Statistical Analysis of a Round-Robin Measurement Survey of Two Candidate Materials for a Seebeck Coefficient Standard Reference Material.

In an effort to develop a Standard Reference Material (SRM™) for Seebeck coefficient, we have conducted a round-robin measurement survey of two candidate materials-undoped Bi2Te3 and Constantan (55 % Cu and 45 % Ni alloy). Measurements were performed in two rounds by twelve laboratories involved in active thermoelectric research using a number of different commercial and custom-built measurement systems and techniques. In this paper we report the detailed statistical analyses on the interlaboratory measurement results and the statistical methodology for analysis of irregularly sampled measurement curves in the interlaboratory study setting. Based on these results, we have selected Bi2Te3 as the prototype standard material. Once available, this SRM will be useful for future interlaboratory data comparison and instrument calibrations.

Triacylglycerol microemulsions stabilized by alkyl ethoxylate surfactants--a basic study. Phase behavior, interfacial tension and microstructure.

As triacylglycerols are the main components of natural fats and oils their solubilization in the form of emulsions or microemulsions was of great interest within the last years. However, systematic studies of their properties along the classical lines of complex fluids science are still missing. In the present paper we focus on the phase behavior, the interfacial tension and the microstructure of systems of type H(2)O/NaCl-triacylglycerol-alkylpolyglycolether (C(i)E(j)). The interfacial tension between water and oil sigma(ab) is high in a microemulsion system containing triolein. Thus, one needs high surfactant mass fractions to formulate a single-phase microemulsion. We show that this is not only true for triolein, but also for saturated long-chained triacylglycerols. The determination of the amphiphilicity factor f(a) and the calculation of the bending rigidities of the amphiphilic film confirm that despite the fact that high surfactant mass fractions and high temperatures are needed to solubilize triolein in a system of type H(2)O/NaCl-triacylglycerol-alkylpolyglycolether (C(i)E(j)), this is still a microemulsion in the narrower sense.

Elastic properties of polyelectrolyte capsules studied by atomic-force microscopy and RICM.

Mechanical properties of polyelectrolyte multilayer capsules were studied using a new method combining atomic-force microscopy and reflection interference contrast microscopy. By measuring the force vs. deformation for poly(styrene sulfonate)/poly(allylamine) capsules the existence of different deformation regimes depending on the applied deformation was shown. The present paper focuses on the small-deformation regime. The elastic response of the deformed capsule was studied as a function of the wall thickness and the capsule size, and showed the theoretically expected variations. The Young modulus obtained from the experiments ranges between 1.3 and 1.9 GPa.

mAChRs in the grasshopper brain mediate excitation by activation of the AC/PKA and the PLC second-messenger pathways.

The species-specific sound production of acoustically communicating grasshoppers can be stimulated by pressure injection of both nicotinic and muscarinic agonists into the central body complex and a small neuropil situated posterior and dorsal to it. To determine the role of muscarinic acetylcholine receptors (mAChRs) in the control of acoustic communication behavior and to identify the second-messenger pathways affected by mAChR-activation, muscarinic agonists and membrane-permeable drugs known to interfere with specific mechanisms of intracellular signaling pathways were pressure injected to identical sites in male grasshopper brains. Repeated injections of small volumes of muscarine elicited stridulation of increasing duration associated with decreased latencies. This suggested an accumulation of excitation over time that is consistent with the suggested role of mAChRs in controlling courtship behavior: to provide increasing arousal leading to higher intensity of stridulation and finally initiating a mating attempt. At sites in the brain where muscarine stimulation was effective, stridulation could be evoked by forskolin, an activator of adenylate cyclase (AC); 8-Br-cAMP-activating protein kinase A (PKA); and 3-isobuty-1-methylxanthine, leading to the accumulation of endogenously generated cAMP through inhibition of phosphodiesterases. This suggested that mAChRs mediate excitation by stimulating the AC/cAMP/PKA pathway. In addition, muscarine-stimulated stridulation was inhibited by 2'-5'-dideoxyadenonsine and SQ 22536, two inhibitors of AC; H-89 and Rp-cAMPS, two inhibitors of PKA; and by U-73122 and neomycin, two agents that inhibit phospholipase C (PLC) by independent mechanisms. Because the inhibition of AC, PKA, or PLC by various individually applied substances entirely suppressed muscarine-evoked stridulation in a number of experiments, activation of both pathways, AC/cAMP/PKA and PLC/IP(3)/diacylglycerine, appeared to be necessary to mediate the excitatory effects of mAChRs. With these studies on an intact "behaving" grasshopper preparation, we present physiological relevance for mAChR-evoked excitation mediated by sequential activation of the AC- and PLC-initiated signaling pathways that has been reported in earlier in vitro studies.

