How Does the Spacer Length of Cationic Gemini Lipids Influence the Lipoplex Formation with Plasmid DNA? Physicochemical and Biochemical Characterizations and their Relevance in Gene Therapy

Lipoplexes formed by the pEGFP-C3 plasmid DNA (pDNA) and lipid mixtures containing cationic gemini surfactant of the 1,2-bis(hexadecyl dimethyl ammonium) Acmes family referred to as C16CnC16, where n = 2 3, 5, or 12, and the zwitterionic helper lipid, 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) have been studied from a wide variety of physical, chemical, and biological standpoints. The study has been carried out using several experimental methods, such as zeta potential, gel electrophoresis, small-angle X-ray scattering (SAXS), cryo-TEM, gene transfection, cell viability/cytotoxicity, and confocal fluorescence microscopy. As reported recently in a communication (J. Am. Chem. Soc. 2011, 133, 18014), the detailed physicochemical and biological studies confirm that, in the presence of the studied series lipid mixtures, plasmid DNA is compacted with a large number of its associated Na+ counterions. This in turn yields a much lower effective negative charge, q(pDNA)(-), a value that has been experimentally obtained for each mixed lipid mixture. Consequently, the cationic lipid (CL) complexes prepared with pDNA and CL/DOPE mixtures to be used in gene transfection require significantly less amount of CL than the one estimated assuming a value of q(DNA)(-) = -2. This drives to a considerably lower cytotoxicity of the gene vector. Depending on the CL molar composition, alpha, of the lipid mixture, and the effective charge ratio of the lipoplex, rho(eff), the reported SAXS data indicate the presence of two or three structures in the same lipoplex, one in the DOPE-rich region, other in the CL-rich region, and another one present at any CL composition. Cryo-TEM and SAXS studies with C16CnC16/DOPE-pDNA lipoplexes indicate that pDNA is localized between the mixed lipid bilayers of lamellar structures within a monolayer of similar to 2 nm. This is consistent with a highly compacted supercoiled pDNA conformation compared with that of linear DNA. Transfection studies were carried out with HEK293T, HeLa, CHO, U343, and H460 cells. The alpha and rho(eff) values for each lipid mixture were optimized on HEK293T cells for transfection, and using these values, the remaining cells were also transfected in absence (-FBS-FBS) and presence (-FBS+FBS) of serum. The transfection efficiency was higher with the CLs of shorter gemini spacers (n = 2 or 3). Each formulation expressed GFP on pDNA transfection and confocal fluorescence microscopy corroborated the results. C16C2C16/DOPE mixtures were the most efficient toward transfection among all the lipid mixtures and, in presence of serum, even better than the Lipofectamine2000, a commercial transfecting agent Each lipid combination was safe and did not show any significant levels of toxicity. Probably, the presence of two coexisting lamellar structures in lipoplexes synergizes the transfection efficiency of the lipid mixtures which are plentiful in the lipoplexes formed by CLs with short spacer (n = 2, 3) than those with the long spacer (n = 5, 12).

Characterization and iodide adsorption behaviour of HDPY+ modified bentonite

Owing to its favourable physical, chemical and rheological properties, densely compacted bentonite or bentonite-sand mix is considered as a suitable buffer material in deep geological repositories to store high level nuclear waste. Iodine-129 is one of the significant nuclides in the high level waste owing to its long half life and poor sorption onto most geologic media. Bentonite by virtue of negatively charged surface has negligible affinity to retain iodide ions. As organo-bentonites are known to retain iodide ions, the present study characterizes hexadecylpyridinium chloride (HDPyCl.H2O) treated bentonite from Barmer India (referred as HDPy+B) for physico-chemical properties, engineering properties and the iodide adsorption behavior of the organo clay. Batch experiments revealed that HDPy+ ions are largely retained (94 % retention) via cation exchange; the ion-exchange process neutralizes the negative surface charge and bridges clay particles leading to reduction in Atterberg limits, clay content and sediment volume. The organo clay retains iodide by Coulombic attraction (at primary sites) and anion exchange (at secondary sites). The free-energy change (Delta G (o) = -25.5 kJ/mol) value indicated that iodide retention by organo clay is favored physical adsorption process. Iodide adsorption capacity of organo clay decreased significantly (85-100 %) on dilution with 50-80 % bentonite. On the other hand, dilution of bentonite with 50 % organo clay caused 58 % reduction in swell potential and 21 % reduction in swell pressure.

