Pressure – volume curves and drought injury

Leaf hygrometers were used to establish pressure-volume curves on detached leaves of four herbaceous species ( Asarum europaeum , L., Hepatica nobilis , Mill., Phyteuma spicatum , L., Pulmonaria officialis , L.). Breakdown of leaf tissues due to drought injury was independently estimated. There was good agreement between the onset of visible symptoms and beginning deviations from the straight-line portion of the pressure-volume curve. Type I transformation of data (plots of water potential vs. reciprocal relative water content) is superior for recognizing deviating data points. The results are discussed in the context of pressure-volume curve methodology; pressure-volume curves are shown to provide a promising tool for estimating and displaying drought tolerance in plants.     

Wirkung der UV-Strahlung auf Allium-Zellen

Ohne Zusammenfassung     

Röntgenstrahlenwirkungen auf keimenden Weizen

Ohne Zusammenfassung     

Beobachtungen über das Vorkommen von Algen an Thermalstandorten auf Island

Zusammenfassung Es werden einige Warmwasserstandorte auf Island geschildert und Listen der in ihnen gefundenen Algen mitgeteilt. An anderem Ort veröffentlichte Versuche über die Wärmeresistenz einiger Grünalgen warmer Bäche Islands (Biebl, 1967) haben gezeigt, daß diese an ihren natürlichen Standorten beinahe die Grenze ihrer Wärmeresistenz und damit ihrer Lebensmöglichkeit erreichen. Chlorophyceen wurden bis zu Temperaturen von 32° C gefunden,Cosmarium botrytis var.mediolaeve auch noch in 35° C warmem Wasser. Diatomeen wurden lebend bei 35° C beobachtet, in den fixierten Proben aber auch noch bei 48° bis 55,5° C gefunden. Vor allemRhopalodia gibberula trat noch in diesen hoch temperierten Wässern auf. Es muß aber offen bleiben, ob diese im fixierten Material gefundenen Diatomeen am Standort noch lebend waren. Vielversprechend erschiene eine Untersuchung der oft eng benachbarten alkalischen und sauren Quelltümpel hinsichtlich der Artenzusammensetzung. So zeigte z. B.Pinnularia microstauron in Wässern mit einem ph von 5, 5–6, 5 bei Temperaturen zwischen 17,5° und 25° C ausgesprochene Massenentwicklung, während in gleich temperierten alkalischen Wässern andere Arten auftraten. Von den Cyanophyceen wurdeMastigodadus laminosus in einem Bereich von 20° C bis 60° C lebend gefunden. Von noch heißerem Wasser überrieselte Kieselsinterablagerungen enthielten zahlreiche verkieselte Blaualgenreste.     

Phylogenetics of <Emphasis Type="Italic">Cucumis</Emphasis> (Cucurbitaceae): Cucumber (<Emphasis Type="Italic">C. sativus</Emphasis>) belongs in an Asian/Australian clade far from melon (<Emphasis Type="Italic">C. melo</Emphasis>)

Background  

Melon, Cucumis melo, and cucumber, C. sativus, are among the most widely cultivated crops worldwide. Cucumis, as traditionally conceived, is geographically centered in Africa, with C. sativus and C. hystrix thought to be the only Cucumis species in Asia. This taxonomy forms the basis for all ongoing Cucumis breeding and genomics efforts. We tested relationships among Cucumis and related genera based on DNA sequences from chloroplast gene, intron, and spacer regions (rbcL, matK, rpl20-rps12, trnL, and trnL-F), adding nuclear internal transcribed spacer sequences to resolve relationships within Cucumis.     

Escherichia coli alpha-haemolysin induces focal leaks in colonic epithelium: a novel mechanism of bacterial translocation

Extraintestinal pathogenic Escherichia coli (ExPEC) are usually harmless colonizer of the intestinal microflora. However, they are capable to translocate and cause life-threatening disease. Translocation of ExPEC isolates was quantified in colonic monolayers. Transepithelial resistance (R(t)) was monitored and local changes in conductivity analysed with conductance scanning. Confocal microscopy visualized the translocation route. Corroboratory experiments were performed on native rat colon. One translocating strain E. coli O4 was identified. This translocation process was associated with an R(t) decrease (36 +/- 1% of initial resistance) beginning only 2 h after inoculation. The sites of translocation were small defects in epithelial integrity (focal leaks) exhibiting highly increased local ion permeability. Translocation was enhanced by preincubation of monolayers with tumour necrosis factor-alpha or interleukin-13. Mutant strains lacking alpha-haemolysin lost the ability to induce focal leaks, while this effect could be restored by re-introducing the haemolysin determinant. Filtrate of a laboratory strain carrying the alpha-haemolysin operon was sufficient for focal leak induction. In native rat colon, E. coli O4 decreased R(t) and immunohistology demonstrated focal leaks resembling those in cell monolayers. E. coli alpha-haemolysin is able to induce focal leaks in colonic cell cultures as well as in native colon. This process represents a novel route of bacterial translocation facilitated by pro-inflammatory cytokines.     

