Dynamics of the cytoskeleton in Amoeba proteus

Fluorescein-labeled muscle actin was microinjected into Amoeba proteus and followed during intracellular redistribution by means of the image-intensification technique. The fully polymerization-competent protein becomes part of the endogenous actomyosin system undergoing dynamic changes over time periods of several hours. Single-frame analysis of long-term sequences enabled the direct demonstration of both the contractile activities and morphological transformations of microfilaments in normally locomoting, immobilized and phagocytozing specimens. In normally locomoting cells the filament layer undergoes continuous changes in spatial distribution depending on the actual pattern of cytoplasmic streaming and cell shape. The highest degree of differentiation is always maintained in the intermediate region between the front and the uroid, thus indicating this segment of the cortex to be the most important site in generating motive force for pseudopodium formation and ameboid movement. In immobilized cells contracted by the application of ruthenium red or relaxed by different anesthetics, the filament layer forms a continuous thick sheath beneath the cell surface or becomes completely disintegrated. In phagocytozing cells the local polymerization of actin at the tip of pseudopodia forming the food-cup and around the nascent phagosome points to a significant participation of the actomyosin system in the process of capturing and constricting prey organisms. Although our results provide clear evidence for the overall importance of motive force generation according to the hydraulic pressure theory, some motile phenomena exist in Amoeba proteus that cannot exclusively be explained by this mechanism.     

Echoortung bei der Fledermaus Chilonycteris rubiginosa

Zusammenfassung Chilonycteris rubiginosa erzeugt in allen Orientierungsituationen dreiteilige Ortungslaute. Im Anfangsteil steigt die Frequenz um etwa 1–2 kHz an. Der folgende Mittelteil hat eine konstante Frequenz von etwa 57 bis 57,6 kHz. Im Endteil fällt die Frequenz um etwa 8 kHz ab. Die Laute werden in Folgen von Lautgruppen ausgesendet.CR erzeugt pro Flügelschlag eine Lautgruppe. Im freien Flug zeigt CR Gruppen mit 2 Lauten von etwa 17–23 msec Dauer. Landende Fledermäuse senden in der Annäherungsphase Gruppen mit einer zunehmenden Zahl immer kürzerer Laute und in der Schlußphase eine längere Gruppe mit vielen kurzen Lauten.Fliegende Tiere senken die Frequenz des konstantfrequenten Mittelteils immer um etwa den Betrag der durch die Fluggeschwindigkeit bedingten Dopplereffekte ab, so daß die von den Tieren gehörte Echofrequenz nahezu konstant in Höhe der vor dem Flug ausgesendeten Frequenz gehalten wird.CR zeigt Kopf- und Ohrbewegungen. Die Ohrbewegungen stehen in Beziehung zur Lautaussendung.

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Echolocation by the bat Chilonycteris rubiginosa

Summary Chilonycteris rubiginosa (CR) produces tripartite ultrasonic sounds in all orientation situations. During the first part the frequency rises by 1–2 kHz. The following middle part has a constant frequency of about 57–57,6 kHz. In the terminal part the frequency decreases by about 8 kHz. The sounds are emitted as a sequence of groups of sounds.In flight they produce per wingbeat one group of sounds at a repetition rate of 10–11 Hz. In free flight CR emits groups of 2 sounds of about 17–23 msec duration. During the approach landing bats emit groups consisting of an increasing number of sounds of decreasing duration. During the terminal phase the group is longer in duration and consists of many short sounds.Flying CR lower the frequency of the middle part by an amount which compensates for Doppler shifts caused by the flight velocity. The frequency heard by the bats is, thus, always kept constant and equal to a frequency which is about 100–150 Hz above the medium frequency emitted before the flight. CR shows head and ear movements. The ear movements are correlated to the sound emission.

     

The role of vanadium in green plants

Cells of Chlorella pyrenoidosa, derived from vanadium free agar slants, respond with great sensitivity to microamounts of vanadium, added as NH₄VO₃ to autotrophic liquid cultures. Between 0.01 and 1 g V per litre nutrient medium (2·10^-10-2·10^-8g-at/l), the algae respond with a continuous increase in dry weight. At higher V-concentrations, further enhancement in biomass is accompanied by a additional increase in chlorophyll content. Maximum V-effect on both parameters was found to be at 500g V/l (10^-5 g-at/l). Dry weight as well as chlorophyll content of Chlorella are decreased by concentrations above 25 mg V/l; 100 mg V/l (2·10^-3 g-at/l) stop growth and cause death of the cells. The toxic threshold for the V-content in the algae was determined to be at 150–200 g V/g (3–4·10^-6 g-at/g) dry weight.Two different pH-optima for a positive vanadium action on dry weight and chlorophyll biosynthesis were established, the first at pH 7, the other in the range pH 7.5–8. Two sites of vanadium action in green algae are discussed.Part I: Arch. Microbiol. 105, 77–82 (1975)     

