Mother Vocal Recognition in Antarctic Fur Seal Arctocephalus gazella Pups: A Two-Step Process

In otariids, mother’s recognition by pups is essential to their survival since females nurse exclusively their own young and can be very aggressive towards non-kin. Antarctic fur seal, Arctocephalus gazella, come ashore to breed and form dense colonies. During the 4-month lactation period, females alternate foraging trips at sea with suckling period ashore. On each return to the colony, females and pups first use vocalizations to find each other among several hundred conspecifics and olfaction is used as a final check. Such vocal identification has to be highly efficient. In this present study, we investigated the components of the individual vocal signature used by pups to identify their mothers by performing playback experiments on pups with synthetic signals. We thus tested the efficiency of this individual vocal signature by performing propagation tests and by testing pups at different playback distances. Pups use both amplitude and frequency modulations to identify their mother’s voice, as well as the energy spectrum. Propagation tests showed that frequency modulations propagated reliably up to 64m, whereas amplitude modulations and spectral content greatly were highly degraded for distances over 8m. Playback on pups at different distances suggested that the individual identification is a two-step process: at long range, pups identified first the frequency modulation pattern of their mother’s calls, and other components of the vocal signature at closer range. The individual vocal recognition system developed by Antarctic fur seals is well adapted to face the main constraint of finding kin in a crowd.     

An efficient model development strategy for bioprocesses based on neural networks in macroscopic balances: Part II

There is a need for efficient modeling strategies which quickly lead to reliable mathematical models that can be applied for design and optimization of (bio)-chemical processes. The serial gray box modeling strategy is potentially very efficient because no detailed knowledge is needed to construct the white box part of the model and because covenient black box modeling techniques like neural networks can be used for the black box part of the model. This paper shows for a typical biochemical conversion how the serial gray box modeling strategy can be applied efficiently to obtain a model with good frequency extrapolation properties. Models with good frequency extrapolation properties can be applied under dynamic conditions that were not present during the identification experiments. For a given application domain of a model, this property can be used to considerably reduce the number of identification experiments. The serial gray box modeling strategy is demonstrated to be successful for the modeling of the enzymatic conversion of penicillin G In the concentration range of 10-100 mM and temperature range of 298-335 K. Frequency extrapolation is shown by using only constant temperatures in the (batch) identification experiments, while the model can be used reliable with varying temperatures during the (batch) validation experiments. No reliable frequency extrapolation properties could be obtained for a black box model, and for a more knowledge-driven white box model reliable frequency extrapolation properties could only be obtained by incorporating more knowledge in the model. Copyright 1999 John Wiley & Sons, Inc.     

MICE: multiple-peak identification, characterization, and estimation

MICE--multiple-peak identification, characterization, and estimation--is a procedure for estimating a lower bound of the number of frequency peaks and for estimating the frequency peak parameters. The leading application is protein structure determination using nuclear magnetic resonance (NMR) experiments. NMR frequency data are multiple-peak data, where each frequency peak corresponds to two connected atoms in the three-dimensional protein structure. We analyze the NMR frequency data through a series of steps: a preliminary step for separating the signal from the background followed by identification of local maxima up to a noise-level-dependent threshold, estimation of the frequency peak parameters using an iterative algorithm, and detection of mixtures of peaks using hypothesis testing.     

传粉导致的转基因白菜与其近缘种属材料间的基因流动

从形态、染色体及分子水平上证实,转基因不结球白菜(Brassica campestris ssp．chinensis var．utilis Tsen et Lee)中编码除草剂Basta抗性的bar基因能在田间条件下,经自然传粉,以较高频率侵入芜菁(B．campestris ssp．rapifera)、结球白菜(B．campestris／sssp．pek／nens／s)和不结球白菜(B．campestrs ssp．ch／nens／s)的基因组中,也能少量侵入同属异种的甘蓝型油菜(B.napus)基因组中;在温室人工辅助授粉条件下,除在上述种中的基因漂移率提高外,bar基因尚能以一定频率侵入同属的黑芥(B．nigra)、埃塞俄比亚芥(B．car／nata)、芥菜(B．juncea)基因组中,但始终未能得到转基因白菜与结球甘蓝(B．oleracea)、萝卜(尺．sativus)的杂种。转基因白菜与十字花科的7种常见杂草经温室人工辅助授粉,也均未得到抗性杂种。     