Stridulatory pattern generation in acridid grasshoppers: metathoracic interneurons in Stenobothrus rubicundus (Germar 1817).

Stridulation was elicited in tethered gomphocerine grasshoppers of the species Stenobothrus rubicundus in order to identify interneurons of the stridulation pattern generator, and describe their morphological and physiological properties. Nine types of such neurons could be found and characterized; eight of those could additionally be compared to corresponding neuron types previously known from other species. As shown in detail for one selected type, the neurons of the stridulation pattern generator are very similar in their anatomical appearance, and possess similar physiological qualities at least in two species with similar stridulation patterns. Stridulation interneurons of species with largely different stridulatory motor patterns have a similar morphology, but show a different activation timing throughout the stridulation. Nevertheless, special properties such as resetting or initiation capability of certain stridulation interneurons seem to be conserved throughout the species. The results suggest that the stridulation pattern generator of different species consists of a uniform set of interneurons that change their activity pattern to produce species-specific song movements.

A role for muscarinic excitation: control of specific singing behavior by activation of the adenylate cyclase pathway in the brain of grasshoppers.

Muscarinic acetylcholine receptors exert slow and prolonged synaptic effects in both vertebrate and invertebrate nervous systems. Through activation of G proteins, they typically decrease intracellular cAMP levels by inhibition of adenylate cyclase or stimulate phospholipase C and the turnover of inositol phosphates. In insects, muscarinic receptors have been credited with two main functions: inhibition of transmitter release from sensory neuron terminals and regulation of the excitability of motoneurons and interneurons. Our pharmacological studies with intact and behaving grasshoppers revealed a functional role for muscarinic acetylcholine receptors as being the basis for specific arousal in defined areas of the brain, underlying the selection and control of acoustic communication behavior. Periodic injections of acetylcholine into distinct areas of the brain elicited songs of progressively increasing duration. Coinjections of the muscarinic receptor antagonist scopolamine and periodic stimulations with muscarine identified muscarinic receptor activation as being the basis for the underlying accumulation of excitation. In contrast to reports from other studies on functional circuits, muscarinic excitation was apparently mediated by activation of the adenylate cyclase pathway. Stimulation of adenylate cyclase with forskolin and of protein kinase A with 8-Br-cAMP mimicked the stimulatory effects of muscarine whereas inhibition of adenylate cyclase with SQ22536 and of protein kinase A with H-89 and Rp-cAMPs suppressed muscarine-stimulated singing behavior. Activation of adenylate cyclase by muscarinic receptors has previously been reported from studies on membrane preparations and heterologous expression systems, but a physiological significance of this pathway remained to be demonstrated in an in vivo preparation.

Pharmacological brain stimulation releases elaborate stridulatory behaviour in gomphocerine grasshoppers--conclusions for the organization of the central nervous control.

Grasshoppers produce a variety of sounds generated by complex movements of the hindlegs. Stridulation, performed in the context of partner finding, mating and rivalry, can be released by pressure injection of cholinergic agonists into the protocerebrum. Particularly stimulation with muscarinic agonists induced long-lasting stridulation that resembled the natural behaviour to an astonishing degree, not only with respect to their temporal structure and right/left coordination, but also to changes in the song sequences according to the progress of courtship stridulation, even including accessory movements of other parts of the body. According to the complexity of their stridulatory behaviour ten gomphocerine species were chosen for this comparative study. The results indicate that the protocerebrum fulfils two important tasks in the control of stridulation: (1) it integrates sensory input relevant to stridulation that represents a certain behavioural situation and internal state of arousal, and (2) it selectively activates and deactivates the thoracic networks that generate the appropriate movement and sound patterns. With the knowledge of the natural behaviour and the accessibility to pharmacological and electrophysiological studies, the cephalic control system for stridulation in grasshoppers appears to be a suitable model for how the brain selects and controls appropriate behaviours for a given situation.

Directional hearing of a grasshopper in the field.