Development and laboratory-scale testing of a fully automated online flow cytometer for drinking water analysis

Accurate and sensitive online detection tools would benefit both fundamental research and practical applications in aquatic microbiology. Here, we describe the development and testing of an online flow cytometer (FCM), with a specific use foreseen in the field of drinking water microbiology. The system incorporated fully automated sampling and fluorescent labeling of bacterial nucleic acids with analysis at 5-min intervals for periods in excess of 24 h. The laboratory scale testing showed sensitive detection (< 5% error) of bacteria over a broad concentration range (1 × 10(3) -1 × 10(6) cells mL(-1) ) and particularly the ability to track both gradual changes and dramatic events in water samples. The system was tested with bacterial pure cultures as well as indigenous microbial communities from natural water samples. Moreover, we demonstrated the possibility of using either a single fluorescent dye (e.g., SYBR Green I) or a combination of two dyes (SYBR Green I and Propidium Iodide), thus broadening the application possibilities of the system. The online FCM approach described herein has considerable potential for routine and continuous monitoring of drinking water, optimization of specific drinking water processes such as biofiltration or disinfection, as well as aquatic microbiology research in general.     

Selective inhibition of dipeptidyl peptidase 4 by targeting a substrate-specific secondary binding site

Dipeptidyl peptidase 4/CD26 (DP4) is a multifunctional serine protease liberating dipeptide from the N-terminus of (oligo)peptides which can modulate the activity of these peptides. The enzyme is involved in physiological processes such as blood glucose homeostasis and immune response. DP4 substrate specificity is characterized in detail using synthetic dipeptide derivatives. The specificity constant k(cat)/K(m) strongly depends on the amino acid in P?-position for proline, alanine, glycine and serine with 5.0 x 10? M?1 s?1, 1.8 x 10? M?1 s?1, 3.6 x 102 M?1 s?1, 1.1 x 102 M?1 s?1, respectively. By contrast, kinetic investigation of larger peptide substrates yields a different pattern. The specific activity of DP4 for neuropeptide Y (NPY) cleavage comprising a proline in P?-position is the same range as the k(cat)/K(m) values of NPY derivatives containing alanine or serine in P?-position with 4 x 10? M?1 s?1, 9.5 x 10? M?1 s?1 and 2.1 x 10? M?1 s?1, respectively. The proposed existence of an additional binding region outside the catalytic center is supported by measurements of peptide substrates with extended chain length. This 'secondary' binding site interaction depends on the amino acid sequence in P?'-P?'-position. Interactions with this binding site could be specifically blocked for substrates of the GRF/glucagon peptide family. By contrast, substrates not belonging to this peptide family and dipeptide derivative substrates that only bind to the catalytic center of DP4 were not inhibited. This more selective inhibition approach allows, for the first time, to distinguish between substrate families by substrate-discriminating inhibitors.     

The Brain Response to Peripheral Insulin Declines with Age: A Contribution of the Blood-Brain Barrier?

ObjectivesIt is a matter of debate whether impaired insulin action originates from a defect at the neural level or impaired transport of the hormone into the brain. In this study, we aimed to investigate the effect of aging on insulin concentrations in the periphery and the central nervous system as well as its impact on insulin-dependent brain activity.MethodsInsulin, glucose and albumin concentrations were determined in 160 paired human serum and cerebrospinal fluid (CSF) samples. Additionally, insulin was applied in young and aged mice by subcutaneous injection or intracerebroventricularly to circumvent the blood-brain barrier. Insulin action and cortical activity were assessed by Western blotting and electrocorticography radiotelemetric measurements.ResultsIn humans, CSF glucose and insulin concentrations were tightly correlated with the respective serum/plasma concentrations. The CSF/serum ratio for insulin was reduced in older subjects while the CSF/serum ratio for albumin increased with age like for most other proteins. Western blot analysis in murine whole brain lysates revealed impaired phosphorylation of AKT (P-AKT) in aged mice following peripheral insulin stimulation whereas P-AKT was comparable to levels in young mice after intracerebroventricular insulin application. As readout for insulin action in the brain, insulin-mediated cortical brain activity instantly increased in young mice subcutaneously injected with insulin but was significantly reduced and delayed in aged mice during the treatment period. When insulin was applied intracerebroventricularly into aged animals, brain activity was readily improved.ConclusionsThis study discloses age-dependent changes in insulin CSF/serum ratios in humans. In the elderly, cerebral insulin resistance might be partially attributed to an impaired transport of insulin into the central nervous system.     