性诱盆与频振灯在棉铃虫成虫动态监测中的作用比较

20 0 1年在南疆棉区做了性信息素水盆诱捕器 (性诱盆 )和频振式杀虫灯 (频振灯 ,3 0W)对棉铃虫成虫诱捕效率的对比试验。结果表明 ,6个监测点平均 ,每日每只性诱盆和频振灯的诱蛾量分别为3 5 .4和 3 2 .3头 ,差异不显著 (P >0 .0 5 )。在 1代成虫盛发的前期和中后期 ,性诱盆相对于频振灯的诱蛾量有先低后高的现象。考虑到经济性和可识别性等因素 ,作者认为在南疆监测棉铃虫成虫动态性诱盆优于频振灯     

A simple vector modification to facilitate oligonucleotide-directed mutagenesis.

We describe a simple modification of commonly used single-stranded cloning vectors that permits the efficient recovery of mutant DNA molecules in oligonucleotide-directed mutagenesis experiments, even when the absolute efficiency of mutagenesis is very low. The modification consists of the insertion of a short synthetic DNA fragment into the vector's polylinker and permits the identification of mutant clones based on a standard chromogenic plate assay for bacterial colonies or phage plaques producing functional beta-galactosidase. Other useful properties of the original vector are retained in the modified version. In vitro mutagenesis reactions are carried out with two oligonucleotides, one to introduce the mutation of interest, and the second to correct a frameshift mutation introduced into the beta-galactosidase gene of the modified vector. We have found that these two sequence changes are closely linked following transformation of an appropriate E. coli strain with the products of the in vitro mutagenesis reaction, and have thereby recovered desired mutations at a frequency of about 50% even when the overall mutagenesis efficiency is less than 1%. By alternately correcting and re-introducing the beta-galactosidase frameshift mutation, we have shown that multiple rounds of mutagenesis can be carried out on the same template with a high efficiency of mutant recovery in each step. Modifications similar or identical to those we describe here should be feasible for most commonly used single-stranded cloning vectors and should increase the usefulness of these vectors by providing an additional option for oligonucleotide-directed mutagenesis to be used in conjunction with or in lieu of other commonly used approaches.     

Leaf senescence and N uptake parameters as selection traits for nitrogen efficiency of oilseed rape cultivars

The cultivation of N-efficient oilseed rape cultivars could contribute to a reduction of the large N balance surpluses of this crop. To facilitate the breeding process of N-efficient cultivars, the identification of secondary plant traits correlating with N efficiency is necessary. The objectives of this study were to investigate leaf senescence and N uptake parameters of oilseed rape cultivars and doubled haploid (DH) lines with contrasting N efficiency in a short-term nutrient solution experiment and to relate these results to their performance in field experiments. In the nutrient solution experiment, genotypes differed in leaf senescence of fully expanded leaves and maximum N uptake rate per unit root length under low N supply. A high maximum N uptake rate seemed to have contributed to delayed leaf senescence by enhancing N accumulation in leaves. Also in the field experiments, genotypes differed in leaf senescence after flowering at limiting N supply. Additionally, the most N-efficient DH line was able to adapt leaf photosynthetic capacity to the low-light conditions in the canopy during flowering. N efficiency (grain yield at limiting N supply) was positively correlated with delayed leaf senescence both in nutrient solution and field experiments. It is concluded that important leaf and root traits of N-efficient cultivars are expressed in short-term nutrient solution experiments, which may facilitate the selection of N-efficient cultivars.     

