Imaging of lactic acid bacteria with AFM--elasticity and adhesion maps and their relationship to biological and structural data

The adhesion of lactic acid bacteria to the intestinal epithelium is one of the most important factors determining probiotic ability of a bacterial strain. Studying bacterial adhesion requires knowledge of the structure and properties of the bacterial surface, which can be studied by atomic force microscopy under native conditions. The observation of the surface topography of bacteria from the species Lactobacillus crispatus, L. helveticus and L. johnsonii shows major differences between bacteria having a crystalline-like protein layer as part of the cell wall and those without such layers. Force volume images calculated into elasticity and adhesion force maps of different bacterial strains show that L. crispatus and L. helveticus have a surface with a homogeneous stiffness with no adhesion events. This is most likely caused by the S-layer, which completely covers the surface of the bacteria. We infer that the absence of adhesion peaks is caused by the semi-crystalline character of such protein layers, in agreement with the results obtained from electron microscopy. Analysis of a number of L. johnsonii strains shows that these bacteria have surface properties which strongly differ from the L. crispatus and L. helveticus strains. For L. johnsonii DMS20533 and L. johnsonii ATCC33200 high adhesion forces are observed, which can be related to a surface rich in polysaccharides. L. johnsonii ATCC332 has lower adhesion forces compared to the other two and, furthermore, the surface topography shows depressions. We suppose that this strain has a surface pattern consisting of crystalline-like proteins alternating with polysaccharide-rich domains. The wide variety in surface properties of lactobacilli could well have wide-ranging implications for food processing and for health benefits.     

大角度弹性波阻抗

弹性波阻抗及相关技术是以二项AVO方程和三项AVO方程为基础的,它的有效性在世界范围内的许多沉积盆地都得到了验证.在过去的几年中,大偏移距的数据采集使得入射角普遍大于30°,因此必须用三项AVO方程来解决储层流体和岩性识别问题.在入射角小于30°时,所有的线性弹性波阻抗方程都能给出可靠的结果,当入射角大于这一角度时,某...     

Differences in Volatile Profiles of Turnip Plants Subjected to Single and Dual Herbivory Above- and Belowground

Plants attacked by herbivorous insects emit volatile organic compounds that are used by natural enemies to locate their host or prey. The composition of the blend is often complex and specific. It may vary qualitatively and quantitatively according to plant and herbivore species, thus providing specific information for carnivorous arthropods. Most studies have focused on simple interactions that involve one species per trophic level, and typically have investigated the aboveground parts of plants. These investigations need to be extended to more complex networks that involve multiple herbivory above- and belowground. A previous study examined whether the presence of the leaf herbivore Pieris brassicae on turnip plants (Brassica rapa subsp. rapa) influences the response of Trybliographa rapae, a specialist parasitoid of the root feeder Delia radicum. It showed that the parasitoid was not attracted by volatiles emitted by plants under simultaneous attack. Here, we analyzed differences in the herbivore induced plant volatile (HIPV) mixtures that emanate from such infested plants by using Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA). This multivariate model focuses on the differences between odor blends, and highlights the relative importance of each compound in an HIPV blend. Dual infestation resulted in several HIPVs that were present in both isolated infestation types. However, HIPVs collected from simultaneously infested plants were not the simple combination of volatiles from isolated forms of above- and belowground herbivory. Only a few specific compounds characterized the odor blend of each type of damaged plant. Indeed, some compounds were specifically induced by root herbivory (4-methyltridecane and salicylaldehyde) or shoot herbivory (methylsalicylate), whereas hexylacetate, a green leaf volatile, was specifically induced after dual herbivory. It remains to be determined whether or not these minor quantitative variations, within the background of more commonly induced odors, are involved in the reduced attraction of the root feeder’s parasitoid. The mechanisms involved in the specific modification of the odor blends emitted by dual infested turnip plants are discussed in the light of interferences between biosynthetic pathways linked to plant responses to shoot or root herbivory.     