Polymorphisms within the Novel Type 2 Diabetes Risk Locus MTNR1B Determine β-Cell Function

Background

Very recently, a novel type 2 diabetes risk gene, i.e., MTNR1B, was identified and reported to affect fasting glycemia. Using our thoroughly phenotyped cohort of subjects at an increased risk for type 2 diabetes, we assessed the association of common genetic variation within the MTNR1B locus with obesity and prediabetes traits, namely impaired insulin secretion and insulin resistance.

Methodology/Principal Findings

We genotyped 1,578 non-diabetic subjects, metabolically characterized by oral glucose tolerance test, for five tagging single nucleotide polymorphisms (SNPs) covering 100% of common genetic variation (minor allele frequency >0.05) within the MTNR1B locus (rs10830962, rs4753426, rs12804291, rs10830963, rs3781638). In a subgroup (N = 513), insulin sensitivity was assessed by hyperinsulinemic-euglycemic clamp, and in a further subgroup (N = 301), glucose-stimulated insulin secretion was determined by intravenous glucose tolerance test. After appropriate adjustment for confounding variables and Bonferroni correction for multiple comparisons, none of the tagging SNPs was reliably associated with measures of adiposity. SNPs rs10830962, rs4753426, and rs10830963 were significantly associated with higher fasting plasma glucose concentrations (p<0.0001) and reduced OGTT- and IVGTT-induced insulin release (p≤0.0007 and p≤0.01, respectively). By contrast, SNP rs3781638 displayed significant association with lower fasting plasma glucose levels and increased OGTT-induced insulin release (p<0.0001 and p≤0.0002, respectively). Moreover, SNP rs3781638 revealed significant association with elevated fasting- and OGTT-derived insulin sensitivity (p≤0.0021). None of the MTNR1B tagging SNPs altered proinsulin-to-insulin conversion.

Conclusions/Significance

In conclusion, common genetic variation within MTNR1B determines glucose-stimulated insulin secretion and plasma glucose concentrations. Their impact on β-cell function might represent the prevailing pathomechanism how MTNR1B variants increase the type 2 diabetes risk.     

Fledermäuse im Windkanal

Zusammenfassung In einem Windkanal wurde für eine Myotis lucifugus die Abhängigkeit der Fluggeschwindigkeit (v_F) und der Geschwindigkeit über Grund (v_G) von der Windgeschwindigkeit (v_W) bestimmt. Die Fledermaus flog bei Windstille mit einer mittleren v_F von etwa 4,5 m/sec. Bei zunehmenden Gegenwinden erhöhte sie v_F und verringerte v_G, um bei v_W=7,7 m/sec für kurze Zeit stationären Flug (v_G=0) zu erreichen. Die Flügelschlagfrequenz lag bei Gegenwinden von 0–7,7 m/sec zwischen 10–11/sec. Bei zunehmenden Rückenwinden wurde der Flug immer mehr dem Rüttelflug ähnlich und die Flügelschlagfrequenz stieg bis 16/sec an. Die v_G blieb nahezu konstant in einem Bereich zwischen 4,5–5 m/sec. Bei Myotis lucifugus, Chilonycteris rubiginosa, Carollia perspillicata und Rhinolophus ferrum-equinum wurde die Flügelstellung während der Lautaussendung ermittelt. Alle Arten erzeugten entweder einen Einzellaut oder eine Gruppe von Lauten pro Flügelschlag.

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Bats in the wind tunnel

Summary In an experimental wind tunnel air speed (v_F) and ground speed (v_G) of a Myotis lucifugus were measured as a function of wind speed (v_W). The bat had a v_F of about 4,5 m/sec in still air. With head winds it increased v_F and lowered v_G to reach stationary flight (v_G=0) at a v_W of 7,7 m/sec. The rate of wing motion remained at about 10–11/sec at head winds from 0–7,7 m/sec. With tail winds the bat changed to a semi-hovering flight with wing beat frequencies rising to about 16/sec and v_F dropping to almost zero at 4–5 m/sec tailwinds. The v_G remained nearly constant between about 4,5–5 m/sec. (Figs. 2 and 3). The wing positions during which orientation sounds were emitted were determined for Myotis lucifugus (Fig. 4), Chilonycteris rubiginosa, Carollia perspillicata and Rhinolophus ferrum-equinum (Fig. 5). All bats emitted either one single sound or a group of sounds per wing beat.