Assessment of the proportion of transgene-bearing sperm by fluorescence in situ hybridization: a novel approach for the detection of germline mosaicism in transgenic male founders

Genetic mosaicism is frequent among transgenic animals produced by pronuclear microinjection. A successful method for the screening of founder animals for germline mosaicism prior to mating would greatly reduce the costs associated with the propagation of the transgenic lines, and improve the efficiency of transgenic livestock production. With this aim, we have devised a simple method to detect integrated transgenes in individual spermatozoa using fluorescence in situ hybridization (FISH). The experiments reported here were undertaken to investigate the efficiency of this FISH-based approach to accurately evaluate the proportion of transgene-bearing sperm and to be applied for the detection of potential germline mosaics. Sperm samples from mice homozygous and hemizygous for a beta-lactoglobulin transgene were analyzed in a first set of experiments. A high hybridization efficiency was achieved, and the proportions of transgene-positive sperm cells in both homozygous (94.8-98.2%) and hemizygous (49.8-51.9%) animals were close to the expected frequencies (100 and 50%, respectively). To evaluate the sensitivity of the assay more directly, simulated mosaic samples with 5, 10, 15, 20 and 40% of transgene-bearing spermatozoa were then prepared and analyzed by FISH. Significant differences in the frequency of transgene-positive sperm were observed between all mosaic samples, indicating that even small deviations (5%) from the expected 50% transgene transmission rate in a founder animal could be reliably detected with our assay. Therefore, the method proposed represents a novel approach for the identification of germline mosaic founder males in livestock transgenic projects and a much more economic and faster alternative to breeding.     

Psychoacoustic sampling as a reliable,non-invasive method to monitor orthopteran species diversity in tropical forests

We evaluated trained listener—based acoustic sampling as a reliable and non-invasive method for rapid assessment of ensiferan species diversity in tropical evergreen forests. This was done by evaluating the reliability of identification of species and numbers of calling individuals using psychoacoustic experiments in the laboratory and by comparing psychoacoustic sampling in the field with ambient noise recordings made at the same time. The reliability of correct species identification by the trained listener was 100 % for 16 out of 20 species tested in the laboratory. The reliability of identifying the numbers of individuals correctly was 100% for 13 out of 20 species. The human listener performed slightly better than the instrument in detecting low frequency and broadband calls in the field, whereas the recorder detected high frequency calls with greater probability. To address the problem of pseudoreplication during spot sampling in the field, we monitored the movement of calling individuals using focal animal sampling. The average distance moved by calling individuals for 17 out of 20 species was less than 1.5 m in half an hour. We suggest that trained listener—based sampling is preferable for crickets and low frequency katydids, whereas broadband recorders are preferable for katydid species with high frequency calls for accurate estimation of ensiferan species richness and relative abundance in an area.     

Phenotyping transgenic wheat for drought resistance

Realistic experimental protocols to screen for drought adaptation in controlled conditions are crucial if high throughput phenotyping is to be used for the identification of high performance lines, and is especially important in the evaluation of transgenes where stringent biosecurity measures restrict the frequency of open field trials. Transgenic DREB1A-wheat events were selected under greenhouse conditions by evaluating survival and recovery under severe drought (SURV) as well as for water use efficiency (WUE). Greenhouse experiments confirmed the advantages of transgenic events in recovery after severe water stress. Under field conditions, the group of transgenic lines did not generally outperform the controls in terms of grain yield under water deficit. However, the events selected for WUE were identified as lines that combine an acceptable yield-even higher yield (WUE-11) under well irrigated conditions-and stable performance across the different environments generated by the experimental treatments.     

Nonlinear broadband doubling of the extraordinary wave frequency in inhomogeneous magnetoactive plasma

The nonlinear resonance doubling of radio wave frequencies in inhomogeneous plasma is studied as applied to the ionosphere under the conditions of the phase synchronism between an extraordinary pump wave and its second harmonic. The synchronism is not related to plasma resonances, but is determined by the magnetic field and plasma electron density in the transparency region. The generation efficiency of the second harmonic of a transversely propagating wave is calculated for a wide frequency band lying higher than the lower hybrid resonance frequency. It is shown that this effect is physically analogous to the generation of the second harmonic of laser radiation in a nonlinear crystal. The generation efficiency of the second harmonic is determined for inhomogeneous ionospheric plasma in which the synchronism condition is satisfied in a limited frequency range. It is shown that this effect can be used for remote nonlinear diagnostics of the upper ionospheric plasma, in which the characteristic size of the synchronism region can reach several kilometers. It is proposed to use a combination of satellite and ground-based ion probes in experiments on transionospheric probing. Even if the frequency of the wave emitted from the satellite is lower than the critical frequency in the ionosphere, the frequency of its second harmonic can exceed the critical frequency, so that it can be recorded by a ground-based ion probe or a specially designed receiver. The reflected second-harmonic signal can also be detected at the satellite by using a broadband radio-frequency spectrometer.     