Abundance-based Classifier for the Prediction of Mass Spectrometric Peptide Detectability Upon Enrichment (PPA)

The function of a large percentage of proteins is modulated by post-translational modifications (PTMs). Currently, mass spectrometry (MS) is the only proteome-wide technology that can identify PTMs. Unfortunately, the inability to detect a PTM by MS is not proof that the modification is not present. The detectability of peptides varies significantly making MS potentially blind to a large fraction of peptides. Learning from published algorithms that generally focus on predicting the most detectable peptides we developed a tool that incorporates protein abundance into the peptide prediction algorithm with the aim to determine the detectability of every peptide within a protein. We tested our tool, “Peptide Prediction with Abundance” (PPA), on in-house acquired as well as published data sets from other groups acquired on different instrument platforms. Incorporation of protein abundance into the prediction allows us to assess not only the detectability of all peptides but also whether a peptide of interest is likely to become detectable upon enrichment. We validated the ability of our tool to predict changes in protein detectability with a dilution series of 31 purified proteins at several different concentrations. PPA predicted the concentration dependent peptide detectability in 78% of the cases correctly, demonstrating its utility for predicting the protein enrichment needed to observe a peptide of interest in targeted experiments. This is especially important in the analysis of PTMs. PPA is available as a web-based or executable package that can work with generally applicable defaults or retrained from a pilot MS data set.Post-translational modification (PTM)¹ of proteins is a key regulatory mechanism in the vast majority of biological processes. Historically, to follow PTMs, site-specific antibodies had to be generated in a time-consuming and laborious process associated with high failure rates. Mass spectrometry (MS) holds enormous promise in PTM analysis as it is currently the only technique that has the ability to both discover, localize, and quantify proteome-wide modifications (1). Recent advances in instrumentation and method optimization makes it possible to detect the complete yeast proteome within one hour (2), an ever increasing proportion of the human proteome (3–6), and more than 10,000 phosphorylation sites in a single MS experiment (7, 8). As a result one of the major publicly available databases (www.phosphosite.org (9)) has curated >200,000 phosphorylation sites.Although the number of proteins and PTMs that can be identified is impressive, many modifications have still not been identified in any MS-based experiment. The identification and quantification of biologically relevant modifications is challenging for three reasons: (1) many proteins of interest are of very low abundance rendering them difficult to detect and quantify; (2) many modifications sites are present at substoichiometric quantities, further reducing their detectability; and (3) as large scale proteomics is based on the detection of peptides after a proteolytic digest, and the detectability of a peptide is determined by its physiochemical properties (10), many peptides from highly abundant proteins are never detected. This is particularly important, as there is a shift in the use of MS-based proteomics from large scale, unbiased, discovery-focused experiments toward directed experiments for accurate and precise quantification of biologically relevant PTMs. Protein and peptide enrichment strategies and/or targeted MS experiments like single reaction monitoring (SRM) (11) have increased the number of detectable peptides; however, both of these methods are laborious, and often not successful, that is, the peptide carrying the modification of interest is still not observed as it is fundamentally very difficult to detect.Protein enrichment is the method choice for most experimentalists, but there is no current way to determine whether this is likely to succeed prior to engaging in lengthy biochemical and/or analytical experiments. In an effort to gauge the chances of success for detecting a particular peptide we sought to develop an algorithm that can predict both the chances of detecting a particular peptide and, more importantly, what enrichment it would take to detect a particular peptide that is not easily detected. Here we present such a tool that predicts the detectability and estimates an enrichment factor, i.e. an increase in signal over the background that is necessary to actually detect a particular peptide. Our algorithm development was motivated by two premises: (1) In silico methods have been developed that focus on the prediction of easily detectable “proteotypic” peptides (peptides that are likely to provide the best detection sensitivity) with good accuracy (12–15). (2) Comprehensive proteome studies have shown that the number of detected peptides per protein, and thus the sequence coverage, varies with protein abundance (which is the basis for spectral counting-based protein quantification (16, 17)). We find that incorporation of protein abundance in a peptide classification tool improves the accuracy of the prediction of peptide detectability allowing us to predict the detectability of all peptides within a protein as well as the amount of enrichment needed to detect a peptide of interest.We used a set of 120 purified in vitro expressed proteins as a training set to develop a prediction tool. We deliver this in the form of a web-based interface that provides information about: (1) the probability of detecting the different tryptic peptides of a protein, and (2) the fold enrichment that would be required to bring a peptide of interest into the detectable range. This tool will help guide researchers in their efforts to monitor particular peptides and their modified cognates by MS, specifically, in prioritizing their efforts toward enriching proteins where they would be likely to be able to detect a peptide or modification of interest.     