Isolation and characterization of biofumigant from leaves of Lantana camara for control of stored grain insect pests

Due to environmental concerns, health hazards to man and the evolution of resistance in insect pests, there have been constant efforts to discover newer insecticides both from natural sources and by chemical synthesis. Natural sources for novel molecules hold promise in view of their eco-friendly nature, selectivity and mammalian safety. We have isolated one natural bioactive molecule from the leaves of Lantana camara named Coumaran, based on various physical-chemical and spectroscopic techniques (IR, H-1 NMR, C-13 NMR and MS). Coumaran is highly toxic and very low concentration is needed for control of stored product insects. This molecule has potent grain protectant potential and caused significant reduction in F1 progeny of all the three species in the treated grain and the progeny was completely suppressed at 30 mu g/l. The differences in germination between the control and treated grains were not significant. The lack of any adverse effect of Coumaran on the seed germination is highly desirable for a grain protectant, becoming a potential source of biofumigant for economical and environmentally friendly pest control strategies against stored grain pests during storage of grains or pulses. (C) 2013 Elsevier B.V. All rights reserved.

Why is the influence of soil macrofauna on soil structure only considered by soil ecologists?

These last twenty years have seen the development of an abundant literature on the influence of soil macrofauna on soil structure. Amongst these organisms, earthworms, termites and ants are considered to play a key role in regulating the physical, chemical and microbiological properties of soils. Due to these influential impacts, soil ecologists consider these soil macro-invertebrates as `soil engineers' and their diversity and abundance are nowadays considered as relevant bioindicators of soil quality by many scientists and policy makers. Despite this abundant literature, the soil engineering concept remains a `preach to the choir' and bioturbation only perceived as important for soil ecologists. We discussed in this article the main mechanisms by which soil engineers impact soil structure and proposed to classify soil engineers with respect to their capacity to produce biostructures and modify them. We underlined the lack of studies considering biostructure dynamics and presented recent techniques in this purpose. We discussed why soil engineering concept is mainly considered by soil ecologists and call for a better collaboration between soil ecologists and soil physicists. Finally, we summarized main challenges and questions that need to be answered to integrate soil engineers activities in soil structure studies. (C) 2014 Elsevier B.V. All rights reserved.

Comparison of Convective and Radiative Heating Modes on the Thermophysical Changes of a Cerium Nitrate Droplet

In this paper, we try to establish the equivalence or similarity in the thermal and physiochemical changes in precursor droplets (cerium nitrate) in convective and radiative fields. The radiative field is created through careful heating of the droplet using a monochromatic light source (CO2 laser). The equivalence is also established for different modes of convection like droplet injected into a high-speed flow and droplet experiencing a convective flow due to acoustic streaming (levitated) only. The thermophysical changes are studied in an aqueous cerium nitrate droplet, and the dissociation of cerium nitrate to ceria is modeled using modified Kramers' reaction rate formulation. It is observed that vaporization, species accumulation, and chemical characteristics obtained in a convectively heated droplet are retained in a radiatively heated droplet by careful adjustment of the laser intensity. The timescales and ceria yield match reasonably well for both the cases. It is also noted that similar conclusions are drawn in both levitated droplet and a nonlevitated droplet.

