Protease inhibitor from Moringa oleifera leaves: Isolation, purification, and characterization

Protease inhibitors have great demand in medicine and biotechnology. We report here the purification and characterization of a protease inhibitor isolated from mature leaf extract of Moringa oleifera that showed maximum inhibitor activity. The protease inhibitor was purified to 41.4-fold by Sephadex G75 and its molecular mass was calculated as 23,600 Da. Inhibitory activity was confirmed by dot-blot and reverse zymogram analyses. Glycine, glutamic acid, alanine, proline and aspartic acid were found as the major amino acids of the inhibitor protein. Maximal activity was recorded at pH 7 and at 40 ◦C. The inhibitor was stable over pH 5–10; and at 50 ◦C for 2 h. Thermostability was promoted by CaCl2, BSA and sucrose. Addition of Zn2+ and Mg2+, SDS, dithiothreitol and -mercaptoethanol enhanced inhibitory activity, while DMSO and H2O2 affected inhibitory activity. Modification of amino acids at the catalytic site by PMSF and DEPC led to an enhancement in the inhibitory activity. Stoichiometry of trypsin–protease inhibitor interaction was 1:1.5 and 0.6 nM of inhibitor effected 50% inhibition. The low Ki value (1.5 nM) obtained indicated scope for utilization of M. oliefera protease inhibitor against serine proteases

Age and growth of Gerres filamentosus (Cuvier, 1829) from Kodungallur, Azhikode Estuary, Kerala

The age and growth, length – weight relationship and relative condition factor of Gerres filamentosus (Cuvier, 1829) from Kodungallur, Azhikode Estuary were studied by examination of 396 specimens collected between May 2008 to October 2008. Here, length frequency method was used to study age and growth in fishes. L∞, K and t 0 obtained from seasonal and non - seasonal growth curves. Gerres filamentosus showed a low mortality rate (Z) 3.702 y-1. G. filamentosus has moderately low K value and long life span. The relation between the total length and weight of G. filamentosus was described as Log W = 1.321+2.5868 log L for males, Log W = 1.467 + 2.7227 log L for females and Log W = 1.481 + 2.7316 log L for sexes combined. The mean relative condition factor (Kn) values ranged from 0.9 to 1.14 for males, 0.89 to 1.11 for females and 0.73 to 1.08 for sexes combined. The length weight relationship and relative condition factor showed that the wellbeing of G. filamentosus were good. The morphometric measurements of various body parts were recorded. The morphometric measurements were found to be nonlinear and there is no significant difference observed between the two sexes.

Effects of post-harvest treatments on the carbohydrate composition of yacon roots in the Peruvian Andes

Yacon (Smallanthus sonchifolius [Poepp. & Endl.] H. Robinson) is an under-exploited native root crop of the Andes, which stores oligofructans (fructo-oligosaccharides, FOS) as its main component of dry matter (DM). FOS are of increasing economic interest because of their low caloric value in human diets and bifidogenic benefits on colon health. Two on-farm experiments were conducted to: (i) determine the effect of shaded, short-term storage at 1990 and 2930 m a.s.l. in the Andean highlands; and (ii) address the effects of a traditional sunlight exposure (‘sunning’) on the carbohydrate composition in the DM of tuberous yacon roots. After a 6-day shade storage FOS concentrations were smaller at the lower (36–48% of DM) than at the higher altitude (39–58% of DM). After 12 days FOS concentrations were nearly equal at both sites (27–39% of DM). The concentration of free sugars (fructose, glucose, sucrose) increased accordingly from 29–34 to 48–52%. During the 6-day sunning experiment FOS concentrations decreased from 50–62 to 29–44% and free sugars increased from 29–34 to 45–51%. The results indicate that partial hydrolysis of oligofructans starts shortly after harvest. Storage in highland environments should wherever possible exploit the cooler temperatures at higher altitudes. Sunning of yacon’s tuberous roots effectively reduces much of the roots’ water content, in this experiment 40%, and thus allows energy to be saved if yacon is processed into dehydrated products.

