The structure of the cochlea in chiroptera. I. Microchiroptera. Emballonuroidea and Rhinolophoidea

The structure of the ears, especially of the cochleae, has been examined in two superfamilies, i.e. Emballonuroidea and Rhinolophoidea of the Microchiroptera. An intra-vitam fixation method was used and gave good histological preparations. The following features of the cochlea have been measured: the general size of the cochlea, the width and thickness of the basilar membrane, the size of the spiral ligament and the height of the cells of Claudius. The interpretation of measurements is discussed in relation to the nature of ultrasonic sounds produced by bats. It is concluded that functional interpretation of specific cochlear modifications cannot be assessed at present.     

Molecules,morphometrics and new fossils provide an integrated view of the evolutionary history of Rhinopomatidae (Mammalia: Chiroptera)

Background  

The Rhinopomatidae, traditionally considered to be one of the most ancient chiropteran clades, remains one of the least known groups of Rhinolophoidea. No relevant fossil record is available for this family. Whereas there have been extensive radiations in related families Rhinolophidae and Hipposideridae, there are only a few species in the Rhinopomatidae and their phylogenetic relationship and status are not fully understood.     

Parallel evolution of auditory genes for echolocation in bats and toothed whales

The ability of bats and toothed whales to echolocate is a remarkable case of convergent evolution. Previous genetic studies have documented parallel evolution of nucleotide sequences in Prestin and KCNQ4, both of which are associated with voltage motility during the cochlear amplification of signals. Echolocation involves complex mechanisms. The most important factors include cochlear amplification, nerve transmission, and signal re-coding. Herein, we screen three genes that play different roles in this auditory system. Cadherin 23 (Cdh23) and its ligand, protocadherin 15 (Pcdh15), are essential for bundling motility in the sensory hair. Otoferlin (Otof) responds to nerve signal transmission in the auditory inner hair cell. Signals of parallel evolution occur in all three genes in the three groups of echolocators--two groups of bats (Yangochiroptera and Rhinolophoidea) plus the dolphin. Significant signals of positive selection also occur in Cdh23 in the Rhinolophoidea and dolphin, and Pcdh15 in Yangochiroptera. In addition, adult echolocating bats have higher levels of Otof expression in the auditory cortex than do their embryos and non-echolocation bats. Cdh23 and Pcdh15 encode the upper and lower parts of tip-links, and both genes show signals of convergent evolution and positive selection in echolocators, implying that they may co-evolve to optimize cochlear amplification. Convergent evolution and expression patterns of Otof suggest the potential role of nerve and brain in echolocation. Our synthesis of gene sequence and gene expression analyses reveals that positive selection, parallel evolution, and perhaps co-evolution and gene expression affect multiple hearing genes that play different roles in audition, including voltage and bundle motility in cochlear amplification, nerve transmission, and brain function.     

The structure of the cochlea in chiroptera. III. Microchiroptera: Phyllostomatoidea

The middle ear and the cochlea have been described in 20 representatives from the superfamily Phyllostomatoidea of the suborder Microchiroptera. Various measurements have been made and the results compared by means of graphs. The ultrasonic pulses used by these bats for echolocation have been recorded and analysed. Generally, the structure of the cochlea does not show extreme modifications as found in some other superfamilies that have previously been discussed.     

Morphological diversity of wing structure in Rhinolophoid bats (Chiroptera,Rhinolophoidea)

The results of studies on the morphological diversity of wing structure in horseshoe bats (Rhinolophoidea) are considered in relation to ecological differentiation within the superfamily. Various skeletal elements of the wing are shown to form several groups of mutually correlated characters. The main groups comprise the characters that account for the size of the wing and play a role in the formation of its outer contours. The distribution of characters shows that they describe different aspects of variation and can provide a fairly complete picture of the morphological and ecological structure of the superfamily. The results obtained by means of multidimensional scaling and clustering methods primarily reflect ecological types of bats and only then characterize their taxonomic relationships. Analysis of variation in structural features of the wing within the superfamily Rhinolophoidea shows that the same wing form in different families may be accounted for by different ratios of digit elements, primarily the lengths of distal and proximal phalanges of digits III and IV.     