An electrophysiological method for making long-term recordings from the tympanal nerve was developed in Chorthippus biguttulus (Gomphocerinae) to gain insight into the ecophysiological constraints of sound localization in acridid grasshoppers. Using this 'biological microphone', the directional dependence of auditory nerve activity was monitored both in the laboratory and in various natural habitats of this species. On gravel and in sparse vegetation, the overall patterns of directionality were found to be very similar to those in the free sound field in the laboratory, regardless of whether the animal was positioned horizontally or vertically. However, the differences between the ipsi- and contralateral sides were smaller in these habitats than in the laboratory. In dense vegetation, the directional patterns were greatly affected by the environment. Moreover, a minimum in nerve activity was not always reached on the contralateral side, as is typical for the free sound field situation. On the basis of these data, predictions can be made about the ability of the animals to determine the correct side of a sound source. In the free sound field of the laboratory, correct lateralizations are expected at all angles of sound incidence between 20 and 160 degrees, a prediction corresponding to the results of behavioural studies. In sparse vegetation, a similar accuracy can be anticipated, whereas on gravel and in dense vegetation directional hearing is expected to be severely degraded, especially if the animal is oriented horizontally. The predictions from our present electrophysiological investigations must now be confirmed by behavioural studies in the field.

Sound emission and the acoustic far field of a singing acridid grasshopper (Omocestus viridulus L.)

An array of eight microphones, all at a distance of 15 cm, was used to make simultaneous recordings of the sounds emitted by courting male acridid grasshoppers of the species Omocestus viridulus. In this species, the movement pattern for sound production differs in the two hindlegs, and in most cases the leg facing the female moves with the larger amplitude. The sonic sound intensity (the total sound in the one-third octave bands with centre frequencies from 5 to 20 kHz) is maximal ipsilateral to the leg stridulating with the larger amplitude (the dominant leg). A spontaneous switch of dominance to the other leg may cause a significant change in the emitted sound power. The sound intensities contralateral to the dominant leg and frontal to the animal are, on average, approximately half (-3 dB) of the ipsilateral value, whereas the mean sound intensities behind and above the singer are approximately one-fifth (-7 dB) of the ipsilateral value. In most singers, the patterns of sound radiation are close to these mean values, but in some singers the radiation patterns are radically different. The sound radiated in various directions differs not only in terms of sound intensity but also with respect to the frequency spectrum, which was studied up to the one-third octave band with a centre frequency of 31.5 kHz. In particular, the ratio between the ultrasonic and sonic components is much smaller in the forward direction than in other directions. This may allow the courted female to hear whether the courting male is oriented directly towards her.

Cholinergic activation of stridulatory behaviour in the grasshopper Omocestus viridulus (L.)

When acetylcholine (ACh) and its agonists are injected into neuropile regions of the protocerebrum and the suboesophageal ganglion of male and female grasshoppers of the species Omocestus viridulus (L.), they elicit stridulation in a pattern no different from that of natural song. Stridulation can even be evoked in mated females which normally do not sing. By choosing suitable ACh agonists, nicotinic and muscarinic ACh receptors can be activated selectively. Activation of nicotinic ACh receptors produces individual song sequences with rapid onset; the stridulation induced by activation of the muscarinic ACh receptors begins after a longer latency, increases slowly in intensity and is maintained for many minutes. The sites within the cephalic ganglia where song can be initiated pharmacologically coincide with regions in which descending stridulatory command neurones arborize.

How well are FREQUENCY SENSITIVITIES OF grasshopper ears tuned to species-specific song spectra?

Grasshoppers of 20 acridid species were examined using spectral analysis, laser vibrometry and electrophysiology to determine whether the song spectra, the best frequencies of tympanal-membrane vibrations and the threshold curves of the tympanal nerves are adapted to one another. The songs of almost all species have a relatively broad-band maximum in the region between 20 and 40 kHz and a narrower peak between 5 and 15 kHz. There are clear interspecific differences in the latter, which are not correlated with the length of the body or of the elytra. At the site of attachment of the low-frequency receptors (a-cells), the tympanal membrane oscillates with maximal amplitude in the region from 5 to 10 kHz. At the attachment site of the high-frequency receptors (d-cells), there is also a maximum in this region as well as another around 15-20 kHz. The tympanal nerve is most sensitive to tones between 5 and 10 kHz, with another sensitivity maximum between 25 and 35 kHz. The species may differ from one another in the position of the low-frequency peaks of the membrane oscillation, of the nerve activity and of the song spectra. No correlation was found between the characteristic frequency of the membrane oscillation and the area of the tympanal membrane. Within a given species, the frequency for maximal oscillation of the membrane at the attachment site of the low-frequency receptors and the frequency for maximal sensitivity of the tympanal nerve are in most cases very close to the low-frequency peak in the song spectrum. In the high-frequency range, the situation is different: here, the position of the peak in the song spectrum is not correlated with the membrane oscillation maximum at the attachment site of the high-frequency receptors, although there is a correlation between the song spectrum and the sensitivity of the tympanal nerve. On the whole, therefore, hearing in acridid grasshoppers is quite well adjusted to the frequency spectra of the songs, partly because the tympanal membrane acts as a frequency filter in the low-frequency range.