Einfluss der Führung des Trocknungsprozesses auf den Trocknungsverlauf und die Produkteigenschaften empfindlicher biologischer Güter

Die Konvektionstrocknung ist eine der ältesten Methoden zur Konservierung von Lebensmitteln. Es ist bekannt, dass Agrarprodukte temperaturempfindlich sind. Bei Überhitzung erleiden sie chemische und physikalische Veränderungen, die ihre Qualität vermindern. In der industriellen Praxis wird die Konvektionstrocknung in der Regel auf Grundlage empirisch ermittelter Werte durchgeführt, welche die Produkttemperatur nicht berücksichtigen. Es ist deshalb nicht sichergestellt, ob der Prozess optimal oder auch nur gut betrieben wird. Inhalt dieser Arbeit ist ein Vergleich der klassischen einstufigen Konvektionstrocknung mit einer Prozessführungsstrategie bei der die Produkttemperatur als Regelgröße eingeführt wird. Anhand der Untersuchung des Trocknungsverhaltens von Äpfeln werden die beiden Verfahren analysiert, die erhaltenen Ergebnisse miteinander verglichen und daraus optimierte Trocknungsbedingungen abgeleitet. Die für dieses Projekt entwickelte Prozessanlage erlaubt die simultane Untersuchung sämtlicher wesentlicher Temperaturen, der Gewichtsveränderung und der optischen Qualitätskriterien. Gleichzeitig ist es möglich, entweder die Lufttemperatur oder die Temperatur des Produktes zur regeln, während die jeweils andere Größe als Messwert erfasst wird. Es kann weiterhin zwischen Durch- und Überströmung gewählt werden.

Eficacia diagnostica del ácido láctico en liquido cefalorraquídeo en meningitis postclipaje de aneurisma cerebral y hemorragia subaracnoidea

La concentración de ácido láctico en LCR en pacientes con sospecha de meningitis postquirúrgica luego de clipaje de aneurisma cerebral y hemorragia subaracnoidea espontánea se midió prospectivamente por un período de tres años. Se analizaron un total de 32 muestras de líquido cefalorraquídeo, se midió la concentración de ácido láctico y se comparó con el cultivo de LCR. Los cultivos fueron positivos en cinco pacientes, con una prevalencia de infección del 15%. Se utilizó un valor umbral de ácido láctico de 4 mmol/L. y se encontró una sensibilidad del 80%, especificidad del 52%, VPP del 23%, VPN del 93%, y likelihood ratio (LHR) positivo de 1,66 con una probabilidad post test de 15% de la concentración del ácido láctico en el diagnóstico de meningitis postquirúrgica en pacientes con hemorragia subaracnoidea aneurismática. La concentración de ácido láctico en LCR tiene un desempeño limitado en el diagnóstico de meningitis postquirúrgica en pacientes con hemorragia subaracnoidea aneurismática.

Estrategias para potencializar el sector turístico en el departamento del Guainía

El departamento del Guainía, comprende una región con un alto potencial de desarrollo económico y social para el país debido a su biodiversidad y a su riqueza cultural. Sin embargo, ha sido un departamento que no ha presentado un desarrollo creciente de su economía debido a factores como: el aislamiento, la dependencia del estado, la limitada inversión en tecnología e infraestructura vial y la falta de capacitación en el capital humano, los cuales incrementa los costos logísticos y dificulta el desarrollo microempresarial. Es así, como en la actualidad la economía de la región se centra en los sectores primarios y en el sector público, los cuales presentan un mínimo valor agregado y de calidad para la zona. Teniendo en cuenta, la situación actual del departamento dentro del plan de desarrollo económico del Guainía y las apuestas productivas para la Amazorinoquia, se plantearon los lineamientos económicos y los ejes transversales de los sectores con mayor potencial de desarrollo para el Guainía. El turismo, el Bio-comercio y los mercados verdes, son los sectores analizados que podrían mejorar las condiciones socioeconómicas del departamento. De este modo, y teniendo en cuenta las condiciones de la región, el sector turístico, es un sector con un alto potencial de desarrollo que impacta a la economía en general y propende por el desarrollo social y el cuidado del medio ambiente. Por ello, la economía debería enfocar sus esfuerzos en el desarrollo de dicho sector, a través de un clúster del sector turístico que integre las iniciativas de los empresarios locales, las instituciones públicas y los gremios, promoviendo la competitividad, la innovación, la generación de sinergia y la promoción del departamento como centro eco-turístico de la Amazorinoquia colombiana.