Evaluating Letter Recognition,Flicker Fusion,and the Talbot-Plateau Law using Microsecond-Duration Flashes

Four experiments examined the ability of respondents to identify letters that were displayed on an LED array with flashes lasting little more than a microsecond. The first experiment displayed each letter with a single, simultaneous flash of all the dots forming the letter and established the relation of flash intensity to the probability of letter identification. The second experiment displayed the letters with multiple flashes at different frequencies to determine the probability that the sequence of flashes would be perceived as fused. The third experiment displayed the letters at a frequency that was above the flicker-fusion frequency, varying flash intensity to establish the amount needed to elicit a given probability of letter identification. The fourth experiment displayed each letter twice, once at a frequency where no flicker was perceived and also with steady light emission. The intensity of each flash was fixed and the steady intensity was varied; respondents were asked to judge whether the fused-flicker display and the steady display appeared to be the same brightness. Steady intensity was about double the average flash intensity where the two conditions were perceived as being equal in brightness. This is at odds with Talbot-Plateau law, which predicts that these two values should be equal. The law was formulated relative to a flash lasting half of each period, so it is surprising that it comes this close to being correct where the flash occupies only a millionth of the total period.     

Sequence and structure space of RNA-binding peptides

Studies of RNA-binding peptides, and recent combinatorial library experiments in particular, have demonstrated that diverse peptide sequences and structures can be used to recognize specific RNA sites. The identification of large numbers of sequences capable of binding to a particular site has provided extensive phylogenetic information used to deduce basic principles of recognition. The high frequency at which RNA-binding peptides are found in large sequence libraries suggests plausible routes to evolve sequence-specific binders, facilitating the design of new binding molecules and perhaps reflecting characteristics of natural evolution.     

传粉导致的转基因白菜与其近缘种属材料间的基因流动

从形态、染色体及分子水平上证实,转基因不结球白菜(Brassica campestris ssp. chinensis var. utilis Tsen et Lee)中编码除草剂Basta抗性的bar基因能在田间条件下,经自然传粉,以较高频率侵入芜菁(B. campestris ssp. rapifera)、结球白菜(B. campestris ssp. pekinensis)和不结球白菜(B. campestris ssp. chinensis)的基因组中,也能少量侵入同属异种的甘蓝型油菜(B. napus)基因组中; 在温室人工辅助授粉条件下,除在上述种中的基因漂移率提高外,bar基因尚能以一定频率侵入同属的黑芥(B. nigra)、埃塞俄比亚芥(B. carinata)、芥菜(B. juncea)基因组中,但始终未能得到转基因白菜与结球甘蓝(B. oleracea)、萝卜(R. sativus)的杂种.转基因白菜与十字花科的7种常见杂草经温室人工辅助授粉,也均未得到抗性杂种.     

Floral organ-specific and constitutive expression of an Arabidopsis thaliana heat-shock HSP18.2:: GUS fusion gene is retained even after homeotic conversion of flowers by mutation

Using the one-step gene disruption technique, we studied the effect of various parameters on the disruption frequency (percentage of homologous integrants) and transformation efficiency (number of transformants per μg of input DNA) of integrative transformation in Schizosaccharomyces pombe. We used suc1 as the target gene for disruption and ura4 as the selectable marker. Our results are as follows. 1) Use of the strong adh1 promoter to drive the expression of ura4 did not affect the disruption frequency but modestly increased the transformation efficiency. 2) The transformation method had a profound effect, with the lithium acetate method yielding both a 10-fold higher disruption frequency compared to the protoplast method and a 5- to 10-fold higher transformation efficiency. 3) The presence of increasing amounts of non-homologous sequences at the ends of the transforming DNA decreased the disruption frequency by up to 5-fold but had no effect on the transformation efficiency. We also describe the use of the sup3–5 allele in an ade6-704 genetic background to discriminate between the products of homologous versus non-homologous integration, thereby promoting the identification of rare homologous integrants.     