Environmental Mechanics Research in China

By recalling mankind's path during past 50 years in the present article, we mainly highlight the significance of environmental issues today. In particular, two major factors leading to environment deterioration in China such as water resources and coal burning are stressed on. Present-day environmental issues are obviously interdisciplinary, of multiple scales and multi-composition in nature. Therefore, a process-based approach for environment research is absolutely necessarily. A series of sub-processes, either physical, chemical or biological, are subsequently analyzed in order to established reasonable parameterization scheme and credible comprehensive model. And we are now in a position to answer questions still open to us, improve existing somewhat empirical engineering approaches and enhance quantitative accuracy in prediction. To illustrate this process-based research approach, three typical examples associated with the Yangtze River Estuary, Loess Plateau and Tenggeli Desert environments have been dealt with respectively. A theoretical model of vertical flow field accounting for runoff and tide interaction has been established to delineate salinity and sediment motion which are responsible for the formation of mouth bar at the outlet and the ecological evolution there. A kinematic wave theory combined with the revised Green-Ampt infiltration formula is applied to the prediction of runoff generation and erosion in three types of erosion region on the Loess Plateau. Three approaches describing water motion in SPAC system in arid areas at different levels have been improved by introducing vegetation sub-models. However, we have found that the formation of a dry sandy layer and biological crust skin are additional primary causes leading to deterioration of water supply and succession of ecological system.

Micro-systems for Optical Protein-Chip

Protein-Chip as micro-assays for the determination of protein interaction, the analysis, the identification and the purification of proteins has large potential applications. The Optical Protein-Chip is able to detect the multi-interaction of proteins and multi-bio-activities of molecules directly and simultaneously with no labeling. The chip is a small matrix on solid substrate containing multi-micro-area prepared by microfabrication with photolithography or soft lithography for surface patterning, and processed with surface modification which includes the physical, chemical, and bio-chemical modifications, etc. The ligand immobilization, such as protein immobilization, especially the oriented immobilization with low steric hindrance and high bio-specific binding activity between ligand and receptor is used to form a sensing surface. Each area of the pattern is corresponding to only one bioactivity. The interval between the areas is non-bioactive and optically extinctive. The affinity between proteins is used to realize non-labeling microassays for the determination of protein identification and protein interaction. The sampling of the chip is non-disturbing, performed with imaging ellipsometry and image processing on a database of proteins.

Proceedings of the PICES/CoML/IPRC Workshop on Impact of Climate Variability on Observation and Prediction of Ecosystem and Biodiversity Changes in the North Pacific

Table of Contents [pdf, 0.09 Mb] Section I - Presentations and Discussions at Plenary Sessions Introduction and Overview of Workshop Objectives [pdf, 0.07 Mb] Plenary Session Presentations [pdf, 2.23 Mb] Reports of the Breakout Group Discussions [pdf, 0.43 Mb] Closing Plenary Discussion and Recommendations [pdf, 0.11 Mb] Section II - Extended Abstracts of Individual Presentations at Breakout Group Sessions Breakout Group 1: Physical/Chemical Oceanography and Climate [pdf, 6.14 Mb] Breakout Group 2: Phytoplankton, Zooplankton, Micronekton and Benthos [pdf, 28.14 Mb] Breakout Group 3: Fish, Squid, Crabs and Shrimps [pdf, 4.30 Mb] Breakout Group 4: Highly Migratory Fishes, Seabirds and Marine Mammals [pdf, 6.27 Mb] Appendix 1. Workshop agenda [pdf, 0.15 Mb] Appendix 2. List of participants [pdf, 0.13 Mb] (Document pdf contains 216 pages)

Test well exploration in the Myakka River Basin area, Florida

In recent years, difficulties encountered in obtaining ground-water supplies with acceptable chemical characteristics in the Myakka River basin area led to the implementation of a test drilling program. Under this program, well drilling and data collection were executed in such a manner that all water-producing zones of the local aquifers, together with the quality and quantity of the water available, were effectively identified. A step-drilling method was utilized which allowed the collection of formation cuttings, water samples, and water-level data, from isolated zones in the well as drilling proceeded. The step drilling procedure is described. The driller's logs, geophysical logs, and chemical quality of water tables are presented.(Document has 66 pages.)