Evolution of the placenta and associated reproductive characters in bats

Recent advances in molecular phylogenetics indicate that the order Chiroptera is monophyletic and that one of four lineages of microbats (Rhinolophoidea) shares a common origin with megabats. Against this background we undertook a comprehensive analysis of placental evolution in bats. We defined a range of characters and character states associated with female reproduction, early development, placentation and the neonate. These were then mapped on a pre-existing hypothesis of bat relationships that represents the current view from molecular studies. Our purpose was threefold. First, on the assumption of bat monophyly, we wished to establish the stem species pattern of extant chiropterans. Secondly, we asked whether there are derived character conditions in support of a common origin for Rhinolophoidea and the megabats. Thirdly, we looked for evolutionary character transformations that characterize higher-level clades within Chiroptera, i.e. the megabats and the four lineages of microbats. The character condition occurring in the last common ancestor of Chiroptera was unequivocal for 21 of the 25 characters included in the analysis. The data did not offer support for a megabat-rhinolophoid clade or the implication that microbats are paraphyletic. However, analysis of early development, placentation and other reproductive parameters resulted in derived character conditions for the megabats as well as for each of the four major lineages of microbats.     

Synchrotron excitation of DNA fluorescence. Decay time evidence for excimer emission at room temperature

The first lifetime measurements of DNA fluorescence are reported. Natural and synthetic DNA have been excited by 1.76 ns pulses of synchrotron ultraviolet radiation (270 nm) and the time profile of the fluorescence has been measured by synchronous single-photon counting. A post-pulse exponentially decaying emission has been observed with a lifetime of 2.9 +/- 0.4 ns for calf thymus DNA and 3.0 +/- 0.3 ns for poly(dA-T); this is most likely an excimer fluorescence.     

Phase transition of dimyristoylphosphatidylglycerol bilayers induced by electric field pulses

The phase transition of dimyristoylphosphatidylglycerol (DMPG) bilayers has been studied by measurements of light scattering under high electric field pulses. Midpoints of phase transitions have been identified by a clear discontinuity of field induced relaxation amplitudes. We show that the phase transition of DMPG suspensions in monovalent salt is virtually independent of the electric field strength up to approx. 35 kV/cm. A shift of the lipid phase by electric field pulses has been observed, however, for DMPG suspensions in the presence of Ca2+ ions. DMPG suspensions exhibit a jump of the phase transition temperature from 17 degrees C at Ca/DMPG molar ratios r less than 1/7 to 32 degrees C at r greater than 1/7. Field pulses of 60 to 100 microseconds applied to DMPG suspensions with Ca2+ at r greater than 1/7 induce discontinuities of relaxation amplitudes in the temperature range 15 to 22 degrees C in addition to the 'standard' one at 32 degrees C, when the electric field strength is above 15 kV/cm. These results indicate that electric field pulses induce a transition from the phase formed at 'high' Ca(2+)- to the one formed at 'low' Ca(2+)-ion concentrations. Our results are consistent with a dissociation field effect on Ca(2+)-lipid complexes which drives the phase transition.     

Closing in on maximum yield of chlorophyll fluorescence using a single multiphase flash of sub‐saturating intensity

Estimation of the maximum chlorophyll fluorescence yield under illumination, or Fm′, by traditional single saturation pulse (SP) methodology is prone to underestimation error because of rapid turnover within photosystem (PS) II. However, measurements of fluorescence yield during several single pulses of variable intensity describes the irradiance dependence of apparent Fm′, from which estimates of Fm′ at infinite irradiance can be derived. While such estimates have been shown to result in valid approximations of Fm′, the need to apply several single pulses limits its applicability. We introduce a novel approach that determines the relationship between apparent Fm′ and variable irradiance within a single ～1 s multiphase flash (MPF). Through experiments and simulations, we demonstrate that the rate of variation in irradiance during an MPF is critical for achieving quasi–steady‐state changes in the proportions of PSII acceptor side redox intermediates and the corresponding fluorescence yields, which are prerequisites for accurately estimating Fm′ at infinite irradiance. The MPF methodology is discussed in the context of improving the accuracy of various parameters derived from chlorophyll fluorescence measurements, such as photochemical and non‐photochemical quenchings and efficiencies. The importance of using MPF methodology for interpreting chlorophyll fluorescence, in particular for integrating fluorescence and gas exchange measurements, is emphasized.     

