourization of paper mill effluent were found to be 1% and 0.2% respectively. During the fungal growth process, the pH of the paper mill effluent decreased from 7.94 to 4.0.

Solubilization of water-insoluble beta-glucan isolated from Ganoderma lucidum.

The fungal beta-D-glucan is a biological response modifier (BRM), but a major obstacle to the clinical utilization of beta-glucan BRMs is thei relative lack of solubility in aqueous media. Water insoluble fungal glucans extracted by alkali from the mycelia of Ganoderma lucidum were sulfated to yield their corresponding water-soluble derivatives. Insoluble glucan is dissolved in methyl sulfoxide and urea, and is partially sulfated with sulfuric acid. The sulfated glucan (SGL) yield prepared from insoluble glucan (IGL) was 85%, the sulfation degree of SGL was about 14.9%, and the solubility of SGL was above 95% in water. The monosugar SGL content was 34.9% alpha-glucose and 35.9% beta-glucose. The mean molecular weight (MW) of SGL was shown as a single peak on Sepharose CL-4B column chromatography, and their MW was approximately 9.3 kDa. The 13C NMR spectrum analysis shows that SGL has a high similarity with the beta-(1-->3)-linked triple-helical control.

Glucose level in the bronchial aspirate as a marker of ventilator associated pneumonia in mechanically ventilated patients

Ventilator-associated pneumonia [VAP] is the leading cause of death amongst hospital-acquired infections. Despite this, the diagnosis of VAP remains challenging and there is a lack of diagnostic standardization. In humans, glucose concentrations are normally low in nasal and bronchial fluid, but are elevated by inflammation or hyperglycaemia. Exhaled breath condensate glucose was reported to be a potential marker of both infection and outcome in patients with acute respiratory distress syndrome [ARDS]. To study the signifcance of detecting glucose in bronchial aspirate in mechaniclly ventilated patients under tight glycemic control and using it as a follow up parameter in patients with VAP versus patients with community acquired pneumonia [CAP]. Also the present work aimed to describe the pattern of changes of C - reactive protein [CRP] in patients with VAP versus those with CAP. Forty patients were enrolled into the present study and were divided into 3 groups: Group I: Thirty critically ill patients requiring intubation and mechanical ventilation were followed up from admission and daily till they fulfilled criteria suggesting development of VAP. Group II: Ten patients with community acquired pneumonia [CAP]. Group III: Ten healthy volunteers constituting the control group. Patients were assessed using the following scoring systems: 1] Assessing the disease severity using the Acute Physiological And Chronic Health Evaluation scoring system [APACHE II] on the day of admission, 2] Daily assessment of disease severity using the Sequential Organ Failure Assessment [SOFA] score, and 3] the Clinical pulmonary infection score [CPIS]. All patients with CAP were evaluated by Pneumonia severity index [PSI] on admission. CRP was assessed on admission, on day 2 and on day 7. All patients were under tight glycemic control to abolish the effect of hyperglycemia on the bronchial aspirate glucose levels. Glucose measurements were performed simultaneously in the blood and bronchial aspirates. Bronchial aspirates were analyzed for glucose on the day of admission and daily. The present study demonstrated a correlation between the presence of glucose in the bronchial aspirates from mechanically ventilated patients and development of VAP. Glucose in the bronchial aspirates preceded the development of VAP in the first 48 hours of mechanical ventilation and steadily increased thereafter. This was later associated with infiltrates on the chest radiographs, increased levels of CRP, with significant CPIS score. In contrast the glucose level in the sputum from patients with CAP was high on admission then steadily decreased till discharge. There was no correlation between patient's blood glucose and the glucose level in the bronchial aspirate. In the present study CRP values in patients with CAP steadily decreased from admission till the patients were discharged. The mean CRP value in the mechanically ventilated patients at day one was statistically significant less than that in patients with CAP. While at the 7[th] day, the mean CRP value in patients with VAP was higher than that in patients with CAP. The mean CRP level in patients with VAP was higher in nonsurvivors than survivors. In critically ill patients kept under tight glycemic control, early detection of glucose in bronchial aspirates may improve the ability of clinicians to early recognize the onset of VAP. This technique is simple, rapid, and inexpensive and following it daily may serve as a marker for infection or clinical resolution. CRP levels in patients with CAP steadily decrease from time of admission till discharge, whereas in patients with VAP, CRP steadily increases from time of VAP development thereafter thus suggesting a role in following such patients as a prognostic marker

Electrochemical studies of glucose oxidase immobilized on glutathione coated gold nanoparticles.

