Apple polyphenols and products formed in the gut differently inhibit survival of human cell lines derived from colon adenoma (LT97) and carcinoma (HT29)

Colorectal tumor risks could be reduced by polyphenol-rich diets that inhibit cell growth. Here, apple polyphenols were studied for effects on the survival of colon adenoma (LT97) and carcinoma-derived (HT29) cell lines. Three apple extracts (AEs) from harvest years 2002-2004 were isolated (AE02, AE03, and AE04) and fermented in vitro with human fecal flora. Extracts and fermentation products were analyzed for polyphenols with HPLC. The cells were treated with AEs (0-850 microg/mL) or fermented AEs (F-AEs, 0-9%), and survival was measured by DNA staining. All AEs contained high amounts of polyphenols (311-534 mg/g) and reduced cell survival (in LT97 > HT29). AE03 was most potent, possibly because it contained more quercetin compounds. Fermentation of AEs resulted in an increase of short chain fatty acids, and polyphenols were degraded. The F-AEs were approximately 3-fold less bioactive than the corresponding AEs, pointing to a loss of chemoprotective properties through fermentation.     

Comparability of and Alternatives to Leaching Tests for the Assessment of the Emission of Inorganic Soil Contamination (11 pp)

Goal, Scope and Background

The suitability of leaching tests for the assessment of soil water concentrations is a controversial issue. The aim of this paper is to review and critically discuss the applicability and comparability of leaching tests in the scope of groundwater risk assessment of inorganic contamination and to discuss soil water sampling methods as alternative.

Main Features

First, the theoretical background of leaching tests and the main leaching controlling parameters (pH, redox, ionic strength, reaction kinetics, surface, and speciation) are discussed. Experience with common batch leaching tests (the German DEV S4 test (S4), saturation soil extraction (SSE), ammonia nitrate extraction (ANE), and pHstatic tests) are compiled and an emphasis is set on the comparability of the results of batch leaching tests. Additionally, the comparability between batch tests and column tests is discussed and comparison studies are reviewed. As a feasible alternative, soil water sampling strategies (the suction cup method and centrifugation) are outlined.

Results and Conclusions

For an expedient application of leaching tests, it is crucial to identify the main release controlling parameters, which can be both site and contaminant specific. Possible controlling parameters are, for example, pH, redox conditions, specific surface area of the investigated material, liquid to solid-ratio, ionic strength, cation exchange capacity, DOC, speciation, temperature and contact time. It depends on the release process of the contaminant in the investigated material, which parameters are influencing the release predominantly. The following questions have to be answered: Is the release process availability controlled or solubility controlled and are there kinetic limitations? Suction cups are particularly useful for long-term monitoring. In contrast, centrifugation is a fast and simple method to sample soil water also at larger and heterogeneous sites. SSE or similar batch tests for coarse material are suitable if the soil water cannot be investigated directly. Contaminant concentrations of the SSE fit best to contaminant concentrations in soil water compared to other leaching tests. Concentrations measured with S4 tests are usually significantly lower than in real soil water due to the unrealistically high liquid to solid-ratio. The ANE is used for the evaluation of plant uptake. It does not represent realistic soil water concentrations. Cationic contaminants are usually significantly higher concentrated in ANE eluates. pHstatic tests provide an evaluation of the pH dependency of contaminant release and buffer capacity of the investigated material. It provides indications to release processes and often explains deviations in the results of different leaching tests. Previous practical experience and fundamental research studies show that a conversion of leaching test results, as is proposed by the (already withdrawn) DIN V 19735, is impossible. Correlations of results from different methods, regarding different kinds of materials and different sites, are not significant. This is due to different leaching processes.

Recommendation

. For short-term risk assessment, soil water should be sampled and investigated directly by suction cups (for monitoring purposes) or centrifugation (for inventory purposes). If this is not possible, the SSE or analogous batch tests for coarse material with a narrow L/S ratio should be applied. A suggestion could be a modified saturation soil extraction where a soil column is saturated and the eluate is removed by suction at the bottom of the column. With this method, an unsaturated column test could follow in the same vessel. In order to assess the long-term leaching behavior, total contents of the investigated material have to be taken into account additionally. Furthermore, it is essential to understand the dominant physical and chemical release processes and to figure out the main leaching controlling parameters. Therefore, the following methods are recommended: pHstatic tests at different pH values provide an insight to leaching processes and possible future leaching scenarios. Batch tests similar to the S4 procedure with different L/S ratios are useful to find out whether the release process is solubility or availability controlled. Additionally, this method allows the determination of maximum solubility and maximum availability, respectively. Furthermore, unsaturated column tests provide an insight into leaching processes and releasable amounts even though they require great experimental effort. Other leaching tests like S4 or ANE are not suitable for the estimation of soil water concentrations.     

Classical swine fever virus replicon particles lacking the Erns gene: a potential marker vaccine for intradermal application

Classical swine fever virus replicon particles (CSF-VRP) deficient for E(rns) were evaluated as a non-transmissible marker vaccine. A cDNA clone of CSFV strain Alfort/187 was used to obtain a replication-competent mutant genome (replicon) lacking the sequence encoding the 227 amino acids of the glycoprotein E(rns) (A187delE(rns)). For packaging of A187delE(rns) into virus particles, porcine kidney cell lines constitutively expressing E(rns) of CSFV were established. The rescued VRP were infectious in cell culture but did not yield infectious progeny virus. Single intradermal vaccination of two pigs with 10(7) TCID(50) of VRP A187delE(rns) elicited neutralizing antibodies, anti-E2 antibodies, and cellular immune responses determined by an increase of IFN-gamma producing cells. No anti-E(rns) antibodies were detected in the vaccinees confirming that this vaccine represents a negative marker vaccine allowing differentiation between infected and vaccinated animals. The two pigs were protected against lethal challenge with the highly virulent CSFV strain Eystrup. In contrast, oral immunization resulted in only partial protection, and neither CSFV-specific antibodies nor stimulated T-cells were found before challenge. These data represent a good basis for more extended vaccination/challenge trials including larger numbers of animals as well as more thorough analysis of virus shedding using sentinel animals to monitor horizontal spread of the challenge virus.     