Peculiarities of detecting pulses of runaway electrons and X-rays generated by high-voltage nanosecond discharges in open atmosphere

Peculiarities of detecting pulses of high-energy runaway electrons and the accompanying X-rays generated by high-voltage nanosecond discharges at high overvoltages in dense gases are discussed. Methods for overcoming difficulties encountered in such measurements are demonstrated. Different techniques for detecting runaway electrons and X-rays generated by discharges in open atmosphere are described, and typical errors that may be done when interpreting measurement results are considered. Experiments with the use of a small-size generator of nanosecond pulses with the idle-running voltage of 250–270 kV and stored energy of ～0.5 J have been carried out. It is shown that, in measuring runaway electron pulses by using an experimental configuration with a grid anode, a major portion of the recorded signal is attributed to electromagnetic disturbances. It is found that X-rays are mainly generated due to the deceleration of runaway electrons in the anode, rather than in gas. The number of runaway electrons with energies ～300 keV does not exceed 10⁹ per shot, and the X-ray dose is 60–200 μR/shot, depending on the anode material.     

Dynamics of the anaerobic process: effects of volatile fatty acids

A complex and fast dynamic response of the anaerobic biogas system was observed when the system was subjected to pulses of volatile fatty acids (VFAs). It was shown that a pulse of specific VFAs into a well-functioning continuous stirred tank reactor (CSTR) system operating on cow manure affected both CH(4) yield, pH, and gas production and that a unique reaction pattern was seen for the higher VFAs as a result of these pulses. In this study, two thermophilic laboratory reactors were equipped with a novel VFA-sensor for monitoring specific VFAs online. Pulses of VFAs were shown to have a positive effect on process yield and the levels of all VFA were shown to stabilize at a lower level after the biomass had been subjected to several pulses. The response to pulses of propionate or acetate was different from the response to butyrate, iso-butyrate, valerate, or iso-valerate. High concentrations of propionate affected the degradation of all VFAs, while a pulse of acetate affected primarily the degradation of iso-valerate or 2-methylbutyrate. Pulses of n-butyrate, iso-butyrate, and iso-valerate yielded only acetate, while degradation of n-valerate gave both propionate and acetate. Product sensitivity or inhibition was shown for the degradation of all VFAs tested. Based on the results, it was concluded that measurements of all specific VFAs are important for control purposes and increase and decrease in a specific VFA should always be evaluated in close relationship to the conversion of other VFAs and the history of the reactor process. It should be pointed out that the observed dynamics of VFA responses were based on hourly measurements, meaning that the response duration was much lower than the hydraulic retention time, which exceeds several days in anaerobic CSTR systems.     

Electrical Impedance Studies on Potato and Alfalfa Tissue

Small pulses of alternating current have been found to be conductedchiefly in the channels of the cell walls of tubers of potato(Solatium tuberosum), the roots and stems of alfalfa (Medicagosativa), and similar plant tissue. Impedance measurements mayusefully detect the state of water relations in these channelswhen the frequency of the measuring A.C. is low (around 60 c/s),when chlori-dized silver probes are used and when the cell-wallresistance is low compared to the cell membranes in series.Activation energies have been calculated for the conductivityof these tissues. Over the temperature and soil moisture rangesinvestigated, the log impedance has been found to be approximatelylinear with the reciprocal of these parameters. Impedance changeshave also been found to accompany endogenous and other changesin the plant. In particular, pulses of light or heat causedchanges of impedance which reflect a change of internal waterrelations and possibly transport. Suitable light and potassiumfertilizer treatments at temperatures around 4 °C enhancedthe impedance of alfalfa and its apparent resistance to frost.     