Glutathione (L-gamma-glutamyl-L-cysteinyl-L-glycine; GSH) forms a surface monolayer on gold nanoparticles by tethering via sulfur bonds (Au:GSH). In the present study, glucose oxidase (GOx; EC 1.1.3.4) was immobilized by covalent chemical coupling reactions on to Au:GSH nanoparticles and the enzyme coupled nanoparticles formed a stable colloid (stable for several weeks) in water. The immobilized enzyme was investigated for electrochemical characteristics to monitor the FAD (prosthetic group of the GOx) redox potentials. Various concentrations of substrate (glucose) were added to check the oxidation characteristics. It was observed that with increase in substrate concentrations, the oxidation rate increased proportionally with the current. The present study demonstrated that GOx was effectively coupled to the gold nanoparticle (Au:GSH). The coupled nanoparticle system could be used in a potential biosensor application. Similarly, other enzymes (e.g., horseradish peroxidase) could be immobilized to the Au:GSH nanoparticles via the peptide arm (GSH) to achieve the desired characteristics needed for a specific application in biosensor.

Response surface optimization of effective medium constituents for the production of alkaline protease from a newly isolated strain of Pseudomonas aeruginosa.

Optimization of the fermentation medium for maximum alkaline protease production was carried out with a new strain of Pseudomonas aeruginosa (B-2). Replacing the protein source/inducer (albumin in place of casein) brought about significant increase in yield after 48 hr of inoculation. Three most effective medium constituents identified by initial screening method of Plackett-Burman were albumin, (NH4)2SO4 and glucose. Central Composite Design (CCD) and Response Surface Methodology (RSM) were used in the design of the experiment and in the analysis of the results. Optimum levels of the effective medium constituents were albumin (6.586%); (NH4)2SO4, 0.164%; and glucose, 6.72%. The alkaline protease production increased from 533460 to 793492 Ul(-1).

Characterization and identification of chitinase producing Streptomyces venezuelae P10.

In an attempt to isolate chitinase producers from soil, a streptomycete strain was found potent using natural chitin as the substrate. Chitinolytic activity was tested directly on agar plates, also with crude enzyme. Chitinase assay showed that the isolate could produce 0.8 U/ml of the enzyme. The morphological, cultural, physiological and biochemical characters of the isolate P10 were studied, and identified as Streptomyces venezuelae P10.

Glucose isomerase de s violaceoniger aspects fondamentaux et appliques

XylA gene of Streptomyces violaceoniger [Drocourt et coll. 1988] code for a D-xlyose isomerase activity which is used as a D-glucose isomerase activity in large scale. This gene is a part of regulated region involved in xylose utilization [Marcel et coll. 1987]. Sequence determination of this region enabled us to characterize xylB gene [Xylulose kinase activity] and xylX gene which is involved in xylA and xylB expression. In order to construct a new strain having a strong and constitutive glucose isomerase activity, a newly isolated strong streptomyces promoter [P1 promoter], has been cloned behind xylA gene. To avoid instability of plasmid and glucose-isomerase activity, the Pl-xylA gene of S. violaceoniger has been integrated into the chromosome, using the integrative vector pTS55. The resultant CBS1 strain has four to five fold higher glucose-isomerase activity in absence of xylose compared to that of strain SV1 fully induced by xylose. In addition, specific glucose-isomerase activity of CBS1 strain increases in the secondary growth phase, in contrast to wild type and SV1 strains