New mono- and dinuclear ruthenium complexes containing the 3,5-bis(2-pyridyl)pyrazole ligand. Synthesis, characterization and applications

Se han sintetizado dos nuevos complejos mononucleares de Ru, con formula [RuCl2(Hbpp)(dmso)2], a partir de la reacción entre [RuCl2(dmso)4] y Hbpp (3,5-bis(2-piridil)pirazola). El hecho que sólo tres de los seis posibles estereoisómeros se obtengan a partir de esta reacción, se ha racionalizado en base a factores estructurales y electrónicos. Estos complejos se han caracterizado de forma estructural, espectroscópica y electroquímica. En acetonitrilo en medio básico, el isómero trans,cis-[RuCl2(Hbpp)(dmso)2] da lugar a procesos de isomerización de enlace de un ligando dmso cuando el Ru(II) se oxida a Ru(III). Las constantes termodinámicas y cinéticas para el proceso se han determinado por voltametria cíclica. La irradiación de trans,cis-[RuCl2(Hbpp)(dmso)2] y cis(out),cis-[RuCl2(Hbpp)(dmso)2] con luz UV o solar da lugar a reacciones de fotosustitución de un ligando dmso por una molécula de acetonitrilo para dar un nuevo compuesto el cual ha sido caracterizado en solución por técnicas espectroscópicas y electroquímicas. Ambos complejos resultan catalizadores útiles en la transferencia de hidrógeno de isopropanol a acetofenona, obteniéndose 1-feniletanol como único producto y un 42.1% de conversión (36.1 ciclos metálicos) a 80ºC con el isómero trans,cis-[RuCl2(Hbpp)(dmso)2], que resulta significativamente más eficaz que el complejo cis(out),cis-[RuCl2(Hbpp)(dmso)2]. La reacción de cis(out),cis-[RuCl2(Hbpp)(dmso)2] con trpy (2,2':6',2"-terpiridina) da lugar a los dos isómeros geométricos del complejo [Ru(Hbpp)(trpy)(Cl)]+, el in y el out. Estos complejos se han aislado y caracterizado por técnicas estructurales, espectroscópicas y electroquímicas. Estos cloro complejos han sido utilizados como precursores para la síntesis de los complejos análogos con ligandos aqua (in,out-[Ru(Hbpp)(trpy)(H2O)]2+) y piridina (in,out-[Ru(Hbpp)(trpy)(py)]2+), los cuales también han sido aislados y caracterizados. Las propiedades ácido-base de los aqua complejos, y del complejo out-py se han estudiado detalladamente por voltametria cíclica y mediante valoraciones espectrofotométricas ácido-base. El tratamiento matemático de los datos así obtenidos nos ha permitido determinar los valores de pKa para los distintos equilibrios de protonación de los complejos en los estados de oxidación II y III. El complejo out-aqua ha demostrado ser un buen catalizador para la oxidación electroquímica del alcohol benzílico, presumiblemente a benzaldehido. La constante de velocidad de segundo orden para el proceso ha sido determinada como 17.1 M-1 s-1, por simulación matemática. El dímero con un puente cloro, [Ru2Cl(bpp)(trpy)2]2+ ha sido preparado por dos rutas sintéticas diferentes. El dímero análogo con un puente acetato se ha obtenido por reacción del cloro dímero con un exceso de acetato sódico. El dímero con dos ligandos aqua [Ru2(bpp)(trpy)2(OH2)2]3+ puede obtenerse por hidrólisis ácida del complejo con un acetato puente o por hidrólisis básica del complejo con un puente cloro. Estos complejos han sido caracterizados por técnicas estructurales, espectroscópicas y electroquímicas. Las soluciones del dímero con dos ligandos aqua en medio ácido resultan inestables a la coordinación de aniones de la solución con el tiempo. Las propiedades ácido-base del dímero con dos aguas coordinadas han sido estudiadas por voltametria cíclica y mediante experimentos de electrólisis a potencial controlado. El pKa para la desprotonación de uno de los ligandos aqua ha sido determinado mediante una valoración espectrofotométrica ácido-base como 6.7. Este valor tan bajo de pKa se atribuye a la formación de la entidad {Ru2O2H3}, favorable termodinámicamente. Los espectros UV-vis para los distintos estados de oxidación del aqua dímero, de RuIIRuII a RuIIIRuIV, han sido obtenidos por oxidación química y electroquímica del complejo. Se han llevado a cabo estudios cinéticos de la oxidación, paso a paso, de RuII,II a RuIV,IV , y se han determinado las constantes de oxidación de segundo orden para los distintos procesos de oxidación. La capacidad del aqua dímero en la oxidación del agua a oxígeno molecular ha sido investigada en solución homogénea utilizando CeIV como oxidante. La evolución de oxígeno se ha demostrado por cromatografia de gases. Se ha obtenido una eficiencia del 73% y 18.6 ciclos catalíticos, cuando 1.83 x 10-6 moles de dímero se han mezclado con un exceso de 100 equivalentes de cerio. El dímero con dos aguas cataliza también la oxidación del agua de forma heterogénea, con el complejo adsorbido sobre una membrana de nafion, aunque la eficiencia es menor. Se ha propuesto un mecanismo intramolecular para la reacción de oxidación del agua. Consiste en la oxidación a 4 electrones del dímero, de RuII,II a RuIV,IV, el cual reacciona con el agua para formar oxígeno y revierte nuevamente al estado de oxidación II,II. Este modelo es consistente con estudios catalíticos de la evolución de oxígeno en función de las concentraciones de cerio y catalizador, llevados a cabo en solución ácida homogénea, que demuestran que la oxidación a 4 electrones del agua se encuentra catalizada por una sola molécula de complejo bajo concentraciones elevadas de cerio. La constante de pseudo-primer-orden para la evolución de oxígeno tiene un valor de 1.4 x 10-2 s-1, que es uno de los valores de constante más elevados obtenidos hasta la fecha. Desafortunadamente, el aqua dímero se desactiva durante el proceso de catálisis dando lugar a una especie naranja, la cual estamos actualmente tratando de caracterizar.