Techniques for spatial analysis and visualization of benthic mapping data: final report

The mapping and geospatial analysis of benthic environments are multidisciplinary tasks that have become more accessible in recent years because of advances in technology and cost reductions in survey systems. The complex relationships that exist among physical, biological, and chemical seafloor components require advanced, integrated analysis techniques to enable scientists and others to visualize patterns and, in so doing, allow inferences to be made about benthic processes. Effective mapping, analysis, and visualization of marine habitats are particularly important because the subtidal seafloor environment is not readily viewed directly by eye. Research in benthic environments relies heavily, therefore, on remote sensing techniques to collect effective data. Because many benthic scientists are not mapping professionals, they may not adequately consider the links between data collection, data analysis, and data visualization. Projects often start with clear goals, but may be hampered by the technical details and skills required for maintaining data quality through the entire process from collection through analysis and presentation. The lack of technical understanding of the entire data handling process can represent a significant impediment to success. While many benthic mapping efforts have detailed their methodology as it relates to the overall scientific goals of a project, only a few published papers and reports focus on the analysis and visualization components (Paton et al. 1997, Weihe et al. 1999, Basu and Saxena 1999, Bruce et al. 1997). In particular, the benthic mapping literature often briefly describes data collection and analysis methods, but fails to provide sufficiently detailed explanation of particular analysis techniques or display methodologies so that others can employ them. In general, such techniques are in large part guided by the data acquisition methods, which can include both aerial and water-based remote sensing methods to map the seafloor without physical disturbance, as well as physical sampling methodologies (e.g., grab or core sampling). The terms benthic mapping and benthic habitat mapping are often used synonymously to describe seafloor mapping conducted for the purpose of benthic habitat identification. There is a subtle yet important difference, however, between general benthic mapping and benthic habitat mapping. The distinction is important because it dictates the sequential analysis and visualization techniques that are employed following data collection. In this paper general seafloor mapping for identification of regional geologic features and morphology is defined as benthic mapping. Benthic habitat mapping incorporates the regional scale geologic information but also includes higher resolution surveys and analysis of biological communities to identify the biological habitats. In addition, this paper adopts the definition of habitats established by Kostylev et al. (2001) as a “spatially defined area where the physical, chemical, and biological environment is distinctly different from the surrounding environment.” (PDF contains 31 pages)

Biological production in the Eastern Pacific Ocean

ENGLISH:The present paper is principally concerned with the geographic distribution of the standing crop and production of phytoplankton at the surface of the eastern Pacific, east of 130°W and between 10°N and 33°S, as reflected by recently collected data. In addition we discuss some of the more obvious, general relationships among thermocline topography, nutrient concentration, and the various trophic levels from primary production to fish production. The limited data do not allow a seasonal study. We have therefore mapped all of the data together regardless of the time of collection, but do not wish to imply that the physical, chemical and biological system is without seasonal or periodic change. SPANISH:Como lo reflejan los datos recientemente recolectados, el presente trabajo está dedicado principalmente a la distribución geográfica de las cosechas estables y a la producción del fitoplancton en la superficie del Pacífico Oriental, al este de los 130°W y entre los 10°N y 33°S. Además discutimos algunas de las relaciones generales más obvias entre la topografía de la termoclina, la concentración de los nutrientes, y los varios niveles tróficos, desde la producción primaria hasta la producción de los peces. Los datos limitados no permiten un estudio estacional. Por lo tanto, hemos combinado todos los datos no tomando en cuenta el tiempo de la recolección, pero no queremos implicar que no existen cambios estacionales o periódicos en el sistema físico, químico y biológico.