Altered circadian rhythm of melatonin concentrations in hypocretin-deficient men

Hypocretin deficiency causes narcolepsy. It is unknown whether melatonin secretion is affected in this sleep disorder. Therefore, in both narcolepsy patients and matched controls, the authors measured plasma melatonin levels hourly for 24 h before and after 5 days of sodium oxybate (SXB) administration. Although mean melatonin concentrations were similar between patients and controls, in narcoleptics the percentage of 24-h melatonin secreted during the daytime was significantly higher, and melatonin secretion exhibited a weaker coupling to sleep. SXB did not affect melatonin secretion. These findings suggest that hypocretin deficiency might disturb both the circadian control of melatonin release and its temporal association with sleep.     

Ambivalent bacteriophages of different species active on Escherichia coli K12 and Salmonella sps. strains

A study was made of several bacteriophages (including phages U2 and LB related to T-even phages of Escherichia coli) that grow both on E. coli K12 and on some Salmonella strains. Such phages were termed ambivalent. T-even ambivalent phages (U2 and LB) are rare and have a limited number of hosts among Salmonella strains. U2 and LB are similar to canonical E. coli-specific T-even phages in morphological type and size of the phage particle and in reaction with specific anti-T4 serum. Phages U2 and LB have identical sets of structural proteins, some of which are similar in size to structural proteins of phages T2 and T4. DNA restriction patterns of phages U2 and LB differ from each other and from those of T2 and T4. Still, DNAs of all four phages have considerable homology. Unexpectedly, phages U2 and LB grown on Salmonella bungori were unstable during centrifugation in a CsCl gradient. Ambivalent bacteriophages were found in species other than T-even phages and were similar in morphotype to lambdoid and other E. coli phages. One of the ambivalent phages was highly similar to well-known Felix01, which is specific for Salmonella. Ambivalent phages can be used to develop a new set for phage typing in Salmonella. An obvious advantage is that ambivalent phages can be reproduced in the E. coli K12 laboratory strain, which does not produce active temperate phages. Consequently, the resulting typing phage preparation is devoid of an admixture of temperate phages, which are common in Salmonella. The presence of temperate phages in phage-typing preparations may cause false-positive results in identifying specific Salmonella strains isolated from the environment or salmonellosis patients. Ambivalent phages are potentially useful for phage therapy and prevention of salmonellosis in humans and animals.     

Feeding enrichment in an opportunistic carnivore: The red fox

In captive carnivores, species-specific behaviour is often restricted by inadequate feeding regimens. Feeding live prey is not feasible in most places and food delivery is often highly predictable in space and time which is considerably different from the situation in the wild. As a result, captive carnivores are often inactive, show little behavioural diversity and are prone to behavioural problems such as stereotypic pacing. Using artificial feeding devices to substitute for natural food resources is a way to address these problems. In a group of four red fox (Vulpes vulpes), we compared a conventional feeding method to four different methods through the use of feeding enrichment that were based on natural foraging strategies of opportunistic carnivores. Feeding enrichments consisted of electronic feeders delivering food unpredictable in time which were successively combined with one of the three additional treatments: a self-service food box (allowing control over access to food), manually scattering food (unpredictable in space), and an electronic dispenser delivering food unpredictably both in space and time. The aim of administering feeding enrichment in this study was to stimulate appetitive (food searching) behaviour and to increase time spent in feeding. Compared to conventional feeding, diversity of behaviour and overall activity were significantly enhanced in the presence of electronic feeders in all four foxes (EF > CON1 = CON2, EF + SF > CON1 = CON2, EF + MS > CON1 = CON2, EF + ED > CON1 = CON2). Behavioural diversity was highest when the foxes had control over access to food (EF + SF), while the manual scattering of food (EF + MS) and the electronic dispenser (EF + ED) enhanced food searching behaviour. These results indicate that in opportunistic carnivores natural foraging and feeding behaviour can be stimulated by simple feeding enrichment strategies, and that foraging behaviour is stimulated most when food delivery is unpredictable both in space and time.