Biodiversity of Collembola in urban soils and the use of Folsomia candida to assess soil 'quality'

The effects of metal contamination on natural populations of Collembola in soils from five sites in the Wolverhampton area ( West Midlands, England) were examined. Analysis revealed that metal concentrations were elevated above background levels at all sites. One location in particular (Ladymoor, a former smelting site) was highly contaminated with Cd, Cu, Pb and Zn at more than 20 times background levels. Biodiversity indices ( Shannon - Weiner, Simpson index, Margalef index, alpha index, species richness, Shaneven ( evenness) and Berger - Parker dominance) were calculated. Of these indices, estimates of species richness and evenness were most effective at highlighting the differences between the Collembola communities. Indeed, the highest number of species were found at the most contaminated site, although the Collembola population also had a comparatively low evenness value, with just two species dominating. The number of individuals per species were allocated into geometric classes and plotted against the cumulative number of species as a percentage. At Ladymoor, there were more geometric classes, and the slope of the line was shallower than at the other four sites. This characteristic is a feature of polluted sites, where a few species are dominant and most species are rare. The Ladymoor soil also had a dominance of Isotomurus palustris, and was the only site in which Ceratophysella denticulata was found. Previous studies have shown that these two species are often found in sites subject to high metal contamination. Survival and reproduction of the "standard'' test springtail, Folsomia candida (Willem), were determined in a 4 week exposure test to soils from all five sites. Mortality was significantly increased in adults and reproduction significantly lower in the Ladymoor soil in comparison to the other four sites. This study has shown that severe metal contamination can be related to the population structure of Collembola in the field, and performance of F. candida ( in soils from such sites) in the laboratory.