Primary production, chlorophyll, and zooplankton volumes in the tropical Eastern Pacific Ocean

ENGLISH: Between 1 October and 17 December 1955 investigations of the physical, chemical and biological oceanography of the Eastern Pacific Ocean in a region bounded approximately by 30° N. latitude, 9° S. latitude, 120° W. longitude and the mainland coast were conducted from the vessels Horizon and Spencer F. Baird of the Scripps Institution of Oceanography of the University of California. These were part of a cooperative operation, designated for convenience by the code name "Eastropic," in which a vessel of the U. S. Fish and Wildlife Service worked, during this same period, further west and a vessel of the Peruvian Navy worked further south, offshore from Peru. A vessel of the California State Fisheries Laboratory also conducted certain sub-surface tuna fishing operations and other studies in the same general region as the Scripps vessels. In addition to carrying out a number of special studies related to particular oceanographic features, the Scripps vessels occupied a considerable number of hydrographic stations. The locations of these stations, at each of which were made net-hauls for zooplankton, are shown in Figure 4 and Tables 2 and 3. At some of the hydrographic stations, and in Some places between stations, there were made from the Spencer F. Baird measurements of chlorophyll "a" and of primary production (by the C14 technique), both in situ and in a shipboard incubator. The purpose of this paper is to report on the results of these biological observations. SPANISH: Entre el 1° de octubre y el 17 de diciembre de 1955, a bordo de los barcos Horizon y Spencer F. Baird) de la Institución Scripps de Oceanografía de la Universidad de California, se hicieron investigaciones sobre la oceanografía física, química y biológica del Océano Pacífico Oriental, en una región limitada aproximadamente por los 30° N. de latitud, 9° S. de latitud, 120° O. de longitud y la costa continental. Estas investigaciones fueron parte de una operación que se realizó cooperativamente y a la que se convino darle el nombre codificado de "Eastropic". En ella, durante el mismo período, una embarcación del Servicio de Pesca y Vida Silvestre de los Estados Unidos (U. S. Fish and Wildlife Service) trabajó más hacia el oeste, y un barco de la armada peruana más hacia el sur, frente a la costa del Perú. También colaboró una nave del Laboratorio de Pesquerías del Estado de California (California State Fisheries Laboratory), realizando algunas operaciones de pesca de atún en aguas subsuperficiales, y otros estudios en la misma región general que recorrieron las embarcaciones de Scripps. Además de efectuar estudios especiales relacionados con las caracteristicas oceanográficas particulares de la región, las naves de Scripps establecieron un buen número de estaciones hidrográficas. La localización de estas estaciones se indica en la Figura 4 y en las Tablas 2 y 3; en cada una de ellas se hicieron rastreos con redes planctónicas para recoger muestras de zooplancton. En algunas de las estaciones hidrográficas, así como en algunos lugares entre estaciones, en el Spencer F. Baird se hicieron mediciones de la clorofila "a" y de la producción primaria (mediante la técnica del C14), tanto in situ como en una incubadora instalada a bordo. El propósito del presente trabajo es dar a conocer los resultados de estas observaciones biológicas. (PDF contains 44 pages.)