Recognizing Words and Reading Sentences with Microsecond Flash Displays

Strings of dots can be used to construct easily identifiable letters, and these in turn can be used to write words and sentences. Prior work found that respondents could identify individual letters when all the dots were simultaneously flashed for an ultra-brief duration. Four of the experiments reported here constructed five-letter words with these dot-letters and a fifth experiment used them to write complete sentences. Respondents were able to recognize individual words that were displayed with a single, simultaneous ultra-brief flash of all the letters. Further, sentences could be efficiently read with a sequence of simultaneous flashes at a frequency that produced perceptual fusion. One experiment determined the frequency range that would produce flicker-fusion. Two experiments established the relation of intensity to probability of recognition with single flashes and with fused-flicker frequencies. Another established the intensities at which flicker-fused and steady displays were judged to be equal in brightness. The final experiment used those flicker-fused and steady intensities to display sentences. The two display conditions were read with equal efficiency, even though the flicker-fused displays provided light stimulation only 0.003% of the time.     

棺头蟋属六种常见蟋蟀鸣声特征分析与种类鉴定(直翅目:蟋蟀总科)

本文对蟋蟀科Ｇｒｙｌｉｄａｅ棺头蟋属Ｌｏｘｏｂｌｅｍｍｕｓ６种常见种类的鸣声特征进行了较为系统的分析研究。从其频域特征和时域特征上明显地显示了种间差异,并将其鸣声特征用于分类。     

High-frequency transformation of human repair-deficient cell lines by an Epstein-Barr virus-based cDNA expression vector.

We constructed a human cDNA expression vector by combining an episomal Epstein-Barr virus (EBV) vector with the expression cassette from the transient-expression vector, pCDM8. This new vector, designated pEBS7, exhibited high-level expression of reporter genes in normal and repair-deficient xeroderma pigmentosum cell lines. Reconstruction experiments indicated that marker genes diluted to a frequency of 10(-5) can be rescued on a single transfection dish. Moreover, derivative cell lines that constitutively express the gene encoding EBV nuclear antigen 1 exhibited a tenfold enhancement in the frequency of rescue of marker genes. The feasibility of preparing large-scale directional or nondirectional cDNA libraries in pEBS7 was demonstrated and reconstruction experiments indicated that marker genes could be rescued from either library with equal efficiency. These results establish a high-efficiency system for the isolation of genes by direct phenotypic selection in human mutant cell lines.     

In situ rat fast skeletal muscle is more efficient at submaximal than at maximal activation levels.

The influence of stimulation frequency on efficiency (= total work output/high-energy phosphate consumption) was studied using in situ medial gastrocnemius muscle tendon complexes of the rat. The muscles performed 20 repeated concentric contractions (2/s) at 34 degrees C. During these repeated contractions, the muscle was stimulated via the severed sciatic nerve with either 60, 90, or 150 Hz. The muscle was freeze-clamped immediately after these contractions, and high-energy phosphate consumption was determined by measuring intramuscular chemical change relative to control muscles. The average values (+/-SD) of efficiency calculated for 60, 90, and 150 Hz were 18.5 +/- 1.5 (n = 7), 18.6 +/- 1.5 (n = 9), and 14.7 +/- 1.3 mJ/micromol phosphate (n = 9). The results indicate that the efficiency of the muscles that were submaximally activated (60 or 90 Hz) was higher (+26%, P < 0.05) than that of those maximally activated (150 Hz). Additional experiments showed that the low efficiency at maximal activation levels is unlikely to be the result of a higher energy turnover by the Ca2+ -ATPase relative to the total energy turnover. Therefore, alternative explanations are discussed.