Effect of cell electroporation on the conductivity of a cell suspension

An increased permeability of a cell membrane during the application of high-voltage pulses results in increased transmembrane transport of molecules that otherwise cannot enter the cell. Increased permeability of a cell membrane is accompanied by increased membrane conductivity; thus, by measuring electric conductivity the extent of permeabilized tissue could be monitored in real time. In this article the effect of cell electroporation caused by high-voltage pulses on the conductivity of a cell suspension was studied by current-voltage measurements during and impedance measurement before and after the pulse application. At the same time the percentage of permeabilized and survived cells was determined and the extent of osmotic swelling measured. For a train of eight pulses a transient increase in conductivity of a cell suspension was obtained above permeabilization threshold in low- and high-conductive medium with complete relaxation in <1 s. Total conductivity changes and impedance measurements showed substantial changes in conductivity due to the ion efflux in low-conductive medium and colloid-osmotic swelling in both media. Our results show that by measuring electric conductivity during the pulses we can detect limit permeabilization threshold but not directly permeabilization level, whereas impedance measurements in seconds after the pulse application are not suitable.     

Role of pulse shape in cell membrane electropermeabilization

The role of the amplitude, number, and duration of unipolar rectangular electric pulses in cell membrane electropermeabilization in vitro has been the subject of several studies. With respect to unipolar rectangular pulses, an improved efficiency has been reported for several modifications of the pulse shape: separate bipolar pulses, continuous bipolar waveforms, and sine-modulated pulses. In this paper, we present the results of a systematic study of the role of pulse shape in permeabilization, cell death, and molecular uptake. We have first compared the efficiency of 1-ms unipolar pulses with rise- and falltimes ranging from 2 to 100 μs, observing no statistically significant difference. We then compared the efficiency of triangular, sine, and rectangular bipolar pulses, and finally the efficiency of sine-modulated unipolar pulses with different percentages of modulation. We show that the results of these experiments can be explained on the basis of the time during which the pulse amplitude exceeds a certain critical value.     

Differences in the electric birefringence of spectrin dimers and tetramers as shown by the fast reversing electric pulse method

The electric birefringence of purified Spectrin has been examined in medium of low ionic strength ai 20°C and for electric fields smaller than 4 × 10⁴ V m^?1. using the reversing electric pulse method. This technique allows study of the permanent and induced dipole electric moment of macromolecules more easily than in measurements using only rectangular pulses. We show that spectrin heterodimers and heterotetramers have different electro-optical properties. The relaxation time of the tetramer (7 μs) is significantly longer than that of the dimer (4.5 μs). Tetramers and dimers have also different polarizability parameters.     

Noninvasive cardiac output estimation: a preliminary study

 The availability of a simple-to-use, automatic measurement system for noninvasive flow estimation is imperative, given the clinical demand for an acceptable noninvasive procedure rather than the standard invasive procedure of thermodilution. A method for calculating cardiac output from noninvasively derived pressure pulses has been developed, and the results of a preliminary evaluation study on post-cardiac surgery patients for whom invasive flow measures were readily available for comparison are provided in this report. The proposed method relies on fast Fourier transform (FFT) analysis of pulses measured externally at the carotid and femoral pressure points. A transfer function of the aorta is computed from digitally filtered pulse measurements, and a tapered model of the aorta is parametrically adapted using a simplex optimization algorithm so that its transfer function matches that derived experimentally. An aortic input impedance term is obtained from the optimized model and utilized along with the carotid pulse (analogous to input voltage) to compute aortic flow. In addition to its automation, attractive features of this method include the requirement for relatively few pulses for analysis as well as considerable resistance to noise artifact. For 59 data records collected from 54 post-cardiac surgery patients, the average flow measurements computed over several pulses compare well with the standard, invasive method of thermodilution. Preliminary results also indicate a strong potential for tracking changes in cardiac output over time, and invite further use of the method in monitoring hemodynamically unstable patients. Received: 18 February 1997 / Accepted in revised form: 12 May 1997     

Flow-injection analysis of catecholamine secretion from bovine adrenal medulla cells on microbeads