Ankyloglossia in an Anatolian Shepherd dog

Ankyloglossia, commonknown as tongue-tie, is a rare congenital oral anomaly in dogs. A complete attachment of the lingual frenulum to the floor of the oral cavity leads to limited mobility of the tongue including problems during eating and swallowing. In humans ankyloglossia is a common anomaly in newborn infants. In our report a 5-month old Anatolian Shepherd dog was surgically treated and full function of the tongue could be achieved with a horizontal-to-vertical frenuloplasty.     

iRhom2 regulation of TACE controls TNF-mediated protection against Listeria and responses to LPS

Innate immune responses are vital for pathogen defense but can result in septic shock when excessive. A key mediator of septic shock is tumor necrosis factor-α (TNFα), which is shed from the plasma membrane after cleavage by the TNFα convertase (TACE). We report that the rhomboid family member iRhom2 interacted with TACE and regulated TNFα shedding. iRhom2 was critical for TACE maturation and trafficking to the cell surface in hematopoietic cells. Gene-targeted iRhom2-deficient mice showed reduced serum TNFα in response to lipopolysaccharide (LPS) and could survive a lethal LPS dose. Furthermore, iRhom2-deficient mice failed to control the replication of Listeria monocytogenes. Our study has identified iRhom2 as a regulator of innate immunity that may be an important target for modulating sepsis and pathogen defense.     

High-resolution protein structure determination by serial femtosecond crystallography

Structure determination of proteins and other macromolecules has historically required the growth of high-quality crystals sufficiently large to diffract x-rays efficiently while withstanding radiation damage. We applied serial femtosecond crystallography (SFX) using an x-ray free-electron laser (XFEL) to obtain high-resolution structural information from microcrystals (less than 1 micrometer by 1 micrometer by 3 micrometers) of the well-characterized model protein lysozyme. The agreement with synchrotron data demonstrates the immediate relevance of SFX for analyzing the structure of the large group of difficult-to-crystallize molecules.     

Synthetic genetic polymers capable of heredity and evolution

Genetic information storage and processing rely on just two polymers, DNA and RNA, yet whether their role reflects evolutionary history or fundamental functional constraints is currently unknown. With the use of polymerase evolution and design, we show that genetic information can be stored in and recovered from six alternative genetic polymers based on simple nucleic acid architectures not found in nature [xeno-nucleic acids (XNAs)]. We also select XNA aptamers, which bind their targets with high affinity and specificity, demonstrating that beyond heredity, specific XNAs have the capacity for Darwinian evolution and folding into defined structures. Thus, heredity and evolution, two hallmarks of life, are not limited to DNA and RNA but are likely to be emergent properties of polymers capable of information storage.     

Heralded entanglement between widely separated atoms

Entanglement is the essential feature of quantum mechanics. Notably, observers of two or more entangled particles will find correlations in their measurement results that cannot be explained by classical statistics. To make it a useful resource, particularly for scalable long-distance quantum communication, the heralded generation of entanglement between distant massive quantum systems is necessary. We report on the creation and analysis of heralded entanglement between spins of two single rubidium-87 atoms trapped independently 20 meters apart. Our results illustrate the viability of an integral resource for quantum information science, as well as for fundamental tests of quantum mechanics.     

Water repellency and pore clogging at early successional stages of microbiotic crusts on inland dunes,Brandenburg, NE Germany

Biological soil crusts play a key role for hydrological processes in many open landscapes. They seal the topsoil and generate surface run-off. Utilization of the mineral substrate at early stages of microbiotic crusts was investigated using scanning electron microscopy (SEM), water repellency indices were measured using the ethanol/water microinfiltrometer method, and steady state water flow was determined on the dry crusts and after 300, 600, 1200 and 1800 s of wetting, thus allowing to follow pore clogging through swelling of extracellular polymeric substances (EPS). It was found that water repellency increases with initial crust development where filamentous cyanobacteria and filamentous green algae were dominating, but decreases as coccal algae, bryophytes and fungi associated with bryophytes emerged. Swelling of EPS took place immediately after wetting, and its influence on steady state water flow was most pronounced when filamentous cyanobacteria and algae partially filled in the matrix pores and enmeshed sand grains, still leaving micropore channels available for free water infiltration, but prior to appearance of coccal algae, bryophytes and associated fungi which formed a dense cover on the surface. It was concluded that a new phase of crust succession was accompanied by easier wettability but slower infiltration. Transition from hydrophobicity to pore clogging as ruling mechanisms causing water run-off may occur during wetting of individual biological crusts, but also during crust succession over time.     

Potential effects of food addition to sediment on test conditions in sediment toxicity tests

Purpose  

Standardized sediment toxicity assays often employ periodic additions of uncontaminated food to sustain energy and growth requirements of the test organisms. Consequently, selective feeding on this uncontaminated food may reduce exposure to sediment particles containing the test substance. To address this issue, some standard guidelines propose to add food to the sediment before spiking with the test substance to account for multiple exposure routes, including ingestion of contaminated food. The present study focused on the influence of different feeding regimens and compositions of the aqueous medium on water quality (ammonia concentrations) and test organism development.