Cellulose microfibril angle in the cell wall of wood fibres

The term microfibril angle (MFA) in wood science refers to the angle between the direction of the helical windings of cellulose microfibrils in the secondary cell wall of fibres and tracheids and the long axis of cell. Technologically, it is usually applied to the orientation of cellulose microfibrils in the S2 layer that makes up the greatest proportion of the wall thickness, since it is this which most affects the physical properties of wood. This review describes the organisation of the cellulose component of the secondary wall of fibres and tracheids and the various methods that have been used for the measurement of MFA. It considers the variation of MFA within the tree and the biological reason for the large differences found between juvenile (or core) wood and mature (or outer) wood. The ability of the tree to vary MFA in response to environmental stress, particularly in reaction wood, is also described. Differences in MFA have a profound effect on the properties of wood, in particular its stiffness. The large MFA in juvenile wood confers low stiffness and gives the sapling the flexibility it needs to survive high winds without breaking. It also means, however, that timber containing a high proportion of juvenile wood is unsuitable for use as high-grade structural timber. This fact has taken on increasing importance in view of the trend in forestry towards short rotation cropping of fast grown species. These trees at harvest may contain 50% or more of timber with low stiffness and therefore, low economic value. Although they are presently grown mainly for pulp, pressure for increased timber production means that ways will be sought to improve the quality of their timber by reducing juvenile wood MFA. The mechanism by which the orientation of microfibril deposition is controlled is still a matter of debate. However, the application of molecular techniques is likely to enable modification of this process. The extent to which these techniques should be used to improve timber quality by reducing MFA in juvenile wood is, however, uncertain, since care must be taken to avoid compromising the safety of the tree.

Biodiversity of Collembola in urban soils and the use of Folsomia candida to assess soil 'quality'

The effects of metal contamination on natural populations of Collembola in soils from five sites in the Wolverhampton area ( West Midlands, England) were examined. Analysis revealed that metal concentrations were elevated above background levels at all sites. One location in particular (Ladymoor, a former smelting site) was highly contaminated with Cd, Cu, Pb and Zn at more than 20 times background levels. Biodiversity indices ( Shannon - Weiner, Simpson index, Margalef index, alpha index, species richness, Shaneven ( evenness) and Berger - Parker dominance) were calculated. Of these indices, estimates of species richness and evenness were most effective at highlighting the differences between the Collembola communities. Indeed, the highest number of species were found at the most contaminated site, although the Collembola population also had a comparatively low evenness value, with just two species dominating. The number of individuals per species were allocated into geometric classes and plotted against the cumulative number of species as a percentage. At Ladymoor, there were more geometric classes, and the slope of the line was shallower than at the other four sites. This characteristic is a feature of polluted sites, where a few species are dominant and most species are rare. The Ladymoor soil also had a dominance of Isotomurus palustris, and was the only site in which Ceratophysella denticulata was found. Previous studies have shown that these two species are often found in sites subject to high metal contamination. Survival and reproduction of the "standard'' test springtail, Folsomia candida (Willem), were determined in a 4 week exposure test to soils from all five sites. Mortality was significantly increased in adults and reproduction significantly lower in the Ladymoor soil in comparison to the other four sites. This study has shown that severe metal contamination can be related to the population structure of Collembola in the field, and performance of F. candida ( in soils from such sites) in the laboratory.