Pharmacological induction and modulation of stridulation in two species of acridid grasshoppers

The influence of neurotransmitters and neuroactive substances on stridulatory behaviour was analysed in two species of acridid grasshoppers (Omocestus viridulus and Chorthippus mollis). Acetylcholine, octopamine, gamma-aminobutyric acid and glutamate were applied by pressure injection (0.5­1.0 nl, 10(-3) mol l-1) into the protocerebrum. All except octopamine were also applied to the metathoracic ganglion by pressure injection or superfusion (1 ml). Injection of acetylcholine into the medial dorsal neuropile of the protocerebrum elicited continuous long-lasting species-specific stridulation in both acridid species. All other substances tested had no effect when injected into the brain. Injection of acetylcholine into the medial dorsal neuropile of the metathoracic ganglion enhanced the amplitude of the stridulatory leg movements elicited by electrical brain stimulation. It did not alter the repetition rate or coordination of the movements in O. viridulus; but it decreased the length of stridulatory cycles in C. mollis. Injection of gamma-aminobutyric acid into the medial dorsal metathoracic neuropile in both species suppressed the stridulatory leg movements ipsilateral to the injection site but did not alter those on the contralateral side. Superfusion of the metathoracic ganglion with gamma-aminobutyric acid suppressed the movements of both hindlegs. Pressure injection of glutamate into the metathoracic ganglion had no effect on the stridulatory leg movements, but superfusion enhanced the stridulatory movements.

Morphology of local "stridulation" interneurons in the metathoracic ganglion of the acridid grasshopper Omocestus viridulus L.

The morphological characteristics of five types of local spiking interneurons in the metathoracic ganglion of the acridid grasshopper Omocestus viridulus L. have been revealed by intracellular injection of the fluorescent dye Lucifer Yellow. All these neurons are active during induced stridulation and discharge in the stridulation rhythm. They are structurally unlike any of the metathoracic local interneurons previously described in the Acrididae. Type 1: Unilateral interneurons in the metathoracic neuromere; soma in anterior position; arborization dorsal, in anterolateral, posterolateral, and midline regions. Type 2: Unilateral interneurons in the metathoracic neuromere; neurite with characteristic hairpin bend; soma in lateral position; arborization through the lateral neuropil region. Type 3: Unilateral interneurons in the metathoracic and abdominal neuromeres; soma anterior at the base of the connective; arborization dorsal, in anterolateral, posterolateral, and midline regions, with ladderlike branches into the abdominal neuromeres. Type 4: Symmetrically bilateral neurons; soma in ventral median position; arborization dorsal, in anterolateral, posterolateral, and midline regions. Ipsilateral branches mostly smooth, contralateral mostly varicose. Type 5: Asymmetrically bilateral neurons, soma in ventral median position; ipsilateral branches mostly smooth, contralateral mostly varicose. The ipsilateral branches in particular penetrate all layers of the ganglion. The main arborizations of interneurons of types 1 and 4, and to some extent those of types 3 and 5, occupy the same region, which extends very far dorsally into the vicinity of the median and lateral tracts. There are additional antero- and posterolateral branches in the metathoracic neuromere, in positions where in principle they can overlap with the arborizations of the motoneurons. The interneurons of type 2, and to some extent type 5, arborize mainly in a lateral region that also includes deeper layers of the ganglion and hence encompasses the projections of mechanoreceptive proprioceptors. The interneurons of types 1, 3, and 4 have so-called "sister" neurons, identical to the typical neurons in their metathoracic arborizations but with additional collaterals that pass into the mesothoracic ganglion.