Characterization of a putative phosphorylation switch: adaptation of SPOT synthesis to analyze PDZ domain regulation mechanisms

Transient macromolecular complexes are often formed by protein-protein interaction domains (e.g., PDZ, SH2, SH3, WW), which are often regulated (positively or negatively) by phosphorylation. To address the in vitro analysis of PDZ domain regulation by such phosphorylation, we improved the inverted peptide method. This method is based on standard SPOT synthesis, followed by inversion of the peptide under acidic conditions to generate the free C termini necessary for PDZ domain ligand recognition. The benefit of the newly introduced acidic conditions is the preservation of the incorporated phosphate group during peptide synthesis. Furthermore, the improved method is more robust and shows an increased signal-to-noise ratio. As representative examples, we used the AF6, ERBIN, and SNA1 (alpha-1-syntrophin) PDZ domains to analyze the influence of ligand-position-dependent phosphorylation. We could clearly demonstrate severe down-regulation by phosphorylation of the PDZ ligand position -2 (<50 %) and slightly less at position -1 ( approximately 50 %). These results are specific and reproducible for all three PDZ domains. Finally, we confirmed the influence of negative regulation by using the protein kinase BCR as the AF6 PDZ domain ligand. For the first time, this approach allows the SPOT synthesis technique to be used to screen large libraries of phosphorylated peptides in vitro. This should ultimately help in the identification of phosphorylation-dependent regulation mechanisms in vivo.     

Identification of IgE Binding to Api g 1‐Derived Peptides

Celery is a frequent cause of food allergy in pollen‐sensitized patients and can induce severe allergic reactions. Clinical symptoms cannot be predicted by skin prick tests (SPTs) or by determining allergen‐specific immunoglobulin E (IgE). Our aim was to identify specific IgE binding peptides by using an array technique. For our study, the sera of 21 patients with positive double‐blind, placebo‐controlled food challenge (DBPCFC) to celery, as well as the sera of 17 healthy patients were used. Additionally, all patients underwent skin tests along with determinations of specific IgE binding. The major allergen of celery Api g 1.0101 (Apium graveolens) was synthesized as an array of overlapping peptides and probed with the patients' sera. We developed an improved immunoassay protocol by investigating peptide lengths, peptide densities, incubation parameters, and readout systems, which could influence IgE binding. Sera of celery‐allergic patients showed binding to three distinct regions of Api g 1.0101. The region including amino acids 100 to 126 of Api g 1.0101 is the most important region for IgE binding. This region caused a fivefold higher binding of IgE from the sera of celery‐allergic patients compared to those of healthy individuals. In particular, one peptide (VLVPTADGGSIC) was recognized by all sera of celery‐allergic patients. In contrast, no binding to this peptide was detected in sera of the healthy controls. Our improved assay strategy allows us to distinguish between celery‐allergic and healthy individuals, but needs to be explored in a larger cohort of well‐defined patients.     

Videos of conspecifics elicit interactive looking patterns and facial expressions in monkeys.

A broader understanding of the neural basis of social behavior in primates requires the use of species-specific stimuli that elicit spontaneous, but reproducible and tractable behaviors. In this context of natural behaviors, individual variation can further inform about the factors that influence social interactions. To approximate natural social interactions similar to those documented by field studies, we used unedited video footage to induce in viewer monkeys spontaneous facial expressions and looking patterns in the laboratory setting. Three adult male monkeys (Macaca mulatta), previously behaviorally and genetically (5-HTTLPR) characterized, were monitored while they watched 10 s video segments depicting unfamiliar monkeys (movie monkeys) displaying affiliative, neutral, and aggressive behaviors. The gaze and head orientation of the movie monkeys alternated between “averted” and “directed” at the viewer. The viewers were not reinforced for watching the movies, thus their looking patterns indicated their interest and social engagement with the stimuli. The behavior of the movie monkey accounted for differences in the looking patterns and facial expressions displayed by the viewers. We also found multiple significant differences in the behavior of the viewers that correlated with their interest in these stimuli. These socially relevant dynamic stimuli elicited spontaneous social behaviors, such as eye-contact induced reciprocation of facial expression, gaze aversion, and gaze following, that were previously not observed in response to static images. This approach opens a unique opportunity to understanding the mechanisms that trigger spontaneous social behaviors in humans and nonhuman primates. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

Seasonal patterns of thermal stratification and primary production in the northern parts of Lake Tanganyika