Bovine adrenal medullary cells have been cultured on microbeads which are placed in a low-volume flow system for measurements of stimulation-response parameters. Electronically controlled stream switching allows stimulation of cells with pulse lengths from 1 s to many minutes; pulses may be repeated indefinitely. Catecholamines secreted are detected by an electrochemical detector downstream from the cells. This flow-injection analysis technique provides a new level of sensitivity and precision for measurement of kinetic parameters of secretion. A manual injection valve allows stimulation by higher levels of stimulant in the presence of constant low levels of stimulant. Such experiments show interesting differences between the effects of K+ and acetylcholine on cells partially desensitized to acetylcholine.     

The effect of multiple pulses of light on the circadian phase response of the rat.

Multiple pulses of light administered to humans have been reported to result in type 0 phase responses. These results suggest the underlying pacemaker to be nonsimple. At present, results with this type of protocol have only been reported for humans. Therefore, multiple pulses of light were administered to rats. Rats were exposed to one, two, three, or four pulses of light for 5 h (1000 lux) at successive 24-h intervals. Results did not suggest a type 0 phase response. Nonetheless, results with a second, third, or fourth light exposure were not fully predictable from a phase response curve derived from a single light pulse.     

Are ecosystem carbon inputs and outputs coupled at short time scales? A case study from adjacent pine and hardwood forests using impulse–response analysis

A number of recent studies have attributed a large proportion of soil respiration (R(soil)) to recently photoassimilated carbon (C). Time lags (tau(PR)) associated with these pulses of photosynthesis and responses of R(soil) have been found on time scales of hours to weeks for different ecosystems, but most studies find evidence for tau(PR) on the order of 1-5 d. We showed that such time scales are commensurate with CO(2) diffusion time scales from the roots to the soil surface, and may thus be independent from photosynthetic pulses. To further quantify the role of physical (i.e. edaphic) and biological (i.e. vegetative) controls on such lags, we investigated tau(PR) at adjacent planted pine (PP) and hardwood (HW) forest ecosystems over six and four measurement years, respectively, using both autocorrelation analysis on automated soil surface flux measurements and their lagged cross-correlations with drivers for and surrogates of photosynthesis. Evidence for tau(PR) on the order of 1-3 d was identified in both ecosystems and using both analyses, but this lag could not be attributed to recently photoassimilated C because the same analysis yielded comparable lags at HW during leaf-off periods. Future efforts to model ecosystem C inputs and outputs in a pulse-response framework must combine measurements of transport in the physical and biological components of terrestrial ecosystems.     

Sound radiation in a cicada: the role of different structures

Sound radiation was studied in males of Tympanistalna gastrica St»l during a spontaneous song with the characteristics of the conspecific calling song, which was elicited as an after effect of brain stimulation. The song contains two different kinds of sound pulses: 1) loud clicks and 2) soft pulses, presenting different spectra.The timbals, abdomen, tympana, folded membranes and opercula were tested as potential radiators of the song. The experiments included: 1) probe microphone measurements of the spectra of loud clicks and soft pulses in several positions around the animal and close to the body surface; 2) measurements of the spectra before and after covering with vaseline different structures that might be relevant to the radiation of the song, and manipulations of the size and shape of the abdominal and thoracic portions of the tracheal air sac; 3) laser vibrometry measurements in different parts of the body, both during singing and external sound stimulation.The data obtained demonstrate that several structures contribute differently to the radiation of clicks and soft pulses: 1) The timbals are the main radiators at frequencies around the dominant spectral peak, 10–11 kHz in clicks and 12–13 kHz in soft pulses; 2) The tympana are important in radiation of frequencies below and above the timbal peak, especially during the generation of soft pulses; 3) The abdomen is more activated during the generation of clicks, and is more important in the radiation of low frequencies around 5 kHz.Manipulations of the body cavities showed that neither the thoracic nor the abdominal portions of the air sac are critical for the song tuning. The large abdominal cavity do not seem to work as a Helmholtz resonator. We found no evidence that resonances inside this cavity should play an important role in enhancing sound radiation in T. gastrica.