Herstellung und Zusammensetzung von asiatischen Fischsaucen

Abstract Fish sauce belongs to the most important condiments in Southeast Asian cuisine. It is a clear, amber to reddish liquid with an intensive smell. Fish sauce is used instead of salt for nearly each meal. Asian fish sauce is made from anchovies and other small fish. For the traditional process whole fresh fish are mixed with salt in the ratio 1:1 to 6:1 in wooden, clay or concrete tanks at tropical temperatures for 6 to 18 months. The liquefaction of the fish tissue is due to the action of endogenous enzymes in fish and exogenous enzymes from bacteria. During the fermentation amino acids, peptides and a lot of other substances are built, which are responsible for the characteristic aroma and flavour of these sauces. You can buy pure fish sauce, diluted fish sauce and fish sauce made from other types of animals like mussels, prawns and squids. In single Asian countries there are different national standards for the quality of fish sauces. In order to get a general idea of these products we have bought 16 fish and two oyster sauces from the retail trade in Hamburg and analyzed them with physical, chemical, sensory and microbiological methods. Kurzfassung Fischsauce gehört zu den wichtigsten Würzsaucen in der südostasiatischen Küche. Es ist eine klare, bernsteinfarbene bis rötlichbraune, sehr intensiv riechende Flüssigkeit. Sie wird anstelle von Salz verwendet und daher fast zu jedem Essen gereicht. Zur Herstellung von Fischsaucen werden hauptsächlich Anchovis und ähnliche kleine Fische verwendet. Bei der traditionellen Herstellung werden die ganzen Fische mit Meersalz in einem Holzfass, Tongefäß oder Betontank im Verhältnis 1:1 bis 6:1 gemischt. Während der anschließenden 6 – 18 Monate dauernden Lagerung bei tropischen Temperaturen bauen sich die Gewebeproteine durch fischeigene Enzyme und Mikroorganismen ab. Bei diesem mehrmonatigen Fermentationsprozess entstehen die für den Geschmack wichtigen Aminosäuren, Peptide und Aromastoffe. Es gibt neben reiner Fischsauce, auch verdünnte Fischsauce und Fischsaucen aus anderen Tieren wie Muscheln, Garnelen und Tintenfische. In den einzelnen asiatischen Ländern gibt es unterschiedliche nationale Qualitätsstandards. Um diese Produktgruppe näher kennen zu lernen, haben wir 16 Fisch- und 2 Austernsaucen aus dem Einzelhandel (Hamburg) mit physikalischen, chemischen, sensorischen und mikrobiologischen Verfahren untersucht.

Biocompatible Bacterial Cellulose-Poly(2-hydroxyethyl methacrylate) Nanocomposite Films

A series of bacterial cellulose-poly(2-hydroxyethyl methacrylate) nanocomposite films was prepared by in situ radical polymerization of 2-hydroxyethyl methacrylate (HEMA), using variable amounts of poly(ethylene glycol) diacrylate (PEGDA) as crosslinker. Thin films were obtained, and their physical, chemical, thermal, and mechanical properties were evaluated. The films showed improved translucency compared to BC and enhanced thermal stability and mechanical performance when compared to poly(2-hydroxyethyl methacrylate) (PHEMA). Finally, BC/PHEMA nanocomposites proved to be nontoxic to human adipose-derived mesenchymal stem cells (ADSCs) and thus are pointed as potential dry dressings for biomedical applications.

Some aspects of upwelling in the Gulf of Panama

ENGLISH: Strong coastal upwelling occurs in the Gulf of Panama regularly each year during the season, from about January through April, when strong northerly winds are blowing offshore. Because of the evident importance of upwelling to the ecology of the Gulf of Panama, we commenced in the fall of 1954 a study of various physical, chemical, and biological phenomena associated therewith. Observations were taken at bi-weekly intervals at a fixed location in the Gulf (approximately 10 miles SE of Taboga Island) to supplement the serial observations of sea level, sea temperature, and winds that have been gathered for many years by the Panama Canal Company. SPANISH: Cado año, en la estación de enero a abril, cuando los vientos del norte soplan vigorosamente frente a la costa, ocurre en el Golfo de Panamá un fuerte afloramiento costanero. Se cree que este afloramiento periódico en el Golfo de Panamá es responsable de la alta productividad biológica que sostiene considerables cantidades de organismos de importancia comercial. Esta región, por ejemplo, es una fuente importante de la especie Cetengraulis mysticetus) pez de carnada para el atún, (Alverson y Shimada, 1957) y mantiene una considerable pesca de camarones llamados langostinos (Burkenroad, Obarrio y Mendoza,1955). (PDF contains 54 pages.)