Thermal stratification in meromictic Lake Tanganyika weakens during the cool, dry and windy season, allowing intrusions of deeper nutrient-rich waters into the upper mixed layer enhancing primary productivity. The current study examined the seasonal influence of thermal structure on the patterns of primary production in Lake Tanganyika. Two sites (Kigoma Bay and Mahale) were sampled on a monthly basis for 1?year. Water temperature and chlorophyll a fluorescence profiles (0–100?m) were measured using a multisonde. Water samples were taken every 20?m (0–100?m) to measure soluble reactive phosphorus (SRP), chlorophyll a and primary production. Pulse Amplitude Modulated Fluorometry was used for the measurements of primary production. The results show that the dry season coincided with higher wind speeds, elevated SRP and some peaks of chlorophyll a and primary production at both sites. During the wet season, high levels of chlorophyll a and primary production coincided with an increase in the euphotic depth, a deep chlorophyll maximum (DCM) and a contribution from metalimnetic areal production at both sites. Our results suggest that the vertical distribution of primary production in Lake Tanganyika is affected by the lake thermal structure, nutrient availability and the extent of the euphotic zone. In Lake Tanganyika, estimates of areal primary production were significantly affected by DCM and required estimation throughout the entire upper mixed layer and the metalimnion which extended to ~89?m.     

p-Cycle protection at the glass fiber level

The cost and complexity of wavelength assignment, wavelength conversion and wavelength-selective switching are always of primary considerations in the design of survivable optical networks. This proposal recognizes that as long as the loss budgets are adequate, entire DWDM wavebands could be restored with no switching or manipulation of individual lightpaths; so that the DWDM layer would never know the break happened. And environments where fiber switching devices are low cost, and ducts are full of dark fibers provide a very low cost alternative to protect an entire DWDM transport layer (or working capacity envelope) against the single largest cause of outage. Yet, while nodes and single DWDM channels may fail, a pre-dominant source of unavailability is the physical damage of optical cables. Thus, with the objectives of reducing the overall real CapEx costs and removing the complexity due to wavelength assignment and wavelength continuity constraints when configuring p-cycles in a fully transparent network context, this paper addresses the subsequent questions: if it is ultimately glass that fails, what if just the glass is directly replaced? More specifically, what if p-cycles were used to rapidly, simply and efficiently provide for the direct replacement of failed fiber sections with whole replacement fibers?     

Nanocapsules: coating for living cells

One of the most promising tools for future applications in science and medicine is the use of nanotechnologies. Especially self-assembly systems, e.g., polyelectrolyte (PE) capsules prepared by means of the layer-by-layer technique with tailored properties, fulfill the requirements for nano-organized systems in a satisfactory manner. The nano-organized shells are suitable as coating for living cells or artificial tissue to prevent immune response. With these shells, material can be delivered to predefined organs. In this paper, some preliminary results are presented, giving a broad overview over the possibilities to use nano-organized capsules. Based on the observations that the cells while duplicating break the capsule a mutant yeast strain (Saccharomyces cerevisiae), which express GFP-tubulin under galactose promotion, was investigated by means of confocal laser scanning microscopy. The measurements reveal an increased surface charge in the region of buds developed prior encapsulation. In order to test the used PE pair for cytotoxicity, germinating conidia of the fungi Neurospora crassa were coated. The investigation with fluorescence microscopy shows a variation in the surface charge for the growing region and the conidium poles. The capsules exhibit interesting properties as valuable tool in science and a promising candidate for application in the field of medicine.     

Semantic activation in action planning.

Four experiments investigated activation of semantic information in action preparation. Participants either prepared to grasp and use an object (e.g., to drink from a cup) or to lift a finger in association with the object's position following a go/no-go lexical-decision task. Word stimuli were consistent to the action goals of the object use (Experiment 1) or to the finger lifting (Experiment 2). Movement onset times yielded a double dissociation of consistency effects between action preparation and word processing. This effect was also present for semantic categorizations (Experiment 3), but disappeared when introducing a letter identification task (Experiment 4). In sum, our findings indicate that action semantics are activated selectively in accordance with the specific action intention of an actor. (PsycINFO Database Record (c) 2011 APA, all rights reserved)