Cellulose microfibril angle in the cell wall of wood fibres

The term microfibril angle (MFA) in wood science refers to the angle between the direction of the helical windings of cellulose microfibrils in the secondary cell wall of fibres and tracheids and the long axis of cell. Technologically, it is usually applied to the orientation of cellulose microfibrils in the S2 layer that makes up the greatest proportion of the wall thickness, since it is this which most affects the physical properties of wood. This review describes the organisation of the cellulose component of the secondary wall of fibres and tracheids and the various methods that have been used for the measurement of MFA. It considers the variation of MFA within the tree and the biological reason for the large differences found between juvenile (or core) wood and mature (or outer) wood. The ability of the tree to vary MFA in response to environmental stress, particularly in reaction wood, is also described. Differences in MFA have a profound effect on the properties of wood, in particular its stiffness. The large MFA in juvenile wood confers low stiffness and gives the sapling the flexibility it needs to survive high winds without breaking. It also means, however, that timber containing a high proportion of juvenile wood is unsuitable for use as high-grade structural timber. This fact has taken on increasing importance in view of the trend in forestry towards short rotation cropping of fast grown species. These trees at harvest may contain 50% or more of timber with low stiffness and therefore, low economic value. Although they are presently grown mainly for pulp, pressure for increased timber production means that ways will be sought to improve the quality of their timber by reducing juvenile wood MFA. The mechanism by which the orientation of microfibril deposition is controlled is still a matter of debate. However, the application of molecular techniques is likely to enable modification of this process. The extent to which these techniques should be used to improve timber quality by reducing MFA in juvenile wood is, however, uncertain, since care must be taken to avoid compromising the safety of the tree.

Shovel ready? An empirical investigation of stalled residential sites.

Drawing upon a national database of unimplemented planning permissions and 18 in-depth case studies, this paper provides both a quantitative and qualitative analysis of the phenomenon of stalled sites in England. The practical and conceptual difficulties of classifying sites as stalled are critically reviewed. From the literature, it is suggested that planning permission may not be implemented due to lack of financial viability, strategic behaviour by landowners and house-builders and other problems associated with the development process. Consistent with poor viability, the analysis of the national database indicates that a substantial proportion of the stalled sites is high density apartment development and/or is located in low house value areas. The case studies suggest that a combination of interlinked issues may need to be resolved before a planning permission can be implemented. These include; the sale of the land to house-builders, re-negotiation of the planning permission and, most importantly, improvement in housing market conditions.

Characterisation of microsatellite markers for fig-pollinating wasps in the Pleistodontes imperialis species complex

We characterised a set of nine polymorphic microsatellite loci for Pleistodontes imperialis sp. 1, the pollinator wasp of Port Jackson fig (Ficus rubiginosa) in south-eastern Australia. Characterisation was performed on 30 female individuals collected from a population in Sydney, Australia. The average number of alleles per locus was 7.33, and eight loci were not in Hardy–Weinberg equilibrium. This was expected as fig wasps are known to be highly inbred. A test of genetic differentiation between two natural populations of P. imperialis sp. 1 (Sydney and Newcastle, Australia – some 120 km apart) yielded a very low FST value of 0.012, suggesting considerable gene flow. Bayesian clustering analysis using TESS 2.3.1, which does not assume Hardy–Weinberg equilibrium, however, indicated potential spatial substructuring between the Sydney and Newcastle populations, as well as within the Sydney population. The described loci were also characterised for two other species in the P. imperialis complex: P. imperialis sp. 2 (Townsville, Australia) and P. imperialis sp. 4 (Brisbane, Australia). Seven and six of the nine loci were polymorphic for P. imperialis sp. 2 and P.imperialis sp. 4, respectively.

Adaptiv stämning : En bedömning av acceptabilitet och tonkvalitetsuppfattning

This thesis presents an adaptive tuning system that can be described as a dynamic Just Intonation tuning system, being compatible with equally tempered instruments. The tuning system is called Hermode Tuning (HMT) and the tuning used as comparison for evaluation is the standardized western tuning, the equal tempered tuning. This study investigates preferences for these two musical tuning systems, depending on whether the tunings are presented on a piano or with woodwind instruments. A listening test was done with students at the Falun Conservatory of Music, including both a vertical listening (intervalls) and a horizontal listening (cadenses and musical compositions) of Hermode tuned musical material. Overall the results showed no significant preferences for either tuning system irrespectively of what instrument it was presented with. The clearest results was that of a misjudged just intonated perfect third on the piano and a preference for an adaptively tuned piano presented in a simple harmonic structure, with a parameter setting of HMT 70%. Materials for comparison was partly taken from Hermode´s own website, but overall the attitude towards these sequenses (using a likert scale of one to five) showed a low expected value. This shows the complexity of the topic and no general conclusions regarding the choice of intonation or tuning system could be done for the presented material.